Researcher Profile and Settings

Affiliation

• Faculty of Medicine　Pathological Science　Pathology

Job Title

• Professor

Degree

• (BLANK)

URL

http://patho2.med.hokudai.ac.jp/

J-Global ID

• 200901003910683567

Research Interests

• cancer stem cell   biomaterial   腫瘍学   実験病理学   細胞生物学   

Research Areas

• Nanotechnology/Materials / Nanobioscience / biomedical engineering
• Life sciences / Experimental pathology
• Life sciences / Human pathology
• Life sciences / Molecular biology
• Life sciences / Neuroanatomy and physiology

Educational Organization

•  Bachelor's degree program　Departments of Medicine, School of Medicine
•  Master's degree program　Graduate School of Medicine
•  Doctoral (PhD) degree program　Graduate School of Medicine

Academic & Professional Experience

• 2008/05 Hokkaido University Graduate School of Medicine
• 2003/04 - 2008/04 同 助教授
• 2000/04 - 2003/03 同 講師
• 1994/04 - 2000/03 北海道大学医学部病理学第２講座 助手
• 1994/10 - 1997/08 ロックフェラー大学留学(花房秀三郎教授)

Education

• 1990/04 - 1994/03  Hokkaido University  Graduate School of Medicine
• 1984/04 - 1990/03  Hokkaido University  School of Medicine

Association Memberships

• 日本脳腫瘍病理学会   日本脳腫瘍学会   日本神経病理学会   日本臨床細胞診学会   日本分子生物学会   日本癌学会   日本病理学会   

Research Activities

Published Papers

• A familial case of extramammary Paget disease: Analysis of whole-exome sequencing

  Takuya Maeda, Teruki Yanagi, Shinya Kitamura, Hiroshi Nishihara, Yusuke Ono, Yusuke Mizukami, Shinya Tanaka, Hideyuki Ujiie

  EJC Skin Cancer 2024/03 [Refereed]
  
• Virological characteristics of the SARS-CoV-2 Omicron XBB.1.5 variant.

  Tomokazu Tamura, Takashi Irie, Sayaka Deguchi, Hisano Yajima, Masumi Tsuda, Hesham Nasser, Keita Mizuma, Arnon Plianchaisuk, Saori Suzuki, Keiya Uriu, Mst Monira Begum, Ryo Shimizu, Michael Jonathan, Rigel Suzuki, Takashi Kondo, Hayato Ito, Akifumi Kamiyama, Kumiko Yoshimatsu, Maya Shofa, Rina Hashimoto, Yuki Anraku, Kanako Terakado Kimura, Shunsuke Kita, Jiei Sasaki, Kaori Sasaki-Tabata, Katsumi Maenaka, Naganori Nao, Lei Wang, Yoshitaka Oda, Terumasa Ikeda, Akatsuki Saito, Keita Matsuno, Jumpei Ito, Shinya Tanaka, Kei Sato, Takao Hashiguchi, Kazuo Takayama, Takasuke Fukuhara

  Nature communications 15 (1) 1176 - 1176 2024/02/08 

  Circulation of SARS-CoV-2 Omicron XBB has resulted in the emergence of XBB.1.5, a new Variant of Interest. Our phylogenetic analysis suggests that XBB.1.5 evolved from XBB.1 by acquiring the S486P spike (S) mutation, subsequent to the acquisition of a nonsense mutation in ORF8. Neutralization assays showed similar abilities of immune escape between XBB.1.5 and XBB.1. We determine the structural basis for the interaction between human ACE2 and the S protein of XBB.1.5, showing similar overall structures between the S proteins of XBB.1 and XBB.1.5. We provide the intrinsic pathogenicity of XBB.1 and XBB.1.5 in hamsters. Importantly, we find that the ORF8 nonsense mutation of XBB.1.5 resulted in impairment of MHC suppression. In vivo experiments using recombinant viruses reveal that the XBB.1.5 mutations are involved with reduced virulence of XBB.1.5. Together, our study identifies the two viral functions defined the difference between XBB.1 and XBB.1.5.
• Targeted demethylation and activation of NLRC5 augment cancer immunogenicity through MHC class I.

  Xin Sun, Toshiyuki Watanabe, Yoshitaka Oda, Weidong Shen, Alaa Ahmad, Ryota Ouda, Paul de Figueiredo, Hidemitsu Kitamura, Shinya Tanaka, Koichi S Kobayashi

  Proceedings of the National Academy of Sciences of the United States of America 121 (6) e2310821121  2024/02/06 

  Impaired expression of MHC (major histocompatibility complex) class I in cancers constitutes a major mechanism of immune evasion. It has been well documented that the low level of MHC class I is associated with poor prognosis and resistance to checkpoint blockade therapies. However, there is lmited approaches to specifically induce MHC class I to date. Here, we show an approach for robust and specific induction of MHC class I by targeting an MHC class I transactivator (CITA)/NLRC5, using a CRISPR/Cas9-based gene-specific system, designated TRED-I (Targeted reactivation and demethylation for MHC-I). The TRED-I system specifically recruits a demethylating enzyme and transcriptional activators on the NLRC5 promoter, driving increased MHC class I antigen presentation and accelerated CD8+ T cell activation. Introduction of the TRED-I system in an animal cancer model exhibited tumor-suppressive effects accompanied with increased infiltration and activation of CD8+ T cells. Moreover, this approach boosted the efficacy of checkpoint blockade therapy using anti-PD1 (programmed cell death protein) antibody. Therefore, targeting NLRC5 by this strategy provides an attractive therapeutic approach for cancer.
• Fibrinous pericarditis secondary to recurrent acute myeloid leukaemia.

  Takao Konishi, Taichi Kimura, Koichiro Minauchi, Shinya Tanaka

  European heart journal. Case reports 7 (11) ytad537  2023/11
• Akaluc bioluminescence offers superior sensitivity to trackin vivodynamics of SARS-CoV-2 infection

  Tomokazu Tamura, Hayato Ito, Shiho Torii, Lei Wang, Rigel Suzuki, Shuhei Tusjino, Akifumi Kamiyama, Yoshitaka Oda, Yuhei Morioka, Saori Suzuki, Kotaro Shirakawa, Kei Sato, Kumiko Yoshimatsu, Yoshiharu Matsuura, Satoshi Iwano, Shinya Tanaka, Takasuke Fukuhara

  2023/10/13 

  Summary Monitoringin vivoviral dynamics can improve our understanding of pathogenicity and tissue tropism. For positive-sense, single-stranded RNA viruses, several studies have attempted to monitor viral kineticsin vivousing reporter genomes. The application of such recombinant viruses can be limited by challenges in accommodating bioluminescent reporter genes in the viral genome. Conventional luminescence also exhibits relatively low tissue permeability and thus less sensitivity for visualizationin vivo. Here we show that unlike NanoLuc bioluminescence, the improved method, termed AkaBLI, allows visualization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Syrian hamsters. By successfully incorporating a codon-optimized Akaluc luciferase gene into the SARS-CoV-2 genome, we visualizedin vivoinfection, including the tissue-specific differences associated with particular variants. Additionally, we could evaluate the efficacy of neutralizing antibodies and mRNA vaccination by monitoring changes in Akaluc signals. Overall, AkaBLI is an effective technology for monitoring viral dynamics in live animals.
• SARS-CoV-2オミクロン株におけるハムスター肺炎モデルの病理組織学的解析

  小田 義崇, 津田 真寿美, 王 磊, 種井 善一, 福原 崇介, 佐藤 佳, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (2) 132 - 132 0300-9181 2023/10
• Lethal complication: Ventricular septal perforation and right ventricular infarction after acute myocardial infarction.

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Shinya Tanaka

  Clinical case reports 11 (10) e7994  2023/10 

  Here, we report a case of ventricular septal perforation complicated with right ventricular infarction after inferior acute myocardial infarction, which was associated with a poor clinical outcome despite the successful surgical treatment.
• Deep vein thrombosis due to left iliac vein compression syndrome complicated by acute pulmonary thromboembolism and cerebral infarction.

  Takao Konishi, Naohiro Funayama, Daisuke Hotta, Shinya Tanaka

  Acta cardiologica 1 - 2 2023/08/29
• Increased Piezo1 expression in myofibroblasts in patients with symptomatic carotid atherosclerotic plaques undergoing carotid endarterectomy: A pilot study.

  Takao Konishi, Kenji Kamiyama, Toshiaki Osato, Tetsuyuki Yoshimoto, Takeshi Aoki, Toshihisa Anzai, Shinya Tanaka

  Vascular 17085381231192380 - 17085381231192380 2023/07/27 

  OBJECTIVES: We aimed to investigate Piezo1 expression in myofibroblasts in symptomatic and asymptomatic patients undergoing carotid endarterectomy and its relationship with atherosclerotic plaque formation. METHODS: This cross-sectional study analyzed carotid plaques of 17 randomly selected patients who underwent carotid endarterectomy from May 2015 to August 2017. In total, 51 sections (the most stenotic lesion, and the sections 5-mm proximal and distal) stained with hematoxylin-eosin and elastica-Masson were examined. Immunohistochemistry was performed using antibodies to Piezo1. The Piezo1 score of a section was calculated semiquantitatively, averaged across 30 randomly selected myofibroblasts in the fibrous cap of the plaque. RESULTS: Of 17 patients (mean age: 74.2 ± 7.1 years), 15 were men, 9 had diabetes mellitus, and 13 had hypertension. Symptomatic patients had higher mean Piezo1 score than asymptomatic patients (1.78 ± 0.23 vs 1.34 ± 0.17, p < .001). Univariate linear regression analyses suggested an association between plaque rupture, thin-cap fibroatheroma and microcalcifications and the Piezo1 score (p = .001, .008, and 0.003, respectively). CONCLUSIONS: Increased Piezo1 expression of myofibroblasts may be associated with atherosclerotic carotid plaque instability. Further study is warranted to support this finding.
• Comparative pathogenicity of SARS-CoV-2 Omicron subvariants including BA.1, BA.2, and BA.5.

  Tomokazu Tamura, Daichi Yamasoba, Yoshitaka Oda, Jumpei Ito, Tomoko Kamasaki, Naganori Nao, Rina Hashimoto, Yoichiro Fujioka, Rigel Suzuki, Lei Wang, Hayato Ito, Yukie Kashima, Izumi Kimura, Mai Kishimoto, Masumi Tsuda, Hirofumi Sawa, Kumiko Yoshimatsu, Yuki Yamamoto, Tetsuharu Nagamoto, Jun Kanamune, Yutaka Suzuki, Yusuke Ohba, Isao Yokota, Keita Matsuno, Kazuo Takayama, Shinya Tanaka, Kei Sato, Takasuke Fukuhara

  Communications biology 6 (1) 772 - 772 2023/07/24 

  The unremitting emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants necessitates ongoing control measures. Given its rapid spread, the new Omicron subvariant BA.5 requires urgent characterization. Here, we comprehensively analyzed BA.5 with the other Omicron variants BA.1, BA.2, and ancestral B.1.1. Although in vitro growth kinetics of BA.5 was comparable among the Omicron subvariants, BA.5 was much more fusogenic than BA.1 and BA.2. Airway-on-a-chip analysis showed that, among Omicron subvariants, BA.5 had enhanced ability to disrupt the respiratory epithelial and endothelial barriers. Furthermore, in our hamster model, in vivo pathogenicity of BA.5 was slightly higher than that of the other Omicron variants and less than that of ancestral B.1.1. Notably, BA.5 gains efficient virus spread compared with BA.1 and BA.2, leading to prompt immune responses. Our findings suggest that BA.5 has low pathogenicity compared with the ancestral strain but enhanced virus spread /inflammation compared with earlier Omicron subvariants.
• A Case of Uterine Tumor Resembling Ovarian Sex Cord Tumor With Prominent Myxoid Features.

  Koki Ise, Zen-Ichi Tanei, Yoshitaka Oda, Satoshi Tanikawa, Hirokazu Sugino, Yusuke Ishida, Masumi Tsuda, Yuko Gotoda, Kunihiko Nishiwaki, Hiroyuki Yanai, Tadashi Hasegawa, Kazuo Nagashima, Shinya Tanaka

  International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists 2023/06/14 

  Uterine tumor resembling ovarian sex cord tumor (UTROSCT) is a rare tumor with low malignant potential that commonly occurs in middle age. Although more than 100 cases have been reported to date, myxoid morphology is not well documented. Here, we present a 75-yr-old woman with abnormal vaginal bleeding, with an 8-cm mass in the uterine corpus detected by irregular, high-intensity signaling on T2-weighted imaging. The uterine mass had a glistening mucinous appearance on gross examination. Microscopically, most of the tumor cells were floating in the myxoid stroma. The tumor cells formed clusters or nests with abundant cytoplasm, while some exhibited trabecular or rhabdoid appearances. Immunohistochemically, tumor cells were positive for pancytokeratin (AE1/AE3), α-smooth muscle actin, CD10, progesterone receptor, and some sex cord markers such as calretinin, inhibin, CD56, steroidogenic factor-1. Electron microscopy demonstrated epithelial and sex cord differentiation. This tumor was negative for JAZF1-JJAZ1 fusion gene that is frequently found in low-grade endometrial stromal sarcoma. Fusion genes related to UTROSCT, including NCOA2/3, were not detected by reverse transcription polymerase chain reaction. The present case suggests that UTROSCT should be included in the differential diagnosis of myxoid uterine tumors.
• Virological characteristics of the SARS-CoV-2 XBB variant derived from recombination of two Omicron subvariants.

  Tomokazu Tamura, Jumpei Ito, Keiya Uriu, Jiri Zahradnik, Izumi Kida, Yuki Anraku, Hesham Nasser, Maya Shofa, Yoshitaka Oda, Spyros Lytras, Naganori Nao, Yukari Itakura, Sayaka Deguchi, Rigel Suzuki, Lei Wang, Mst Monira Begum, Shunsuke Kita, Hisano Yajima, Jiei Sasaki, Kaori Sasaki-Tabata, Ryo Shimizu, Masumi Tsuda, Yusuke Kosugi, Shigeru Fujita, Lin Pan, Daniel Sauter, Kumiko Yoshimatsu, Saori Suzuki, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Yuki Yamamoto, Tetsuharu Nagamoto, Gideon Schreiber, Katsumi Maenaka, Takao Hashiguchi, Terumasa Ikeda, Takasuke Fukuhara, Akatsuki Saito, Shinya Tanaka, Keita Matsuno, Kazuo Takayama, Kei Sato

  Nature communications 14 (1) 2800 - 2800 2023/05/16 

  In late 2022, SARS-CoV-2 Omicron subvariants have become highly diversified, and XBB is spreading rapidly around the world. Our phylogenetic analyses suggested that XBB emerged through the recombination of two cocirculating BA.2 lineages, BJ.1 and BM.1.1.1 (a progeny of BA.2.75), during the summer of 2022. XBB.1 is the variant most profoundly resistant to BA.2/5 breakthrough infection sera to date and is more fusogenic than BA.2.75. The recombination breakpoint is located in the receptor-binding domain of spike, and each region of the recombinant spike confers immune evasion and increases fusogenicity. We further provide the structural basis for the interaction between XBB.1 spike and human ACE2. Finally, the intrinsic pathogenicity of XBB.1 in male hamsters is comparable to or even lower than that of BA.2.75. Our multiscale investigation provides evidence suggesting that XBB is the first observed SARS-CoV-2 variant to increase its fitness through recombination rather than substitutions.
• Convergent evolution of SARS-CoV-2 Omicron subvariants leading to the emergence of BQ.1.1 variant.

  Jumpei Ito, Rigel Suzuki, Keiya Uriu, Yukari Itakura, Jiri Zahradnik, Kanako Terakado Kimura, Sayaka Deguchi, Lei Wang, Spyros Lytras, Tomokazu Tamura, Izumi Kida, Hesham Nasser, Maya Shofa, Mst Monira Begum, Masumi Tsuda, Yoshitaka Oda, Tateki Suzuki, Jiei Sasaki, Kaori Sasaki-Tabata, Shigeru Fujita, Kumiko Yoshimatsu, Hayato Ito, Naganori Nao, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Yuki Yamamoto, Tetsuharu Nagamoto, Jin Kuramochi, Gideon Schreiber, Akatsuki Saito, Keita Matsuno, Kazuo Takayama, Takao Hashiguchi, Shinya Tanaka, Takasuke Fukuhara, Terumasa Ikeda, Kei Sato

  Nature communications 14 (1) 2671 - 2671 2023/05/11 

  In late 2022, various Omicron subvariants emerged and cocirculated worldwide. These variants convergently acquired amino acid substitutions at critical residues in the spike protein, including residues R346, K444, L452, N460, and F486. Here, we characterize the convergent evolution of Omicron subvariants and the properties of one recent lineage of concern, BQ.1.1. Our phylogenetic analysis suggests that these five substitutions are recurrently acquired, particularly in younger Omicron lineages. Epidemic dynamics modelling suggests that the five substitutions increase viral fitness, and a large proportion of the fitness variation within Omicron lineages can be explained by these substitutions. Compared to BA.5, BQ.1.1 evades breakthrough BA.2 and BA.5 infection sera more efficiently, as demonstrated by neutralization assays. The pathogenicity of BQ.1.1 in hamsters is lower than that of BA.5. Our multiscale investigations illuminate the evolutionary rules governing the convergent evolution for known Omicron lineages as of 2022.
• 肺腫瘍血栓性微小血管症(PTTM)により急激な転帰を辿った転移性膀胱癌の2症例

  宮田 遥, 大澤 崇宏, 山田 修平, 細川 智加, 星 達也, 坪内 駿, 松本 隆児, 安部 崇重, 中里 信一, 岩崎 沙理, 谷口 浩二, 谷川 聖, 田中 伸哉, 篠原 信雄

  泌尿器外科 医学図書出版(株) 36 (5) 428 - 428 0914-6180 2023/05
• 家族性乳房外パジェット病における遺伝子変異解析結果

  前田 拓哉, 柳 輝希, 西原 広史, 小野 裕介, 水上 裕輔, 田中 伸哉, 氏家 英之

  日本皮膚科学会雑誌 (公社)日本皮膚科学会 133 (5) 1350 - 1350 0021-499X 2023/05
• てんかんを発症した72歳女性の右側頭葉内側部病変

  種井 善一, 浅野目 卓, 小野 裕介, 小田 義崇, 王 磊, 津田 真寿美, 佐藤 憲市, 水上 裕輔, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 40 (Suppl.) 126 - 126 1433-7398 2023/05
• 拡張型心筋症を発症したEmery-Dreifuss型筋ジストロフィーの一剖検例

  佐々木 美羽, 種井 善一, 松島 理明, 石垣 隆弘, 桑原 健, 小田 義嵩, 谷川 聖, 津田 真寿美, 矢部 一郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 381 - 381 0300-9181 2023/03
• 脳腫瘍に対する病理研究者としての克服戦略 ハイドロゲルによる髄膜腫幹細胞の探索(Strategies for overcoming the brain tumors presented by pathology researchers Search for meningioma stem cells using hydrogel)

  小田 義崇, 津田 真寿美, 湯澤 明夏, 王 磊, 鈴鹿 淳, ハビバ・ウンマ, 種井 善一, モーウリン・クリスチアン, グン 剣萍, 田中 伸哉

  日本病理学会会誌 112 (1) 184 - 184 0300-9181 2023/03
• 難病克服を目指す、ヒト死後脳研究の成果と未来 ブレインバンクから分かってきた高齢者のレビー病理

  種井 善一, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 208 - 208 0300-9181 2023/03
• TKIs耐性膠芽腫細胞の特性と耐性メカニズムの解析

  津田 真寿美, 王 磊, 小田 義崇, 谷川 聖, 種井 善一, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 278 - 278 0300-9181 2023/03
• ハイドロゲルを用いた中皮腫幹細胞の創出および治療標的分子の探索

  加藤 万里絵, 杉野 弘和, 津田 真寿美, 王 磊, 種井 善一, 小田 義崇, 谷川 聖, グン 剣萍, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 332 - 332 0300-9181 2023/03
• 72歳女性のMultinodular and Vacuolating Neuronal Tumor of the cerebrumの1例

  寺島 祐樹, 種井 善一, 浅野目 卓, 黒田 花音, 小田 義崇, 谷川 聖, 王 磊, 津田 真寿美, 佐藤 憲市, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 375 - 375 0300-9181 2023/03
• JCVとCMVの脳幹部重複感染症例におけるウイルスの局在解析

  黒田 花音, 種井 善一, 岡崎 ななせ, 工藤 彰彦, 阿部 恵, 寺島 祐樹, 谷川 聖, 津田 真寿美, 矢部 一郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 375 - 375 0300-9181 2023/03
• 慢性血栓塞栓性肺高血圧症の一剖検例

  岸本 佳子, 種井 善一, 青木 健志, 加藤 万里絵, 小田 義崇, 谷川 聖, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 378 - 378 0300-9181 2023/03
• Becker型筋ジストロフィーの兄弟剖検症例の病理組織学的検討

  宮本 裕也, 種井 善一, 谷川 聖, 小田 義崇, 津田 真寿美, 加納 崇裕, 横田 卓, 矢部 一郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 378 - 378 0300-9181 2023/03
• 電子顕微鏡的検討を行ったラブドイド髄膜腫の一症例

  戸田 壮太郎, 種井 善一, 京野 里虹, 寺島 祐樹, 谷川 聖, 小田 義崇, 王 磊, 津田 真寿美, 瀬尾 善宣, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 380 - 380 0300-9181 2023/03
• 髄膜腫の骨化におけるEpithelial-mesenchymal transitionの関与についての検討

  京野 里虹, 種井 善一, 寺島 祐樹, 小田 義崇, 谷川 聖, 王 磊, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 381 - 381 0300-9181 2023/03
• FFPE検体の質量分析による肺小細胞癌の脳転移関連分子の解析

  江端 美織, 何 錦涛, 小田 義崇, 谷川 聖, 王 磊, 津田 真寿美, 種井 善一, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 383 - 383 0300-9181 2023/03
• 進行性核上性麻痺にTDP43 pathologyを合併した一剖検例

  鍵谷 豪太, 種井 善一, 谷川 聖, 小田 義崇, 王 磊, 津田 真寿美, 大槻 美佳, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 379 - 379 0300-9181 2023/03
• 拡張型心筋症を発症したEmery-Dreifuss型筋ジストロフィーの一剖検例

  佐々木 美羽, 種井 善一, 松島 理明, 石垣 隆弘, 桑原 健, 小田 義嵩, 谷川 聖, 津田 真寿美, 矢部 一郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 112 (1) 381 - 381 0300-9181 2023/03
• Combination therapy with bevacizumab and a CCR2 inhibitor for human ovarian cancer: An in vivo validation study.

  Tianyue Zhai, Takashi Mitamura, Lei Wang, Shimpei I Kubota, Masaaki Murakami, Shinya Tanaka, Hidemichi Watari

  Cancer medicine 12 (8) 9697 - 9708 2023/02/22 

  BACKGROUND: Anti-angiogenic therapy with bevacizumab (BEV), an anti-VEGF antibody, plays a critical role in the treatment of ovarian cancer. However, despite an encouraging initial response, most tumors become resistant to BEV over time, and a new strategy that enables sustainable treatment using BEV is therefore needed. METHODS: To overcome the resistance to BEV in patients with ovarian cancer, we performed a validation study of combination therapy with BEV (10 mg/kg) and the CCR2 inhibitor BMS CCR2 22 (20 mg/kg) (BEV/CCR2i) using 3 consecutive patient-derived xenografts (PDXs) of immunodeficient mice. RESULTS: BEV/CCR2i demonstrated a significant effect of growth suppression in the BEV-resistant serous PDX and BEV-sensitive serous PDX compared with BEV (30.4% after the second cycle and 15.5% after the first cycle, respectively), and treatment cessation did not attenuate this effect. Tissue clearing and immunohistochemistry with an anti-α-SMA antibody suggested that BEV/CCR2i suppressed angiogenesis from the host mice more than BEV. In addition, human CD31 immunohistochemistry revealed that BEV/CCR2i decreased microvessels originating from the patients to a significantly greater degree than BEV. Regarding the BEV-resistant clear cell PDX, the effect of BEV/CCR2i was unclear during the first five cycles, but the following two cycles of increased-dose BEV/CCR2i (CCR2i 40 mg/kg) significantly suppressed tumor growth compared with BEV (28.3%) by inhibiting the CCR2B-MAPK pathway. CONCLUSIONS: BEV/CCR2i showed a sustained anticancer immunity-independent effect in human ovarian cancer that was more significant in serous carcinoma than in clear cell carcinoma.
• 鞍上部に生じた毛様細胞性星細胞腫により続発性副腎皮質機能低下症を呈した1例

  関 萌花, 亀田 啓, 宮本 麻唯子, 小野 翼, 宮 愛香, 野本 博司, 曹 圭龍, 中村 昭伸, 山口 秀, 田中 伸哉, 三好 秀明, 渥美 達也

  日本内分泌学会雑誌 (一社)日本内分泌学会 98 (4) 718 - 718 0029-0661 2023/02
• FAM83Hの発現低下はケラチン分布の変化を介して皮膚有棘細胞癌の遊走と浸潤を促進する

  得地 景子, 北村 真也, 前田 拓哉, 氏家 英之, 柳 輝希, 渡部 昌, 畠山 鎮次, 加納 里志, 田中 伸哉

  日本皮膚科学会雑誌 (公社)日本皮膚科学会 133 (2) 265 - 265 0021-499X 2023/02
• Multimodality imaging approach to an adult case with cor triatriatum sinister.

  Takao Konishi, Naohiro Funayama, Daisuke Hotta, Shinya Tanaka

  Cardiology journal 30 (6) 1057 - 1058 2023
• Generation of Fetal Intestinal Organoids and Their Maturation into Adult Intestinal Cells

  Masamichi Imajo, Akira Hirota, Shinya Tanaka

  Methods in Molecular Biology 2650 133 - 140 1064-3745 2023 

  During embryonic development, the gut tube undergoes massive morphological changes from the simple tube structure composed of the pseudostratified epithelium into the mature intestinal tract composed of the columnar epithelium and characterized by the unique crypt-villus structures. In mice, maturation of fetal gut precursor cells into adult intestinal cells starts around embryonic day (E) 16.5, during which adult intestinal stem cells and their differentiated progenies are generated. In contrast to adult intestinal cells that form budding organoids containing both the crypt-like and villus-like regions, fetal intestinal cells can be cultured as simple spheroid-shaped organoids that show a uniform proliferation pattern. The fetal intestinal spheroids can undergo spontaneous maturation into adult budding organoids that contain intestinal stem cells and differentiated cells, including enterocytes, goblet, enteroendocrine, and Paneth cells, recapitulating intestinal cell maturation in vitro. Here, we provide detailed methods for establishment of fetal intestinal organoids and their differentiation into adult intestinal cells. These methods enable in vitro recapitulation of intestinal development and would be useful to reveal mechanisms that regulate the transition from fetal to adult intestinal cells.
• Geometrical analysis identified morphological features of hydrogel-induced cancer stem cells in synovial sarcoma model cells

  Zannatul Ferdous, Jean-Emmanuel Clément, Jian Ping Gong, Shinya Tanaka, Tamiki Komatsuzaki, Masumi Tsuda

  Biochemical and Biophysical Research Communications 642 41 - 49 0006-291X 2023/01 [Refereed][Not invited]
   

  Cancer stem cells (CSCs) has been a key target to cure cancer patients completely. Although many CSC markers have been identified, they are frequently cancer type-specific and those expressions are occasionally variable, which becomes an obstacle to elucidate the characteristics of the CSCs. Here we scrutinized the relationship between stemness elevation and geometrical features of single cells. The PAMPS hydrogel was utilized to create the CSCs from mouse myoblast C2C12 and its synovial sarcoma model cells. qRT-PCR analysis confirmed the significant increase in expression levels of Sox2, Nanog, and Oct3/4 on the PAMPS gel, which was higher in the synovial sarcoma model cells. Of note, the morphological heterogeneity was appeared on the PAMPS gel, mainly including flat spreading, elongated spindle, and small round cells, and the Sox2 expression was highest in the small round cells. To examine the role of morphological differences in the elevation of stemness, over 6,400 cells were segmented along with the Sox2 intensity, and 12 geometrical features were extracted at single cell level. A nonlinear mapping of the geometrical features by using uniform manifold approximation and projection (UMAP) clearly revealed the existence of relationship between morphological differences and the stemness elevation, especially for C2C12 and its synovial sarcoma model on the PAMPS gel in which the small round cells possess relatively high Sox2 expression on the PAMPS gel, which supports the strong relationship between morphological changes and the stemness elevation. Taken together, these geometrical features can be useful for morphological profiling of CSCs to classify and distinguish them for understanding of their role in disease progression and drug discovery.
• Monitoring fusion kinetics of viral and target cell membranes in living cells using a SARS-CoV-2 spike-protein-mediated membrane fusion assay

  Hesham Nasser, Ryo Shimizu, Jumpei Ito, Akatsuki Saito, Kei Sato, Terumasa Ikeda, Keita Matsuno, Naganori Nao, Hirofumi Sawa, Mai Kishimoto, Shinya Tanaka, Masumi Tsuda, Lei Wang, Yoshikata Oda, Marie Kato, Zannatul Ferdous, Hiromi Mouri, Kenji Shishido, Takasuke Fukuhara, Tomokazu Tamura, Rigel Suzuki, Hayato Ito, Daichi Yamasoba, Izumi Kimura, Naoko Misawa, Keiya Uriu, Yusuke Kosugi, Shigeru Fujita, Mai Suganami, Mika Chiba, Ryo Yoshimura, So Nakagawa, Jiaqi Wu, Akifumi Takaori-Kondo, Kotaro Shirakawa, Kayoko Nagata, Yasuhiro Kazuma, Ryosuke Nomura, Yoshihito Horisawa, Yusuke Tashiro, Yugo Kawai, Takashi Irie, Ryoko Kawabata, MST Monira Begum, Otowa Takahashi, Kimiko Ichihara, Takamasa Ueno, Chihiro Motozono, Mako Toyoda, Yuri L. Tanaka, Erika P. Butlertanaka, Maya Shofa, Kazuo Takayama, Rina Hashimoto, Sayaka Deguchi, Takao Hashiguchi, Tateki Suzuki, Kanako Kimura, Jiei Sasaki, Yukari Nakajima, Kaori Tabata

  STAR Protocols 3 (4) 101773 - 101773 2666-1667 2022/12
• ERK MAP Kinase Signaling Regulates RAR Signaling to Confer Retinoid Resistance on Breast Cancer Cells

  Akira Hirota, Jean-Emmanuel Clément, Satoshi Tanikawa, Takayuki Nonoyama, Tamiki Komatsuzaki, Jian Ping Gong, Shinya Tanaka, Masamichi Imajo

  Cancers MDPI AG 14 (23) 5890 - 5890 2022/11/29 [Refereed][Not invited]
   

  Retinoic acid (RA) and its synthetic derivatives, retinoids, have been established as promising anticancer agents based on their ability to regulate cell proliferation and survival. Clinical trials, however, have revealed that cancer cells often acquire resistance to retinoid therapy. Therefore, elucidation of underlying mechanisms of retinoid resistance has been considered key to developing more effective use of retinoids in cancer treatment. In this study, we show that constitutive activation of ERK MAP kinase signaling, which is often caused by oncogenic mutations in RAS or RAF genes, suppresses RA receptor (RAR) signaling in breast cancer cells. We show that activation of the ERK pathway suppresses, whereas its inhibition promotes, RA-induced transcriptional activation of RAR and the resultant upregulation of RAR-target genes in breast cancer cells. Importantly, ERK inhibition potentiates the tumor-suppressive activity of RA in breast cancer cells. Moreover, we also reveal that suppression of RAR signaling and activation of ERK signaling are associated with poor prognoses in breast cancer patients and represent hallmarks of specific subtypes of breast cancers, such as basal-like, HER2-enriched and luminal B. These results indicate that ERK-dependent suppression of RAR activity underlies retinoid resistance and is associated with cancer subtypes and patient prognosis in breast cancers.
• Virological characteristics of the SARS-CoV-2 Omicron BA.2.75 variant.

  Akatsuki Saito, Tomokazu Tamura, Jiri Zahradnik, Sayaka Deguchi, Koshiro Tabata, Yuki Anraku, Izumi Kimura, Jumpei Ito, Daichi Yamasoba, Hesham Nasser, Mako Toyoda, Kayoko Nagata, Keiya Uriu, Yusuke Kosugi, Shigeru Fujita, Maya Shofa, Mst Monira Begum, Ryo Shimizu, Yoshitaka Oda, Rigel Suzuki, Hayato Ito, Naganori Nao, Lei Wang, Masumi Tsuda, Kumiko Yoshimatsu, Jin Kuramochi, Shunsuke Kita, Kaori Sasaki-Tabata, Hideo Fukuhara, Katsumi Maenaka, Yuki Yamamoto, Tetsuharu Nagamoto, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Takamasa Ueno, Gideon Schreiber, Akifumi Takaori-Kondo, Kotaro Shirakawa, Hirofumi Sawa, Takashi Irie, Takao Hashiguchi, Kazuo Takayama, Keita Matsuno, Shinya Tanaka, Terumasa Ikeda, Takasuke Fukuhara, Kei Sato

  Cell host & microbe 30 (11) 1540 - 1555 2022/11/09 

  The SARS-CoV-2 Omicron BA.2.75 variant emerged in May 2022. BA.2.75 is a BA.2 descendant but is phylogenetically distinct from BA.5, the currently predominant BA.2 descendant. Here, we show that BA.2.75 has a greater effective reproduction number and different immunogenicity profile than BA.5. We determined the sensitivity of BA.2.75 to vaccinee and convalescent sera as well as a panel of clinically available antiviral drugs and antibodies. Antiviral drugs largely retained potency, but antibody sensitivity varied depending on several key BA.2.75-specific substitutions. The BA.2.75 spike exhibited a profoundly higher affinity for its human receptor, ACE2. Additionally, the fusogenicity, growth efficiency in human alveolar epithelial cells, and intrinsic pathogenicity in hamsters of BA.2.75 were greater than those of BA.2. Our multilevel investigations suggest that BA.2.75 acquired virological properties independent of BA.5, and the potential risk of BA.2.75 to global health is greater than that of BA.5.
• Intraoperative rapid immunohistochemistry with noncontact antibody mixing for undiagnosed pulmonary tumors.

  Kazuhiro Imai, Hiroshi Nanjo, Wataru Shigeeda, Tamotsu Sugai, Tomoo Ito, Yoshimasa Maniwa, Shinogu Takashima, Hajime Saito, Naoki Yanagawa, Yugo Tanaka, Takefumi Doi, Yuko Hiroshima, Kyoko Nomura, Mishie Tanino, Shinya Tanaka, Yoshihiro Minamiya

  Cancer science 114 (2) 702 - 711 2022/10/25 

  Knowledge of the histologic type and primary origin of pulmonary tumors is essential when preparing a surgical strategy. Intraoperative diagnosis of hematoxylin and eosin (H&E)-stained frozen sections is the gold standard, but reliable pathology requires time-consuming immunohistochemistry (IHC) to distinguish among histological types/organ origins and to analyze molecular status. The aim of this study was to evaluate the clinical reliability of a new rapid-IHC technique for intraoperative diagnosis of pulmonary tumors. In total, 169 patients with undiagnosed pulmonary tumors were enrolled in a multicenter prospective observational study. At three institutes, pulmonary tumor samples were collected through core needle biopsy and/or surgery to determine surgical strategies. Using a new device for rapid IHC, we applied a high-voltage, low-frequency alternating current (AC) field, which mixes the available antibody as the voltage is switched on/off. Rapid IHC can provide tumor histologic type/origin diagnoses within 20 min, as opposed to the 3-6 h required for conventional IHC. No false diagnoses of malignancy were rendered in any of the cases when using simple H&E staining. With H&E staining alone, the overall definitive diagnosis rate, the rate of defined tumor origin, and the rate of determined histological type were 76.92%, 85.80%, and 90.53%, respectively. When rapid IHC was added, those rates were significantly improved to 88.76%, 94.67%, and 91.72%, respectively. By providing prompt and accurate intraoperative histological/molecular analysis, rapid IHC driven by AC mixing could serve as an effective clinical tool guiding the surgical strategy for undiagnosed pulmonary tumors.
• Force-triggered rapid microstructure growth on hydrogel surface for on-demand functions

  Qifeng Mu, Kunpeng Cui, Zhi Jian Wang, Takahiro Matsuda, Wei Cui, Hinako Kato, Shotaro Namiki, Tomoko Yamazaki, Martin Frauenlob, Takayuki Nonoyama, Masumi Tsuda, Shinya Tanaka, Tasuku Nakajima, Jian Ping Gong

  Nature Communications 13 (1) 2022/10/20 

  Abstract Living organisms share the ability to grow various microstructures on their surface to achieve functions. Here we present a force stamp method to grow microstructures on the surface of hydrogels based on a force-triggered polymerisation mechanism of double-network hydrogels. This method allows fast spatial modulation of the morphology and chemistry of the hydrogel surface within seconds for on-demand functions. We demonstrate the oriented growth of cells and directional transportation of water droplets on the engineered hydrogel surfaces. This force-triggered method to chemically engineer the hydrogel surfaces provides a new tool in addition to the conventional methods using light or heat, and will promote the wide application of hydrogels in various fields.
• An autopsy case report of adult-onset Krabbe disease: Comparison with an infantile-onset case.

  Miu Sasaki, Miori Ebata, Zen-Ichi Tanei, Yoshitaka Oda, Akiko Hamauchi, Satoshi Tanikawa, Hirokazu Sugino, Yusuke Ishida, Takenori Abe, Nobutaka Arai, Kazuya Sako, Shinya Tanaka

  Pathology international 2022/10/06 

  Krabbe disease is a lysosomal storage disease caused by a deficiency of the galactocerebrosidase (GALC) enzyme, which leads to demyelination of the central and peripheral nervous systems. Almost all patients with Krabbe disease are infants, and this is the first report of adult-onset cases that describe pathological findings. Here, we present two autopsy cases: a 73-year-old female and a 2-year-old male. The adult-onset case developed symptoms in her late thirties and was diagnosed by the identification of GALC D528N and L634S mutations and by T2-weighted magnetic resonance imaging; she had increased signal in the white matter along the pyramidal tract to the bilateral precentral gyrus, as well as from the triangular part to the posterior horn of the lateral ventricle. Microscopically, Klüver-Barrera staining was pale in the white matter of the precentral gyrus and occipito-thalamic radiation, and a few globoid cells were observed. The GALC mutations that were identified in the present adult-onset case do not completely inactivate GALC enzyme activity, resulting in focal demyelination of the brain.
• SARS-CoV-2変異株を用いたハムスター肺炎モデルの病理組織学的解析

  小田 義崇, 津田 真寿美, 王 磊, 谷川 聖, 種井 善一, 佐藤 佳, 福原 崇介, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (2) 113 - 113 0300-9181 2022/10
• Analysis of clinicopathological features and NAB2-STAT6 fusion variants of meningeal solitary fibrous tumor with ectopic salivary gland components in the cerebellopontine angle.

  Takahiro Shirakura, Yuichi Yamada, Satoshi Nakata, Bunsho Asayama, Yoshinobu Seo, Satoshi Tanikawa, Takayuki Kato, Nobukazu Komoribayashi, Naohiko Kubo, Nobuhiro Monma, Naoki Okura, Shinya Tanaka, Yoshinao Oda, Junko Hirato, Hideaki Yokoo, Sumihito Nobusawa

  Virchows Archiv : an international journal of pathology 481 (6) 913 - 923 2022/09/02 

  Solitary fibrous tumors (SFTs) are rare mesenchymal tumors that can occur at any location. Since the identification of specific NAB2-STAT6 fusion in SFTs, the fusion gene variants, NAB2 exon 4-STAT6 exon 2/3 and NAB2 exon 5/6/7-STAT6 exon 16/17/18, have been reported to be associated with clinicopathological features, and the latter variant is predominant in meningeal SFTs. SFTs developing in the salivary glands are rare, and more rarely, those involving ectopic salivary glands (ESGs) have been reported in the cerebellopontine angle (CPA); however, their characteristics remain not well understood. In this study, we performed a clinicopathological and molecular analysis of 3 cases of meningeal SFT with ESGs. All cases presented with an extra-axial mass in the CPA, which is a rarer location for intracranial ESGs compared to the sellar region. Histologically, except for the presence of ESGs, there was no significant difference between current cases and ordinary SFTs. The ESGs demonstrated no cellular atypia, and although the spindle tumor cells were immunopositive for STAT6, the ESGs were negative in all cases, supporting that the ESGs are non-neoplastic components. In 1 case, ESGs were found only in the primary tumor and disappeared in recurrence/dissemination. Of note, molecular analysis identified NAB2 exon 4-STAT6 exon 2 in all cases. In conclusion, our results suggest that ESGs particularly in the CPA may be associated with SFTs and that meningeal SFTs with ESGs may be associated with the minor fusion variant of NAB2-STAT6 in the intracranial lesions.
• ハイドロゲルを用いた髄膜腫がん幹細胞マーカーの検索(Identification of novel stemness marker of meningioma by using hydrogel)

  小田 義崇, 津田 真寿美, 湯澤 明夏, 王 磊, 谷川 聖, 種井 善一, グン 剣萍, 田中 伸哉

  日本癌学会総会記事 81回 P - 2111 0546-0476 2022/09
• Virological characteristics of the SARS-CoV-2 Omicron BA.2 subvariants including BA.4 and BA.5

  Izumi Kimura, Daichi Yamasoba, Tomokazu Tamura, Naganori Nao, Tateki Suzuki, Yoshitaka Oda, Shuya Mitoma, Jumpei Ito, Hesham Nasser, Jiri Zahradnik, Keiya Uriu, Shigeru Fujita, Yusuke Kosugi, Lei Wang, Masumi Tsuda, Mai Kishimoto, Hayato Ito, Rigel Suzuki, Ryo Shimizu, M.S.T. Monira Begum, Kumiko Yoshimatsu, Kanako Terakado Kimura, Jiei Sasaki, Kaori Sasaki-Tabata, Yuki Yamamoto, Tetsuharu Nagamoto, Jun Kanamune, Kouji Kobiyama, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Kotaro Shirakawa, Akifumi Takaori-Kondo, Jin Kuramochi, Gideon Schreiber, Ken J. Ishii, Takao Hashiguchi, Terumasa Ikeda, Akatsuki Saito, Takasuke Fukuhara, Shinya Tanaka, Keita Matsuno, Kei Sato

  Cell 0092-8674 2022/09
• Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike.

  Daichi Yamasoba, Izumi Kimura, Hesham Nasser, Yuhei Morioka, Naganori Nao, Jumpei Ito, Keiya Uriu, Masumi Tsuda, Jiri Zahradnik, Kotaro Shirakawa, Rigel Suzuki, Mai Kishimoto, Yusuke Kosugi, Kouji Kobiyama, Teppei Hara, Mako Toyoda, Yuri L Tanaka, Erika P Butlertanaka, Ryo Shimizu, Hayato Ito, Lei Wang, Yoshitaka Oda, Yasuko Orba, Michihito Sasaki, Kayoko Nagata, Kumiko Yoshimatsu, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Jin Kuramochi, Motoaki Seki, Ryoji Fujiki, Atsushi Kaneda, Tadanaga Shimada, Taka-Aki Nakada, Seiichiro Sakao, Takuji Suzuki, Takamasa Ueno, Akifumi Takaori-Kondo, Ken J Ishii, Gideon Schreiber, Hirofumi Sawa, Akatsuki Saito, Takashi Irie, Shinya Tanaka, Keita Matsuno, Takasuke Fukuhara, Terumasa Ikeda, Kei Sato

  Cell 185 (12) 2103 - 2115 2022/05/02 [Refereed][Not invited]
   

  Soon after the emergence and global spread of the SARS-CoV-2 Omicron lineage BA.1, another Omicron lineage, BA.2, began outcompeting BA.1. The results of statistical analysis showed that the effective reproduction number of BA.2 is 1.4-fold higher than that of BA.1. Neutralization experiments revealed that immunity induced by COVID vaccines widely administered to human populations is not effective against BA.2, similar to BA.1, and that the antigenicity of BA.2 is notably different from that of BA.1. Cell culture experiments showed that the BA.2 spike confers higher replication efficacy in human nasal epithelial cells and is more efficient in mediating syncytia formation than the BA.1 spike. Furthermore, infection experiments using hamsters indicated that the BA.2 spike-bearing virus is more pathogenic than the BA.1 spike-bearing virus. Altogether, the results of our multiscale investigations suggest that the risk of BA.2 to global health is potentially higher than that of BA.1.
• Psammomatous meningioma周囲に脳実質石灰化を認めた一例

  岡崎 ななせ, 谷川 聖, 種井 善一, 津田 真寿美, 大澤 崇宏, 松野 吉宏, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 39 (Suppl.) 124 - 124 1433-7398 2022/05
• Psammomatous meningioma周囲に脳実質石灰化を認めた一例

  岡崎 ななせ, 谷川 聖, 種井 善一, 津田 真寿美, 大澤 崇宏, 松野 吉宏, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 39 (Suppl.) 124 - 124 1433-7398 2022/05
• ゲノム解析により膵管内乳頭粘液性腫瘍併存癌の初期像と診断し得た1例

  佐藤 允洋, 唐崎 秀則, 富永 素矢, 坂本 淳, 木村 圭介, 太田 智之, 田中 伸哉, 水上 裕輔

  日本消化器病学会雑誌 (一財)日本消化器病学会 119 (4) 368 - 376 0446-6586 2022/04 

  症例は70歳代男性.膵頭体部の分枝型IPMNの経過観察中に膵炎を繰り返し,嚢胞径の増大を認め切除した.嚢胞性病変はすべて病理学的に低異型IPMNであり,病変間に連続性はなく,これらと離れた膵切除断端に上皮内癌を認めた.遺伝子解析の結果,上皮内癌はIPMNとは異なるクローンに由来する併存癌の初期像と考えられた.併存癌の早期診断のためには,病理学的解析に加えゲノム異常を明らかにすることが重要と考えられた.(著者抄録)
• Clinical and radiological findings of glioblastomas harboring a BRAF V600E mutation.

  Yukitomo Ishi, Shigeru Yamaguchi, Michinari Okamoto, Ryosuke Sawaya, Shogo Endo, Hiroaki Motegi, Shunsuke Terasaka, Zen-Ichi Tanei, Kanako C Hatanaka, Shinya Tanaka, Miki Fujimura

  Brain tumor pathology 39 (3) 162 - 170 2022/04/01 

  The aim of this study was to analyze the clinical and radiological characteristics of glioblastomas (GBMs) harboring a BRAF mutation. Sequencing analysis of BRAF, IDH1/2, and TERT promoters was performed on GBM samples of patients older than 15 years. The clinical, pathological, and radiological data of patients were retrospectively reviewed. Patients were classified into three groups according to their BRAF and IDH1/2 status: BRAF group, IDH group, and BRAF/IDH-wild-type (WT) group. Among 179 GBM cases, we identified nine cases with a BRAF mutation and nine with IDH mutation. The WT group had 161 cases. Age at onset in the BRAF group was significantly lower compared to the WT group and was similar to the IDH group. In cases with negative IDH1-R132H staining and age < 55 years, 15.2% were BRAF-mutant cases. Similar to the IDH group, overall survival of the BRAF group was significantly longer compared with the WT group. Among nine cases in the BRAF group, three cases had hemorrhagic onset and prior lesions were observed in two cases. In conclusion, age < 55 years, being IDH1-R132H negative, with hemorrhagic onset or the presence of prior lesions are factors that signal recommendation of BRAF analysis for adult GBM patients.
• Hydroxyapatite-hybridized double-network hydrogel surface enhances differentiation of bone marrow-derived mesenchymal stem cells to osteogenic cells.

  Takuma Kaibara, Lei Wang, Masumi Tsuda, Takayuki Nonoyama, Takayuki Kurokawa, Norimasa Iwasaki, Jian Ping Gong, Shinya Tanaka, Kazunori Yasuda

  Journal of biomedical materials research. Part A 110 (4) 747 - 760 2022/04 

  Recently, we have developed a hydroxyapatite (HAp)-hybridized double-network (DN) hydrogel (HAp/DN gel), which can robustly bond to the bone tissue in the living body. The purpose of this study is to clarify whether the HAp/DN gel surface can differentiate the bone marrow-derived mesenchymal stem cells (MSCs) to osteogenic cells. We used the MSCs which were harvested from the rabbit bone marrow and cultured on the polystyrene (PS) dish using the autogenous serum-supplemented medium. First, we confirmed the properties of MSCs by evaluating colony forming unit capacity, expression of MSC markers using flow cytometry, and multidifferential capacity. Secondly, polymerase chain reaction analysis demonstrated that the HAp/DN gel surface significantly enhanced mRNA expression of the eight osteogenic markers (TGF-β1, BMP-2, Runx2, Col-1, ALP, OPN, BSP, and OCN) in the cultured MSCs at 7 days than the PS surfaces (p < 0.0001), while the DN gel and HAp surfaces provided no or only a slight effect on the expression of these markers except for Runx2. Additionally, the alkaline phosphatase activity was significantly higher in the cells cultured on the HAp/DN gel surface than in the other three material surfaces (p < 0.0001). Thirdly, when the HAp/DN gel plug was implanted into the rabbit bone marrow, MSC marker-positive cells were recruited in the tissue generated around the plug at 3 days, and Runx2 and OCN were highly expressed in these cells. In conclusion, this study demonstrated that the HAp/DN gel surface can differentiate the MSCs into osteogenic cells.
• 細胞外基質の電位変化に伴うJCウイルス増殖の制御

  谷川 聖, 野々山 貴行, 津田 真寿美, 王 磊, 種井 善一, Gong Jian Ping, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 263 - 263 0300-9181 2022/03
• 脊髄capillary hemangiomaの病理像

  種井 善一, 津田 真寿美, 小田 義崇, 谷川 聖, 杉野 弘和, 大竹 安史, 今村 博幸, 小柳 泉, 飛騨 一利, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 264 - 264 0300-9181 2022/03
• びまん性大細胞型B細胞性リンパ腫の化学療法中に突然死した1剖検例

  加藤 万里絵, 種井 善一, 小島 圭祐, 太田 秀一, ウンマ・ハビバ, 小田 義崇, 谷川 聖, 杉野 弘和, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 308 - 308 0300-9181 2022/03
• 内頸動脈瘤術後に急激な意識障害をきたした神経線維腫症1型の一剖検例

  京野 里虹, 種井 善一, 岡崎 ななせ, 長内 俊也, 小田 義崇, 谷川 聖, 杉野 弘和, 津田 真寿美, 藤村 幹, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 356 - 356 0300-9181 2022/03
• SLEとGood症候群の治療経過中に脳幹障害を来たした一例

  黒田 花音, 種井 善一, 岡崎 ななせ, 工藤 彰彦, 阿部 恵, 小田 義崇, 谷川 聖, 杉野 弘和, 矢部 一郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 356 - 356 0300-9181 2022/03
• 甲状腺乳頭癌多発肺転移後に生じた拘束性換気障害の一剖検例

  鍵谷 豪太, 種井 善一, 若林 健人, 堀井 洋志, 小田 義崇, 谷川 聖, 杉野 弘和, 谷野 美智枝, 今野 哲, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 357 - 357 0300-9181 2022/03
• 細胞外基質の電位変化に伴うJCウイルス増殖の制御

  谷川 聖, 野々山 貴行, 津田 真寿美, 王 磊, 種井 善一, Gong Jian Ping, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 263 - 263 0300-9181 2022/03
• 脊髄capillary hemangiomaの病理像

  種井 善一, 津田 真寿美, 小田 義崇, 谷川 聖, 杉野 弘和, 大竹 安史, 今村 博幸, 小柳 泉, 飛騨 一利, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 264 - 264 0300-9181 2022/03
• びまん性大細胞型B細胞性リンパ腫の化学療法中に突然死した1剖検例

  加藤 万里絵, 種井 善一, 小島 圭祐, 太田 秀一, ウンマ・ハビバ, 小田 義崇, 谷川 聖, 杉野 弘和, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 308 - 308 0300-9181 2022/03
• 内頸動脈瘤術後に急激な意識障害をきたした神経線維腫症1型の一剖検例

  京野 里虹, 種井 善一, 岡崎 ななせ, 長内 俊也, 小田 義崇, 谷川 聖, 杉野 弘和, 津田 真寿美, 藤村 幹, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 356 - 356 0300-9181 2022/03
• SLEとGood症候群の治療経過中に脳幹障害を来たした一例

  黒田 花音, 種井 善一, 岡崎 ななせ, 工藤 彰彦, 阿部 恵, 小田 義崇, 谷川 聖, 杉野 弘和, 矢部 一郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 356 - 356 0300-9181 2022/03
• 甲状腺乳頭癌多発肺転移後に生じた拘束性換気障害の一剖検例

  鍵谷 豪太, 種井 善一, 若林 健人, 堀井 洋志, 小田 義崇, 谷川 聖, 杉野 弘和, 谷野 美智枝, 今野 哲, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 111 (1) 357 - 357 0300-9181 2022/03
• Attenuated fusogenicity and pathogenicity of SARS-CoV-2 Omicron variant.

  Rigel Suzuki, Daichi Yamasoba, Izumi Kimura, Lei Wang, Mai Kishimoto, Jumpei Ito, Yuhei Morioka, Naganori Nao, Hesham Nasser, Keiya Uriu, Yusuke Kosugi, Masumi Tsuda, Yasuko Orba, Michihito Sasaki, Ryo Shimizu, Ryoko Kawabata, Kumiko Yoshimatsu, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Hirofumi Sawa, Terumasa Ikeda, Takashi Irie, Keita Matsuno, Shinya Tanaka, Takasuke Fukuhara, Kei Sato

  Nature 603 (7902) 700 - 705 2022/03 [Refereed]
   

  The emergence of the Omicron variant of SARS-CoV-2 is an urgent global health concern1. In this study, our statistical modelling suggests that Omicron has spread more rapidly than the Delta variant in several countries including South Africa. Cell culture experiments showed Omicron to be less fusogenic than Delta and than an ancestral strain of SARS-CoV-2. Although the spike (S) protein of Delta is efficiently cleaved into two subunits, which facilitates cell-cell fusion2,3, the Omicron S protein was less efficiently cleaved compared to the S proteins of Delta and ancestral SARS-CoV-2. Furthermore, in a hamster model, Omicron showed decreased lung infectivity and was less pathogenic compared to Delta and ancestral SARS-CoV-2. Our multiscale investigations reveal the virological characteristics of Omicron, including rapid growth in the human population, lower fusogenicity and attenuated pathogenicity.
• Engineering of an electrically charged hydrogel implanted into a traumatic brain injury model for stepwise neuronal tissue reconstruction

  Satoshi Tanikawa, Yuki Ebisu, Tomáš Sedlačík, Shingo Semba, Takayuki Nonoyama, Akira Hirota, Taiga Takahashi, Kazushi Yamaguchi, Masamichi Imajo, Hinako Kato, Takuya Nishimura, Zen-ichi Tanei, Masumi Tsuda, Tomomi Nemoto, Jian Ping Gong, Shinya Tanaka

  2022/02/19 

  Abstract Neural regeneration is extremely difficult to achieve. In traumatic brain injuries, the loss of brain parenchyma volume hinders neural regeneration. In this study, neuronal tissue engineering was performed by using electrically charged hydrogels composed of cationic and anionic monomers in a 1:1 ratio (C1A1 hydrogel), which served as an effective scaffold for the attachment of neural stem cells (NSCs). In the 3D environment of porous C1A1 hydrogels engineered by the cryogelation technique, NSCs differentiated into neuroglial cells. The C1A1 porous hydrogel was implanted into brain defects in a mouse traumatic damage model. The VEGF-immersed C1A1 porous hydrogel promoted host-derived vascular network formation together with the infiltration of macrophages/microglia and astrocytes into the gel. Furthermore, the stepwise transplantation of GFP-labeled NSCs supported differentiation to glial and neuronal cells. Therefore, this two-step method for neural regeneration may become a new approach for therapeutic brain tissue reconstruction after brain damage in the future. One Sentence Summary Brain tissue reconstruction using charged hydrogel and stepwise NCS injection
• Novel rapid immunohistochemistry using an alternating current electric field identifies Rac and Cdc42 activation in human colon cancer FFPE tissues.

  Masumi Tsuda, Runa Horio, Lei Wang, Tomoko Takenami, Jun Moriya, Jun Suzuka, Hirokazu Sugino, Zenichi Tanei, Mishie Tanino, Shinya Tanaka

  Scientific reports 12 (1) 1733 - 1733 2022/02/02 

  It is important to determine the activation status of Rac and Cdc42 in cancer tissues for the prediction of metastasis and patient prognosis. However, it has been impossible to detect their spatial activation on formalin-fixed paraffin embedded (FFPE) surgical specimens thus far. Here, we established a novel detection technique for activated Rac/Cdc42 in human colon cancer FFPE tissues by using a p21-activated kinase (PAK)-Rac binding domain (RBD) detection probe fused with glutathione S-transferase (GST), designated GST-PAK-RBD, and novel rapid-immunohistochemistry (R-IHC) systems using noncontact alterating-current electric field mixing, although there is a technical limitation in that it may not distinguish between Rac members and Cdc42. In 50 cases of colon cancer, various activation patterns of Rac/Cdc42 were observed, which were designated plasma membrane, cytoplasm, mixed pattern, and polarized distribution. The activity was striking in the invasive fronts of tumors and significantly correlated with tumor invasion properties evaluated by TNM classification. Of note, in tissue microarray (TMA) samples, 29 of 33 cases demonstrated higher Rac1/Cdc42 activity in the tumor area than the corresponding normal mucosa. In addition, positive correlations were detected between Rac/Cdc42 activity and clinicopathological factors such as venous and lymphatic vessel invasion. These results suggest that understanding Rac and Cdc42 activations in cancer tissues would be valuable as an option for molecular therapy as personalized medicine.
• Enhanced fusogenicity and pathogenicity of SARS-CoV-2 Delta P681R mutation.

  Akatsuki Saito, Takashi Irie, Rigel Suzuki, Tadashi Maemura, Hesham Nasser, Keiya Uriu, Yusuke Kosugi, Kotaro Shirakawa, Kenji Sadamasu, Izumi Kimura, Jumpei Ito, Jiaqi Wu, Kiyoko Iwatsuki-Horimoto, Mutsumi Ito, Seiya Yamayoshi, Samantha Loeber, Masumi Tsuda, Lei Wang, Seiya Ozono, Erika P Butlertanaka, Yuri L Tanaka, Ryo Shimizu, Kenta Shimizu, Kumiko Yoshimatsu, Ryoko Kawabata, Takemasa Sakaguchi, Kenzo Tokunaga, Isao Yoshida, Hiroyuki Asakura, Mami Nagashima, Yasuhiro Kazuma, Ryosuke Nomura, Yoshihito Horisawa, Kazuhisa Yoshimura, Akifumi Takaori-Kondo, Masaki Imai, Shinya Tanaka, So Nakagawa, Terumasa Ikeda, Takasuke Fukuhara, Yoshihiro Kawaoka, Kei Sato

  Nature 602 (7896) 300 - 306 2022/02 

  During the current coronavirus disease 2019 (COVID-19) pandemic, a variety of mutations have accumulated in the viral genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and, at the time of writing, four variants of concern are considered to be potentially hazardous to human society1. The recently emerged B.1.617.2/Delta variant of concern is closely associated with the COVID-19 surge that occurred in India in the spring of 2021 (ref. 2). However, the virological properties of B.1.617.2/Delta remain unclear. Here we show that the B.1.617.2/Delta variant is highly fusogenic and notably more pathogenic than prototypic SARS-CoV-2 in infected hamsters. The P681R mutation in the spike protein, which is highly conserved in this lineage, facilitates cleavage of the spike protein and enhances viral fusogenicity. Moreover, we demonstrate that the P681R-bearing virus exhibits higher pathogenicity compared with its parental virus. Our data suggest that the P681R mutation is a hallmark of the virological phenotype of the B.1.617.2/Delta variant and is associated with enhanced pathogenicity.
• Germline PRDM1 Variant rs2185379 in Long-Term Recurrence-Free Survivors of Advanced Ovarian Cancer.

  Takashi Mitamura, Tianyue Zhai, Kanako C Hatanaka, Yutaka Hatanaka, Toraji Amano, Lei Wang, Shinya Tanaka, Hidemichi Watari

  Pharmacogenomics and personalized medicine 15 977 - 984 2022 

  PURPOSE: To identify the germline genetic characteristics of long-term recurrence-free survivors that can be applied to establishing a new strategy for curing advanced cancer, we investigated the whole-genome single nucleotide variants of ovarian cancer patients. PATIENTS AND METHODS: DNA specimens were obtained from rare long-term recurrence-free survivors with FIGO stage III-IV ovarian cancer with no recurrence for 8-23 years after primary treatments for a whole-genome analysis of approximately 660,000 single nucleotide variants. We then established a mouse model with a notable gene alteration by CRISPR/Cas9 to confirm the biological role. RESULTS: The long-term recurrence-free survivors more frequently had germline heterozygous variant rs2185379 of the PRDM1 gene exon than patients with early recurrence (6.8-fold, P=0.013) and the general population. In the mouse model, primary intraperitoneal disseminated tumors of allograft ID8 were significantly smaller in the germline heterozygous rs2185379 group than in the wild-type group (57.4% decrease, P=0.008). Immunohistochemistry showed that the area of distribution of infiltrating T lymphocytes with positive CD8 staining was significantly increased in the germline heterozygous rs2185379 group in comparison to the wild-type group. CONCLUSION: Germline heterozygous rs2185379 in PRDM1 is correlated with an excellent prognosis and can be used to establish a new strategy for treating advanced ovarian cancer.
• Neoatherosclerosis with silent plaque rupture in a saphenous vein graft causing no re-flow phenomenon assessed by optical coherence tomography and histopathology.

  Takao Konishi, Yuki Takahashi, Sho Kazui, Yutaro Yasui, Kohei Saiin, Seiichiro Naito, Sakae Takenaka, Yoshifumi Mizuguchi, Atsushi Tada, Yuta Kobayashi, Kazunori Omote, Takuma Sato, Kiwamu Kamiya, Toshiyuki Nagai, Shinya Tanaka, Toshihisa Anzai

  Cardiology journal 29 (4) 718 - 719 2022
• Vulnerable plaque derived from aspirated thrombi in recurrent acute coronary syndrome with familial hypercholesterolemia despite intensive lipid-lowering statin therapy.

  Hiroyuki Natsui, Takao Konishi, Kohei Saiin, Youji Tamaki, Tomoya Sato, Sakae Takenaka, Atsushi Tada, Yoshifumi Mizuguchi, Yuta Kobayashi, Takuma Sato, Rui Kamada, Kiwamu Kamiya, Toshiyuki Nagai, Shinya Tanaka, Toshihisa Anzai

  Cardiology journal 29 (2) 362 - 363 2022
• Aberrant expression of MYD88 via RNA-controlling CNOT4 and EXOSC3 in colonic mucosa impacts generation of colonic cancer.

  Masumi Tsuda, Misa Noguchi, Tsuyoshi Kurai, Yuji Ichihashi, Koki Ise, Lei Wang, Yusuke Ishida, Mishie Tanino, Satoshi Hirano, Masahiro Asaka, Shinya Tanaka

  Cancer science 112 (12) 5100 - 5113 2021/12 

  In 2020, the worldwide incidence and mortality of colorectal cancer (CRC) were third and second, respectively. As the 5-y survival rate is low when CRC is diagnosed at an advanced stage, a reliable method to predict CRC susceptibility is important for preventing the onset and development and improving the prognosis of CRC. Therefore, we focused on the normal colonic mucosa to investigate changes in gene expression that may induce subsequent genetic alterations that induce malignant transformation. Comprehensive gene expression profiling in the normal mucosa adjacent to colon cancer (CC) compared with tissue from non-colon cancer patients was performed. PCR arrays and qRT-PCR revealed that the expression of 5 genes involved in the immune response, including MYD88, was increased in the normal mucosa of CC patients. The expression levels of MYD88 were strikingly increased in precancerous normal mucosa specimens, which harbored no somatic mutations, as shown by immunohistochemistry. Microarray analysis identified 2 novel RNA-controlling molecules, EXOSC3 and CNOT4, that were significantly upregulated in the normal mucosa of CC patients and were clearly visualized in the nuclei. Forced expression of EXOSC3 and CNOT4 in human colonic epithelial cells increased the expression of IFNGR1, MYD88, NFκBIA, and STAT3 and activated ERK1/2 and JNK in 293T cells. Taken together, these results suggested that, in the inflamed mucosa, EXOSC3- and CNOT4-mediated RNA stabilization, including that of MYD88, may trigger the development of cancer and can serve as a potential predictive marker and innovative treatment to control cancer development.
• Genome-wide CRISPR screen identifies CDK6 as a therapeutic target in Adult T-cell leukemia/lymphoma.

  Takashi Ishio, Sarvesh Kumar, Joji Shimono, Anusara Daenthanasanmak, Sigrid Dubois, Yuquan Lin, Bonita R Bryant, Michael N Petrus, Emmanuel Bachy, Da Wei Huang, Yandan Yang, Patrick L Green, Hiroo Hasegawa, Michiyuki Maeda, Hideki Goto, Tomoyuki Endo, Takashi Yokota, Kanako C Hatanaka, Yutaka Hatanaka, Shinya Tanaka, Yoshihiro Matsuno, Yibin Yang, Satoshi Hashino, Takanori Teshima, Thomas A Waldmann, Louis M Staudt, Masao Nakagawa

  Blood 139 (10) 1541 - 1556 2021/11/24 

  Adult T-cell leukemia/lymphoma (ATLL) is an aggressive T-cell malignancy with a poor prognosis with current therapy. Here we report genome-wide CRISPR-Cas9 screening of ATLL models, which identified CDK6, CCND2, BATF3, JUNB, STAT3, and IL10RB as genes that are essential for the proliferation and/or survival of ATLL cells. As a single agent, the CDK6 inhibitor palbociclib induced cell cycle arrest and apoptosis in ATLL models with wild type TP53. ATLL models that had inactivated TP53 genetically were relatively resistant to palbociclib owing to compensatory CDK2 activity, and this resistance could be reversed by APR-246, a small molecule activator of mutant TP53. The CRISPR-Cas9 screen further highlighted the dependence of ATLL cells on mTORC1 signaling. Treatment of ATLL cells with palbociclib in combination with mTORC1 inhibitors was synergistically toxic irrespective of the TP53 status. This work defines CDK6 as a novel therapeutic target for ATLL and supports the clinical evaluation of palbociclib in combination with mTORC1 inhibitors in this recalcitrant malignancy.
• Loss of FAM83H promotes cell migration and invasion in cutaneous squamous cell carcinoma via impaired keratin distribution.

  Keiko Tokuchi, Shinya Kitamura, Takuya Maeda, Masashi Watanabe, Shigetsugu Hatakeyama, Satoshi Kano, Shinya Tanaka, Hideyuki Ujiie, Teruki Yanagi

  Journal of dermatological science 104 (2) 112 - 121 2021/11 

  BACKGROUNDS: FAM83H is essential for amelogenesis, but recent reports implicate that FAM83H is involved in the tumorigenesis. We previously clarified that TRIM29 binds to FAM83H to regulate keratin distribution and squamous cell migration. However, little is known about FAM83H in normal/malignant skin keratinocytes. OBJECTIVE: To investigate the expression of FAM83H in cutaneous squamous cell carcinoma (SCC) and its physiological function. METHODS: Immunohistochemical analysis and RT-PCR of human SCC tissues were performed. Next, we examined the effect of FAM83H knockdown/overexpression in SCC cell lines using cell proliferation, migration, and invasion assay. To investigate the molecular mechanism, immunoprecipitation of FAM83H was examined. Further, Immunofluorescence staining was performed. Finally, we examined the correlation between the expressions of FAM83H and the keratin distribution. RESULTS: FAM83H expression was lower in SCC lesions than in normal epidermis and correlated with differentiation grade. The mRNA expression levels of FAM83H in SCC tumors were also lower than in normal epidermis. The knockdown of FAM83H enhanced SCC cell migration and invasion, whereas the overexpression of FAM83H led to decreases in both. Furthermore, the knockdown of FAM83H enhanced the cancer cell metastasis in vivo. FAM83H formed a complex with TRIM29 and keratins. The knockdown of FAM83H altered keratin distribution and solubility. Clinically, the loss of FAM83H correlates with an altered keratin distribution. CONCLUSION: Our findings reveal a critical function for FAM83H in regulating keratin distribution, as well as in the migration/invasion of cutaneous SCC, suggesting that FAM83H could be a crucial molecule in the tumorigenesis of cutaneous SCC.
• ハイドロゲルを用いた新規髄膜腫治療標的分子の検討(Analysis of novel therapeutic target for meningioma using hydrogel)

  小田 義崇, 津田 真寿美, 湯澤 明夏, 王 磊, Umma Habiba, 杉野 弘和, 種井 善一, グン 剣萍, 田中 伸哉

  日本病理学会会誌 110 (2) 110 - 110 0300-9181 2021/10
• 非流暢/失文法型原発性進行性失語(naPPA)を呈したPick病

  大槻 美佳, 谷川 聖, 中川 賀嗣, 廣谷 真, 江口 克紀, 白井 慎一, 岩田 育子, 松島 理明, 脇田 雅大, 芳野 正修, 大嶌 祐貴, 水島 慶一, 田中 伸哉, 佐々木 秀直, 矢部 一郎

  日本神経心理学会総会プログラム・予稿集 日本神経心理学会 45回 112 - 112 2021/09
• Fast in vivo fixation of double network hydrogel to bone by monetite surface hybridization

  Takayuki Nonoyama, Lei Wang, Ryuji Kiyama, Naohiro Kashimura, Kazunori Yasuda, Shinya Tanaka, Takayuki Kurokawa, Jian Ping Gong

  JOURNAL OF THE CERAMIC SOCIETY OF JAPAN 129 (9) 584 - 589 1882-0743 2021/09 

  Double network (DN) hydrogels, possessing biocompatibility, low sliding friction, high strength and toughness, are promising as artificial cartilages for next-generation joint disease treatment. For such application, a fast and robust fixation of DN hydrogel to bone tissue in vivo is indispensable. However, bonding the DN hydrogel that contains similar to 90 wt % of water to bone is a grand challenge since glues do not work on hydrated surfaces. Recently, we reported that a DN hydrogel of its subsurface hybridized with low crystalline hydroxyapatite (HAp) can achieve robust fixation to bone after 4 weeks implantation in rabbit knees, owing to the HAp-induced osteogenesis penetration into the hydrogel matrix. For clinical application, achieving a quick fixation at the early stage of implantation remains as a next subject. In this study, instead of HAp, we hybridized calcium monohydrogen phosphate (monetite), which is a HAp precursor calcium phosphate salt, in the subsurface of the DN hydrogel and we observed an increase in the pushout resistance of the DN hydrogel to bone after 1 week implantation, prior to the HAp-induced osteogenesis penetration. In physiological environment, the monetite hybridized in the subsurface of the DN gel spontaneously dissolved to calcium and phosphate ions and then recrystallized to more stable HAp. We consider that the HAp formed in the boundary between the gel and the bone forms physical interlocking that significantly enhances the frictional resistance against the pushout force. The fast temporally pre-fixation to the bone by monetite surface hybridization makes one step closer to the clinical application of the DN gels as artificial cartilages. (C) 2021 The Ceramic Society of Japan. All rights reserved.
• ハイドロキシアパタイト複合化ダブルネットワークゲル上での骨髄間葉系幹細胞の骨分化能評価

  甲斐原 拓真, 王 磊, 津田 真寿美, 野々山 貴行, 黒川 孝幸, きょう 剣萍, 岩崎 倫政, 田中 伸哉, 安田 和則

  日本整形外科学会雑誌 (公社)日本整形外科学会 95 (8) S1561 - S1561 0021-5325 2021/08
• Arid5a Promotes Immune Evasion by Augmenting Tryptophan Metabolism and Chemokine Expression.

  Gyanu Parajuli, Murat Tekguc, James B Wing, Ari Hashimoto, Daisuke Okuzaki, Takeshi Hirata, Atsushi Sasaki, Takahide Itokazu, Haruka Handa, Hirokazu Sugino, Yoshihiro Nishikawa, Hozaifa Metwally, Yuzo Kodama, Shinya Tanaka, Hisataka Sabe, Toshihide Yamashita, Shimon Sakaguchi, Tadamitsu Kishimoto, Shigeru Hashimoto

  Cancer immunology research 9 (8) 862 - 876 2021/08 

  The acquisition of mesenchymal traits leads to immune evasion in various cancers, but the underlying molecular mechanisms remain unclear. In this study, we found that the expression levels of AT-rich interaction domain-containing protein 5a (Arid5a), an RNA-binding protein, were substantially increased in mesenchymal tumor subtypes. The deletion of Arid5a in tumor cell lines enhanced antitumor immunity in immunocompetent mice, but not in immunodeficient mice, suggesting a role for Arid5a in immune evasion. Furthermore, an Arid5a-deficient tumor microenvironment was shown to have robust antitumor immunity, as manifested by suppressed infiltration of granulocytic myeloid-derived suppressor cells and regulatory T cells. In addition, infiltrated T cells were more cytotoxic and less exhausted. Mechanistically, Arid5a stabilized Ido1 and Ccl2 mRNAs and augmented their expression, resulting in enhanced tryptophan catabolism and an immunosuppressive tumor microenvironment. Thus, our findings demonstrate the role of Arid5a beyond inflammatory diseases and suggest Arid5a as a promising target for the treatment of immunotolerant malignant tumors.See related Spotlight by Van den Eynde, p. 854.
• The CD44/COL17A1 pathway promotes the formation of multilayered, transformed epithelia.

  Kei Kozawa, Miho Sekai, Kenji Ohba, Shoko Ito, Hiroaki Sako, Takeshi Maruyama, Mai Kakeno, Takanobu Shirai, Keisuke Kuromiya, Tomoko Kamasaki, Koki Kohashi, Shinya Tanaka, Susumu Ishikawa, Nanami Sato, Shota Asano, Hironori Suzuki, Nobuyuki Tanimura, Yohei Mukai, Noriko Gotoh, Mishie Tanino, Shinya Tanaka, Ken Natsuga, Tomoyoshi Soga, Tomonori Nakamura, Yukihiro Yabuta, Mitinori Saitou, Takahiro Ito, Kenkyo Matsuura, Makoto Tsunoda, Toyone Kikumori, Tadashi Iida, Yasuyuki Mizutani, Yuki Miyai, Kozo Kaibuchi, Atsushi Enomoto, Yasuyuki Fujita

  Current biology : CB 31 (14) 3086 - 3097 2021/07/26 

  At the early stage of cancer development, oncogenic mutations often cause multilayered epithelial structures. However, the underlying molecular mechanism still remains enigmatic. By performing a series of screenings targeting plasma membrane proteins, we have found that collagen XVII (COL17A1) and CD44 accumulate in RasV12-, Src-, or ErbB2-transformed epithelial cells. In addition, the expression of COL17A1 and CD44 is also regulated by cell density and upon apical cell extrusion. We further demonstrate that the expression of COL17A1 and CD44 is profoundly upregulated at the upper layers of multilayered, transformed epithelia in vitro and in vivo. The accumulated COL17A1 and CD44 suppress mitochondrial membrane potential and reactive oxygen species (ROS) production. The diminished intracellular ROS level then promotes resistance against ferroptosis-mediated cell death upon cell extrusion, thereby positively regulating the formation of multilayered structures. To further understand the functional role of COL17A1, we performed comprehensive metabolome analysis and compared intracellular metabolites between RasV12 and COL17A1-knockout RasV12 cells. The data imply that COL17A1 regulates the metabolic pathway from the GABA shunt to mitochondrial complex I through succinate, thereby suppressing the ROS production. Moreover, we demonstrate that CD44 regulates membrane accumulation of COL17A1 in multilayered structures. These results suggest that CD44 and COL17A1 are crucial regulators for the clonal expansion of transformed cells within multilayered epithelia, thus being potential targets for early diagnosis and preventive treatment for precancerous lesions.
• Comprehensive molecular profiling of pulmonary pleomorphic carcinoma.

  Masaaki Nagano, Shinji Kohsaka, Takuo Hayashi, Toshihide Ueno, Shinya Kojima, Aya Shinozaki-Ushiku, Shigeki Morita, Masumi Tsuda, Shinya Tanaka, Toshiya Shinohara, Yuko Omori, Fumiko Sugaya, Hiroaki Kato, Yoshiaki Narita, Jun Nakajima, Kenji Suzuki, Kazuya Takamochi, Hiroyuki Mano

  NPJ precision oncology 5 (1) 57 - 57 2021/06/22 

  Information regarding the molecular features of pulmonary pleomorphic carcinoma (PPC) is insufficient. Here, we performed next-generation sequencing to determine the genomic and transcriptomic profiles of PPC. We sequenced the DNAs and RNAs of 78 specimens from 52 patients with PPC. We analyzed 15 PPC cases to identify intratumoral differences in gene alterations, tumor mutation burden (TMB), RNA expression, and PD-L1 expression between epithelial and sarcomatoid components. The genomic alterations of six cases of primary tumors and corresponding metastatic tumors were analyzed. KRAS mutations (27%) were the most common driver mutations, followed by EGFR (8%), and MET (8%) mutations. Epithelial and sarcomatoid components shared activating driver mutations, and there were no significant differences in CD274 expression or TMB between the two components. However, PD-L1 was highly expressed in the sarcomatoid component of several cases compared with the epithelial component. Primary and metastatic tumors shared oncogenic mutations among genes such as KRAS and TP53, and additional alterations including NOTCH4 mutations were specifically identified in the metastatic regions. Our data suggest that therapies targeting activating driver mutations may be effective for patients with PPC and that immune checkpoint inhibitors of PPC may be recommended after careful assessment of PD-L1 expression in each epithelial and sarcomatoid component.
• Improved Mitochondria! Function by Luseogliflozin Prevents Pancreatic Beta-Cell Damage

  Yuki Yamauchi, Akinobu Nakamura, Takashi Yokota, Kiyohiko Takahashi, Shinichiro Kawata, Kazuhisa Tsuchida, Kazuno Omori, Hiroshi Nomoto, Hiraku Kameda, Kyuyong Cho, Toshihisa Anzai, Shinya Tanaka, Yasuo Terauchi, Hideaki Miyoshi, Tatsuya Atsumi

  DIABETES 70 0012-1797 2021/06
• Loss of H3K27 trimethylation is frequent in IDH1-R132H but not in non-canonical IDH1/2 mutated and 1p/19q codeleted oligodendroglioma: a Japanese cohort study.

  Umma Habiba, Hirokazu Sugino, Roumyana Yordanova, Koki Ise, Zen-Ichi Tanei, Yusuke Ishida, Satoshi Tanikawa, Shunsuke Terasaka, Ken-Ichi Sato, Yuuta Kamoshima, Masahiko Katoh, Motoo Nagane, Junji Shibahara, Masumi Tsuda, Shinya Tanaka

  Acta neuropathologica communications 9 (1) 95 - 95 2021/05/21 

  Oligodendrogliomas are defined by mutation in isocitrate dehydrogenase (NADP(+)) (IDH)1/2 genes and chromosome 1p/19q codeletion. World Health Organisation diagnosis endorses testing for 1p/19q codeletion to distinguish IDH mutant (Mut) oligodendrogliomas from astrocytomas because these gliomas require different treatments and they have different outcomes. Several methods have been used to identify 1p/19q status; however, these techniques are not routinely available and require substantial infrastructure investment. Two recent studies reported reduced immunostaining for trimethylation at lysine 27 on histone H3 (H3K27me3) in IDH Mut 1p/19q codeleted oligodendroglioma. However, the specificity of H3K27me3 immunostaining in this setting is controversial. Therefore, we developed an easy-to-implement immunohistochemical surrogate for IDH Mut glioma subclassification and evaluated a validated adult glioma cohort. We screened 145 adult glioma cases, consisting of 45 IDH Mut and 1p/19q codeleted oligodendrogliomas, 30 IDH Mut astrocytomas, 16 IDH wild-type (Wt) astrocytomas, and 54 IDH Wt glioblastomas (GBMs). We compared immunostaining with DNA sequencing and fluorescent in situ hybridization analysis and assessed differences in H3K27me3 staining between oligodendroglial and astrocytic lineages and between IDH1-R132H and non-canonical (non-R132H) IDH1/2 Mut oligodendroglioma. A loss of H3K27me3 was observed in 36/40 (90%) of IDH1-R132H Mut oligodendroglioma. In contrast, loss of H3K27me3 was never seen in IDH1-R132L or IDH2-mutated 1p/19q codeleted oligodendrogliomas. IDH Mut astrocytoma, IDH Wt astrocytoma and GBM showed preserved nuclear staining in 87%, 94%, and 91% of cases, respectively. A high recursive partitioning model predicted probability score (0.9835) indicated that the loss of H3K27me3 is frequent to IDH1-R132H Mut oligodendroglioma. Our results demonstrate H3K27me3 immunohistochemical evaluation to be a cost-effective and reliable method for defining 1p/19q codeletion along with IDH1-R132H and ATRX immunostaining, even in the absence of 1p/19q testing.
• Difference in the malignancy between RAS and GLI1-transformed astrocytes is associated with frequency of p27KIP1-positive cells in xenograft tissues.

  Ken Sasai, Kouichi Tabu, Takashi Saito, Yukio Matsuba, Takaomi C Saido, Shinya Tanaka

  Pathology, research and practice 223 153465 - 153465 2021/05/05 

  We demonstrate that the introduction of GLI1 is sufficient for immortalized human astrocytes to be transformed whereas FOXM1 fails to induce malignant transformation, suggesting differences between GLI1 and FOXM1 in terms of transforming ability despite both transcription factors being overexpressed in malignant gliomas. Moreover, in investigations of mechanisms underlying relatively less-malignant features of GLI1-transformed astrocytes, we found that p27KIP1-positive cells were frequently observed in xenografts derived from GLI1-transformed astrocytes compared to those from RAS-transformed cells. As shRNA-mediated knockdown of p27KIP1 accelerates tumor progression of GLI1-transformed astrocytes, downregulation of p27KIP1 contributes to malignant features of transformed astrocytes. We propose that the models using immortalized/transformed astrocytes are useful to identify the minimal and most crucial set of changes required for glioma formation.
• 脊髄capillary hemangiomaの臨床病理学的特徴

  種井 善一, 津田 真寿美, 谷川 聖, 杉野 弘和, 石田 雄介, 大竹 安史, 今村 博幸, 小柳 泉, 飛騨 一利, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 38 (Suppl.) 119 - 119 1433-7398 2021/05
• Severe infection including disseminated herpes zoster triggered by subclinical Cushing's disease: a case report.

  Yuki Yamauchi, Hiraku Kameda, Kazuno Omori, Michio Tani, Kyu Yong Cho, Akinobu Nakamura, Hideaki Miyoshi, Shinya Tanaka, Tatsuya Atsumi

  BMC endocrine disorders 21 (1) 84 - 84 2021/04/27 

  BACKGROUND: Subclinical Cushing's disease (SCD) is defined by corticotroph adenoma-induced mild hypercortisolism without typical physical features of Cushing's disease. Infection is an important complication associated with mortality in Cushing's disease, while no reports on infection in SCD are available. To make clinicians aware of the risk of infection in SCD, we report a case of SCD with disseminated herpes zoster (DHZ) with the mortal outcome. CASE PRESENTATION: An 83-year-old Japanese woman was diagnosed with SCD, treated with cabergoline in the outpatient. She was hospitalized for acute pyelonephritis, and her fever gradually resolved with antibiotics. However, herpes zoster appeared on her chest, and the eruptions rapidly spread over the body. She suddenly went into cardiopulmonary arrest and died. Autopsy demonstrated adrenocorticotropic hormone-positive pituitary adenoma, renal abscess, and DHZ. CONCLUSIONS: As immunosuppression caused by SCD may be one of the triggers of severe infection, the patients with SCD should be assessed not only for the metabolic but also for the immunodeficient status.
• Distinct TERT promoter C228T and C250T mutations in a patient with an oligodendroglioma: A case report.

  Yukitomo Ishi, Hiromi Okada, Michinari Okamoto, Hiroaki Motegi, Shinya Tanaka, Tomoko Mitsuhashi, Shigeru Yamaguchi

  Neuropathology : official journal of the Japanese Society of Neuropathology 41 (3) 236 - 242 2021/04/26 

  The majority of oligodendroglial tumors harbor mutations in the telomerase reverse transcriptase (TERT) gene (TERT) promoter and the isocitrate dehydrogenase 1/2 (IDH1/2) gene (IDH1/2), as well as 1p/19q codeletion. Generally, TERT promoter mutations, C250T and C228T, are mutually exclusive. We present a case of oligodendroglioma harboring both C250T and C228T mutations in TERT promoter. A 38-year-old man presented with grand mal seizures and underwent a resection surgery for a left frontal lobe tumor. He was pathologically diagnosed as having oligodendroglioma and was carefully observed. At 48 years of age, he underwent another resection surgery due to tumor regrowth, with the pathological diagnosis of anaplastic oligodendroglioma. Genetic analysis of the initial tumor specimen revealed IDH1 R132H mutation and both C250T and C228T mutations in TERT promoter. Using mutation-specific primers, two mutations were considered to be distributed in different alleles. In the tumor specimen obtained during the second surgery, IDH1 R132H mutation was detected to be similar to that of the initial specimen; however, only C228T mutation was detected in TERT promoter. The 1p/19q codeletion was detected in both the initial and recurrent tumor specimens. According to the sequencing data from the two tumor specimens, although TERT promoter mutation has been considered to be an early genetic event in the tumorigenesis of oligodendroglial tumors, it is likely that the C250T and C228T mutations in TERT promoter are subclonally distributed in the same tumor specimen of the present case.
• Correlation between endothelial CXCR7 expression and clinicopathological factors in oral squamous cell carcinoma.

  Misa Yanagiya, Randa I H Dawood, Nako Maishi, Yasuhiro Hida, Chisaho Torii, Dorcas A Annan, Hiroshi Kikuchi, Aya Yanagawa Matsuda, Tetsuya Kitamura, Yoichi Ohiro, Masanobu Shindoh, Shinya Tanaka, Yoshimasa Kitagawa, Kyoko Hida

  Pathology international 71 (6) 383 - 391 2021/03/30 

  Oral squamous cell carcinoma (OSCC) impairs functionality and sensuousness resulting in poor quality of life. Biomarkers can predict disease trajectory and lead to effective treatments. Transcriptomics have identified genes that are upregulated in tumor endothelial cells (TECs) compared with normal endothelial cells (NECs). Among them, chemokine receptor 7 (CXCR7) is highly expressed in TECs of several cancers and involved in angiogenesis of TECs. However, levels of CXCR7 in OSCC blood vessels have not been fully investigated. In this study, we analyzed the correlation between CXCR7 expression in TECs and clinicopathological factors in OSCC. Immunohistochemistry for CXCR7 and CD34 was performed on 59 OSCC tissue specimens resected between 1996 and 2008 at Hokkaido University Hospital. CXCR7 expression in blood vessels was evaluated by the ratio of CXCR7+/CD34+ blood vessels. CXCR7 expression was 42% and 19% in tumor and non-tumor parts, respectively, suggesting that CXCR7 expression is higher in TECs than in NECs. CXCR7 expression in TECs correlated with advanced T-stage and cancer stage. Overall survival and disease-free survival rates were higher in low-expressing CXCR7 patients than in high-expressing. These results suggest that CXCR7 expression in blood vessels may be a useful diagnostic and prognostic marker for OSCC patients.
• Rapid reprogramming of tumour cells into cancer stem cells on double-network hydrogels.

  Jun Suzuka, Masumi Tsuda, Lei Wang, Shinji Kohsaka, Karin Kishida, Shingo Semba, Hirokazu Sugino, Sachiyo Aburatani, Martin Frauenlob, Takayuki Kurokawa, Shinya Kojima, Toshihide Ueno, Yoshihiro Ohmiya, Hiroyuki Mano, Kazunori Yasuda, Jian Ping Gong, Shinya Tanaka

  Nature biomedical engineering 5 (8) 914 - 925 2021/03/29 

  Cancer recurrence can arise owing to rare circulating cancer stem cells (CSCs) that are resistant to chemotherapies and radiotherapies. Here, we show that a double-network hydrogel can rapidly reprogramme differentiated cancer cells into CSCs. Spheroids expressing elevated levels of the stemness genes Sox2, Oct3/4 and Nanog formed within 24 h of seeding the gel with cells from any of six human cancer cell lines or with brain cancer cells resected from patients with glioblastoma. Human brain cancer cells cultured on the double-network hydrogel and intracranially injected in immunodeficient mice led to higher tumorigenicity than brain cancer cells cultured on single-network gels. We also show that the double-network gel induced the phosphorylation of tyrosine kinases, that gel-induced CSCs from primary brain cancer cells were eradicated by an inhibitor of the platelet-derived growth factor receptor, and that calcium channel receptors and the protein osteopontin were essential for the regulation of gel-mediated induction of stemness in brain cancer cells.
• A Rare Combination: Cardiac Myxoma and Aortic Stenosis.

  Takao Konishi, Daisuke Hotta, Shinya Tanaka, Toshihisa Anzai

  Internal medicine (Tokyo, Japan) 60 (6) 961 - 962 2021/03/15
• High-grade neuroepithelial tumor with BCL6 corepressor-alteration presenting pathological and radiological calcification: A case report.

  Yukitomo Ishi, Ai Shimizu, Emi Takakuwa, Minako Sugiyama, Michinari Okamoto, Hiroaki Motegi, Shinsuke Hirabayashi, Yuko Cho, Akihiro Iguchi, Atsushi Manabe, Sumihito Nobusawa, Shinya Tanaka, Shigeru Yamaguchi

  Pathology international 71 (5) 348 - 354 2021/03/13 

  A 5-year-old girl presented with headache and vomiting. Head computed tomography and magnetic resonance imaging showed a right frontal lobe tumor with marked calcification. The patient underwent resection surgery with suspicion of anaplastic ependymoma, and the tumor was gross totally removed. Pathological examination revealed areas of dense tumor cells with a high nucleocytoplasmic ratio and myxoid areas consisting of tumor cells with a round-shaped nucleus and eosinophilic cytoplasm. Perivascular pseudorosette, necrosis, circumscribed growth, and microcalcification were also observed. Immunohistochemistry demonstrated negative staining for glial fibrillary protein and epithelial membrane antigen. Diagnosis of a high-grade neuroepithelial tumor (HGNET) with BCL6 corepressor (BCOR) alteration was made based on pathological findings and internal tandem duplication in the exon 15 of BCOR. Although calcification on radiological and pathological examination is not typical, it would be essential to recognize that calcification could appear in HGNET-BCOR.
• 成人型Krabbe病の一剖検例

  佐々木 美羽, 江端 美織, 小田 義崇, 濱内 朗子, 谷川 聖, 種井 善一, 杉野 弘和, 石田 雄介, 佐光 一也, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 110 (1) 379 - 379 0300-9181 2021/03
• Involvement of BMP and Wnt Signals Leadingto Epithelial-Mesenchymal Transition in Colon Adenocarcinoma with Heterotopic Ossification.

  Naho Katono, Masumi Tsuda, Jun Suzuka, Yoshitaka Oda, Lei Wang, Zen-Ichi Tanei, Mishie Tanino, Takanobu Ohata, Eisuke Nagabuchi, Yusuke Ishida, Shunsuke Kimura, Toshihiko Iwanaga, Shinya Tanaka

  Annals of clinical and laboratory science 51 (2) 271 - 276 2021/03 

  Here we present the case of a 73-year-old male with rectal adenocarcinoma with heterotopic ossification (HO). Cancer-associated HO in the digestive system is rare. Thus, the precise mechanism and clinicopathological significance of HO have not yet been defined. To clarify the molecular mechanisms of HO, we analyzed the expression levels of signaling molecules related to epithelial-mesenchymal transition (EMT) that lead to ossification in the tumor cells discriminating the ossified area (HO-area) and non-ossified area (non-HO area). Expression levels of BMP4 were elevated in both areas, whereas BMP2 was specifically increased in the HO-area by qPCR. EMT-related molecules such as Snail and Slug were especially higher in the HO-area. By immunohistochemistry, the expression of Smad4, nuclear staining of β-catenin, and the phosphorylated form of GSK-3β were detectable in both areas, and GSK-3β was highly phosphorylated in the HO-area. The tumor growth rate was extremely high, with the Ki-67 labeling index at 90%. In the HO-area, osteoblasts with alkaline phosphatase expression were distributed surrounding the tumor cells. This is the first demonstration of the involvement of EMT in HO of colon cancer through BMP/SMAD and WNT/β-catenin signaling pathways, which are especially prominent in the HO-area leading to the osteogenic property.
• Synthetic poly(2-acrylamido-2-methylpropanesulfonic acid) gel induces chondrogenic differentiation of ATDC5 cells via a novel protein reservoir function.

  Shingo Semba, Nobuto Kitamura, Masumi Tsuda, Keiko Goto, Sadamu Kurono, Yoshihiro Ohmiya, Takayuki Kurokawa, Jian Ping Gong, Kazunori Yasuda, Shinya Tanaka

  Journal of biomedical materials research. Part A 109 (3) 354 - 364 2021/03 [Refereed][Not invited]
   

  We previously demonstrated that a synthetic negatively charged poly(2-acrylamido-2-methylpropanesulfonic acid) (PAMPS) gel induced chondrogenic differentiation of ATDC5 cells. In this study, we clarified the underlying molecular mechanism, in particular, focusing on the events that occurred at the interface between the gel and the cells. Gene expression profiling revealed that the expression of extracellular components was enhanced in the ATDC5 cells that were cultured on the PAMPS gel, suggesting that extracellular proteins secreted from the ATDC5 cells might be adsorbed in the PAMPS gel, thereby contributing to the induction of chondrogenic differentiation. Therefore, we created "Treated-PAMPS gel," which adsorbed various proteins secreted from the cultured ATDC5 cells during 7 days. Proteomic analysis identified 27 proteins, including extracellular matrix proteins such as Types I, III, and V collagens and thrombospondin (THBS) in the Treated-PAMPS gel. The Treated-PAMPS gel preferentially induced expression of chondrogenic markers, namely, aggrecan and Type II collagen, in the ATDC5 cells compared with the untreated PAMPS gel. Addition of recombinant THBS1 to the ATDC5 cells significantly enhanced the PAMPS-induced chondrogenic differentiation, whereas knockdown of THBS1 completely abolished this response. In conclusion, we demonstrated that the PAMPS gel has the potential to induce chondrogenic differentiation through novel reservoir functions, and the adsorbed THBS plays a significant role in the induction.
• Ependymoma-like tumor with mesenchymal differentiation harboring C11orf95-NCOA1/2 or -RELA fusion: A hitherto unclassified tumor related to ependymoma.

  Ran Tomomasa, Yasuhito Arai, Reika Kawabata-Iwakawa, Kohei Fukuoka, Yoshiko Nakano, Natsuko Hama, Satoshi Nakata, Nozomi Suzuki, Yukitomo Ishi, Shinya Tanaka, Jun A Takahashi, Yoshiaki Yuba, Mitsutaka Shiota, Atsushi Natsume, Michihiro Kurimoto, Yoshiki Shiba, Mikiko Aoki, Kazuki Nabeshima, Toshiyuki Enomoto, Tooru Inoue, Junya Fujimura, Akihide Kondo, Takashi Yao, Naoki Okura, Takanori Hirose, Atsushi Sasaki, Masahiko Nishiyama, Koichi Ichimura, Tatsuhiro Shibata, Junko Hirato, Hideaki Yokoo, Sumihito Nobusawa

  Brain pathology (Zurich, Switzerland) 31 (3) e12943  2021/02/12 

  Recurrent fusion genes involving C11orf95, C11orf95-RELA, have been identified only in supratentorial ependymomas among primary CNS tumors. Here, we report hitherto histopathologically unclassifiable high-grade tumors, under the tentative label of "ependymoma-like tumors with mesenchymal differentiation (ELTMDs)," harboring C11orf95-NCOA1/2 or -RELA fusion. We examined the clinicopathological and molecular features in five cases of ELTMDs. Except for one adult case (50 years old), all cases were in children ranging from 1 to 2.5 years old. All patients presented with a mass lesion in the cerebral hemisphere. Histologically, all cases demonstrated a similar histology with a mixture of components. The major components were embryonal-appearing components forming well-delineated tumor cell nests composed of small uniform cells with high proliferative activity, and spindle-cell mesenchymal components with a low- to high-grade sarcoma-like appearance. The embryonal-appearing components exhibited minimal ependymal differentiation including a characteristic EMA positivity and tubular structures, but histologically did not fit with ependymoma because they lacked perivascular pseudorosettes, a histological hallmark of ependymoma, formed well-delineated nests, and had diffuse and strong staining for CAM5.2. Molecular analysis identified C11orf95-NCOA1, -NCOA2, and -RELA in two, one, and two cases, respectively. t-distributed stochastic neighbor embedding analysis of DNA methylation data from two cases with C11orf95-NCOA1 or -NCOA2 and a reference set of 380 CNS tumors revealed that these two cases were clustered together and were distinct from all subgroups of ependymomas. In conclusion, although ELTMDs exhibited morphological and genetic associations with supratentorial ependymoma with C11orf95-RELA, they cannot be regarded as ependymoma. Further analyses of more cases are needed to clarify their differences and similarities.
• Prognostic value of admission serum magnesium in acute myocardial infarction complicated by malignant ventricular arrhythmias.

  Yoshifumi Mizuguchi, Takao Konishi, Toshiyuki Nagai, Tomoya Sato, Sakae Takenaka, Atsushi Tada, Yuta Kobayashi, Hirokazu Komoriyama, Yoshiya Kato, Kazunori Omote, Takuma Sato, Kiwamu Kamiya, Shingo Tsujinaga, Hiroyuki Iwano, Kenjiro Kikuchi, Shinya Tanaka, Toshihisa Anzai

  The American journal of emergency medicine 44 100 - 105 2021/02/05 

  OBJECTIVES: Although electrolyte abnormalities are related to worse clinical outcomes in patients with acute myocardial infarction (AMI), little is known about the association between admission serum magnesium level and adverse events in AMI patients complicated by out-of-hospital cardiac arrest presenting with malignant ventricular arrhythmias (OHCA-MVA). We investigated the prognostic value of serum magnesium level on admission in these patients. METHODS: We retrospectively analyzed the data of 165 consecutive reperfused AMI patients complicated with OHCA-MVA between April 2007 and February 2020 in our university hospital. Serum magnesium concentration was measured on admission. The primary outcome was in-hospital death. RESULTS: Fifty-four patients (33%) died during hospitalization. Higher serum magnesium level was significantly related to in-hospital death (Fine & Gray's test; p < 0.001). In multivariable logistic regression analyses, serum magnesium level on admission was independently associated with in-hospital death (hazard ratio 2.68, 95% confidence interval 1.24-5.80) even after adjustment for covariates. Furthermore, the incidences of cardiogenic shock necessitating an intra-aortic balloon pump (p = 0.005) or extracorporeal membrane oxygenation (p < 0.001), tracheal intubation (p < 0.001) and persistent vegetative state (p = 0.002) were significantly higher in patients with higher serum magnesium level than in those with lower serum magnesium level. CONCLUSIONS: In reperfused AMI patients complicated by OHCA-MVA, admission serum magnesium level might be a potential surrogate marker for predicting in-hospital death.
• Isotope Microscopic Observation of Osteogenesis Process Forming Robust Bonding of Double Network Hydrogel to Bone.

  Takayuki Nonoyama, Lei Wang, Masumi Tsuda, Yuki Suzuki, Ryuji Kiyama, Kazunori Yasuda, Shinya Tanaka, Kousuke Nagata, Ryosuke Fujita, Naoya Sakamoto, Noriyuki Kawasaki, Hisayoshi Yurimoto, Jian Ping Gong

  Advanced healthcare materials 10 (3) e2001731  2021/02 

  Tough double network (DN) hydrogels are promising substitutes of soft supporting tissues such as cartilage and ligaments. For such applications, it is indispensable to robustly fix the hydrogels to bones with medically feasible methods. Recently, robustly bonding the DN hydrogels to defected bones of rabbits in vivo has been proved successful. The low crystalline hydroxyapatite (HAp) of calcium-phosphate-hydroxide salt coated on the surface layer of the DN hydrogels induced spontaneous osteogenesis penetrating into the semi-permeable hydrogels to form a gel/bone composite layer. In this work, the 44 Ca isotope-doped HAp/DN hydrogel is implanted in a defect of rabbit femoral bone and the dynamic osteogenesis process at the gel/bone interface is analyzed by tracing the calcium isotope ratio using isotope microscopy. The synthetic HAp hybridized on the surface layer of DN gel dissolves rapidly in the first two weeks by inflammation, and then the immature bone with a gradient structure starts to form in the gel region, reutilizing the dissolved Ca ions. These results reveal, for the first time, that synthetic HAp is reutilized for osteogenesis. These facts help to understand the lifetime of bone absorbable materials and to elucidate the mechanism of spontaneous, non-toxic, but strong fixation of hydrogels to bones.
• Cardiac-specific loss of mitoNEET expression is linked with age-related heart failure.

  Takaaki Furihata, Shingo Takada, Naoya Kakutani, Satoshi Maekawa, Masaya Tsuda, Junichi Matsumoto, Wataru Mizushima, Arata Fukushima, Takashi Yokota, Nobuyuki Enzan, Shouji Matsushima, Haruka Handa, Yoshizuki Fumoto, Junko Nio-Kobayashi, Toshihiko Iwanaga, Shinya Tanaka, Hiroyuki Tsutsui, Hisataka Sabe, Shintaro Kinugawa

  Communications biology 4 (1) 138 - 138 2021/01/29 

  Heart failure (HF) occurs frequently among older individuals, and dysfunction of cardiac mitochondria is often observed. We here show the cardiac-specific downregulation of a certain mitochondrial component during the chronological aging of mice, which is detrimental to the heart. MitoNEET is a mitochondrial outer membrane protein, encoded by CDGSH iron sulfur domain 1 (CISD1). Expression of mitoNEET was specifically downregulated in the heart and kidney of chronologically aged mice. Mice with a constitutive cardiac-specific deletion of CISD1 on the C57BL/6J background showed cardiac dysfunction only after 12 months of age and developed HF after 16 months; whereas irregular morphology and higher levels of reactive oxygen species in their cardiac mitochondria were observed at earlier time points. Our results suggest a possible mechanism by which cardiac mitochondria may gradually lose their integrity during natural aging, and shed light on an uncharted molecular basis closely related to age-associated HF.
• Takotsubo syndrome in association with acute myocardial infarction: diagnostic caveats and clinical implications. Authors' reply.

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Shinya Tanaka, Toshihisa Anzai

  Kardiologia polska 79 (1) 96 - 96 2021/01/25
• The effectiveness of drug-coated balloons for two dissimilar calcific lesions assessed by near-infrared spectroscopy intravascular ultrasound and optical coherence tomography.

  Takao Konishi, Kohei Saiin, Youji Tamaki, Hiroyuki Natsui, Tomoya Sato, Sakae Takenaka, Atsushi Tada, Yoshifumi Mizuguchi, Yuta Kobayashi, Hirokazu Komoriyama, Yoshiya Kato, Takuma Sato, Rui Kamada, Kiwamu Kamiya, Toshiyuki Nagai, Shinya Tanaka, Toshihisa Anzai

  Cardiology journal 28 (5) 794 - 795 2021
• Lewy pathology of the esophagus correlates with the progression of Lewy body disease: a Japanese cohort study of autopsy cases.

  Zen-Ichi Tanei, Yuko Saito, Shinji Ito, Tomoyasu Matsubara, Atsuko Motoda, Mikihiro Yamazaki, Yasuhiro Sakashita, Ito Kawakami, Masako Ikemura, Shinya Tanaka, Renpei Sengoku, Tomio Arai, Shigeo Murayama

  Acta neuropathologica 141 (1) 25 - 37 2021/01 

  Lewy body disease (LBD) is a spectrum of progressive neurodegenerative disorders characterized by the wide distribution of Lewy bodies and neurites in the central and peripheral nervous system (CNS, PNS). Clinical diagnoses include Parkinson's disease (PD), dementia with Lewy bodies, or pure autonomic failure. All types of LBD are accompanied by non-motor symptoms (NMSs) including gastrointestinal dysfunctions such as constipation. Its relationship to Lewy body-related α-synucleinopathy (Lewy pathology) of the enteric nervous system (ENS) is attracting attention because it can precede the motor symptoms. To clarify the role of ENS Lewy pathology in disease progression, we performed a clinicopathological study using the Brain Bank for Aging Research in Japan. Five-hundred and eighteen cases were enrolled in the study. Lewy pathology of the CNS and PNS, including the lower esophagus as a representative of the ENS, was examined via autopsy findings. Results showed that one-third of older people (178 cases, 34%) exhibited Lewy pathology, of which 78 cases (43.8%) exhibited the pathology in the esophagus. In the esophageal wall, Auerbach's plexus (41.6%) was most susceptible to the pathology, followed by the adventitia (33.1%) and Meissner's plexus (14.6%). Lewy pathology of the esophagus was significantly associated with autonomic failures such as constipation (p < 0.0001) and among PNS regions, correlated the most with LBD progression (r = 0.95, p < 0.05). These findings suggest that the propagation of esophageal Lewy pathology is a predictive factor of LBD.
• Brain-Derived Neurotrophic Factor Improves Impaired Fatty Acid Oxidation Via the Activation of Adenosine Monophosphate-Activated Protein Kinase-ɑ - Proliferator-Activated Receptor-r Coactivator-1ɑ Signaling in Skeletal Muscle of Mice With Heart Failure.

  Junichi Matsumoto, Shingo Takada, Takaaki Furihata, Hideo Nambu, Naoya Kakutani, Satoshi Maekawa, Wataru Mizushima, Ippei Nakano, Arata Fukushima, Takashi Yokota, Shinya Tanaka, Haruka Handa, Hisataka Sabe, Shintaro Kinugawa

  Circulation. Heart failure 14 (1) e005890  2021/01 

  BACKGROUND: We recently reported that treatment with rhBDNF (recombinant human brain-derived neurotrophic factor) improved the reduced exercise capacity of mice with heart failure (HF) after myocardial infarction (MI). Since BDNF is reported to enhance fatty acid oxidation, we herein conducted an in vivo investigation to determine whether the improvement in exercise capacity is due to the enhancement of the fatty acid oxidation of skeletal muscle via the AMPKα-PGC1α (adenosine monophosphate-activated protein kinase-ɑ-proliferator-activated receptor-r coactivator-1ɑ) axis. METHODS: MI and sham operations were conducted in C57BL/6J mice. Two weeks postsurgery, we randomly divided the MI mice into groups treated with rhBDNF or vehicle for 2 weeks. AMPKα-PGC1α signaling and mitochondrial content in the skeletal muscle of the mice were evaluated by Western blotting and transmission electron microscopy. Fatty acid β-oxidation was examined by high-resolution respirometry using permeabilized muscle fiber. BDNF-knockout mice were treated with 5-aminoimidazole-4-carboxamide-1-beta-d-riboruranoside, an activator of AMPK. RESULTS: The rhBDNF treatment significantly increased the expressions of phosphorylated AMPKα and PGC1α protein and the intermyofibrillar mitochondrial density in the MI mice. The lowered skeletal muscle mitochondrial fatty acid oxidation was significantly improved in the rhBDNF-treated MI mice. The reduced exercise capacity and mitochondrial dysfunction of the BDNF-knockout mice were improved by 5-aminoimidazole-4-carboxamide-1-beta-d-riboruranoside. CONCLUSIONS: Beneficial effects of BDNF on the exercise capacity of mice with HF are mediated through an enhancement of fatty acid oxidation via the activation of AMPKα-PGC1α in skeletal muscle. BDNF may become a therapeutic option to improve exercise capacity as an alternative or adjunct to exercise training.
• CMKLR1は肝癌幹細胞標的治療のための候補分子である

  谷 道夫, 津田 真寿美, 鈴鹿 淳, 王 磊, 武冨 紹信, 田中 伸哉

  日本癌学会総会記事 79回 PJ14 - 1 0546-0476 2020/10
• CMKLR1は肝癌幹細胞標的治療のための候補分子である

  谷 道夫, 津田 真寿美, 鈴鹿 淳, 王 磊, 武冨 紹信, 田中 伸哉

  日本癌学会総会記事 79回 PJ14 - 1 0546-0476 2020/10
• Chitin-Based Double-Network Hydrogel as Potential Superficial Soft-Tissue-Repairing Materials

  Junchao Huang, Martin Frauenlob, Yuki Shibata, Lei Wang, Tasuku Nakajima, Takayuki Nonoyama, Masumi Tsuda, Shinya Tanaka, Takayuki Kurokawa, Jian Ping Gong

  Biomacromolecules 1525-7797 2020/09/27
• Loss of dynamin-related protein 1 (Drp1) does not affect epidermal development or UVB-induced apoptosis but does accelerate UVB-induced carcinogenesis.

  Teruki Yanagi, Shinya Kitamura, Keisuke Imafuku, Asuka Suto, Takuya Maeda, Shinya Tanaka, Hiromi Sesaki, Riichiro Abe, Hiroshi Shimizu

  Journal of dermatological science 99 (2) 109 - 118 2020/08 

  BACKGROUND: Mitochondrial morphology is controlled by fission and fusion. Dynamin-related protein 1 (Drp1, dynamin-1-like protein (Dnml1)) regulates mitochondrial fission, which is associated with cell division and apoptosis. We previously reported that DRP1 is indispensable for cell growth in cutaneous squamous cell carcinoma. However, little is known about Drp1 in normal epidermis/keratinocytes. OBJECTIVES: We investigated the function of Drp1 in normal epidermis/keratinocytes. METHODS: Epidermis-specific Drp1 knockout (EKO) mice were analyzed. RESULTS: Epidermal development in the EKO mice were indistinguishable from those in the wild-type (WT) mice. Ultrastructural analysis and immunohistochemistry revealed that the mitochondria of keratinocytes in the EKO mice were neither elongated nor constricted. Drp1 knockdown did not diminish the cell growth of normal human keratinocytes. Both in vivo and in vitro, UVB-induced apoptosis in the EKO epidermis and keratinocytes did not differ from that in the WT mice. In chronic UVB-irradiation, the loss of Drp1 sensitized the epidermis to the development of skin tumors. Clinically, DRP1 is expressed more highly in sun-exposed skin than in non-exposed skin in individuals under age 40, but not in those over age 60. CONCLUSION: EKO mice demonstrate that Drp1 is dispensable for the development and apoptosis of the epidermis. Drp1 plays critical roles in malignant tumors; thus, the molecular machinery of mitochondrial dynamics involving Drp1 could be a novel therapeutic target for malignant keratinocytic lesions. On the other hand, the anti-tumorigenic role of Drp1 in chronic UVB-induced carcinogenesis need to be further investigated.
• バイオマテリアルによる肝癌幹細胞の新規誘導法の開発とその解析

  谷 道夫, 津田 真寿美, 鈴鹿 淳, 王 磊, 杉野 弘和, 谷川 聖, 石田 雄介, グン 剣萍, 田中 伸哉, 武冨 紹信

  日本外科学会定期学術集会抄録集 (一社)日本外科学会 120回 DP - 6 2020/08
• バイオマテリアルによる肝癌幹細胞の新規誘導法の開発とその解析

  谷 道夫, 津田 真寿美, 鈴鹿 淳, 王 磊, 杉野 弘和, 谷川 聖, 石田 雄介, グン 剣萍, 田中 伸哉, 武冨 紹信

  日本外科学会定期学術集会抄録集 (一社)日本外科学会 120回 DP - 6 2020/08
• Acute anterior myocardial infarction complicated by takotsubo syndrome: the value of multimodality imaging.

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Shinya Tanaka, Toshihisa Anzai

  Kardiologia polska 78 (10) 1055 - 1056 2020/07/07 [Refereed][Not invited]
  
• Prognostic role of H3K27M mutation, histone H3K27 methylation status, and EZH2 expression in diffuse spinal cord gliomas.

  Yukitomo Ishi, Soichiro Takamiya, Toshitaka Seki, Kazuyoshi Yamazaki, Kazutoshi Hida, Kanako C Hatanaka, Yusuke Ishida, Yoshitaka Oda, Shinya Tanaka, Shigeru Yamaguchi

  Brain tumor pathology 37 (3) 81 - 88 2020/07 [Refereed][Not invited]
   

  The objective of this study is to clarify clinical significance of the H3F3A K27M mutation (H3K27M) and analyze the correlation between H3K27M, H3K27me3 status, and EZH2 expression and prognosis in spinal cord gliomas. Patients with spinal cord diffuse glioma regardless of World Health Organization (WHO) grade underwent genetic analysis for H3F3A, HIST1H3B, TERT promoter, IDH1/2, and BRAF. H3K27me3 status and EZH2 expression were analyzed through immunohistochemistry. Thereafter, the association between H3K27M, H3K27me3 status, and EZH2 expression and prognosis was retrospectively analyzed using the log-rank test. A total of 26 cases, 5 with WHO grade 4, 9 with grade 3, and 12 with grade 2 glioma, were analyzed. Although WHO grade 2 cases tended to present favorable overall survival, the difference was not statistically significant. H3K27M, which was detected in four grade 4 cases (80%) and three grade 3 cases (33%), was not associated with prognosis among grade 3 and 4 cases. Among WHO grade 2-4 cases, the combination of retained H3K27me3 and negative EZH2 expression was correlated with favorable overall survival (p = 0.03). The combination of H3K27me3 status and EZH2 expression was considered as a potential prognostic marker in WHO grade 2-4 diffuse spinal cord gliomas.
• Integrin α4 Mediates ATDC5 Cell Adhesion to Negatively Charged Synthetic Polymer Hydrogel Leading to Chondrogenic Differentiatio

  Daiki Hashimoto, Shingo Semba, Masumi Tsuda, Takayuki Kurokawa, Nobuto Kitamura, Kazunori Yasuda, Jian Ping Gong, Shinya Tanaka

  Biochemical and Biophysical Research Communications 528 (1) 120 - 126 2020/05 [Refereed][Not invited]
   

  Negatively charged synthetic hydrogels have been known to facilitate various cellular responses including cell adhesion, proliferation, and differentiation; however, the molecular mechanism of hydrogel-dependent control of cell behavior remains unclear. Recently, we reported that negatively charged poly(2-acrylamido-2-methylpropanesulfonic acid) (PAMPS) gel induces chondrogenic differentiation of ATDC5 cells via novel protein reservoir function. In this study, we identified the cell adhesion molecules binding to PAMPS gels that act as mechanoreceptors. First, we performed a pull-down assay by particle gels using cell membrane proteins of ATDC5, and found that multiple membrane proteins bound to the PAMPS gel, whereas the uncharged poly(N,N'-dimethylacrylamide) gel as control did not bind to any membrane proteins. Western blot analysis indicated differential binding of integrin (ITG) isoforms to the PAMPS gel, in which the α4 isoform, but not α5 and αv, efficiently bound to the PAMPS gel. ITG α4 knockdown decreased cell spreading of ATDC5 on PAMPS gels, whereas the enhanced expression increased the behavior. Furthermore, ITG α4 depletion suppressed PAMPS gel-induced expression of bone morphogenic protein (BMP) 4 contributing to chondrogenic differentiation, in concordance with the reduction of ERK activation. These results demonstrated that membrane protein binding to PAMPS gels occurred in a charge-dependent manner, and that ITG α4 plays a crucial role in cell spreading on PAMPS gels and acts as a mechanoreceptor triggering cellular signaling leads to chondrogenic differentiation.
• Signaling adaptor protein Crk is involved in malignant feature of pancreatic cancer associated with phosphorylation of c-Met.

  Satoko Uemura, Lei Wang, Masumi Tsuda, Jun Suzuka, Satoshi Tanikawa, Hirokazu Sugino, Toru Nakamura, Tomoko Mitsuhashi, Satoshi Hirano, Shinya Tanaka

  Biochemical and biophysical research communications 524 (2) 378 - 384 2020/04/02 

  Signaling adaptor protein Crk has been shown to play an important role in various human cancers. Crk links tyrosine kinases and guanine nucleotide exchange factors (GEFs) such as C3G and Dock180 to activate small G-proteins Rap and Rac, respectively. In pancreatic cancer, various molecular targeted therapies have provided no significant therapeutic benefit for the patients so far due to constitutive activation of KRAS by frequent KRAS mutation. Therefore, the establishment of novel molecular targeted therapy in KRAS-independent manner is required. Here, we investigated a potential of Crk as a therapeutic target in pancreatic cancer. Immunohistochemistry on human pancreatic cancer specimens revealed that the patients with high expression of Crk had a worse prognosis than those with low expression. We established Crk-knockdown pancreatic cancer cells by siRNA using PANC-1, AsPC-1, and MIA PaCa-2 cells, which showed decreased cell proliferation, invasion, and adhesion. In Crk-knockdown pancreatic cancer cells, the decrease of c-Met phosphorylation was observed. In the orthotopic xenograft model, Crk depletion prolonged survival of mice significantly. Thus, signaling adaptor protein Crk is involved in malignant potential of pancreatic cancer associated with decrease of c-Met phosphorylation, and Crk can be considered to be a potential therapeutic molecular target.
• Cisplatin Relocalizes RNA Binding Protein HuR and Enhances the Oncolytic Activity of E4orf6 Deleted Adenovirus

  Umma Habiba, Elora Hossain, Aya Yanagawa-Matsuda, Abu Faem Mohammad Almas Chowdhury, Masumi Tsuda, Asad-uz- Zaman, Shinya Tanaka, Fumihiro Higashino

  Cancers 12 (4) 809 - 809 2020/03/27 [Refereed]
   

  The combination of adenoviruses and chemotherapy agents is a novel approach for human cancer therapeutics. A meticulous analysis between adenovirus and chemotherapeutic agents can help to design an effective anticancer therapy. Human antigen R (HuR) is an RNA binding protein that binds to the AU-rich element (ARE) of specific mRNA and is involved in the export and stabilization of ARE-mRNA. Our recent report unveiled that the E4orf6 gene deleted oncolytic adenovirus (dl355) replicated for certain types of cancers where ARE-mRNA is stabilized. This study aimed to investigate whether a combined treatment of dl355 and Cis-diamminedichloroplatinum (CDDP) can have a synergistic cell-killing effect on cancer cells. We confirmed the effect of CDDP in nucleocytoplasmic HuR shuttling. In vitro and in vivo experiments showed the enhancement of cancer cell death by apoptosis induction and a significant reduction in tumor growth following combination treatment. These results suggested that combination therapy exerted a synergistic antitumor activity by upregulation of CDDP induced cytoplasmic HuR, which led to ARE mRNA stabilization and increased virus proliferation. Besides, the enhanced cell-killing effect was due to the activation of the intrinsic apoptotic pathway. Therefore, the combined treatment of CDDP and dl355 could represent a rational approach for cancer therapy.
• Emery-Dreifuss型筋ジストロフィーに伴う心不全の1剖検例

  五味川 龍, 石田 雄介, 桑原 健, 石垣 隆弘, 谷川 聖, 小田 義崇, 王 磊, 杉野 弘和, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 109 (1) 492 - 492 0300-9181 2020/03
• 前立腺癌のGleason pattern評価のためのSemantic segmentationモデル及びRaspberry Pi端末を用いた応用

  遠田 建, 伊勢 昂生, 石田 雄介, 桑原 健, 谷川 聖, 小田 義崇, 王 磊, 杉野 弘和, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 109 (1) 496 - 496 0300-9181 2020/03
• 新規変異BRAFV601K変異を認める良性脳腫瘍(毛様体性星細胞腫)の一例

  榎枝 未紗, 小田 義崇, 津田 真寿美, 飛弾 一利, 杉野 弘和, 谷川 聖, 鈴鹿 淳, 王 磊, 石田 雄介, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 109 (1) 497 - 497 0300-9181 2020/03
• MM2 cortical form of sporadic Creutzfeldt-Jakob disease without progressive dementia and akinetic mutism: A case deviating from current diagnostic criteria.

  Ikuko Takahashi-Iwata, Ichiro Yabe, Akihiko Kudo, Katsuya Eguchi, Masahiro Wakita, Shinichi Shirai, Masaaki Matsushima, Takanobu Toyoshima, Susumu Chiba, Satoshi Tanikawa, Shinya Tanaka, Katsuya Satoh, Tetsuyuki Kitamoto, Hidenao Sasaki

  Journal of the neurological sciences 412 116759 - 116759 2020/02/24 [Refereed][Not invited]
  
• Pineoblastoma segregates into molecular sub-groups with distinct clinico-pathologic features: a Rare Brain Tumor Consortium registry study.

  Bryan K Li, Alexandre Vasiljevic, Christelle Dufour, Fupan Yao, Ben L B Ho, Mei Lu, Eugene I Hwang, Sridharan Gururangan, Jordan R Hansford, Maryam Fouladi, Sumihito Nobusawa, Annie Laquerriere, Marie-Bernadette Delisle, Jason Fangusaro, Fabien Forest, Helen Toledano, Palma Solano-Paez, Sarah Leary, Diane Birks, Lindsey M Hoffman, Alexandru Szathmari, Cécile Faure-Conter, Xing Fan, Daniel Catchpoole, Li Zhou, Kris Ann P Schultz, Koichi Ichimura, Guillaume Gauchotte, Nada Jabado, Chris Jones, Delphine Loussouarn, Karima Mokhtari, Audrey Rousseau, David S Ziegler, Shinya Tanaka, Scott L Pomeroy, Amar Gajjar, Vijay Ramaswamy, Cynthia Hawkins, Richard G Grundy, D Ashley Hill, Eric Bouffet, Annie Huang, Anne Jouvet

  Acta neuropathologica 139 (2) 223 - 241 2020/02 [Refereed][Not invited]
   

  Pineoblastomas (PBs) are rare, aggressive pediatric brain tumors of the pineal gland with modest overall survival despite intensive therapy. We sought to define the clinical and molecular spectra of PB to inform new treatment approaches for this orphan cancer. Tumor, blood, and clinical data from 91 patients with PB or supratentorial primitive neuroectodermal tumor (sPNETs/CNS-PNETs), and 2 pineal parenchymal tumors of intermediate differentiation (PPTIDs) were collected from 29 centres in the Rare Brain Tumor Consortium. We used global DNA methylation profiling to define a core group of PB from 72/93 cases, which were delineated into five molecular sub-groups. Copy number, whole exome and targeted sequencing, and miRNA expression analyses were used to evaluate the clinico-pathologic significance of each sub-group. Tumors designated as group 1 and 2 almost exclusively exhibited deleterious homozygous loss-of-function alterations in miRNA biogenesis genes (DICER1, DROSHA, and DGCR8) in 62 and 100% of group 1 and 2 tumors, respectively. Recurrent alterations of the oncogenic MYC-miR-17/92-RB1 pathway were observed in the RB and MYC sub-group, respectively, characterized by RB1 loss with gain of miR-17/92, and recurrent gain or amplification of MYC. PB sub-groups exhibited distinct clinical features: group 1-3 arose in older children (median ages 5.2-14.0 years) and had intermediate to excellent survival (5-year OS of 68.0-100%), while Group RB and MYC PB patients were much younger (median age 1.3-1.4 years) with dismal survival (5-year OS 37.5% and 28.6%, respectively). We identified age < 3 years at diagnosis, metastatic disease, omission of upfront radiation, and chr 16q loss as significant negative prognostic factors across all PBs. Our findings demonstrate that PB exhibits substantial molecular heterogeneity with sub-group-associated clinical phenotypes and survival. In addition to revealing novel biology and therapeutics, molecular sub-grouping of PB can be exploited to reduce treatment intensity for patients with favorable biology tumors.
• Cerebral embolization from left atrial myxoma causing takotsubo cardiomyopathy complicated with congestive heart failure.

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Shinya Tanaka

  Cardiology journal 27 (4) 439 - 440 2020
• Histopathologically confirmed very late stent thrombosis associated with stent fracture after implantation of first-generation drug eluting stent.

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Yuta Kobayashi, Hirokazu Komoriyama, Yoshiya Kato, Kazunori Omote, Takuma Sato, Kiwamu Kamiya, Toshiyuki Nagai, Shinya Tanaka, Toshihisa Anzai

  Cardiology journal 27 (2) 204 - 205 2020 [Refereed][Not invited]
  
• Role of Dimerized C16orf74 in Aggressive Pancreatic Cancer: A Novel Therapeutic Target.

  Toshihiro Kushibiki, Toru Nakamura, Masumi Tsuda, Takahiro Tsuchikawa, Koji Hontani, Kazuho Inoko, Mizuna Takahashi, Toshimichi Asano, Keisuke Okamura, Soichi Murakami, Yo Kurashima, Yuma Ebihara, Takehiro Noji, Yoshitsugu Nakanishi, Kimitaka Tanaka, Nako Maishi, Katsunori Sasaki, Woong-Ryeon Park, Toshiaki Shichinohe, Kyoko Hida, Shinya Tanaka, Satoshi Hirano

  Molecular cancer therapeutics 19 (1) 187 - 198 2020/01 [Refereed][Not invited]
   

  Over the past 30 years, the therapeutic outcome for pancreatic ductal adenocarcinoma (PDAC) has remained stagnant due to the lack of effective treatments. We performed a genome-wide analysis to identify novel therapeutic targets for PDAC. Our analysis showed that Homo sapiens chromosome 16 open reading frame 74 (C16orf74) was upregulated in most patients with PDAC and associated with poor prognosis. Previously, we demonstrated that C16orf74 interacts with the catalytic subunit alpha of protein phosphatase 3 and plays an important role in PDAC invasion. However, the pathophysiologic function of C16orf74 is still unclear. In this study, through the analysis of C16orf74 interaction, we demonstrate a new strategy to inhibit the growth and invasion of PDAC. C16orf74 exists in the homodimer form under the cell membrane and binds integrin αVβ3 and is also involved in invasion by activating Rho family (Rac1) and MMP2. Considering that this dimeric form was found to be involved in the function of C16orf74, we designed an 11R-DB (dimer block) cell-permeable dominant-negative peptide that inhibits the dimer form of C16orf74. 11R-DB suppressed invasion and proliferation of PDAC cell lines by inhibiting phosphorylation of Akt and mTOR and also by inactivation of MMP2. 11R-DB also showed antitumor effects in an orthotopic xenograft model and peritoneal metastasis model. Thus, this study demonstrates that dimerized C16orf74, present in the cell membrane, is involved in pancreatic cancer invasion and proliferation. In addition, the C16orf74 dimer block cell-permeable peptide (11R-DB) has a potent therapeutic effect on PDAC in vitro and in vivo.
• Spinal rosette-forming glioneuronal tumor: A case report.

  Shuji Hamauchi, Mishie Tanino, Kazutoshi Hida, Toru Sasamori, Shunsuke Yano, Shinya Tanaka

  Medicine 98 (49) e18271  2019/12 [Refereed][Not invited]
   

  RATIONALE: Rosette-forming glioneuronal tumor (RGNT) is a rare tumor which has been first reported as the fourth ventricle tumor by Komori et al and is classified as a distinct clinicopathological entity by the WHO Classification of Tumors of the Central Nervous System as in 2007. Although RGNTs were reported to occur in both supratentorial and inflatentorial sites, only 4 case reports of spinal RGNT have been demonstrated. PATIENT CONCERNS: A 37-year-old female presenting with slowly progressing right-sided clumsiness. Cervical magnetic resonance imaging revealed a spinal intramedullary tumor between the C2 and C5 levels. DIAGNOSES: Pathological analysis showed unique biphasic cellular architecture consisting of perivascular pseudorosettes dominantly with few neurocytic rosettes and diffuse astrocytoma component. The tumor cells composed of perivascular pseudorosettes showed positivity for both synaptophysin and glial markers such as GFAP and Olig2. Therefore, the diagnosis of RGNT was made. INTERVENTIONS: Gross total resection of the tumor was achieved. No adjuvant chemotherapy nor radiotherapy was conducted after operation. OUTCOMES: At 2 years after the operation, no recurrence was observed. LESSONS: Although RGNT arising from the spinal cord is extremely rare, we need to consider the tumor as a differential diagnosis for intramedullary spinal cord tumors.
• Cancer gene profiling explores the possible precision medicine for diffuse-type gastric adenocarcinoma.

  Marin Ishikawa, Hideyuki Hayashi, Naoya Sakamoto, Shinya Tanaka, Hiroshi Nishihara

  Medical oncology (Northwood, London, England) 37 (1) 10 - 10 2019/11/25 [Refereed][Not invited]
   

  Diffuse type gastric cancer (DGC), a pathological subtype, is one of the most common malignant solid tumors, and mortality of this tumor is not negligible, especially in early-onset cancer patients. In fact, affirmative personalized treatments based on gene profile have not been established yet. The aim of this study was to provide the possible genotype-matched treatment for DGC through comprehensive examination of genomic variants and analysis of clinicopathological characteristics. We retrospectively studied 23 formalin-fixed, paraffin-embedded samples of patients diagnosed as DGC between January 2003 and December 2015 at the Department of Cancer Pathology, Hokkaido University Graduate School of Medicine. The cases were divided into two groups: early-onset (< 50 years old) and elderly-onset (≥ 50 years old) DGC groups. We performed targeted genomic sequencing using a 163 cancer-related gene panel. The sequencing data were analyzed using an original bioinformatics pipeline called GenomeJack and were clinicopathologically evaluated. Intestinal metaplasia and atrophy were highly observed in the adjacent non-cancerous mucosa in the elderly-onset DGC group compared with those in the early-onset DGC group. The number of somatic variants was significantly higher in the elderly-onset DGC group than in the early-onset DGC group. Fifteen patients (65.2%) harbored at least one genomic alteration of the potential target for genotype-matched treatment. In addition, one patient with hypermutation phenotype was diagnosed as Lynch syndrome due to MLH1 mutation, suggesting the sensitivity for the treatment with immune checkpoint inhibitors. Not only does our study demonstrated the potential utility of the targeted genomic sequencing approach for making informed therapeutic decisions, but it also sheds light on DGC pathogenesis and progression.
• Autopsy findings in the early stage of amyotrophic lateral sclerosis with "dropped head" syndrome.

  Satoshi Tanikawa, Mishie Tanino, Lei Wang, Marin Ishikawa, Masaya Miyazaki, Masumi Tsuda, Yasuko Orba, Hirofumi Sawa, Kotarou Matoba, Nishio Nakamura, Kazuo Nagashima, William W Hall, Shinya Tanaka

  Neuropathology : official journal of the Japanese Society of Neuropathology 39 (5) 374 - 377 2019/10 [Refereed][Not invited]
   

  Dropped head syndrome (DHS) has been rarely observed in amyotrophic lateral sclerosis (ALS), and the neuropathological findings of this condition have almost never been described. The identification of transactivation response DNA-binding protein 43 kDa (TDP-43), which binds to RNA/DNA has provided a new method for studying ALS and frontotemporal lobar degeneration (FTLD). Post-mortem examination of an adult sudden death case of a 71-year-old patient who complained of DHS exhibited severe loss of anterior motor neurons in the cervical cord (C4-6). Loss of nerve fibers of the anterior roots was striking compared with posterior roots, together with marked neurogenic atrophy of posterior muscles semispinalis cervicis. Bunina bodies were found in large neurons of Betz giant cells, but not in the motor neurons of spinal cords, or neurons of bulbar regions. Phosphorylated TDP-43 (p-TDP-43)-positive structures were detected in the residual neurons of the cervical, thoracic and lumber cords, hypoglossal nucleus, cerebellar dentate nucleus and parahippocampal cortex, together with ubiquitin-positive inclusions. Phosphorylated Tau positive structures in neuronal cytoplasm were found in the amygdala, entorhinal cortex and parahippocampal cortex, some of which co-expressed p-TDP-43. The medial zone of cervical cords may be the first onset site, and that is the cause of head drop in the early stage of ALS. In spite of detailed examination, the direct cause of sudden death was not verified. This autopsy report revealed the relation of DHS which is a rare clinical manifestation of ALS, and neuropathological findings.
• デジタルPCRによるBRAF遺伝子変異検査は甲状腺乳頭癌のFNA正診率を向上させる(BRAF mutation testing by dPCR improves the diagnosis accuracy of FNA for the papillary thyroid carcinoma)

  北崎 アリサ, 駒林 優樹, 小野 裕介, 木村 太一, 田中 伸哉, 長嶋 和郎, 水上 裕輔

  日本癌学会総会記事 78回 J - 1010 0546-0476 2019/09
• Comparative genome-wide analysis of gastric adenocarcinomas with hyperplastic polyp components.

  Yoshiyasu Takayama, Yusuke Ono, Yusuke Mizukami, Hideaki Itoh, Nozomi Nakajima, Hideo Arai, Shinya Tanaka, Sumihito Nobusawa, Hideaki Yokoo, Yasuhiro Onozato

  Virchows Archiv : an international journal of pathology 475 (3) 383 - 389 2019/09 [Refereed][Not invited]
   

  Gastric hyperplastic polyps are common and generally regarded as benign lesions, whereas gastric adenocarcinomas infrequently occur from gastric hyperplastic polyps. Although gastric hyperplastic polyps have received a lot of attention because of their association with malignant transformation, it remains unclear whether gastric hyperplastic polyps are neoplastic lesions that have sporadic genetic changes similar to colorectal hyperplastic polyps. We performed genome-wide analyses of two gastric adenocarcinomas with hyperplastic polyp components. The interface between "adenocarcinoma" and "hyperplastic polyp" components was fairly sharp, and the adenocarcinoma components had copy number alterations and TP53 mutations, whereas the hyperplastic polyp components had only single nucleotide polymorphisms, which were also found in adenocarcinoma components. We did not detect any somatic changes in the hyperplastic polyp components, even in genome-wide analyses, which was in contrast to the adenocarcinoma components. However, due to the small number of cases examined herein, further genetic analyses of more cases are needed.
• A 41 Year-Old Woman with a Mass in the Posterior Cranial Fossa.

  Bunsho Asayama, Yoshinobu Seo, Yoshimaru Ozaki, Satoshi Tanikawa, Takanori Hirose, Shinya Tanaka, Hirohiko Nakamura

  Brain pathology (Zurich, Switzerland) 29 (5) 699 - 700 2019/09 [Refereed][Not invited]
  
• Modulation and Characterization of the Double Network Hydrogel Surface-bulk Transition

  Martin Frauenlob, Daniel R. King, Honglei Guo, Seiichiro Ishihara, Masumi Tsuda, Takayuki Kurokawa, Hisashi Haga, Shinya Tanaka, Jian Ping Gong

  Macromolecules 52 (17) 6704 - 6713 2019/08 [Refereed][Not invited]
  
• Obstructive sleep apnea is associated with increased coronary plaque instability: an optical frequency domain imaging study.

  Takao Konishi, Yusuke Kashiwagi, Naohiro Funayama, Tadashi Yamamoto, Hironori Murakami, Daisuke Hotta, Shinya Tanaka

  Heart and vessels 34 (8) 1266 - 1279 2019/08 [Refereed][Not invited]
   

  Obstructive sleep apnea (OSA) is associated with coronary artery disease (CAD) and with an increased risk for myocardial infarction, stroke or death due to cardiovascular disease. Optical frequency-domain imaging (OFDI) is a useful modality for evaluating the characteristics of atherosclerotic plaque. The purpose of the study was to use OFDI to investigate the association of OSA with coronary plaque characteristics in patients undergoing percutaneous coronary intervention (PCI). We retrospectively analyzed OFDI data for coronary artery plaques from 15 patients with OSA and 35 non-OSA patients treated between October 2015 and October 2018. Plaque morphology was evaluated for 70 lesions, including 21 from patients with OSA and 49 from non-OSA patients. Compared with the non-OSA group, patients with OSA had significantly higher prevalences of thinned cap fibroatheroma (TCFA) (67% vs. 35%, P = 0.014) and microchannels (86% vs. 55%, P = 0.014); a significantly higher mean lipid index (1392 ± 982 vs. 817 ± 699, P = 0.021), macrophage grade (8.4 ± 6.4 vs. 4.8 ± 4.5, P = 0.030), and maximum number of microchannels (1.5 ± 1.0 vs. 0.7 ± 0.7, P = 0.001); and a significantly lower mean minimum fibrous cap thickness (69.4 ± 28.7 vs. 96.1 ± 51.8 μm, P = 0.008). This OFDI analysis suggests that OSA is associated with unstable plaque characteristics in patients with CAD. More intensive medical management for stabilization of coronary atherosclerotic plaque is required in patients with OSA.
• Exosomes containing ErbB2/CRK induce vascular growth in premetastatic niches and promote metastasis of bladder cancer.

  Kazuhiko Yoshida, Masumi Tsuda, Ryuji Matsumoto, Shingo Semba, Lei Wang, Hirokazu Sugino, Mishie Tanino, Tsunenori Kondo, Kazunari Tanabe, Shinya Tanaka

  Cancer science 110 (7) 2119 - 2132 2019/07 [Refereed][Not invited]
   

  Locally advanced and metastatic invasive bladder cancer (BC) has a poor prognosis, and no advanced therapies beyond cisplatin-based combination chemotherapy have been developed. Therefore, it is an urgent issue to elucidate the underlying mechanisms of tumor progression and metastasis of invasive BC for the development of new therapeutic strategies. Here, we clarified a novel role of exosomes containing ErbB2 and CRK in a formation of premetastatic niches and subsequent metastases. CRK adaptors were overexpressed in invasive UM-UC-3 BC cells. In an orthotopic xenograft model, metastases to lung, liver, and bone of UM-UC-3 cells were completely abolished by CRK elimination. Mass spectrometry analysis identified that ErbB2 was contained in UM-UC-3-derived exosomes in a CRK-dependent manner; the exosomes significantly increased proliferation and invasion properties of low-grade 5637 BC cells and HUVECs through FAK and PI3K/AKT signaling pathways. In athymic mice educated with UM-UC-3-derived exosomes, i.v. implanted UM-UC-3 cells were trapped with surrounding PKH67-labeled exosomes in lung and led to development of lung metastasis with disordered vascular proliferation. In contrast, exosomes derived from CRK-depleted BC cells failed to induce these malignant features. Taken together, we showed that CRK adaptors elevated the expression of ErbB2/3 in BC cells, and these tyrosine kinase/adaptor units were transferred from host BC cells to metastatic recipient cells by exosomes, leading to vascular leakiness and proliferation and contributing to the formation of distant metastasis. Thus, CRK intervention with ErbB2/3 blockade might be a potent therapeutic strategy for patients with ErbB2 overexpressing advanced and metastatic BC.
• Eicosapentaenoic acid therapy is associated with decreased coronary plaque instability assessed using optical frequency domain imaging.

  Takao Konishi, Daisuke Sunaga, Naohiro Funayama, Tadashi Yamamoto, Hironori Murakami, Daisuke Hotta, Masanori Nojima, Shinya Tanaka

  Clinical cardiology 42 (6) 618 - 628 2019/06 [Refereed][Not invited]
   

  BACKGROUND: The relationship between eicosapentaenoic acid (EPA) therapy and coronary plaque stability assessed by optical frequency domain imaging (OFDI) has not been thoroughly described. HYPOTHESIS: EPA therapy is associated with decreased plaque instability in patients undergoing percutaneous coronary intervention (PCI) using OFDI. METHODS: Data on coronary artery plaques from 121 patients who consecutively underwent PCI between October 2015 and July 2018 were retrospectively analyzed. Of these patients, 109 were untreated (no-EPA group), whereas 12 were treated with EPA (EPA group). Each plaque's morphological characteristics were analyzed using OFDI. RESULTS: We used 1:4 propensity score matching for patients who received or did not receive EPA therapy before PCI. Baseline characteristics were balanced between both groups (age, sex, body mass index, diabetes mellitus, hypertension, dyslipidemia, chronic kidney disease, smoking, previous PCI or coronary artery bypass grafting, previous myocardial infarction, prior statin use, acute coronary syndrome, hemoglobin A1c level, low-density lipoprotein cholesterol concentration, triglyceride concentration, and high-density lipoprotein cholesterol concentration). OFDI data from 60 patients were analyzed in this study. The EPA group had significantly lower mean lipid index (818 ± 806 vs 1574 ± 891) and macrophage grade (13.5 ± 5.9 vs 19.3 ± 7.4) but higher mean minimum fibrous cap thickness (109.2 ± 55.7 vs 81.6 ± 36.4 μm) than the no-EPA group (P = 0.010, 0.019, and 0.040, respectively). Multiple logistic regression analyses showed that prior EPA use was independently associated with lower lipid index and macrophage grade (P = 0.043 and 0.024, respectively). CONCLUSION: This OFDI analysis suggests that EPA therapy is associated with decreased plaque instability in patients undergoing PCI.
• 中枢神経原発移植後リンパ増殖性疾患の一例

  浅野目 卓, 佐藤 憲市, 尾崎 義丸, 杉尾 啓徳, 石田 祐樹, 中村 博彦, 石田 雄介, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 36 (Suppl.) 115 - 115 1433-7398 2019/05
• バイオマテリアルによる肝癌幹細胞の新規誘導法

  谷 道夫, 津田 真寿美, 鈴鹿 淳, 王 磊, 杉野 弘和, 谷川 聖, 石田 雄介, グン 剣萍, 田中 伸哉, 武冨 紹信

  日本外科学会定期学術集会抄録集 (一社)日本外科学会 119回 PS - 6 2019/04
• 病理IT化、情報化とAI研究 病理診断と人工知能 スタートアップから脳腫瘍への応用まで

  石田 雄介, 桑原 健, 小田 義崇, 谷川 聖, 杉野 弘和, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 108 (1) 192 - 192 0300-9181 2019/04
• 小児下顎骨に生じたnodular fasciitisの1例

  石田 雄介, 伊勢 昂生, 小田 義崇, 谷川 聖, 杉野 弘和, 嶋崎 康相, 藤森 真樹, 松田 彩, 北村 哲也, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 108 (1) 340 - 340 0300-9181 2019/04
• 初回治療から18年後に生じた多臓器進展を伴うホジキンリンパ腫の一剖検例

  五味川 龍, 杉野 弘和, 白鳥 聡一, 石田 雄介, 王 磊, 畑中 佳奈子, 松野 吉宏, 豊嶋 崇徳, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 108 (1) 453 - 453 0300-9181 2019/04
• 解剖で偶然見つかった無症候性胸髄内神経鞘腫の一例

  有田 梨乃, 谷川 聖, 津田 真寿美, 石田 雄介, 杉野 弘和, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 108 (1) 459 - 459 0300-9181 2019/04
• 髄膜腫におけるTERTプロモーター遺伝子変異の検討

  久世 瑞穂, 小田 義崇, 津田 真寿美, 湯澤 明夏, 谷川 聖, 杉野 弘和, 石田 雄介, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 108 (1) 463 - 463 0300-9181 2019/04
• 次世代シーケンサーを用いた肺原発多形癌の遺伝子変異解析

  長野 匡晃, 高阪 真路, 牛久 綾, 林 大久生, 田中 伸哉, 津田 真寿美, 篠原 敏也, 大森 優子, 菅谷 文子, 加藤 弘明, 成田 吉明, 高持 一矢, 鈴木 健司, 中島 淳, 間野 博行

  日本呼吸器外科学会雑誌 (NPO)日本呼吸器外科学会 33 (3) RO1 - 3 0919-0945 2019/04
• Comprehensive assay for the molecular profiling of cancer by target enrichment from formalin-fixed paraffin-embedded specimens.

  Kohsaka S, Tatsuno K, Ueno T, Nagano M, Shinozaki-Ushiku A, Ushiku T, Takai D, Ikegami M, Kobayashi H, Kage H, Ando M, Hata K, Ueda H, Yamamoto S, Kojima S, Oseto K, Akaike K, Suehara Y, Hayashi T, Saito T, Takahashi F, Takahashi K, Takamochi K, Suzuki K, Nagayama S, Oda Y, Mimori K, Ishihara S, Yatomi Y, Nagase T, Nakajima J, Tanaka S, Fukayama M, Oda K, Nangaku M, Miyazono K, Miyagawa K, Aburatani H, Mano H

  Cancer science 110 (4) 1464 - 1479 1347-9032 2019/04 [Refereed][Not invited]
   

  Tumor molecular profiling is becoming a standard of care for patients with cancer, but the optimal platform for cancer sequencing remains undetermined. We established a comprehensive assay, the Todai OncoPanel (TOP), which consists of DNA and RNA hybridization capture-based next-generation sequencing panels. A novel method for target enrichment, named the junction capture method, was developed for the RNA panel to accurately and cost-effectively detect 365 fusion genes as well as aberrantly spliced transcripts. The TOP RNA panel can also measure the expression profiles of an additional 109 genes. The TOP DNA panel was developed to detect single nucleotide variants and insertions/deletions for 464 genes, to calculate tumor mutation burden and microsatellite instability status, and to infer chromosomal copy number. Clinically relevant somatic mutations were identified in 32.2% (59/183) of patients by prospective TOP testing, signifying the clinical utility of TOP for providing personalized medicine to cancer patients.
• Formation and Disordered Degradation of Neutrophil Extracellular Traps in Necrotizing Lesions of Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis.

  Sakiko Masuda, Mayu Nonokawa, Emika Futamata, Yuka Nishibata, Sari Iwasaki, Takahiro Tsuji, Yutaka Hatanaka, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Tamihiro Kawakami, Tatsuya Atsumi, Akihiro Ishizu

  The American journal of pathology 189 (4) 839 - 846 0002-9440 2019/04 [Refereed][Not invited]
   

  Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is characterized by the production of ANCAs and systemic necrotizing vasculitis in small vessels. Disordered regulation of neutrophil extracellular traps (NETs) is critically involved in the pathogenesis of AAV. NETs are web-like DNA decorated with antimicrobial proteins; they are extruded from activated neutrophils. The principal degradation factor of NETs in vivo is DNase I; however, NETs resistant to DNase I can persist in tissues and can lead to the production of ANCAs. Deposition of NETs has been demonstrated in glomerular crescents and necrotizing vasculitis in AAV. Here, the amount of NETs in formalin-fixed, paraffin-embedded tissue sections was examined, and the results for AAV were compared with the results for diseases that should be distinguished from AAV. NETs were more abundant in necrotizing vasculitis of AAV than in non-ANCA-associated vasculitis, or in granulomatous angiitis. Pulmonary granulomas in AAV and non-ANCA-associated diseases were further studied. The amount of NETs was significantly greater in necrotizing granulomas of AAV than in granulomas of sarcoidosis without necrosis. Although NETs were formed in necrotizing granulomas of tuberculosis equivalently to those formed in AAV, they were more susceptible to degradation by DNase I than were NETs in AAV. The formation and disordered degradation of NETs in necrotizing lesions are characteristics of AAV and are possibly related to its pathogenesis.
• Practical procedures for the integrated diagnosis of astrocytic and oligodendroglial tumors.

  Yukihiko Sonoda, Hideaki Yokoo, Shinya Tanaka, Manabu Kinoshita, Mitsutoshi Nakada, Hiroshi Nishihara

  Brain tumor pathology 36 (2) 56 - 62 2019/04 [Refereed][Not invited]
   

  The publication of the 2016 World Health Organization Classification of Tumors of the Central Nervous System (2016 WHO CNS) represented a major change in the classification of brain tumors. However, many pathologists in Japan cannot diagnose astrocytic or oligodendroglial tumors according to the 2016 WHO CNS due to financial or technical problems. Therefore, the Japan Society of Brain Tumor Pathology established a committee for molecular diagnosis to facilitate the integrated diagnosis of astrocytic and oligodendroglial tumors in Japan. We created three levels of diagnoses: Level 1 was defined as simple histopathological diagnosis using hematoxylin and eosin staining and routine cell lineage-based immunostaining. Level 2 was defined as immunohistochemical diagnosis using immunohistochemical examinations using R132H mutation-specific IDH1, ATRX, and/or p53 antibodies. Level 3 was defined as molecular diagnosis, such as diagnosis based on 1p/19q status or the mutation status of the IDH1 and IDH2 genes. In principle, astrocytic and oligodendroglial tumors should be diagnosed based on the 2016 WHO CNS and/or cIMPACT-NOW criteria; however, the findings obtained through our diagnostic flowchart can be added to the histological diagnosis in parentheses. This classification system would be helpful for pathologists with limited resources.
• Autopsy report of a late delayed radiation injury after a period of 45 years.

  Satoshi Tanikawa, Yasutaka Kato, Mishie Tanino, Shunsuke Terasaka, Yasuo Kurokawa, Nobutaka Arai, Kazuo Nagashima, Shinya Tanaka

  Neuropathology : official journal of the Japanese Society of Neuropathology 39 (2) 106 - 110 0919-6544 2019/04 [Refereed][Not invited]
   

  For delayed radiation injury, image analysis has considerably advanced, but neuropathological findings are still required to establish diagnosis. A patient who had received radiation therapy for pineal germinoma at age 14 developed neurological and psychiatric abnormalities after 15 years as a late delayed radiation injury. Autopsy at age 59 revealed diffuse changes in the white matter consisting in order of severity of myelin pallor, demyelination, and necrosis which were characterized by a lack of glial reaction. The cerebral cortex was relatively well preserved. As delayed radiation injuries, hyalinous changes in the vascular wall, angiomatous lesions and, fresh and old petechial hemorrhages were found. Moreover, vascular changes associated with arteriosclerosis were also present. Furthermore, a focal glial nodule was detected which was considered to be a new radiation-induced neoplasia. These findings suggest that late delayed radiation injury may slowly develop over 30 years and may involve damage to neuroglial stem cell compensation. It is also evident that arteriosclerotic changes and newly induced neoplasia may develop in delayed radiation injury cases.
• Pathways of Progression From Intraductal Papillary Mucinous Neoplasm to Pancreatic Ductal Adenocarcinoma Based on Molecular Features.

  Yuko Omori, Yusuke Ono, Mishie Tanino, Hidenori Karasaki, Hiroshi Yamaguchi, Toru Furukawa, Katsuro Enomoto, Jun Ueda, Atsuko Sumi, Jin Katayama, Miho Muraki, Kenzui Taniue, Kuniyuki Takahashi, Yoshiyasu Ambo, Toshiya Shinohara, Hiroshi Nishihara, Junpei Sasajima, Hiroyuki Maguchi, Yusuke Mizukami, Toshikatsu Okumura, Shinya Tanaka

  Gastroenterology 156 (3) 647 - 661 0016-5085 2019/02 [Refereed][Not invited]
   

  BACKGROUND & AIMS: Intraductal papillary mucinous neoplasms (IPMNs) are regarded as precursors of pancreatic ductal adenocarcinomas (PDAs), but little is known about the mechanism of progression. This makes it challenging to assess cancer risk in patients with IPMNs. We investigated associations of IPMNs with concurrent PDAs by genetic and histologic analyses. METHODS: We obtained 30 pancreatic tissues with concurrent PDAs and IPMNs, and 168 lesions, including incipient foci, were mapped, microdissected, and analyzed for mutations in 18 pancreatic cancer-associated genes and expression of tumor suppressors. RESULTS: We determined the clonal relatedness of lesions, based on driver mutations shared by PDAs and concurrent IPMNs, and classified the lesions into 3 subtypes. Twelve PDAs contained driver mutations shared by all concurrent IPMNs, which we called the sequential subtype. This subset was characterized by less diversity in incipient foci with frequent GNAS mutations. Eleven PDAs contained some driver mutations that were shared with concurrent IPMNs, which we called the branch-off subtype. In this subtype, PDAs and IPMNs had identical KRAS mutations but different GNAS mutations, although the lesions were adjacent. Whole-exome sequencing and methylation analysis of these lesions indicated clonal origin with later divergence. Ten PDAs had driver mutations not found in concurrent IPMNs, called the de novo subtype. Expression profiles of TP53 and SMAD4 increased our ability to differentiate these subtypes compared with sequencing data alone. The branch-off and de novo subtypes had substantial heterogeneity among early clones, such as differences in KRAS mutations. Patients with PDAs of the branch-off subtype had a longer times of disease-free survival than patients with PDAs of the de novo or the sequential subtypes. CONCLUSIONS: Detailed histologic and genetic analysis of PDAs and concurrent IPMNs identified 3 different pathways by which IPMNs progress to PDAs-we call these the sequential, branch-off, and de novo subtypes. Subtypes might be associated with clinical and pathologic features and be used to select surveillance programs for patients with IPMNs.
• Immunohistochemical Analysis of Cerebral Intraparenchymal Choroid Plexus Tumor: Case Report.

  Masahito Katoh, Yutaka Sawamura, Shinya Tanaka, Takamitsu Fujimaki, Shigehisa Hirose, Toshimitsu Aida

  Journal of neurological surgery. Part A, Central European neurosurgery 80 (1) 53 - 57 2019/01 [Refereed][Not invited]
   

  BACKGROUND:  It is very rare for a choroid plexus tumor to occur intraparenchymally in the absence of a relation to the choroid plexus. CLINICAL PRESENTATION:  A case of cerebral intraparenchymal choroid plexus tumor in a 30-year-old woman presenting with left hemiparesis is described. Brain magnetic resonance imaging depicted a large cystic mass in the right frontal lobe. Tumor resection was performed by right frontal craniotomy. No connection with the choroid plexus was observed during the operation. Histologically, the tumor exhibited a glandular structure with a papillary pattern suggesting a neoplasm of epithelial origin. Immunohistochemical analyses revealed the tumor as an atypical choroid plexus papilloma. CONCLUSION:  Immunohistochemical findings, especially regarding Kir7.1, are very important for the differential diagnosis of cerebral intraparenchymal choroid plexus tumors from metastatic tumors. The present case reveals that an atypical choroid plexus papilloma can occur intraparenchymally without an association with the choroid plexus. Intraparenchymal atypical choroid plexus papillomas may have previously been diagnosed incorrectly as metastatic adenocarcinomas of unknown origin.
• Multimodal imaging of right coronary artery to left ventricle fistula complicated by large coronary aneurysm.

  Takao Konishi, Tadashi Yamamoto, Masato Hayakawa, Shizuko Iwasa, Hiroyuki Tsukui, Shinya Tanaka

  Cardiology journal 26 (1) 93 - 94 2019 [Refereed][Not invited]
  
• Woman in her 50s with shortness of breath on exertion.

  Konishi T, Murakami H, Tanaka S

  Heart (British Cardiac Society) 105 (2) 110 - 110 1355-6037 2019/01 [Refereed][Not invited]
   

  CLINICAL INTRODUCTION: A 59-year-old woman visited an outpatient cardiology clinic due to shortness of breath on exertion. Physical examination showed no significant abnormality of vital signs. A III/VI systolic murmur was heard on the fourth intercostal space at the right sternal border. The majority of laboratory tests were normal. Chest X-ray showed a curved vessel shadow (figure 1A). Initial transthoracic echocardiography showed abnormal blood flow into the inferior vena cava (IVC) in the subxiphoid long axis view (figure 1B) and mild right heart dilatation (online supplementary figure 1). Transoesophageal echocardiography showed severe tricuspid regurgitation (online supplementary figure 2).heartjnl;105/2/110/F1F1F1Figure 1(A) Chest X-ray. (B) Colour Doppler image in the subxiphoid long axis view.DC1SP110.1136/heartjnl-2018-313655.supp1Supplementary data DC2SP210.1136/heartjnl-2018-313655.supp2Supplementary data QUESTION: What is the most likely underlying disease for the patient's shortness of breath on exertion?Pulmonary arteriovenous fistula.Pulmonary arterial hypertension.Lung cancer.Partial anomalous pulmonary venous connection.Isolated tricuspid regurgitation.
• "Integrated diagnosis" of pilocytic astrocytoma: Molecular diagnostic procedure for an unusual case.

  Ishida Y, Tsuda M, Sawamura Y, Fujii K, Murai H, Horiuchi N, Orba Y, Sawa H, Hall WW, Nagashima K, Tanaka S

  Pathology international 68 (12) 694 - 699 1320-5463 2018/12 [Refereed][Not invited]
   

  A 24 year-old female presented with a mass lesion in the right temporal lobe. This case was difficult to diagnose using histological and immunological methods and therefore molecular analyses were applied to provide a definitive diagnosis. The tumor was well-demarcated, partially cystic, and irregularly-enhanced on gadolinium-enhanced T1-weighted magnetic resonance images. Pathologically, a large part of the tumor consisted of cells with fine cytoplasmic processes on a myxoid and mucinous background. Cells formed a microcystic structure around the mucinous tissue. Numerous eosinophilic granular bodies, but not Rosenthal fibers, were present. The solid and compact regions of the tumor were composed of fasciculation of dense fibrous glial tissues and occasional multinucleated giant cells. Tumor cells and their fragmented cytoplasmic processes were positively stained with GFAP, while eosinophilic granular bodies were both positive and negative. Xanthomatous changes were not detected and the reticulin fibers were restricted to vascular tissues. The MIB1 index was scored as approximately 10%. In molecular analyses of BRAF, the KIAA1549-BRAF (K16-B9) fusion gene was detected in all tumor regions, whereas BRAF V600E mutation was not detected by either conventional Sanger sequencing or the Eprobe-PCR method. Based on the results of the molecular analyses, this case was diagnosed as pilocytic astrocytoma.
• Stabilization of symptomatic carotid atherosclerotic plaques by statins: a clinico-pathological analysis.

  Konishi T, Funayama N, Yamamoto T, Hotta D, Nomura R, Nakagaki Y, Murahashi T, Kamiyama K, Yoshimoto T, Aoki T, Tanaka S

  Heart and vessels 33 (11) 1311 - 1324 0910-8327 2018/11 [Refereed][Not invited]
  
• Micro patterning of hydroxyapatite by soft lithography on hydrogels for selective osteoconduction.

  Kiyama R, Nonoyama T, Wada S, Semba S, Kitamura N, Nakajima T, Kurokawa T, Yasuda K, Tanaka S, Gong JP

  Acta biomaterialia 81 60 - 69 1742-7061 2018/11 [Refereed][Not invited]
   

  Mechanically robust hydrogels are promising biomaterials as artificial supportive tissue. These applications require selective and robust bonding of the hydrogels to living tissue. Recently, we achieved strong in vivo bone bonding of a tough double network (DN) hydrogel, a potential material for use as artificial cartilage and tendon, by hybridizing osteoconductive hydroxyapatite (HAp) in the gel surface layer. In this work, we report micro patterning of HAp at the surface of the DN hydrogel for selective osteoconduction. Utilizing the dissolution of HAp in an acidic environment, the soft lithography technique using an acid gel stamp was adopted to form a high-resolution HAp pattern on the gel. The HAp-patterned gel showed well-regulated migration and adhesion of cells in vitro. Moreover, the HAp-patterned gel showed selective and robust bonding to the rabbit bone tissue in vivo. This HAp soft lithography technique allows for simple and quick preparation of tailor-made osteoconductive hydrogels and can be applied to other hydrogels for selective bone bonding. STATEMENT OF SIGNIFICANCE: Hydrogels, preserving large amount of water, have been studied for next-generation artificial soft tissues. However, fixation of hydrogels to living tissue was unsolved issue for clinical application. Recently, we achieved robust bonding of a tough double network gel to bone in vivo by coating of osteoconductive hydroxyapatite in the gel surface layer. For further progress for practical use, we report the micro patterning of HAp at the surface of the DN hydrogel by using soft lithography technique, to perform selective bonding to only objective area without unnecessary coalescence. The HAp lithography technique is simple, quick and non-toxic method to prepare tailor-made osteoconductive hydrogels and has universality of species of hydrogels.
• Brain-Derived Neurotrophic Factor Improves Limited Exercise Capacity in Mice With Heart Failure.

  Junichi Matsumoto, Shingo Takada, Shintaro Kinugawa, Takaaki Furihata, Hideo Nambu, Naoya Kakutani, Masaya Tsuda, Arata Fukushima, Takashi Yokota, Shinya Tanaka, Hidehisa Takahashi, Masashi Watanabe, Shigetsugu Hatakeyama, Masaki Matsumoto, Keiichi I Nakayama, Yutaro Otsuka, Hisataka Sabe, Hiroyuki Tsutsui, Toshihisa Anzai

  Circulation 138 (18) 2064 - 2066 0009-7322 2018/10/30 [Refereed][Not invited]
  
• Genetic mutation analysis of the malignant transformation of sinonasal inverted papilloma by targeted amplicon sequencing.

  Yasukawa S, Kano S, Hatakeyama H, Nakamaru Y, Takagi D, Mizumachi T, Suzuki M, Suzuki T, Nakazono A, Tanaka S, Nishihara H, Homma A

  International journal of clinical oncology 23 (5) 835 - 843 1341-9625 2018/10 [Refereed][Not invited]
  
• 馬尾原発の髄外性形質細胞腫の1例

  小田 義崇, 杉野 弘和, 小柳 泉, 谷川 聖, 石田 雄介, 津田 真寿美, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 35 (Suppl.) 177 - 177 1433-7398 2018/09
• Astroblastoma: a distinct tumor entity characterized by alterations of the X chromosome and MN1 rearrangement

  Takanori Hirose, Sumihito Nobusawa, Kazuhiko Sugiyama, Vishwa J. Amatya, Naomi Fujimoto, Atsushi Sasaki, Yoshiki Mikami, Akiyoshi Kakita, Shinya Tanaka, Hideaki Yokoo

  Brain Pathology 28 (5) 684 - 694 1015-6305 2018/09 [Refereed][Not invited]
   

  © 2017 International Society of Neuropathology Astroblastoma is a rare, enigmatic tumor of the central nervous system (CNS) which shares some clinicopathologic aspects with other CNS tumors, especially ependymoma. To further clarify the nature of astroblastoma, we performed clinicopathologic and molecular genetic studies on eight cases of astroblastoma. The median age of the patients was 14.5 years, ranging from 5 to 60 years, and seven of the patients were female. All tumors arose in the cerebral hemisphere and radiologically appeared to be well-bordered, nodular tumors often associated with cystic areas and contrast-enhancement. Six of the seven patients with prognosis data survived without recurrences during the follow-up periods ranging from six to 76 months. One patient had multiple recurrences and died six years later. All tumors exhibited salient microscopic features, such as being well demarcated from the surrounding brain tissue, perivascular arrangement of epithelioid tumor cells (represented by “astroblastic” pseudorosettes, trabecular alignment, and pseudopapillary patterns), and hyalinized blood vessels. Immunoreactivity for GFAP, S-100 protein, Olig2, and EMA was variably demonstrated in all tumors, and IDH1 R132H and L1CAM were negative. Array comparative genomic hybridization revealed numerous heterozygous deletions on chromosome X in the four tumors studied, and break-apart fluorescence in situ hybridization demonstrated rearrangement of MN1 in five tumors with successful testing. The characteristic clinicopathologic and genetic findings support the idea that astroblastoma is distinct from other CNS tumors, in particular, ependymoma. In addition, MN1 rearrangement and aberrations of chromosome X may partly be involved in the pathogenesis of astroblastoma.
• Disseminated fusariosis emerged from prolonged local genital infection after cord blood transplantation

  Kohei Okada, Tomoyuki Endo, Daigo Hashimoto, Tomoyuki Saga, Takahide Ara, Reiki Ogasawara, Atsushi Yasumoto, Makoto Ibata, Mutsumi Takahata, Akio Shigematsu, Takeshi Kondo, Yasunori Muraosa, Toshifumi Nomura, Hiromi Kanno-Okada, Satoshi Hashino, Shinya Tanaka, Katsuhiko Kamei, Takanori Teshima

  Journal of Infection and Chemotherapy 24 (8) 660 - 663 1437-7780 2018/08/01 [Refereed][Not invited]
   

  Disseminated fusariosis (DF) is a rare life threatening fungal infection in immunocompromised hosts. We herein report a case of a fatal DF mimicking varicella zoster virus (VZV) infection that was emerged from a localized genital infection during cord blood transplantation (CBT) in a patient with severe aplastic anemia (SAA). The patient developed an ulcer following small painful vesicles mimics herpes simplex virus infection (HSV) on the glans penis before CBT, but a Fusarium species was identified. Despite administration of voriconazole, liposomal amphotericin B and granulocyte transfusion, the lesion was extended to extensive skin looked like VZV infection and the patients died after CBT. Massive fusarium infiltration was detected in multiple organs at autopsy. A genetic analysis of the mold identified Fusarium solani after his death. It should be noted that in patients with fusarium infection, localized and disseminated lesions of fusarium infection sometimes mimic HSV and VZV infections, which hampers an early diagnosis.
• The prognostic improvement of add-on bevacizumab for progressive disease during concomitant temozolomide and radiation therapy in the patients with glioblastoma and anaplastic astrocytoma.

  Shigeru Yamaguchi, Yukitomo Ishi, Hiroaki Motegi, Michinari Okamoto, Hiroyuki Kobayashi, Kenji Hirata, Yoshitaka Oda, Shinya Tanaka, Shunsuke Terasaka, Kiyohiro Houkin

  Journal of neurosurgical sciences 64 (6) 502 - 508 0390-5616 2018/07/09 [Refereed][Not invited]
   

  BACKGROUND: Although newly diagnosed high-grade glioma patients in Japan can receive bevacizumab (BEV) as first-line chemotherapy, randomized clinical trials have not shown a survival benefit for BEV for these patients. In this study, we investigated whether selective add-on BEV for patients with newly diagnosed glioblastoma (GBM) and anaplastic astrocytoma (AA) improves prognosis, in cases where tumors were continuously growing during radiotherapy concomitant with temozolomide (TMZ). METHODS: We conducted a retrospective survey of the overall survival (OS) of patients with GBM/AAs who were treated in our institution between 2006 and 2016. Patients whose tumors were continuously growing regardless of radiotherapy were categorized as the "progressive" group; remaining patients were categorized as the "non-progressive" group. Since 2013, patients in the "progressive" group received add-on BEV therapy with the Stupp regimen during or just after radiotherapy. RESULTS: Of 151 GBM/AA patients, 34 (22.5%) were categorized in the "progressive" group. Median OSs of the "progressive" and "non-progressive" groups were 13.2 months and 25.3 months, respectively (P < 0.001). Twelve patients in the "progressive" group received add-on BEV therapy, and their median OS was 20.2 months; whereas for the remaining 22 patients in the "progressive" group who were treated before the BEV era, their median OS was 10.5 months. In the "progressive" group, add-on BEV significantly extended OS (P = 0.018) and was the lone clinical factor of better prognosis. CONCLUSIONS: We found that, for patients with GBM/AAs whose tumors were continuously growing during radiotherapy, add-on BEV treatment resulted in survival benefits.
• Tumor budding and human chorionic gonadotropin-β expression correlate with unfavorable patient outcome in colorectal carcinoma.

  Yuji Konishi, Futoshi Kawamata, Hiroshi Nishihara, Shigenori Homma, Yasutaka Kato, Masumi Tsuda, Shinji Kohsaka, Takahiro Einama, Cheng Liu, Tadashi Yoshida, Akihisa Nagatsu, Mishie Tanino, Shinya Tanaka, Hideki Kawamura, Toshiya Kamiyama, Akinobu Taketomi

  Medical oncology (Northwood, London, England) 35 (7) 104 - 104 1357-0560 2018/06/11 [Refereed][Not invited]
   

  Tumor budding is thought to represent a manifestation of epithelial-to-mesenchymal transition (EMT) and it has been correlated with poor patient outcomes in colorectal cancer (CRC). Our group recently demonstrated that human chorionic gonadotropin-β (hCGβ) modulates EMT in CRC. In the current study, based on the likely relationships between tumor budding and hCGβ expression, we examined their clinicopathologic significance in CRC. Twenty-eight of 80 (35.0%) CRC showed tumor budding. Tumor budding significantly correlated with lymph node metastasis (P < 0.01), pathologic stage (P < 0.01), lymphatic invasion (P = 0.044), and vascular invasion (P = 0.013). Thirteen of 80 (16.3%) CRC were hCGβ positive on immunohistochemistry. More tumor buds were present in the hCGβ-positive cases (P < 0.01), and tumor budding was significantly correlated with hCGβ positivity (P < 0.01). Cases with both tumor budding and hCGβ expression had the poorest prognosis compared with all other groups (P < 0.01). In conclusion, tumor budding and hCGβ expression are closely associated with EMT, and they are independent prognostic factors in CRC. They identify patients with an "EMT phenotype" who may respond to targeted molecular therapies.
• Tough and Self-Recoverable Thin Hydrogel Membranes for Biological Applications

  Ya Nan Ye, Martin Frauenlob, Lei Wang, Masumi Tsuda, Tao Lin Sun, Kunpeng Cui, Riku Takahashi, Huijie Zhang, Tasuku Nakajima, Takayuki Nonoyama, Takayuki Kurokawa, Shinya Tanaka, Jian Ping Gong

  Advanced Functional Materials 28 (31) 1801489  1616-301X 2018/06 [Refereed][Not invited]
   

  Tough and self-recoverable hydrogel membranes with micrometer-scale thickness are promising for biomedical applications, which, however, rarely be realized due to the intrinsic brittleness of hydrogels. In this work, for the first time, by combing noncovalent DN strategy and spin-coating method, we successfully fabricated thin (thickness: 5-100 mu m), yet tough (work of extension at fracture: 10(5)-10(7) J m(-3)) and 100% self-recoverable hydrogel membranes with high water content (62-97 wt%) in large size (approximate to 100 cm(2)). Amphiphilic triblock copolymers, which form physical gels by self-assembly, were used for the first network. Linear polymers that physically associate with the hydrophilic midblocks of the first network, were chosen for the second network. The inter-network associations serve as reversible sacrificial bonds that impart toughness and self-recovery properties on the hydrogel membranes. The excellent mechanical properties of these obtained tough and thin gel membranes are comparable, or even superior to many biological membranes. The in vitro and in vivo tests show that these hydrogel membranes are biocompatible, and postoperative nonadhesive to neighboring organs. The excellent mechanical and biocompatible properties make these thin hydrogel membranes potentially suitable for use as biological or postoperative antiadhesive membranes.
• Relationship between left main and left anterior descending arteries bifurcation angle and coronary artery calcium score in chronic kidney disease: A 3-dimensional analysis of coronary computed tomography

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Shinya Tanaka

  PLoS ONE 13 (6) e0198566  1932-6203 2018/06/01 [Refereed][Not invited]
   

  Background: A high coronary artery calcium score (CACS) predicts a poor prognosis in patients with coronary artery disease. We examined the relationship between the bifurcation angle and the CACS of the left main (LM) and left anterior descending (LAD) arteries in patients suffering from chronic kidney disease (CKD). Methods: We analyzed the data of 121 patients who underwent coronary computed tomography between October 2014 and June 2015 and whose estimated glomerular filtration rate (eGFR) was > 60 ml/min/1.73 m2. The LM-LAD bifurcation angle was measured by 3-dimensional coronary computed tomography. The CACS of the LM-LAD arteries was also calculated. We excluded stent recipients and patient who had undergone coronary artery bypass graft surgery. Results: In the overall sample, the mean ± standard deviation (range) LM-LAD bifurcation angle was 35.9 ± 11.4 (6.8-79.4) and mean CACS was 227 ± 351 (0 to 1,695). The mean LM-LAD arteries angle was 40.3 ± 10.0 in 39 patients whose CACS was 200, versus 33.8 ± 11.6 in 82 patients with CACS > 200 (p = 0.003). A weak, but positive correlation (r = 0.269, p = 0.003) was observed between the LM-LAD arteries angle and CACS of the LM-LAD arteries. By multiple variable analysis, hemoglobin A1c, triglycerides, eGFR and the LM-LAD arteries angle were independent predictors of a high CACS of the LM-LAD arteries. Conclusion: In patients with CKD, a wide LM-LAD arteries angle was associated with a high CACS of the LM-LAD arteries. The prognostic value of this observation warrants further evaluation.
• 新規技術を用いた新しい研究アプローチ 高分子ハイドロゲルによる癌幹細胞へのリプログラミング誘導技術

  津田 真寿美, 鈴鹿 淳, 王 磊, 仙葉 慎吾, 油谷 幸代, 黒川 孝幸, 近江谷 克裕, 安田 和則, グン 剣萍, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 217 - 217 0300-9181 2018/04
• 神経再生工学における両電荷を有するハイドロゲルの開発

  谷川 聖, 仙葉 慎吾, 津田 真寿美, 王 磊, 谷野 美智枝, 石田 雄介, 杉野 弘和, 鈴鹿 淳, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 329 - 329 0300-9181 2018/04
• 悪性神経膠腫におけるIDH1遺伝子変異による放射線照射後変化の解析(Analysis of the effect of IDH1 mutation on the radiosensitivity in malignant glioma)

  北崎 アリサ, 谷野 美智枝, 九笹 めい, 杉野 弘和, 王 磊, 石田 雄介, 津田 真寿美, 五十嵐 香織, 曽我 朋義, 田中 伸哉

  日本病理学会会誌 107 (1) 379 - 379 0300-9181 2018/04
• 脳腫瘍組織像の画像解析と遺伝子プロファイルに対応したDeep-Learning法の応用

  石田 雄介, 杉野 弘和, 谷野 美智枝, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 380 - 380 0300-9181 2018/04
• 悪性中皮腫におけるOTUB1の役割

  九笹 めい, 谷野 美智枝, 北崎 アリサ, 杉野 弘和, 石田 雄介, 王 磊, 津田 真寿美, 高澤 啓, 平野 博嗣, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 406 - 406 0300-9181 2018/04
• 中枢神経系に生じたメトトレキサート関連リンパ増殖性疾患の一例

  杉野 弘和, 佐藤 憲市, 笠井 康弘, 孫 慧, 石田 雄介, 谷野 美智枝, 津田 真寿美, 松野 吉宏, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 449 - 449 0300-9181 2018/04
• 脳死肝移植後に感染源不明の敗血症を繰り返し死亡した一例の死後画像および病理解剖所見

  伊勢 昂生, 山下 たんぽぽ, 石田 雄介, 桑原 健, 川村 典生, 菊池 穏香, 杉野 弘和, 谷野 美智枝, 津田 真寿美, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 530 - 530 0300-9181 2018/04
• 末梢性T細胞リンパ腫の剖検時に見出された硬化性胸腺腫の一例

  飯田 圭祐, 植田 沙也加, 杉野 弘和, 曾澤 佳昭, 谷野 美智枝, 石田 雄介, 王 磊, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 107 (1) 533 - 533 0300-9181 2018/04
• 肝転移をきたした直腸原発胎児消化管型癌と考えられた1例(A case of liver metastasis from rectal adenocarcinoma with enteroblastic differentiation(AED))

  山野 三紀, 旭 よう, 沢田 尭史, 喜納 政哉, 益子 博幸, 瀧山 晃弘, 後藤田 裕子, 田中 伸哉, 八尾 隆史, 樋野 興夫

  日本病理学会会誌 107 (1) 487 - 487 0300-9181 2018/04
• Tough and Self‐Recoverable Thin Hydrogel Membranes for Biological Applications

  Ye YN, Frauenlob M, Wang L, Tsuda M, Sun TL, Cui K, Takahashi R, Ahang HJ, Nakajima T, Nonoyama T, Kurokawa T, Tanaka S, Gong JP

  Advanced Functional Materials 29 (1801489) 1 - 11 2018/03 [Refereed][Not invited]
  
• Mutations in bassoon in individuals with familial and sporadic progressive supranuclear palsy-like syndrome.

  Ichiro Yabe, Hiroaki Yaguchi, Yasutaka Kato, Yasuo Miki, Hidehisa Takahashi, Satoshi Tanikawa, Shinichi Shirai, Ikuko Takahashi, Mari Kimura, Yuka Hama, Masaaki Matsushima, Shinsuke Fujioka, Takahiro Kano, Masashi Watanabe, Shin Nakagawa, Yasuyuki Kunieda, Yoshio Ikeda, Masato Hasegawa, Hiroshi Nishihara, Toshihisa Ohtsuka, Shinya Tanaka, Yoshio Tsuboi, Shigetsugu Hatakeyama, Koichi Wakabayashi, Hidenao Sasaki

  Scientific reports 8 (1) 819 - 819 2018/01/16 [Refereed][Not invited]
   

  Clinical diagnosis of progressive supranuclear palsy (PSP) is sometimes difficult because various phenotypes have been identified. Here, we report a mutation in the bassoon (BSN) gene in a family with PSP-like syndrome. Their clinical features resembled not only those of PSP patients but also those of individuals with multiple system atrophy and Alzheimer's disease. The neuropathological findings showed a novel three + four repeat tauopathy with pallido-luysio-nigral degeneration and hippocampal sclerosis. Whole-exome analysis of this family identified a novel missense mutation in BSN. Within the pedigree, the detected BSN mutation was found only in affected individuals. Further genetic analyses were conducted in probands from four other pedigrees with PSP-like syndrome and in 41 sporadic cases. Three missense mutations in BSN that are very rarely listed in databases of healthy subjects were found in four sporadic cases. Western blot analysis of tau following the overexpression of wild-type or mutated BSN revealed the possibility that wild-type BSN reduced tau accumulation, while mutated BSN lost this function. An association between BSN and neurological diseases has not been previously reported. Our results revealed that the neurodegenerative disorder associated with the original proband's pedigree is a novel tauopathy, differing from known dementia and parkinsonism syndromes, including PSP.
• Adaptor protein CRK promotes tumor progression and metastasis of bladder cancer by regulating ErbB2 in exosome

  Tsuda Masumi, Yoshida Kazuhiko, Matsumoto Ryuji, Kondo Tsunenori, Shinohara Nobuo, Tanaka Shinya

  CANCER SCIENCE 109 121  1349-7006 2018/01 [Refereed][Not invited]
  
• Chorionic Gonadotropin-β Modulates Epithelial-Mesenchymal Transition in Colorectal Carcinoma Metastasis.

  Futoshi Kawamata, Hiroshi Nishihara, Shigenori Homma, Yasutaka Kato, Masumi Tsuda, Yuji Konishi, Lei Wang, Shinji Kohsaka, Cheng Liu, Tadashi Yoshida, Mishie Tanino, Shinya Tanaka, Hideki Kawamura, Toshiya Kamiyama, Akinobu Taketomi

  The American journal of pathology 188 (1) 204 - 215 0002-9440 2018/01 [Refereed][Not invited]
   

  Ectopic production of free β human chorionic gonadotropin (hCGβ) has been associated with aggressive behavior in non-trophoblastic tumors. hCGβ shares common evolutionary sequences with transforming growth factor-β (TGF-β), which represents a major driving force of epithelial-to-mesenchymal transition (EMT). In this study, we examined the biological roles of hCGβ during EMT and its clinical significance in colorectal cancer (CRC) progression. Eighty CRC specimens and 54 preoperative serum samples were analyzed. hCGβ-overexpressing human CRC cell lines were examined for invasiveness and tumorigenicity, and the expression of EMT-associated genes was investigated. In human CRC, histologic hCGβ positivity [13/80 (16.3%)] was lower than serologic hCGβ positivity [13/54 (24.1%)]. However, it was significantly correlated with several clinicopathological features and unfavorable outcome (P < 0.05). hCGβ-overexpressing cell lines had increased invasiveness, migratory ability, and metastatic potential in mice (P < 0.01). Western blot, PCR, and microarray analyses showed hCGβ altered expression of EMT-related genes, including E-cadherin, phosphorylated SMAD2, SNAIL, and TWIST. hCGβ-induced SNAIL and TWIST overexpression levels were reversible by type I and type II TGF-β receptor inhibitors (P < 0.05). hCGβ thus induces EMT via the TGF-β signaling pathway, and it may represent a molecular target in CRC treatment.
• A clinicopathological analysis of six autopsy cases of sudden unexpected death due to infectious aortitis in patients with aortic tears

  Marin Ishikawa, Mishie Ann Tanino, Masaya Miyazaki, Taichi Kimura, Yusuke Ishida, Lei Wang, Masumi Tsuda, Hiroshi Nishihara, Kazuo Nagashima, Shinya Tanaka

  Internal Medicine 57 (10) 1375 - 1380 1349-7235 2018 [Refereed][Not invited]
   

  Objective Cardiovascular disease is a leading cause of sudden unexpected death even in hospitalized patients. Infectious aortitis is a rare disease that has the potential to cause aortic tears and hemorrhage followed by sudden death. The aim of this study was to reveal the clinicopathological features of infectious aortitis that are related to sudden unexpected death. Methods We retrospectively reviewed 1,310 autopsy cases over 15 years and selected the cases involving patients who died suddenly due to aortic tears. We analyzed the clinical information and pathological findings. Results One hundred thirty-three of 1,310 cases (10.2%) were autopsied under the clinical diagnosis of unexpected sudden death. Aortic tears were identified in 33 cases (2.5%) and infectious aortitis was diagnosed in 6 (18.2%) of these cases. All cases involved male patients (middle-aged to elderly) with risk factors for atherosclerosis (i.e., hypertension). The laboratory data showed continuous leukocytosis and C-reactive protein elevation, even during the improvement phase, in patients with pre-existing infectious disease. The autopsy findings revealed three types of aortic tears (aneurysms, dissections and penetrating atherosclerotic ulcers with moderate to severe atherosclerosis), and the infiltration of numerous neutrophils at the site of rupture. Gram-positive bacteria were detected in four cases and Gram-negative bacteria were detected in two cases. Discussion We demonstrated that sudden unexpected death caused by infectious aortitis rarely occurred in hospitalized patients, even in the recovery phase of the preceding infectious disease. We therefore recommend that clinicians pay attention to infectious aortitis in patients with infectious disease, particularly elderly patients with atherosclerotic disease, even those who are in the improvement phase. Conclusion Unexpected sudden death by infectious aortitis in the recovery phase of antecedent infection.
• Coexistence of primary colorectal follicular lymphoma and multiple myeloma: a case report.

  Saito M, Miyashita K, Miura Y, Ogasawara R, Kanaya M, Izumiyama K, Mori A, Kondo T, Tanaka M, Morioka M, Tanaka S

  International journal of general medicine 11 363 - 367 2018 [Refereed][Not invited]
  
• miR-23a promotes invasion of glioblastoma via HOXD10-regulated glial-mesenchymal transition.

  Kazuhiro Yachi, Masumi Tsuda, Shinji Kohsaka, Lei Wang, Yoshitaka Oda, Satoshi Tanikawa, Yusuke Ohba, Shinya Tanaka

  Signal transduction and targeted therapy 3 33 - 33 2095-9907 2018 [Refereed][Not invited]
   

  Glioblastoma is the most aggressive and invasive brain tumor and has a poor prognosis; elucidating the underlying molecular mechanisms is essential to select molecular targeted therapies. Here, we investigated the effect of microRNAs on the marked invasiveness of glioblastoma. U373 glioblastoma cells were infected with 140 different microRNAs from an OncomiR library, and the effects of the invasion-related microRNAs and targeted molecules were investigated after repeated Matrigel invasion assays. Screening of the OncomiR library identified miR-23a as a key regulator of glioblastoma invasion. In six glioblastoma cell lines, a positive correlation was detected between the expression levels of miR-23a and invasiveness. A luciferase reporter assay demonstrated that homeobox D10 (HOXD10) was a miR-23a-target molecule, which was verified by high scores from both the PicTar and miRanda algorithms. Forced expression of miR-23a induced expression of invasion-related molecules, including uPAR, RhoA, and RhoC, and altered expression of glial-mesenchymal transition markers such as Snail, Slug, MMP2, MMP9, MMP14, and E-cadherin; however, these changes in expression levels were reversed by HOXD10 overexpression. Thus, miR-23a significantly promoted invasion of glioblastoma cells with polarized formation of focal adhesions, while exogenous HOXD10 overexpression reversed these phenomena. Here, we identify miR-23a-regulated HOXD10 as a pivotal regulator of invasion in glioblastoma, providing a novel mechanism for the aggressive invasiveness of this tumor and providing insight into potential therapeutic targets.
• Pathological Quantification of Carotid Artery Plaque Instability in Patients Undergoing Carotid Endarterectomy.

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Tohru Morita, Daisuke Hotta, Ryota Nomura, Yusuke Nakagaki, Takeo Murahashi, Kenji Kamiyama, Tetsuyuki Yoshimoto, Takeshi Aoki, Hiroshi Nishihara, Shinya Tanaka

  Circulation journal : official journal of the Japanese Circulation Society 82 (1) 258 - 266 1346-9843 2017/12/25 [Refereed][Not invited]
   

  BACKGROUND: Unstable atherosclerotic carotid plaques cause cerebral thromboemboli and ischemic events. However, this instability has not been pathologically quantified, so we sought to quantify it in patients undergoing carotid endarterectomy (CEA).Methods and Results:Carotid plaques were collected during CEA from 67 symptomatic and 15 asymptomatic patients between May 2015 and August 2016. The specimens were stained with hematoxylin-eosin and elastica-Masson. Immunohistochemistry was performed using an endothelial-specific antibody to CD31, CD34 and PDGFRβ. The histopathological characteristics of the plaques were studied. By multiple-variable logistic regression analysis, plaque instability correlated with the presence of plaque rupture [odds ratio (OR), 9.75; P=0.013], minimum fibrous cap thickness (OR per 10 μm 0.70; P=0.025), presence of microcalcifications in the fibrous cap (OR 7.82; P=0.022) and intraplaque microvessels (OR 1.91; P=0.043). Receiver-operating characteristics analyses showed that these factors combined into a single score diagnosed symptomatic carotid plaques in patients with carotid artery stenosis with a high level of accuracy (area under the curve 0.92; 95% confidence interval 0.85-0.99 vs. asymptomatic). CONCLUSIONS: This analysis of carotid plaque instability strongly suggested that the diagnostic scoring of carotid plaque instability improves the understanding and treatment of carotid artery disease in patients undergoing CEA.
• ハイドロゲルを用いた癌幹細胞新規誘導法の開発

  鈴鹿 淳, 津田 真寿美, 王 磊, 仙葉 愼吾, 油谷 幸代, 黒川 孝幸, 近江谷 克裕, 安田 和則, きょう 剣萍, 田中 伸哉

  生命科学系学会合同年次大会 生命科学系学会合同年次大会運営事務局 2017年度 [1P - 0964] 2017/12
• The Anti-tumor Effect of Cabozantinib on Ovarian Clear Cell Carcinoma In Vitro and In Vivo

  Makiko Nakatani, Hidemichi Watari, Takashi Mitamura, Lei Wang, Yutaka Hatanaka, Kanako C. Hatanaka, Kohei Honda, Toshiyuki Nomura, Hiroshi Nishihara, Shinya Tanaka, Noriaki Sakuragi

  ANTICANCER RESEARCH 37 (11) 6125 - 6132 0250-7005 2017/11 [Refereed][Not invited]
   

  Background: Several reports have shown that the overexpression of the MET proto-oncogene, receptor tyrosine kinase (MET), was more frequently observed in clear cell carcinoma (CCC) than in non-CCC. We evaluated the antitumor activity of cabozantinib, that targets MET. Materials and Methods: A gene expression analysis of tumors from human ovarian cancers was carried out by transcriptome sequencing. An in vitro 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide assay (MTT assay) and in vivo experiments were performed to determine the activity of cabozantinib. Results: The MET levels were higher in tumors with CCC than high-grade serous carcinoma (2.2-fold). Cabozantinib inhibited cell viability and phosphorylation of AKT and MAPK under the treatment of hepatocyte growth factor in RMG-I CCC cells. The tumors removed from mice given cabozantinib of 10 mg/kg weighed 70% less than control on day 15, and the immunohistochemical reactivity of phosphorylated MET was reduced compared with control mice. Conclusion: Cabozantinib contributes to tumor reduction, and phosphorylated MET represents an attractive target of CCC.
• Interferon-inducible Mx1 protein is highly expressed in renal tissues from treatment-naïve lupus nephritis, but not in those under immunosuppressive treatment.

  Shimizu Y, Yasuda S, Kimura T, Nishio S, Kono M, Ohmura K, Shimamura S, Kono M, Fujieda Y, Kato M, Oku K, Bohgaki T, Fukasawa Y, Tanaka S, Atsumi T

  Modern rheumatology 28 (4) 1 - 9 1439-7595 2017/11 [Refereed][Not invited]
  
• Persistent homology index as a robust quantitative measure of immunohistochemical scoring

  Akihiro Takiyama, Takashi Teramoto, Hiroaki Suzuki, Katsushige Yamashiro, Shinya Tanaka

  SCIENTIFIC REPORTS 7 (1) 14002  2045-2322 2017/10 [Refereed][Not invited]
   

  Immunohistochemical data (IHC) plays an important role in clinical practice, and is typically gathered in a semi-quantitative fashion that relies on some degree of visual scoring. However, visual scoring by a pathologist is inherently subjective and manifests both intra-observer and inter-observer variability. In this study, we introduce a novel computer-aided quantification methodology for immunohistochemical scoring that uses the algebraic concept of persistent homology. Using 8 bit grayscale image data derived from 90 specimens of invasive ductal carcinoma of the breast, stained for the replicative marker Ki-67, we computed homology classes. These were then compared to nuclear grades and the Ki-67 labeling indices obtained by visual scoring. Three metrics for IHC staining were newly defined: Persistent Homology Index (PHI), center coordinates of positive and negative groups, and the sum of squares within groups (WSS). This study demonstrates that PHI, a novel index for immunohistochemical labeling using persistent homology, can produce highly similar data to that generated by a pathologist using visual evaluation. The potential benefits associated with our novel technology include both improved quantification and reproducibility. Since our method reflects cellularity and nuclear atypia, it carries a greater quantity of biologic data compared to conventional evaluation using Ki-67.
• 悪性中皮腫におけるOTUB1の発現

  九笹 めい, 谷野 美智枝, 北崎 アリサ, 杉野 弘和, 石田 雄介, 王 磊, 津田 真寿美, 高澤 啓, 平野 博嗣, 田中 伸哉

  日本癌学会総会記事 76回 P - 3277 0546-0476 2017/09
• 多形黄色星細胞腫におけるBRAF遺伝子変異(BRAF V600E)とp16の発現の検討

  谷野 美智枝, 南條 博, 津田 真寿美, 杉野 弘和, 王 磊, 石田 雄介, 田中 伸哉

  日本癌学会総会記事 76回 P - 3317 0546-0476 2017/09
• 悪性神経膠腫においてIDH1遺伝子変異は放射線照射後のアポトーシスを亢進する

  北崎 アリサ, 谷野 美智枝, 九笹 めい, 杉野 弘和, 王 磊, 石田 雄介, 仙葉 慎吾, 津田 真寿美, 五十嵐 香織, 曽我 朋義, 田中 伸哉

  日本癌学会総会記事 76回 P - 3323 0546-0476 2017/09
• Notch 1 regulates invasion and metastasis of head and neck squamous cell carcinoma by inducing EMT through c-Myc

  Naoya Inamura, Taichi Kimura, Lei Wang, Hiroko Yanagi, Masumi Tsuda, Mishie Tanino, Hiroshi Nishihara, Satoshi Fukuda, Shinya Tanaka

  AURIS NASUS LARYNX 44 (4) 447 - 457 0385-8146 2017/08 [Refereed][Not invited]
   

  Objective: As 50% of patients of head and neck squamous carcinoma (HNSCC) exhibit poor prognosis, the identification of new therapeutic targets is required. Recently, there have been several reports about the correlation between Notch1 and HNSCC, but the precise mechanism is still obscure. Therefore, in this study, we examined the involvement of Notch I in HNSCC by using HNSCC cell lines and surgical specimens. Methods: To investigate the role of Notch1 in HNSCC, we examined the effect of Notch inhibitor DAPT on cell growth, invasion, and tumorigenicity using five HNSCC cell lines in vitro and in vivo. We further examined that the correlation with Notch expression and clinical prognostic factors was evaluated by using 101 HNSCC surgical specimens. Results: DAPT reduced the nuclear expression of Notch and c-Myc and repressed cell growth, EMT-dependent cell invasion in vitro, and tumorigenicity in vivo. An overexpression of Myc enhanced EMT with an increase of Snail and vimentin together with decreased levels of E-cadherin in HSC3 cells. Finally, we discovered that Notch expression was well correlated with MIB-1 index and lymph node metastases. Conclusion: We discovered that Notch1 was strongly correlated with HNSCC growth, invasion, and metastases. Therefore, Notch1 might be a new therapeutic target and a predictive marker of proliferation and metastasis of HNSCC. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
• Disseminated toxoplasmosis after hematopoietic stem cell transplantation showing unusual magnetic resonance images

  Takahiro Tateno, Masahiro Onozawa, Junichi Hashiguchi, Takashi Ishio, Sayaka Yuzawa, Satomi Matsuoka, Mizuha Kosugi-Kanaya, Kohei Okada, Souichi Shiratori, Hideki Goto, Taichi Kimura, Junichi Sugita, Masao Nakagawa, Daigo Hashimoto, Kaoru Kahata, Katsuya Fujimoto, Tomoyuki Endo, Takeshi Kondo, Shinya Tanaka, Satoshi Hashino, Takanori Teshima

  TRANSPLANT INFECTIOUS DISEASE 19 (4) 1398-2273 2017/08 [Refereed][Not invited]
   

  We herein report a patient who had disseminated toxoplasmosis after hematopoietic stem cell transplantation showing atypical clinical presentation and neuroimaging. Parkinsonism symptoms such as muscle rigidity, bradykinesia, tremor, and postural instability were initial manifestations. Magnetic resonance imaging showed diffuse symmetrical lesions of bilateral basal ganglia lacking ringed enhancement. Post-mortem analysis revealed multiple tachyzoites of Toxoplasma gondii in the basal ganglia, mid brain, cerebellum, and cardiac muscle.
• Novel method of rapid immunohistochemistry based on alternating current electric field (Histo-Tek R-IHC) and its application for clinical and research field

  森谷 純, 谷野 美智枝, 津田 真寿美, 田中 伸哉

  生体の科学 金原一郎記念医学医療振興財団 ; 1949- 68 (4) 365 - 370 0370-9531 2017/07 [Not refereed][Not invited]
  
• Regulation of endothelial Fas expression as a mechanism of promotion of vascular integrity by mural cells in tumors.

  Ryosuke Kamei, Hiroyoshi Y Tanaka, Takao Kawano, Chiharu Morii, Sayaka Tanaka, Hiroshi Nishihara, Caname Iwata, Mitsunobu R Kano

  Cancer science 108 (5) 1080 - 1088 1347-9032 2017/05 [Not refereed][Not invited]
   

  Angiogenesis is a multi-step process that culminates in vascular maturation whereby nascent vessels stabilize to become functional, and mural cells play an essential role in this process. Recent studies have shown that mural cells in tumors also promote and maintain vascular integrity, with wide-reaching clinical implications including the regulation of tumor growth, metastases, and drug delivery. Various regulatory signaling pathways have been hitherto implicated, but whether regulation of Fas-dependent apoptotic mechanisms is involved has not yet been fully investigated. We first compared endothelial FAS staining in human pancreatic ductal adenocarcinomas and colon carcinomas and show that the latter, characterized by lower mural cell coverage of tumor vasculature, demonstrated higher expression of FAS than the former. Next, in an in vitro coculture system of MS-1 and 10T1/2 cells as endothelial and mural cells respectively, we show that mural cells decreased endothelial Fas expression. Then, in an in vivo model in which C26 colon carcinoma cells were inoculated together with MS-1 cells alone or with the further addition of 10T1/2 cells, we demonstrate that mural cells prevented hemorrhage. Finally, knockdown of endothelial Fas sufficiently recapitulated the protection against hemorrhage seen with the addition of mural cells. These results together suggest that regulation of endothelial Fas signaling is involved in the promotion of vascular integrity by mural cells in tumors.
• Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes

  Shunsuke Kon, Kojiro Ishibashi, Hiroto Katoh, Sho Kitamoto, Takanobu Shirai, Shinya Tanaka, Mihoko Kajita, Susumu Ishikawa, Hajime Yamauchi, Yuta Yako, Tomoko Kamasaki, Tomohiro Matsumoto, Hirotaka Watanabe, Riku Egami, Ayana Sasaki, Atsuko Nishikawa, Ikumi Kameda, Takeshi Maruyama, Rika Narumi, Tomoko Morita, Yoshiteru Sasaki, Ryosuke Enoki, Sato Honma, Hiromi Imamura, Masanobu Oshima, Tomoyoshi Soga, Jun-ichi Miyazaki, Michael R. Duchen, Jin-Min Nam, Yasuhito Onodera, Shingo Yoshioka, Junichi Kikuta, Masaru Ishii, Masamichi Imajo, Eisuke Nishida, Yoichiro Fujioka, Yusuke Ohba, Toshiro Sato, Yasuyuki Fujita

  NATURE CELL BIOLOGY 19 (5) 530 - + 1465-7392 2017/05 [Refereed][Not invited]
   

  Recent studies have revealed that newly emerging transformed cells are often apically extruded from epithelial tissues. During this process, normal epithelial cells can recognize and actively eliminate transformed cells, a process called epithelial defence against cancer (EDAC). Here, we show that mitochondrial membrane potential is diminished in RasV12-transformed cells when they are surrounded by normal cells. In addition, glucose uptake is elevated, leading to higher lactate production. The mitochondrial dysfunction is driven by upregulation of pyruvate dehydrogenase kinase 4 (PDK4), which positively regulates elimination of RasV12-transformed cells. Furthermore, EDAC from the surrounding normal cells, involving filamin, drives the Warburg-effect-like metabolic alteration. Moreover, using a cell-competition mouse model, we demonstrate that PDK-mediated metabolic changes promote the elimination of RasV12-transformed cells from intestinal epithelia. These data indicate that non-cell-autonomous metabolic modulation is a crucial regulator for cell competition, shedding light on the unexplored events at the initial stage of carcinogenesis.
• PCTAIRE1/CDK16/PCTK1 is overexpressed in cutaneous squamous cell carcinoma and regulates p27 stability and cell cycle

  Teruki Yanagi, Hiroo Hata, Eri Mizuno, Shinya Kitamura, Keisuke Imafuku, Shinichi Nakazato, Lei Wang, Hiroshi Nishihara, Shinya Tanaka, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGICAL SCIENCE 86 (2) 149 - 157 0923-1811 2017/05 [Refereed][Not invited]
   

  Background: PCTAIRE1 (also known as cyclin-dependent kinase 16 (Cdk16) and PCTK1) is a Cdk family protein that has been implicated in spermatogenesis. We recently revealed the function of PCTAIRE1 in the tumorigenesis of malignancies, including breast and prostate cancers; however, the tumorigenic function of PCTAIRE1 in cutaneous squamous cell carcinoma (SCC) remains unclear. Objective: In this study, we investigated the role of PCTAIRE1 in the tumorigenesis of cutaneous SCCs. Methods and results: In cutaneous/oral SCC A431, DJM-1, HSC-3 cells, PCTAIRE1 gene-knockdown was found to diminish cell proliferation as assessed by cell counting and clonogenic assays. FACS analyses of annexin V-PI staining and DNA content showed PCTAIRE1 knockdown to cause G2/M arrest followed by apoptosis. The depletion of PCTAIRE1 was found to lead to the accumulation of tumor suppressor p27 and down-regulation of c-Myc. In tumor xenografts of A431 cells, the conditional knockdown of PCTAIRE1 restores p27 protein expression and suppresses tumor growth. Clinically, in primary tumors from patients with SCC, PCTAIRE1 is more highly expressed in malignant lesions than in adjacent normal epidermis. Conversely, expression levels of p27 are significantly lower in SCC than in normal epidermis. Conclusions: Our findings reveal a crucial function for PCTAIRE1 in regulating p27, c-Myc levels and tumor growth in cutaneous SCC cells, suggesting that PCTAIRE1 could be a novel target for skin tumor treatment. (C) 2017 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
• 無気肺および胸水貯留、骨盤内腫瘍、多発リンパ節転移を来した原発不明腫瘍の剖検例

  石田 雄介, 谷川 聖, 杉村 拓也, 大森 優子, 篠原 敏也, 竹浪 智子, 漆戸 万紗那, 森谷 純, 谷野 美智枝, 田中 伸哉

  日本臨床細胞学会雑誌 (公社)日本臨床細胞学会 56 (Suppl.1) 393 - 393 0387-1193 2017/04
• THE ADAPTOR PROTEIN CRK-INDUCED ERBB2 EXPRESSION PROMOTES TUMOR PROGRESSION AND METASTASIS OF BLADDER CANCER VIA EXOSOMES

  Kazuhiko Yoshida, Masumi Tsuda, Ryuji Matsumoto, Shingo Semba, Taichi Kimura, Mishie Tanino, Hiroshi Nishihara, Tsunenori Kondo, Kazunari Tanabe, Shinya Tanaka

  JOURNAL OF UROLOGY 197 (4) E1175 - E1175 0022-5347 2017/04 [Refereed][Not invited]
  
• Hypoxic glucose metabolism in glioblastoma as a potential prognostic factor.

  Takuya Toyonaga, Shigeru Yamaguchi, Kenji Hirata, Kentaro Kobayashi, Osamu Manabe, Shiro Watanabe, Shunsuke Terasaka, Hiroyuki Kobayashi, Naoya Hattori, Tohru Shiga, Yuji Kuge, Shinya Tanaka, Yoichi M Ito, Nagara Tamaki

  European journal of nuclear medicine and molecular imaging 44 (4) 611 - 619 1619-7070 2017/04 [Refereed][Not invited]
   

  PURPOSE: Metabolic activity and hypoxia are both important factors characterizing tumor aggressiveness. Here, we used F-18 fluoromisonidazole (FMISO) and F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) to define metabolically active hypoxic volume, and investigate its clinical significance in relation to progression free survival (PFS) and overall survival (OS) in glioblastoma patients. EXPERIMENTAL DESIGN: Glioblastoma patients (n = 32) underwent FMISO PET, FDG PET, and magnetic resonance imaging (MRI) before surgical intervention. FDG and FMISO PET images were coregistered with gadolinium-enhanced T1-weighted MR images. Volume of interest (VOI) of gross tumor volume (GTV) was manually created to enclose the entire gadolinium-positive areas. The FMISO tumor-to-normal region ratio (TNR) and FDG TNR were calculated in a voxel-by-voxel manner. For calculating TNR, standardized uptake value (SUV) was divided by averaged SUV of normal references. Contralateral frontal and parietal cortices were used as the reference region for FDG, whereas the cerebellar cortex was used as the reference region for FMISO. FDG-positive was defined as the FDG TNR ≥1.0, and FMISO-positive was defined as FMISO TNR ≥1.3. Hypoxia volume (HV) was defined as the volume of FMISO-positive and metabolic tumor volume in hypoxia (hMTV) was the volume of FMISO/FDG double-positive. The total lesion glycolysis in hypoxia (hTLG) was hMTV × FDG SUVmean. The extent of resection (EOR) involving cytoreduction surgery was volumetric change based on planimetry methods using MRI. These factors were tested for correlation with patient prognosis. RESULTS: All tumor lesions were FMISO-positive and FDG-positive. Univariate analysis indicated that hMTV, hTLG, and EOR were significantly correlated with PFS (p = 0.007, p = 0.04, and p = 0.01, respectively) and that hMTV, hTLG, and EOR were also significantly correlated with OS (p = 0.0028, p = 0.037, and p = 0.014, respectively). In contrast, none of FDG TNR, FMISO TNR, GTV, HV, patients' age, or Karnofsky performance scale (KPS) was significantly correlated with PSF or OS. The hMTV and hTLG were found to be independent factors affecting PFS and OS on multivariate analysis. CONCLUSIONS: We introduced hMTV and hTLG using FDG and FMISO PET to define metabolically active hypoxic volume. Univariate and multivariate analyses demonstrated that both hMTV and hTLG are significant predictors for PFS and OS in glioblastoma patients.
• 皮膚悪性黒色腫に対するオプジーボ投与後に出現し免疫染色にてS-100陰性を呈した転移性脳腫瘍の1例

  石田 雄介, 高橋 達郎, 佐藤 行真, 池田 正起, 守田 玲菜, 武井 英博, 木村 太一, 津田 真寿美, 谷野 美智枝, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 106 (1) 477 - 477 0300-9181 2017/03
• 急性前骨髄球性白血病(APL)から播種性血管内凝固症候群(DIC)および意識障害を来して死亡した1剖検例

  勝尾 知尋, 中川 恵, 石田 雄介, 高橋 達郎, 下埜 城嗣, 武井 英博, 木村 太一, 谷野 美智枝, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 106 (1) 515 - 515 0300-9181 2017/03
• Pathologically dissimilar acute stent thromboses in a metal allergic patient

  Takao Konishi, Daisuke Hotta, Naohiro Funayama, Tadashi Yamamoto, Hiroshi Nishihara, Shinya Tanaka

  CORONARY ARTERY DISEASE 28 (2) 175 - 176 0954-6928 2017/03 [Refereed][Not invited]
  
• Gene dosage effect in spinocerebellar ataxia type 6 homozygotes: A clinical and neuropathological study

  Kazumasa Soga, Kinya Ishikawa, Tokuro Furuya, Tadatsune Iida, Tetsuo Yamada, Noboru Ando, Kiyobumi Ota, Hiromi Kanno-Okada, Shinya Tanaka, Masayuki Shintaku, Yoshinobu Eishi, Hidehiro Mizusawa, Takanori Yokota

  JOURNAL OF THE NEUROLOGICAL SCIENCES 373 321 - 328 0022-510X 2017/02 [Refereed][Not invited]
   

  Spinocerebellar ataxia type 6 (SCA6) is an autosomal dominant neurodegenerative disorder. However, it remains unclear whether SCA6 shows a gene dosage effect, defined by earlier age-of-onset in homozygotes than heterozygotes. Herein, we retrospectively analyzed four homozygous SCA6 subjects from our single institution cohort of 120 SCA6 subjects. We also performed a neuropathological investigation into an SCA6 individual with compound heterozygous expansions. In the 116 heterozygotes, there was an inverse correlation of age-of-onset with the number of CAG repeats in the expanded allele, and with the total number of CAG repeats, in both normal and expanded alleles. The age-of-onset in the four homozygotes was within the 95% confidence interval of the age-of onset versus the repeat-lengths correlations determined in the 116 heterozygotes. Nevertheless, all homozygotes had earlier onset than their parents, and showed rapid disease progression. Neuropathology revealed neuronal loss, as well as alpha 1A-calcium channel protein aggregates in Purkinje cells, a few alpha 1A-calcium channel protein aggregates in the neocortex and basal ganglia, and neuronal loss in Clarke's column and the globus pallidus not seen in heterozygotes. These data suggest a mild clinical and neuropathological gene dosage effect in SCA6 subjects. (C) 2016 Elsevier B.V. All rights reserved.
• Clinicopathological evaluation of Sox10 expression in diffuse-type gastric adenocarcinoma

  Marin Kato, Hiroshi Nishihara, Hideyuki Hayashi, Taichi Kimura, Yusuke Ishida, Lei Wang, Masumi Tsuda, Mishie Ann Tanino, Shinya Tanaka

  MEDICAL ONCOLOGY 34 (1) 8  1357-0560 2017/01 [Refereed][Not invited]
   

  Sox10, one of the transcription factors, regulates Wnt/beta-catenin signaling in diverse developmental processes in normal tissues. Sox10 is also expressed in variable solid tumors such as breast cancer, salivary tumor, hepatocellular carcinoma, ovarian tumor, nasopharyngeal carcinoma, prostate cancer, and digestive cancer. The role of Sox10 during tumorigenesis is still controversial, especially in digestive cancers; thus, we performed clinicopathological evaluation of Sox10 expression in 41 cases of diffuse-type gastric adenocarcinoma (DGA). We examined the expression of Sox10 by immunohistochemical staining and real-time quantitative reverse transcriptase PCR and evaluated the correlation between Sox10 expression and clinicopathological factors. A low-level expression of Sox10 was significantly associated with high-level venous invasion by immunohistochemical evaluation, while it was significantly associated with high-level lymphatic permeation when analyzed by real-time PCR assay. Survival analysis of 41 cases indicated that high level of vascular permeation was a statistically poor prognostic factor, suggesting that derogation of Sox10 would lead to unfavorable patients' outcome through the acceleration of vascular invasion. In this study, we revealed the clinical benefit of evaluation of Sox10 expression to predict the risk of vascular permeation which yields patients' poor prognosis in DGA.
• Prognostic Value of Eosinophil to Leukocyte Ratio in Patients with ST-Elevation Myocardial Infarction Undergoing Primary Percutaneous Coronary Intervention

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Toru Morita, Daisuke Hotta, Hiroshi Nishihara, Shinya Tanaka

  JOURNAL OF ATHEROSCLEROSIS AND THROMBOSIS 24 (8) 827 - 840 1340-3478 2017 [Refereed][Not invited]
   

  Aim: Leukocyte profile has been related to clinical outcome in patients with ST-segment elevation (STE) myocardial infarction (MI). However, whether eosinophil to leukocyte ratio (ELR) predicts clinical outcome in patients who have undergone primary percutaneous coronary intervention (PCI) remains unclear. Therefore, we examined the prognostic value of ELR in this patient population. Methods: We retrospectively analyzed the data of 331 consecutive patients who underwent primary PCI for STEMI between January 2009 and March 2015. All leukocyte types were counted and ELR was calculated for all patients 24 h after hospital admission. The primary study endpoint was major adverse cardiac events (MACEs) within up to one year of follow-up duration. Results: MACEs including cardiac deaths in 9.4% of the patients, MI in 1.5%, and target lesion or vessel revascularization in 10.3%, occurred within one year in 68 patients (20.5%). The mean ELR was significantly lower in patients with MACEs than in patients without MACEs (0.20 +/- 0.51 vs. 0.49 +/- 0.66, respectively; p < 0.001). An ELR < 0.1 at 24 h was identified as the best cut-off value for mortality prediction. Multivariate analysis identified that an ELR < 0.1 (odds ratio [OR] 0.38; 95% confidence interval [CI] 0.22-0.67; p < 0.001) and chronic kidney disease (OR 2.38; CI 1.334.24; p < 0.003) are independent predictors of MACEs. Conclusion: In primary PCI patients with STEMI, ELR at 24 h was an independent predictor of MACEs in addition to the usual coronary risk factors and commonly used biomarkers.
• A case of giant cell-rich solitary fibrous tumor in the external auditory canal

  Sayaka Yuzawa, Satoshi Tanikawa, Isamu Kunibe, Hiroshi Nishihara, Kazuo Nagashima, Shinya Tanaka

  PATHOLOGY INTERNATIONAL 66 (12) 701 - 705 1320-5463 2016/12 [Refereed][Not invited]
   

  We present a rare case of giant cell-rich solitary fibrous tumor (SFT) arising at the left external auditory canal in a 31-year-old woman. The tumor was well-circumscribed and composed of spindle-shaped cells with abundant collagenous bands. Scattered multinucleate giant cells were observed, some of which lined pseudovascular spaces. Although a focal mild-hypercellular area was observed, mitoses were rare and necrosis was absent. Interstitial mast cells were scattered, especially in the hypercellular area. Immunohistochemically, CD34, vimentin, and Bcl-2 presented diffuse positivity. Moreover, both mononuclear spindle cells and multinucleate cells showed nuclear STAT6 positivity, while NAB2-STAT6 fusion gene could not be detected by reverse transcription polymerase chain reaction using formalin-fixed specimen. These findings suggest the pathological diagnosis of giant cell-rich SFT, previously known as giant cell angiofibroma, which is a rare variant of SFT with multinucleate giant cells and occurs predominantly in orbital region. Although giant cell-rich SFTs of extra-orbital sites have been reported, to our knowledge, this is the first case arising in the external auditory canal. Giant cell-rich SFT should be considered as a differential diagnosis of spindle cell lesion with multinucleate giant cells, and STAT6 immunohistochemistry should be performed to distinguish this rare tumor from other mesenchymal neoplasms.
• MicroRNA expression profiles of multiple system atrophy from formalin-fixed paraffin-embedded samples

  Koichi Wakabayashi, Fumiaki Mori, Akiyoshi Kakita, Hitoshi Takahashi, Shinya Tanaka, Jun Utsumi, Hidenao Sasaki

  NEUROSCIENCE LETTERS 635 117 - 122 0304-3940 2016/12 [Refereed][Not invited]
   

  MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression. Recently, we have shown that informative miRNA data can be derived from archived formalin-fixed paraffin-embedded (FFPE) samples from postmortem cases of amyotrophic lateral sclerosis and normal controls. miRNA analysis has now been performed on FFPE samples from affected brain regions in patients with multiple system atrophy (MSA) and the same areas in neurologically normal controls. We evaluated 50 samples from patients with MSA (n =13) and controls (n=13). Twenty-six samples were selected for miRNA analysis on the basis of the criteria reported previously: (i) a formalin fixation time of less than 4 weeks, (ii) a total RNA yield per sample of more than 500 ng, and (iii) sufficient quality of the RNA electrophoresis pattern. These included 11 cases of MSA and 5 controls. Thus, the success rate for analysis of RNA from FFPE samples was 52% (26 of 50). For MSA, a total of 395 and 383 miRNAs were identified in the pons and cerebellum, respectively; 5 were up-regulated and 33 were down-regulated in the pons and 5 were up-regulated and 18 were down-regulated in the cerebellum. Several miRNAs down-regulated in the pons (miR-129-2-3p and miR-129-5p) and cerebellum (miR-129-2-3p, miR-129-5p and miR-132-3p) had already been identified in frozen cerebellum from MSA patients. These findings suggest that archived FFPE postmortem samples can be a valuable source for miRNA profiling in MSA. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
• Severe mitral regurgitation due to mitral leaflet aneurysm diagnosed by three-dimensional transesophageal echocardiography: a case report

  Takao Konishi, Naohiro Funayama, Tadashi Yamamoto, Daisuke Hotta, Kenjiro Kikuchi, Katsumi Ohori, Hiroshi Nishihara, Shinya Tanaka

  BMC CARDIOVASCULAR DISORDERS 16 (1) 1471-2261 2016/11 [Not refereed][Not invited]
   

  Background: A small mitral valve aneurysm (MVA) presenting as severe mitral regurgitation (MR) is uncommon. Case presentation: A 47-year-old man with a history of hypertension complained of exertional chest discomfort. A transthoracic echocardiogram (TTE) revealed the presence of MR and prolapse of the posterior leaflet. A 6mm in diameter MVA, not clearly visualized by TTE, was detected on the posterior leaflet on a three-dimensional (3D) transesophageal echocardiography (TEE). The patient underwent uncomplicated triangular resection of P2 and mitral valve annuloplasty, and was discharged from postoperative rehabilitation, 2 weeks after the operation. Histopathology of the excised leaflet showed myxomatous changes without infective vegetation or signs of rheumatic heart disease. Conclusions: A small, isolated MVA is a cause of severe MR, which might be overlooked and, therefore, managed belatedly. 3D TEE was helpful in imaging its morphologic details.
• Aldo-keto reductase 1C1 induced by interleukin-1β mediates the invasive potential and drug resistance of metastatic bladder cancer cells.

  Ryuji Matsumoto, Masumi Tsuda, Kazuhiko Yoshida, Mishie Tanino, Taichi Kimura, Hiroshi Nishihara, Takashige Abe, Nobuo Shinohara, Katsuya Nonomura, Shinya Tanaka

  Scientific reports 6 34625 - 34625 2016/10/04 [Refereed][Not invited]
   

  In treating bladder cancer, determining the molecular mechanisms of tumor invasion, metastasis, and drug resistance are urgent to improving long-term patient survival. One of the metabolic enzymes, aldo-keto reductase 1C1 (AKR1C1), plays an essential role in cancer invasion/metastasis and chemoresistance. In orthotopic xenograft models of a human bladder cancer cell line, UM-UC-3, metastatic sublines were established from tumors in the liver, lung, and bone. These cells possessed elevated levels of EMT-associated markers, such as Snail, Slug, or CD44, and exhibited enhanced invasion. By microarray analysis, AKR1C1 was found to be up-regulated in metastatic lesions, which was verified in metastatic human bladder cancer specimens. Decreased invasion caused by AKR1C1 knockdown suggests a novel role of AKR1C1 in cancer invasion, which is probably due to the regulation of Rac1, Src, or Akt. An inflammatory cytokine, interleukin-1β, was found to increase AKR1C1 in bladder cancer cell lines. One particular non-steroidal anti-inflammatory drug, flufenamic acid, antagonized AKR1C1 and decreased the cisplatin-resistance and invasion potential of metastatic sublines. These data uncover the crucial role of AKR1C1 in regulating both metastasis and drug resistance; as a result, AKR1C1 should be a potent molecular target in invasive bladder cancer treatment.
• Genetic landscape of meningioma

  Sayaka Yuzawa, Hiroshi Nishihara, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 33 (4) 237 - 247 1433-7398 2016/10 [Refereed][Not invited]
   

  Meningioma is the most common intracranial tumor, arising from arachnoid cells of the meninges. Monosomy 22 and inactivating mutations of NF2 are well-known genetic alterations of meningiomas. More recently, mutations in TRAF7, AKT1, KLF4, SMO, and PIK3CA were identified by next-generation sequencing. We here reviewed 553 meningiomas for the mutational patterns of the six genes. NF2 aberration was observed in 55 % of meningiomas. Mutations of TRAF7, AKT1, KLF4, PIK3CA, and SMO were identified in 20, 9, 9, 4.5, and 3 % of cases, respectively. Altogether, 80 % of cases harbored at least one of the genetic alterations in these genes. NF2 alterations and mutations of the other genes were mutually exclusive with a few exceptions. Clinicopathologically, tumors with mutations in TRAF7/AKT1 and SMO shared specific features: they were located in the anterior fossa, median middle fossa, or anterior calvarium, and most of them were meningothelial or transitional meningiomas. TRAF7/KLF4 type meningiomas showed different characteristics in that they occurred in the lateral middle fossa and median posterior fossa as well as anterior fossa and median middle fossa, and contained a secretory meningioma component. We also discuss the mutational hotspots of these genes and other genetic/cytogenetic alterations contributing to tumorigenesis or progression of meningiomas.
• 合成PAMPSゲルはタンパク質レザバーとして機能し、軟骨細胞ATDC5細胞に軟骨形成分化を誘導する(Synthetic PAMPS gel functions as protein reservoir, which induces a chondrogenic differentiation of chondrocytic ATDC5 cells)

  仙葉 愼吾, 後藤 佳子, 北村 信人, 黒野 定, 近江谷 克裕, 津田 真寿美, 黒川 孝幸, グン・チェンピン, 田中 伸哉, 安田 和則

  日本生化学会大会プログラム・講演要旨集 89回 [1T18 - 405)] 2016/09
• BRAF V600E変異検索およびKIAA1549-BRAF融合遺伝子検索により診断に至った若年成人発症毛様細胞性星細胞腫

  石田 雄介, 津田 真寿美, 澤村 豊, 村井 宏, 堀内 成好, 長嶋 和郎, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 105 (2) 75 - 75 0300-9181 2016/09
• Dipeptidyl peptidase-4 inhibitor improved exercise capacity and mitochondrial biogenesis in mice with heart failure via activation of glucagon-like peptide-1 receptor signalling

  Shingo Takada, Yoshihiro Masaki, Shintaro Kinugawa, Junichi Matsumoto, Takaaki Furihata, Wataru Mizushima, Tomoyasu Kadoguchi, Arata Fukushima, Tsuneaki Homma, Masashige Takahashi, Shinichi Harashima, Shouji Matsushima, Takashi Yokota, Shinya Tanaka, Koichi Okita, Hiroyuki Tsutsui

  CARDIOVASCULAR RESEARCH 111 (4) 338 - 347 0008-6363 2016/09 [Refereed][Not invited]
   

  Aims Exercise capacity is reduced in heart failure (HF) patients, due mostly to skeletal muscle abnormalities including impaired energy metabolism, mitochondrial dysfunction, fibre type transition, and atrophy. Glucagon-like peptide-1 (GLP-1) has been shown to improve exercise capacity in HF patients. We investigated the effects of the administration of a dipeptidyl peptidase (DPP)-4 inhibitor on the exercise capacity and skeletal muscle abnormalities in an HF mouse model after myocardial infarction (MI). Methods and resultsMI was created in male C57BL/6J mice by ligating the left coronary artery, and a sham operation was performed in other mice. The mice were then divided into two groups according to the treatment with or without a DPP-4 inhibitor, MK-0626 [1 mg/kg body weight (BW)/day] provided in the diet. Four weeks later, the exercise capacity evaluated by treadmill test was revealed to be limited in the MI mice, and it was ameliorated in the MI + MK-0626 group without affecting the infarct size or cardiac function. The citrate synthase activity, mitochondrial oxidative phosphorylation capacity, supercomplex formation, and their quantity were reduced in the skeletal muscle from the MI mice, and these decreases were normalized in the MI + MK-0626 group, in association with the improvement of mitochondrial biogenesis. Immunohistochemical staining also revealed that a shift toward the fast-twitch fibre type in the MI mice was also reversed by MK-0626. Favourable effects of MK-0626 were significantly inhibited by treatment of GLP-1 antagonist, Exendin-(9-39) (150 pmol/kg BW/min, subcutaneous osmotic pumps) in MI + MK-0626 mice. Similarly, exercise capacity and mitochondrial function were significantly improved by treatment of GLP-1 agonist, Exendin-4 (1 nmol/kg/BW/h, subcutaneous osmotic pumps). ConclusionsaEuro integral A DPP-4 inhibitor may be a novel therapeutic agent against the exercise intolerance seen in HF patients by improving the mitochondrial biogenesis in their skeletal muscle.
• Analysis of NAB2-STAT6 Gene Fusion in 17 Cases of Meningeal Solitary Fibrous Tumor/Hemangiopericytoma Review of the Literature

  Sayaka Yuzawa, Hiroshi Nishihara, Lei Wang, Masumi Tsuda, Taichi Kimura, Mishie Tanino, Shinya Tanaka

  AMERICAN JOURNAL OF SURGICAL PATHOLOGY 40 (8) 1031 - 1040 0147-5185 2016/08 [Refereed][Not invited]
   

  Solitary fibrous tumor/hemangiopericytoma (SFT/HPC) is a mesenchymal tumor that can affect virtually any region of the body. SFT/HPC of the thoracic cavity and soft tissue has been histologically considered a single biological entity termed SFT; in fact, NAB2-STAT6 gene fusion was recently identified in both diseases. In contrast, meningeal SFT and HPC still need to be investigated in detail with regard to gene fusion variants. The aim of this study was to verify the frequency of NAB2-STAT6 fusion and the relationship between fusion variants and clinicopathologic findings of SFT/HPC, especially meningeal SFT/HPC. We examined the NAB2-STAT6 fusion by reverse transcription polymerase chain reaction with 4 cases of meningeal SFT and 13 cases of meningeal HPC. NAB2-STAT6 fusion transcripts were identified in 12 of 17 cases, including NAB2ex6-STAT6ex17 (4/17, 24%), NAB2ex6-STAT6ex16 and NAB2ex4-STAT6ex2 (3/17, 18%, respectively), and NAB2ex5-STAT6ex16 (2/17, 12%). Three cases showed a pseudopapillary pattern, and 2 of them carried NAB2ex6-STAT6ex17. In addition, our meta-analysis revealed that the major fusion variant in meningeal SFT/HPC was NAB2ex6-STAT6ex16/17 (29/54, 54%), which was also common in soft tissue and intraperitoneum/retroperitoneum but rare in thoracic SFT. Fusion variant significantly correlated with age and histologic diagnosis in meningeal SFT/HPC but not with prognosis. Our results represented that meningeal SFT and HPC were in a single biological spectrum with NAB2-STAT6 gene fusion as was nonmeningeal SFT and further confirmed the organ-specific tumorigenic process and morphologic differences on the basis of fusion variants in meningeal SFT/HPC.
• Identification and analysis of CXCR4-positive synovial sarcoma-initiating cells

  T. Kimura, L. Wang, K. Tabu, M. Tsuda, M. Tanino, A. Maekawa, H. Nishihara, H. Hiraga, T. Taga, Y. Oda, S. Tanaka

  ONCOGENE 35 (30) 3932 - 3943 0950-9232 2016/07 [Not refereed][Not invited]
   

  Synovial sarcoma accounts for almost 10% of all soft tissue sarcomas, and its prognosis is poor with 5-year survival rates at 36%. Thus, new treatments and therapeutic targets for synovial sarcoma are required. Tumor-initiating cells have been defined by the ability for self-renewal and multipotent differentiation, and they exhibit higher tumorigenic capacity, chemoresistance and radiation resistance, expecting to be a new therapeutic target. In synovial sarcoma, the presence of such stemness remains largely unclear; thus, we analyzed whether synovial sarcoma possessed tumor-initiating cells and explored specific markers, and we discovered that synovial sarcoma cell lines possessed heterogeneity by way of containing a sphere-forming subpopulation highly expressing NANOG, OCT4 and SOX2. By expression microarray analysis, CXCR4 was identified to be highly expressed in the sphere subpopulation and correlated with stem-cell-associated markers. Inhibition of CXCR4 suppressed the cell proliferation of synovial sarcoma cell lines in vitro. The tumor-initiating ability of CXCR4-positive cells was demonstrated by xenograft propagation assay. CXCR4-positive cells showed higher tumorigenicity than negative ones and possessed both self-renewal and multipotent differentiation ability. Immunohistochemical analysis of 39 specimens of synovial sarcoma patients revealed that CXCR4 strongly correlated with poor prognosis of synovial sarcoma. Thus, we conclude that CXCR4 is the marker of synovial sarcoma-initiating cells, a new biomarker for prognosis and a new potential therapeutic target.
• Clinical impact of targeted amplicon sequencing for meningioma as a practical clinical-sequencing system

  Sayaka Yuzawa, Hiroshi Nishihara, Shigeru Yamaguchi, Hiromi Mohri, Lei Wang, Taichi Kimura, Masumi Tsuda, Mishie Tanino, Hiroyuki Kobayashi, Shunsuke Terasaka, Kiyohiro Houkin, Norihiro Sato, Shinya Tanaka

  MODERN PATHOLOGY 29 (7) 708 - 716 0893-3952 2016/07 [Refereed][Not invited]
   

  Recent genetic analyses using next-generation sequencers have revealed numerous genetic alterations in various tumors including meningioma, which is the most common primary brain tumor. However, their use as routine laboratory examinations in clinical applications for tumor genotyping is not cost effective. To establish a clinical sequencing system for meningioma and investigate the clinical significance of genotype, we retrospectively performed targeted amplicon sequencing on 103 meningiomas and evaluated the association with clinicopathological features. We designed amplicon-sequencing panels targeting eight genes including NF2 (neurofibromin 2), TRAF7, KLF4, AKT1, and SMO. Libraries prepared with genomic DNA extracted from PAXgenefixed paraffin-embedded tissues of 103 meningioma specimens were sequenced using the Illumina MiSeq. NF2 loss in some cases was also confirmed by interphase-fluorescent in situ hybridization. We identified NF2 loss and/or at least one mutation in NF2, TRAF7, KLF4, AKT1, and SMO in 81 out of 103 cases (79%) by targeted amplicon sequencing. On the basis of genetic status, we categorized meningiomas into three genotype groups: NF2 type, TRAKLS type harboring mutation in TRAF7, AKT1, KLF4, and/or SMO, and 'not otherwise classified' type. Genotype significantly correlated with tumor volume, tumor location, and magnetic resonance imaging findings such as adjacent bone change and heterogeneous gadolinium enhancement, as well as histopathological subtypes. In addition, multivariate analysis revealed that genotype was independently associated with risk of recurrence. In conclusion, we established a rapid clinical sequencing system that enables final confirmation of meningioma genotype within 7 days turnaround time. Our method will bring multiple benefits to neuropathologists and neurosurgeons for accurate diagnosis and appropriate postoperative management.
• Preclinical Evidence of Anti-Tumor Activity Induced by EZH2 Inhibition in Human Models of Synovial Sarcoma

  Satoshi Kawano, Alexandra R. Grassian, Masumi Tsuda, Sarah K. Knutson, Natalie M. Warholic, Galina Kuznetsov, Shanqin Xu, Yonghong Xiao, Roy M. Pollock, Jesse S. Smith, Kevin K. Kuntz, Scott Ribich, Yukinori Minoshima, Junji Matsui, Robert A. Copeland, Shinya Tanaka, Heike Keilhack

  PLOS ONE 11 (7) e0158888  1932-6203 2016/07 [Refereed][Not invited]
   

  The catalytic activities of covalent and ATP-dependent chromatin remodeling are central to regulating the conformational state of chromatin and the resultant transcriptional output. The enzymes that catalyze these activities are often contained within multiprotein complexes in nature. Two such multiprotein complexes, the polycomb repressive complex 2 (PRC2) methyltransferase and the SWItch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeler have been reported to act in opposition to each other during development and homeostasis. An imbalance in their activities induced by mutations/deletions in complex members (e.g. SMARCB1) has been suggested to be a pathogenic mechanism in certain human cancers. Here we show that preclinical models of synovial sarcoma-a cancer characterized by functional SMARCB1 loss via its displacement from the SWI/SNF complex through the pathognomonic SS18-SSX fusion protein-display sensitivity to pharmacologic inhibition of EZH2, the catalytic subunit of PRC2. Treatment with tazemetostat, a clinicalstage, selective and orally bioavailable small-molecule inhibitor of EZH2 enzymatic activity reverses a subset of synovial sarcoma gene expression and results in concentration-dependent cell growth inhibition and cell death specifically in SS18-SSX fusion-positive cells in vitro. Treatment of mice bearing either a cell line or two patient-derived xenograft models of synovial sarcoma leads to dose-dependent tumor growth inhibition with correlative inhibition of trimethylation levels of the EZH2-specific substrate, lysine 27 on histone H3. These data demonstrate a dependency of SS18-SSX-positive, SMARCB1-deficient synovial sarcomas on EZH2 enzymatic activity and suggests the potential utility of EZH2-targeted drugs in these genetically defined cancers.
• Pineal parenchymal tumor with marked cytologic pleomorphism : Is there a correlation with the malignancy grade?

  Tamio Ito, Kenichi Sato, Yoshimaru Ozaki, Takii Asanome, Hirohiko Nakamura, Shinya Tanaka, Taichi Kimura, Hiromi Kanno

  Neurological Surgery 44 (6) 481 - 487 1882-1251 2016/06/01 [Refereed][Not invited]
   

  Introduction: In benign pineal parenchymal tumors (PPTs), namely, pineocytoma (PC) and PPT of intermediate differentiation (PPTID), cytologic pleomorphism has occasionally been found however, it is controversial as to whether the presence of pleomorphic cells leads to upgrading of tumors. We experienced a rare case of pleomorphic PPT in an elderly woman and compared it with a retrospective series of 12 PPTs (PC : 3, PPTID : 6, pineoblastoma [PB] : 3) to evaluate the correlation between pleomorphism and the malignancy grade. Case and materials: A 76-year-old woman presented with gradual cognitive deterioration and gait disturbance. Gadolinium-enhanced magnetic resonance imaging (Gd-MRI) revealed a small, enhanced tumor in the pineal gland with marked hydrocephalus. Endoscopic tumor biopsy and third ventriculostomy were performed simultaneously. The tumor was soft, pinkish, and slightly hemorrhagic. After the biopsy, the patient underwent gamma knife radiosurgery. Pathological findings: The PPT presented with areas of tumor cells forming pineocytomatous rosettes and areas of giant and multinucleated cells with hyperchromatic nuclei. Neither mitosis nor necrosis was observed. The tumor cells were positive for synaptophysin (SYN) and neurofilament (NF), but negative for glial fibrillary acidic protein (GFAP) and oligodendrocyte lineage transcription factor 2 (Olig2). The MIB-1 labeling index (LI) was 8.1 %. There was no difference in the MIB-1 LI between pleomorphic and non-pleomorphic areas. All the 12 PPTs were immunopositive for the neuronal markers SYN and NF. The MIB-1 LI was 0% in PC, 3.5% in PPTID, and 10.5% in PB. The proliferative potential was correlated with the WHO grade. From these findings, the final diagnosis of this pleomorphic case was PPTID grade II, not PC, because the MIB-1 LI was relatively high, even though some tumor cells were forming pineocytomatous rosettes. Conclusion: Although cytologic pleomorphism in PPTs is generally considered not to be correlated with the malignancy grade, the final pathological diagnosis should be determined while considering the proliferative potential.
• Novel signaling collaboration between TGF-beta and adaptor protein Crk facilitates EMT in human lung cancer

  Aiman Z. Elmansuri, Mishie A. Tanino, Roshan Mahabir, Lei Wang, Taichi Kimura, Hiroshi Nishihara, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Masumi Tsuda, Shinya Tanaka

  ONCOTARGET 7 (19) 27094 - 27107 1949-2553 2016/05 [Refereed][Not invited]
   

  The signaling adaptor protein Crk has been shown to play an important role in various human cancers. However, its regulatory machinery is not clear. Here, we demonstrated that Crk induced EMT in A549 human lung adenocarcinoma cells through differential regulation of Rac1/Snail and RhoA/Slug, leading to decreased expression of E-cadherin and increased N-cadherin, fibronectin, and MMP2 expression. Cancer cells with mesenchymal features produced TGF-beta and also increased the levels of TGF-beta receptor. TGF-beta increased the endogenous levels of Crk and also augmented Crk-dependent expression of Snail and Slug, and conversely TGF-beta receptor inhibitor suppressed the levels of Snail and Slug. Overexpression of Crk was observed at the invasive front of human lung cancer tissues and was significantly associated with poor prognosis. Thus, TGF-beta and Crk collaborate to form a positive feedback loop to facilitate EMT, which may lead to the malignancy of human cancers possibly being affected by their microenvironment.
• PAMPSゲルによるATDC5細胞のインスリン非依存性軟骨分化誘導シグナルの解明

  後藤 佳子, 北村 信人, 和田 進, 安田 和則, 木村 太一, 津田 真寿美, 田中 伸哉, 仙葉 慎吾, 黒川 孝幸, グン・チェンピン

  北海道整形災害外科学会雑誌 北海道整形災害外科学会 57 (2) 287 - 287 1343-3873 2016/04
• Synthetic PAMPS gel activates BMP/Smad signaling pathway in ATDC5 cells, which plays a significant role in the gel-induced chondrogenic differentiation

  Keiko Goto, Taichi Kimura, Nobuto Kitamura, Shingo Semba, Yoshihiro Ohmiya, Sachiyo Aburatani, Satoko Matsukura, Masumi Tsuda, Takayuki Kurokawa, Jian Ping Gong, Shinya Tanaka, Kazunori Yasuda

  JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A 104 (3) 734 - 746 1549-3296 2016/03 [Refereed][Not invited]
   

  The purposes of this study were to identify signaling pathways that were specifically activated in ATDC5 cells cultured on poly (2-acrylamido-2-methylpropanesulfonic acid) (PAMPS) gel in insulin-free maintenance medium and to evaluate the significance of the determined signaling pathways in the chondrogenic differentiation induced by this gel. In this study, ATDC5 cells cultured on PAMPS gel using the maintenance medium without insulin (PAMPS Culture) were compared with cells cultured on polystyrene using the differentiation medium containing insulin (PS-I Culture). The microarray analysis, Western blot analysis, and real-time PCR analysis demonstrated that the TGF-/BMP signaling pathway was significantly enhanced at Days 1, 2, and 3 in the PAMPS Culture when compared with the PS-I Culture. Inhibition of the BMP type-I receptor reduced the phosphorylation level of Smad1/5 and expression of type-2 collagen and aggrecan mRNA in the cells accompanied by a reduction in cell aggregation at Day 13 in the PAMPS Culture. The inhibition of the TGF-/BMP signaling pathway significantly inhibited the chondrogenic differentiation induced by the PAMPS gel. The present study demonstrated that synthetic PAMPS gel activates the TGF-/BMP/Smad signaling pathway in the ATDC5 cells in the absence of insulin, and that this activation plays a significant role in the chondrogenic differentiation induced by PAMPS gel. (c) 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 734-746, 2016.
• Genetic interaction of hnRNPA2B1 and DNAJB6 in a Drosophila model of multisystem proteinopathy

  Songqing Li, Peipei Zhang, Brian D. Freibaum, Nam Chul Kim, Regina-Maria Kolaitis, Amandine Molliex, Anderson P. Kanagaraj, Ichiro Yabe, Mishie Tanino, Shinya Tanaka, Hidenao Sasaki, Eric D. Ross, J. Paul Taylor, Hong Joo Kim

  HUMAN MOLECULAR GENETICS 25 (5) 936 - 950 0964-6906 2016/03 [Refereed][Not invited]
   

  Adult-onset inherited myopathies with similar pathological features, including hereditary inclusion body myopathy (hIBM) and limb-girdle muscular dystrophy (LGMD), are a genetically heterogeneous group of muscle diseases. It is unclear whether these inherited myopathies initiated by mutations in distinct classes of genes are etiologically related. Here, we exploit a genetic model system to establish a mechanistic link between diseases caused by mutations in two distinct genes, hnRNPA2B1 and DNAJB6. Hrb98DE and mrj are the Drosophila melanogaster homologs of human hnRNPA2B1 and DNAJB6, respectively. We introduced disease-homologous mutations to Hrb98DE, thus capturing mutation-dependent phenotypes in a genetically tractable model system. Ectopic expression of the disease-associated mutant form of hnRNPA2B1 or Hrb98DE in fly muscle resulted in progressive, age-dependent cytoplasmic inclusion pathology, as observed in humans with hnRNPA2B1-related myopathy. Cytoplasmic inclusions consisted of hnRNPA2B1 or Hrb98DE protein in association with the stress granule marker ROX8 and additional endogenous RNA-binding proteins (RBPs), suggesting that these pathological inclusions are related to stress granules. Notably, TDP-43 was also recruited to these cytoplasmic inclusions. Remarkably, overexpression of MRJ rescued this phenotype and suppressed the formation of cytoplasmic inclusions, whereas reduction of endogenous MRJ by a classical loss of function allele enhanced it. Moreover, wildtype, but not disease-associated, mutant forms of MRJ interacted with RBPs after heat shock and prevented their accumulation in aggregates. These results indicate both genetic and physical interactions between disease-linked RBPs and DNAJB6/mrj, suggesting etiologic overlap between the pathogenesis of hIBM and LGMD initiated by mutations in hnRNPA2B1 and DNAJB6.
• Diagnostic Value of Liquid-Based Cytology With Fine Needle Aspiration Specimens for Cervical Lymphadenopathy

  Nobuyuki Bandoh, Takashi Goto, Toshiaki Akahane, Natsumi Ohnuki, Tomomi Yamaguchi, Hajime Kamada, Yasuaki Harabuchi, Shinya Tanaka, Hiroshi Nishihara

  DIAGNOSTIC CYTOPATHOLOGY 44 (3) 169 - 176 8755-1039 2016/03 [Refereed][Not invited]
   

  Background: Cervical lymphadenopathy is a symptom that is frequently seen among outpatients, and it is important to differentiate malignant lesions from reactive lymphoid hyperplasia. Fine needle aspiration (FNA) cytology has been widely used for the diagnosis of cervical lymphadenopathy. However, some limitations of the diagnostic accuracy using conventional smear (CS) cytology have been pointed out. The diagnostic value of liquid-based cytology (LBC) with FNA specimens has not yet been fully proven. Methods: Forty-two patients with cervical lymphadenopathy who underwent FNA with CS cytology from 2007 to 2011 and 123 patients who underwent FNA with LBC utilizing LBCPREP2 (TM) from 2011 to 2015 were studied. Diagnostic values were compared between the CS and the LBC groups. Results: Of the total 165 patients representing the combined CS and LBC groups, 81 (49.1%) were diagnosed as benign lymph node and 84 (50.9%) were malignant diseases including 37 (22.4%) of metastatic carcinoma except for thyroid carcinoma, 30 (18.2%) of metastatic thyroid carcinoma, and 17 (10.3%) of malignant lymphoma. The overall statistical values including sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the CS were 75%, 100%, 100%, 78.9%, and 87.1%, respectively, whereas those values for LBC were 91.2%, 100%, 100%, 90.7%, and 95.3%, respectively. The sensitivity of LBC for malignant diseases tended to be higher than that of CS cytology (p = 0.081). Conclusion: LBC with FNA specimens from cervical lymphadenopathy is a useful and reliable method for the diagnosis of malignant diseases, especially of metastatic carcinomas, due to its increased sensitivity compared with CS cytology. (C) 2016 Wiley Periodicals, Inc.
• Receptor activator of NF-kappa B ligand induces cell adhesion and integrin alpha 2 expression via NF-kappa B in head and neck cancers

  Tamaki Yamada, Masumi Tsuda, Takanori Wagatsuma, Yoichiro Fujioka, Mari Fujioka, Aya O. Satoh, Kosui Horiuchi, Shinya Nishide, Asuka Nanbo, Yasunori Totsuka, Hisashi Haga, Shinya Tanaka, Masanobu Shindoh, Yusuke Ohba

  SCIENTIFIC REPORTS 6 23545  2045-2322 2016/03 [Refereed][Not invited]
   

  Cellular interactions with the extracellular matrix play critical roles in tumor progression. We previously reported that receptor activator of NF-kappa B ligand (RANKL) specifically facilitates head and neck squamous cell carcinoma (HNSCC) progression in vivo. Here, we report a novel role for RANKL in the regulation of cell adhesion. Among the major type I collagen receptors, integrin alpha 2 was significantly upregulated in RANKL-expressing cells, and its knockdown suppressed cell adhesion. The mRNA abundance of integrin alpha 2 positively correlated with that of RANKL in human HNSCC tissues. We also revealed that RANK-NF-kappa B signaling mediated integrin alpha 2 expression in an autocrine/paracrine manner. Interestingly, the amount of active integrin beta 1 on the cell surface was increased in RANKL-expressing cells through the upregulation of integrin alpha 2 and endocytosis. Moreover, the RANK-integrin alpha 2 pathway contributed to RANKL-dependent enhanced survival in a collagen gel and inhibited apoptosis in a xenograft model, demonstrating an important role for RANKL-mediated cell adhesion in three-dimensional environments.
• Attenuation of ligand-induced activation of angiotensin II type 1 receptor signaling by the type 2 receptor via protein kinase C

  Takayuki Inuzuka, Yoichiro Fujioka, Masumi Tsuda, Mari Fujioka, Aya O. Satoh, Kosui Horiuchi, Shinya Nishide, Asuka Nanbo, Shinya Tanaka, Yusuke Ohba

  SCIENTIFIC REPORTS 6 21613  2045-2322 2016/02 [Refereed][Not invited]
   

  Angiotensin II (AII) type 2 receptor (AT2R) negatively regulates type 1 receptor (AT1R) signaling. However, the precise molecular mechanism of AT2R-mediated AT1R inhibition remains poorly understood. Here, we characterized the local and functional interaction of AT2R with AT1R. AT2R colocalized and formed a complex with AT1R at the plasma membrane, even in the absence of AII. Upon AII stimulation, the spatial arrangement of the complex was modulated, as confirmed by Forster resonance energy transfer (FRET) analysis, followed by AT2R internalization along with AT1R. AT2R internalization was specifically observed only in the presence of AT1R; AT2R alone could not be internalized. The AT1R-specific inhibitor losartan completely inhibited both the conformational change and the internalization of AT2R with AT1R, whereas the AT2R-specific inhibitor PD123319 partially hindered these phenomena, demonstrating that the activation of both receptors was indispensable for these effects. In addition, treatment with the protein kinase C (PKC) inhibitors inhibited the ligand-dependent accumulation of AT2R but not that of AT1R in the endosomes. A mutation in the putative phosphorylation sites of AT2R also abrogated the co-internalization of ATR2 with AT1R and the inhibitory effect of ATR2 on AT1R. These data suggest that AT2R inhibits ligand-induced AT1R signaling through the PKC-dependent pathway.
• EXO1 homozygous deletion suppresses the hydroxyurea sensitivity in anaplastic meningioma with extracranial metastases

  Sodai Yoshimura, Takashi Ohta, Kotaro Makita, Shun Yamamuro, Yushi Ochai, Koichiro Sumi, Katsunori Shijo, Atsuo Yoshino, Taku Homma, Masahiko Sugitani, Sayaka Yuzawa, Hiroshi Nishihara, Sinya Tanaka

  INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE 9 (9) 18618 - 18625 1940-5901 2016 [Not refereed][Not invited]
   

  Background: Anaplastic meningioma is exceedingly rare, and its clinicopathological features are distinctive. Distant meningioma metastases have previously been reported to be very rare. The most common site of metastasis is the lung, which accounts for 61% of all meningioma metastases. The standard therapy for meningiomas is total resection and/or radiation therapy. Case Report: A 71-year-old man was admitted with headache and left hemiparesis. Magnetic resonance imaging (MRI) revealed mass lesions at the right frontal convexity and left occipital lobe. Following surgery, pathological examinations demonstrated an anaplastic meningioma. At 3 months after the operation and radiation therapy, lung tumor was removed. The pathological findings for the lung tumor resembled those of the brain tumor. A diagnosis of metastatic lung tumor from anaplastic meningioma was made. At 2 months after the lung surgery, MRI disclosed focal recurrence in the frontal convexity area and progression into the cavernous sinus. Abdominal CT detected a new metastatic lesion in the patient's liver. He underwent adjuvant reirradiation consisting of whole brain radiotherapy. In addition, he received chemotherapy using angiogenesis receptor (bevacizumab). Chemotherapy with hydroxyurea was initiated after 2 courses of the bevacizumab chemotherapy, because his hepatic lesion had become more aggressive. The intracranial mass was reduced by 50% at 8 weeks after initiation of the hydroxyurea chemotherapy. However, there was no significant effect on the liver and lung metastases. Analysis and Conclusion: Extracranial metastases have been estimated to occur in only 0.1% of all meningiomas and most often in association with anaplastic meningiomas. While total resection and/or radiation treatment provide the standard therapy for meningiomas, use of chemotherapy against meningiomas has been limited to distant metastases or based on out of surgical criteria. Several reports have demonstrated that hydroxyurea provides on effective chemotherapy for meningiomas including grade I cases. Hydroxyurea halts meningioma cell growth through arrest of the S-phase of the cell cycle, thus inducing apoptosis. Our analysis revealed EXO1 homozygous deletion at exons 8 and 10 in the patient's extracranial metastatic meningioma. The findings suggested that EXO1 homozygous deletion may be associated with suppression of hydroxyurea sensitivity in anaplastic meningiomas. Hydroxyurea should be a justified therapeutic adaptation in cases of anaplastic meningioma without EOX1 homozygous deletion.
• Rapid immunocytochemistry based on alternating current electric field using squash smear preparation of central nervous system tumors

  Jun Moriya, Mishie Ann Tanino, Tomoko Takenami, Tomoko Endoh, Masana Urushido, Yasutaka Kato, Lei Wang, Taichi Kimura, Masumi Tsuda, Hiroshi Nishihara, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 33 (1) 13 - 18 1433-7398 2016/01 [Refereed][Not invited]
   

  The role of intraoperative pathological diagnosis for central nervous system (CNS) tumors is crucial for neurosurgery when determining the surgical procedure. Especially, treatment of carmustine (BCNU) wafers requires a conclusive diagnosis of high-grade glioma proven by intraoperative diagnosis. Recently, we demonstrated the usefulness of rapid immunohistochemistry (R-IHC) that facilitates antigen-antibody reaction under alternative current (AC) electric field in the intraoperative diagnosis of CNS tumors; however, a higher proportion of water and lipid in the brain parenchyma sometimes leads to freezing artifacts, resulting in poor quality of frozen sections. On the other hand, squash smear preparation of CNS tumors for cytology does not affect the frozen artifacts, and the importance of smear preparation is now being re-recognized as being better than that of the tissue sections. In this study, we established the rapid immunocytochemistry (R-ICC) protocol for squash smears of CNS tumors using AC electric field that takes only 22 min, and demonstrated its usefulness for semi-quantitative Ki-67/MIB-1 labeling index and CD 20 by R-ICC for intraoperative diagnosis. R-ICC by AC electric field may become a substantial tool for compensating R-IHC and will be applied for broad antibodies in the future.
• A case of cerebral astroblastoma with rhabdoid features: a cytological, histological, and immunohistochemical study

  Sayaka Yuzawa, Hiroshi Nishihara, Mishie Tanino, Taichi Kimura, Jun Moriya, Yuuta Kamoshima, Kazuo Nagashima, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 33 (1) 63 - 70 1433-7398 2016/01 [Refereed][Not invited]
   

  Astroblastoma is a rare neuroepithelial neoplasm of unknown origin, usually occurring in children and young adults. Here we report a case of astroblastoma with uncommon features in an 18-year-old female. The tumor was a well-circumscribed cystic and solid mass with marked gadolinium enhancement in the right frontal lobe. Cytological examination showed polarized monopolar cells with diminished cohesiveness. Tumor cells possessed eccentric round to oval nuclei with abundant eosinophilic cytoplasm, sometimes having cytoplasmic processes. Histopathologically, the tumor showed perivascular pseudorosettes with prominent vascular sclerosis. Foam cells were frequently infiltrated around blood vessels and among tumor cells. In some areas, a solid growth pattern of plump tumor cells with abundant inclusion-like eosinophilic cytoplasm showing rhabdoid appearance was observed. The immunohistochemical study revealed strong and diffuse positivity for vimentin and epithelial membrane antigen. Tumor cells were focally positive for glial fibrillary acidic protein and cytokeratin AE1/AE3. Nuclear immunoreactivity for INI1 protein was evident. The Ki-67 labeling index was 10.8 %. This tumor was finally diagnosed as low-grade astroblastoma and the patient had no evidence of recurrence without postoperative radiotherapy or chemotherapy during the last 6 months of follow-up. This report describes novel cytological, histopathological, and immunohistochemical features of the rare tumor.
• Chondroma arising from the spinal dura mater at the thoracic level: A case report with molecular analysis

  Masaya Miyazaki, Keita Yashiro, Mishie Tanino, Shinya Tanaka, Yasunori Fujioka

  PATHOLOGY RESEARCH AND PRACTICE 212 (9) 838 - 841 0344-0338 2016 [Refereed][Not invited]
   

  Chondromas are benign tumors that can be found at several sites in the body, while those arising from the dura mater are extremely rare. Among them, although chondromas arising from the cranial dura mater have been reported, those arising from the spinal dura mater have not been reported in the literature to date. A 66-year-old woman presented with right-sided continuous backache which she had developed recently. After detailed examinations, an epidural tumor at the thoracic level was detected. The patient underwent surgery and a total en-bloc resection was accomplished. From the clinical and pathological findings, the tumor was revealed as chondroma arising from the spinal dura mater. A recent comprehensive study has identified the isocitrate dehydrogenase I (IDH1) and IDH2 mutations in conventional central and periosteal cartilaginous tumors, and it has subsequently been analyzed in intracranial chondrosarcoma and chondroma, including chondroma of the convexity dura mater. Herein, we describe a novel case of chondroma arising from the spinal dura mater, with mutation analysis of IDH1 and IDH2 both of which revealed wild-type. (C) 2016 Elsevier GmbH. All rights reserved.
• Neuropeptide signaling through neurokinin-1 and neurokinin-2 receptors augments antigen presentation by human dendritic cells.

  Junya Ohtake, Shun Kaneumi, Mishie Tanino, Takuto Kishikawa, Satoshi Terada, Kentaro Sumida, Kazutaka Masuko, Yosuke Ohno, Toshiyuki Kita, Sadahiro Iwabuchi, Toshiya Shinohara, Yoshinori Tanino, Tamiko Takemura, Shinya Tanaka, Hiroya Kobayashi, Hidemitsu Kitamura

  The Journal of allergy and clinical immunology 136 (6) 1690 - 1694 0091-6749 2015/12 [Refereed][Not invited]
  
• PAMPSゲルはATDC5細胞の軟骨分化においてBMP/Smadシグナル経路を活性化する

  後藤 佳子, 北村 信人, 木村 太一, 仙葉 愼吾, 黒川 孝幸, Gong Jian Ping, 田中 伸哉, 安田 和則

  日本整形外科学会雑誌 (公社)日本整形外科学会 89 (8) S1708 - S1708 0021-5325 2015/09
• Clinicopathological study of pilomyxoid-spectrum astrocytomas: An analysis of the BRAF gene. Report of two cases

  Tamio Ito, Kenichi Sato, Mitsuteru Oikawa, Hironori Sugio, Taku Asanome, Yoshimaru Ozaki, Hirohiko Nakamura, Shinya Tanaka, Masumi Tsuda, Kazuo Nagashima

  Neurological Surgery 43 (9) 825 - 833 1882-1251 2015/09/01 [Refereed][Not invited]
   

  In contrast to pilocytic astrocytomas (PAs), pilomyxoid astrocytomas (PMAs) demonstrate monophasic piloid cells with angiocentric distribution and a more aggressive clinical course. Recently, several reports have described combined histological features of both subtypes accordingly, these were termed intermediate pilomyxoid tumors (IPTs). The KIAA1549-BRAF fusion gene has been found in approximately 70% of PAs, but is reportedly rare in PMAs. We describe a clinicopathological study of two patients with pilomyxoid-spectrum astrocytoma (PMSA). Case 1 was of a 29-year-old man who presented with a generalized seizure. Gadolinium-magnetic resonance imaging (Gd-MRI) demonstrated a less enhanced tumor in the left temporal lobe. Case 2 was of a 9-year-old boy who presented with headache. Gd-MRI revealed an irregularly enhanced tumor in the left cerebellum. In Case 1, the tumor showed monomorphous bipolar cells in a myxoid background and angiocentric arrangement therefore, the diagnosis was PMA. In Case 2, part of the tumor had a myxoid, angiocentric pattern characteristic of PMA the other part had a biphasic pattern characteristic of PA. PMA and PA were mixed in a 7:3 ratio therefore, IPT was diagnosed. No BRAF V600E mutations were found by immunohistochemistry and sequencing in either case. Three major KIAA1549-BRAF fusion subtypes were analyzed by quantitative reverse transcription polymerase chain reaction (RT-PCR) and sequencing. No fusions were found in Case 1. However, K16-B9 fusion was identified in Case 2, and this fusion was more prevalent in the PA component than in the PMA component. In summary, no BRAF V600E mutations were found in PMSAs, but KIAA1549-BRAF fusion was identified in IPT, particularly in the PA component.
• Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy

  Jun-ichi Furukawa, Masumi Tsuda, Kazue Okada, Taichi Kimura, Jinhua Piao, Shinya Tanaka, Yasuro Shinohara

  PLOS ONE 10 (7) e0128300  1932-6203 2015/07 [Refereed][Not invited]
   

  Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, HRasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. More than 160 glycans derived from three major classes of cell surface glycoconjugates (N-and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio-to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, a2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo) lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response.
• Tyr724 phosphorylation of ELMO1 by Src is involved in cell spreading and migration via Rac1 activation

  Yoshinori Makino, Masumi Tsuda, Yusuke Ohba, Hiroshi Nishihara, Hirofumi Sawa, Kazuo Nagashima, Shinya Tanaka

  CELL COMMUNICATION AND SIGNALING 13 35  1478-811X 2015/07 [Refereed][Not invited]
   

  Background: The complex of Dock180/ELMO1 that functions as a bipartite guanine nucleotide exchange factor for Rac is essential for diverse physiological and pathological processes of cells such as cell migration, phagocytosis, and invasion of cancer cells. Among the Src-family tyrosine kinases (SFKs), it has been reported that Hck directly phosphorylates ELMO1, regulating phagocytosis by promoting activation of Rac1; however, the involvement of other SFKs in ELMO1 phosphorylation has remained unknown. Here, we identified novel tyrosine (Y) residues of ELMO1 phosphorylated by SFKs, and examined the effects on Rac1 activity, cell adhesion, spreading, and cell motility on extracellular matrix (ECM). Results: In this study, we unveiled that Src and Fyn can induce tyrosine phosphorylation of ELMO1 in in vivo and in vitro phosphorylation assays. Mutational analyses identified both Y720 and Y724 residues of ELMO1 as Src-mediated phosphorylation sites, preferentially on Y724. Single substitution of Y724 to Phe abrogated Rac1 activation triggered by Src. To elucidate the biological function of pY724, we established NIH3T3 cells stably expressing wild-type ELMO1 or its Y724F mutant together with Dock180. Among them, Y724-deficient cells exhibited a depletion of Rac1 activity with diminished phosphorylation of ELMO1 even upon the ECM-stimulation. It is noteworthy that NIH3T3 cells with ELMO1 Y724F were strikingly defective to promote cell spreading on fibronectin-coated dish, concomitantly exhibiting immature assemblies of actin stress fibers and focal adhesions. Eventually, ELMO1 Y724F significantly impaired cell migration. Conclusion: These results define that Src-mediated Y724 phosphorylation in ELMO1 plays a critical role for cell spreading via activation of Rac1, leading to promotion of cell migration. As the overexpression and/or hyperactivation of Src have been shown in a wide variety of human cancers, Src-mediated phosphorylation of Y724 in ELMO1 may regulate cancer cell adhesion to the ECM, invasion into surrounding tissues, and subsequent distant metastasis.
• Prognostic value of volume-based measurements on (11)C-methionine PET in glioma patients.

  Kentaro Kobayashi, Kenji Hirata, Shigeru Yamaguchi, Osamu Manabe, Shunsuke Terasaka, Hiroyuki Kobayashi, Tohru Shiga, Naoya Hattori, Shinya Tanaka, Yuji Kuge, Nagara Tamaki

  European journal of nuclear medicine and molecular imaging 42 (7) 1071 - 80 1619-7070 2015/06 [Refereed][Not invited]
   

  PURPOSE: (11)C-methionine (MET) PET is an established diagnostic tool for glioma. Studies have suggested that MET uptake intensity in the tumor is a useful index for predicting patient outcome. Because MET uptake is known to reflect tumor expansion more accurately than MRI, we aimed to elucidate the association between volume-based tumor measurements and patient prognosis. METHODS: The study population comprised 52 patients with newly diagnosed glioma who underwent PET scanning 20 min after injection of 370 MBq MET. The tumor was contoured using a threshold of 1.3 times the activity of the contralateral normal cortex. Metabolic tumor volume (MTV) was defined as the total volume within the boundary. Total lesion methionine uptake (TLMU) was defined as MTV times the mean standardized uptake value (SUVmean) within the boundary. The tumor-to-normal ratio (TNR), calculated as the maximum standardized uptake value (SUVmax) divided by the contralateral reference value, was also recorded. All patients underwent surgery (biopsy or tumor resection) targeting the tissue with high MET uptake. The Kaplan-Meier method was used to estimate the predictive value of each measurement. RESULTS: Grade II tumor was diagnosed in 12 patients (3 diffuse astrocytoma, 2 oligodendroglioma, and 7 oligoastrocytoma), grade III in 18 patients (8 anaplastic astrocytoma, 6 anaplastic oligodendroglioma, and 4 anaplastic oligoastrocytoma), and grade IV in 22 patients (all glioblastoma). TNR, MTV and TLMU were 3.1 ± 1.2, 51.6 ± 49.9 ml and 147.7 ± 153.3 ml, respectively. None of the three measurements was able to categorize the glioma patients in terms of survival when all patients were analyzed. However, when only patients with astrocytic tumor (N = 33) were analyzed (i.e., when those with oligodendroglial components were excluded), MTV and TLMU successfully predicted patient outcome with higher values associated with a poorer prognosis (P < 0.05 and P < 0.01, respectively), while the predictive ability of TNR did not reach statistical significance (P = NS). CONCLUSION: MTV and TLMU may be useful for predicting outcome in patients with astrocytic tumor.
• Adaptor protein CRK induces epithelial-mesenchymal transition and metastasis of bladder cancer cells through HGF/c-Met feedback loop.

  Ryuji Matsumoto, Masumi Tsuda, Lei Wang, Nako Maishi, Takashige Abe, Taichi Kimura, Mishie Tanino, Hiroshi Nishihara, Kyoko Hida, Yusuke Ohba, Nobuo Shinohara, Katsuya Nonomura, Shinya Tanaka

  Cancer science 106 (6) 709 - 17 1347-9032 2015/06 [Refereed][Not invited]
   

  We have previously reported that an adaptor protein CRK, including CRK-I and CRK-II, plays essential roles in the malignant potential of various aggressive human cancers, suggesting the validity of targeting CRK in molecular targeted therapy of a wide range of cancers. Nevertheless, the role of CRK in human bladder cancer with marked invasion, characterized by distant metastasis and poor prognosis, remains obscure. In the present study, immunohistochemistry indicated a striking enhancement of CRK-I/-II, but not CRK-like, in human bladder cancer tissues compared to normal urothelium. We established CRK-knockdown bladder cancer cells using 5637 and UM-UC-3, which showed a significant decline in cell migration, invasion, and proliferation. It is noteworthy that an elimination of CRK conferred suppressed phosphorylation of c-Met and the downstream scaffold protein Gab1 in a hepatocyte growth factor-dependent and -independent manner. In epithelial-mesenchymal transition-related molecules, E-cadherin was upregulated by CRK elimination, whereas N-cadherin, vimentin, and Zeb1 were downregulated. A similar effect was observed following treatment with c-Met inhibitor SU11274. Depletion of CRK significantly decreased cell proliferation of 5637 and UM-UC-3, consistent with reduced activity of ERK. An orthotopic xenograft model with bioluminescent imaging revealed that CRK knockdown significantly attenuated not only tumor volume but also the number of circulating tumor cells, resulted in a complete abrogation of metastasis. Taken together, this evidence uncovered essential roles of CRK in invasive bladder cancer through the hepatocyte growth factor/c-Met/CRK feedback loop for epithelial-mesenchymal transition induction. Thus, CRK might be a potent molecular target in bladder cancer, particularly for preventing metastasis, leading to the resolution of clinically longstanding critical issues.
• HIGH ALDO-KETO REDUCTASE 1C1 EXPRESSION IN METASTATIC BLADDER CANCER CELLS ASSOCIATED WITH INVASIVE POTENTIAL AND DRUG RESISTANCE

  Ryuji Matsumoto, Masumi Tsuda, Takashige Abe, Satoru Maruyama, Kunihiko Tsuchiya, Naoto Miyajima, Nobuo Shinohara, Shinya Tanaka

  JOURNAL OF UROLOGY 193 (4) E535 - E535 0022-5347 2015/04 [Refereed][Not invited]
  
• 肺高血圧症における血管病変の形態学的・免疫組織学的変化

  谷野 美智枝, 辻野 一三, 石田 雄介, 加藤 容崇, 王 磊, 木村 太一, 西原 広史, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 104 (1) 311 - 311 0300-9181 2015/03
• Gastritis cystica profundaを背景にした重複胃癌の免疫組織学的検討

  志藤 茜, 杢 里花, 湯澤 明夏, 石川 麻倫, 加藤 容崇, 石田 雄介, 木村 太一, 谷野 美智枝, 西原 広史, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 104 (1) 513 - 513 0300-9181 2015/03
• Adult classical glioblastoma with a BRAF V600E mutation

  Yoshinobu Takahashi, Toshiaki Akahane, Takahiro Sawada, Hidetoshi Ikeda, Akira Tempaku, Shigeru Yamauchi, Hiroshi Nishihara, Shinya Tanaka, Kazumi Nitta, Wataru Ide, Ikuo Hashimoto, Hajime Kamada

  WORLD JOURNAL OF SURGICAL ONCOLOGY 13 (1) 1477-7819 2015/03 [Not refereed][Not invited]
   

  The B-Raf proto-oncogene serine/threonine kinase (B-Raf) is a member of the Raf kinase family. The BRAF V600E mutation occurs frequently in certain brain tumors such as pleomorphic xanthoastrocytoma, ganglioglioma, and pilocytic astrocytoma, and less frequently in epithelioid and giant cell glioblastoma. BRAF V600E mutation in these cases has been canonically detected using Sanger sequencing or immunohistochemistry but not with next-generation sequencing (NGS). Moreover, to our knowledge, there is no detailed report of the BRAF V600E mutation in an adult glioblastoma with classical histologic features (c-GBM). Therefore, we performed NGS analysis to determine the mutational status of BRAF of 13 glioblastomas (GBMs) (11 primary and 2 secondary cases) and detected one tumor harboring the BRAF V600E mutation. We report here the detection of the BRAF V600E mutation in a patient with c-GBM and describe the patient's clinical course as well as the results of histopathological analysis.
• 骨転移をきたした小脳膠芽腫の1例

  湊川 英樹, 森 崇, 鬼丸 力也, 小野寺 俊輔, 白土 博樹, 東海林 菊太郎, 吉田 道春, 小林 浩之, 寳金 清博, 菅野 宏美, 田中 伸哉

  Japanese Journal of Radiology (公社)日本医学放射線学会 33 (Suppl.) 4 - 4 1867-1071 2015/02
• Rapid immunohistochemistry based on alternating current electric field for intraoperative diagnosis of brain tumors

  Mishie Tanino, Toshio Sasajima, Hiroshi Nanjo, Shiori Akesaka, Masami Kagaya, Taichi Kimura, Yusuke Ishida, Masaya Oda, Masataka Takahashi, Taku Sugawara, Toshiaki Yoshioka, Hiroshi Nishihara, Yoichi Akagami, Akiteru Goto, Yoshihiro Minamiya, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 32 (1) 12 - 19 1433-7398 2015/01 [Refereed][Not invited]
   

  Rapid immunohistochemistry (R-IHC) can contribute to the intraoperative diagnosis of central nervous system (CNS) tumors. We have recently developed a new IHC method based on an alternating current electric field to facilitate the antigen-antibody reaction. To ensure the requirement of R-IHC for intraoperative diagnosis, 183 cases of CNS tumors were reviewed regarding the accuracy rate of diagnosis without R-IHC. The diagnostic accuracy was 90.7 % (168/183 cases) in which definitive diagnoses were not provided in 17 cases because of the failure of glioma grading and differential diagnosis of lymphoma and glioma. To establish the clinicopathological application, R-IHC for frozen specimens was compared with standard IHC for permanent specimens. 33 gliomas were analyzed, and the Ki-67/MIB-1 indices of frozen specimens by R-IHC were consistent with the grade and statistically correlated with those of permanent specimens. Thus, R-IHC provided supportive information to determine the grade of glioma. For discrimination between glioma and lymphoma, R-IHC was able to provide clear results of CD20 and Ki-67/MIB-1 in four frozen specimens of CNS lymphoma as well as standard IHC. We conclude that the R-IHC for frozen specimens can provide important information for intraoperative diagnosis of CNS tumors.
• Rapid immunohistochemistry based on alternating current electric field for intraoperative diagnosis of brain tumors (vol 32, pg 12, 2015)

  Mishie Tanino, Toshio Sasajima, Hiroshi Nanjo, Shiori Akesaka, Masami Kagaya, Taichi Kimura, Yusuke Ishida, Masaya Oda, Masataka Takahashi, Taku Sugawara, Toshiaki Yoshioka, Hiroshi Nishihara, Yoichi Akagami, Akiteru Goto, Yoshihiro Minamiya, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 32 (1) 20 - 21 1433-7398 2015/01 [Refereed][Not invited]
  
• Usefulness of C-11-methionine-positron emission tomography for the diagnosis of progressive multifocal leukoencephalopathy

  Shinichi Shirai, Ichiro Yabe, Takahiro Kano, Yuka Shimizu, Toru Sasamori, Kazunori Sato, Makoto Hirotani, Takayuki Nonaka, Ikuko Takahashi, Masaaki Matsushima, Naoya Minami, Kazuo Nakamichi, Masayuki Saijo, Kanako C. Hatanaka, Tohru Shiga, Shinya Tanaka, Hidenao Sasaki

  JOURNAL OF NEUROLOGY 261 (12) 2314 - 2318 0340-5354 2014/12 [Refereed][Not invited]
   

  Progressive multifocal leukoencephalopathy (PML) is a subacute demyelinating disease of the brain caused by the JC virus that occurs mainly in immunocompromised patients. The prognosis is very poor. As the lesion looks like non- specific leukoencephalopathy, making a diagnosis at the early stage is very difficult. We report three PML cases in which there was a mismatch between C-11-methionine-positron emission tomography (MET-PET) uptake and F-18-fluorodeoxyglucose-positron emission tomography (FDG-PET) uptake. All three cases demonstrated the hyper-uptake of MET around the white matter lesions and hypo-uptake of FDG inside the lesions. We speculate that the infection had ended inside the white matter lesions of these patients, while JC virus infection was ongoing around the lesions, resulting in the increase of methionine metabolism, and the glucose metabolism was reduced or intermediate because inflammatory cells infiltrate PML lesions rarely. Two patients who were diagnosed and treated with mefloquine while the JC virus was at a low level in the cerebrospinal fluid are still alive. We suggest the usefulness of MET-PET for the early diagnosis of PML and early treatment with mefloquine.
• Pathology of frontotemporal dementia with limb girdle muscular dystrophy caused by a DNAJB6 mutation

  Ichiro Yabe, Mishie Tanino, Hiroaki Yaguchi, Akihiro Takiyama, Huaying Cai, Hiromi Kanno, Ikuko Takahashi, Yukiko K. Hayashi, Masashi Watanabe, Hidehisa Takahashi, Shigetsugu Hatakeyama, Shinya Tanak, Hidenao Sasaki

  CLINICAL NEUROLOGY AND NEUROSURGERY 127 10 - 12 0303-8467 2014/12 [Refereed][Not invited]
  
• SS18-SSX-regulated miR-17 promotes tumor growth of synovial sarcoma by inhibiting p21WAF1/CIP1

  Yusuke Minami, Shinji Kohsaka, Masumi Tsuda, Kazuhiro Yachi, Nobuaki Hatori, Mishie Tanino, Taichi Kimura, Hiroshi Nishihara, Akio Minami, Norimasa Iwasaki, Shinya Tanaka

  CANCER SCIENCE 105 (9) 1152 - 1159 1347-9032 2014/09 [Refereed][Not invited]
   

  MicroRNA (miRNA) can function as tumor suppressors or oncogenes, and also as potential specific cancer biomarkers; however, there are few published studies on miRNA in synovial sarcomas, and their function remains unclear. We transfected the OncomiR miRNA Precursor Virus Library into synovial sarcoma Fuji cells followed by a colony formation assay to identify miRNAs to confer an aggressive tumorigenicity, and identified miR-17-5p from the large colonies. MiR-17 was found to be induced by a chimeric oncoprotein SS18-SSX specific for synovial sarcoma, and all examined cases of human synovial sarcoma expressed miR-17, even at high levels in several cases. Overexpression of miR-17 in synovial sarcoma cells, Fuji and HS-SYII, increased colony forming ability in addition to cell growth, but not cell motility and invasion. Tumor volume formed in mice in vivo was significantly increased by miR-17 overexpression with a marked increase of MIB-1 index. According to PicTar and Miranda algorithms, which predicted CDKN1A (p21) as a putative target of miR-17, a luciferase assay was performed and revealed that miR-17 directly targets the 3-UTR of p21 mRNA. Indeed, p21 protein level was remarkably decreased by miR-17 overexpression in a p53-independent manner. It is noteworthy that miR-17 succeeded in suppressing doxorubicin-evoked higher expression of p21 and conferred the drug resistance. Meanwhile, introduction of anti-miR-17 in Fuji and HS-SYII cells significantly decreased cell growth, consistent with rescued expression of p21. Taken together, miR-17 promotes the tumor growth of synovial sarcomas by post-transcriptional suppression of p21, which may be amenable to innovative therapeutic targeting in synovial sarcoma.
• Epiregulin enhances tumorigenicity by activating the ERK/MAPK pathway in glioblastoma

  Shinji Kohsaka, Kunihiko Hinohara, Lei Wang, Tatsunori Nishimura, Masana Urushido, Kazuhiro Yachi, Masumi Tsuda, Mishie Tanino, Taichi Kimura, Hiroshi Nishihara, Noriko Gotoh, Shinya Tanaka

  NEURO-ONCOLOGY 16 (7) 960 - 970 1522-8517 2014/07 [Refereed][Not invited]
   

  Glioblastoma multiforme (GBM) is one of the most aggressive human tumors, and the establishment of an effective therapeutic reagent is a pressing priority. Recently, it has been shown that the tumor tissue consists of heterogeneous components and that a highly aggressive population should be the therapeutic target. Through a single subcutaneous passage of GBM cell lines LN443 and U373 in mice, we have developed highly aggressive variants of these cells named LN443X, U373X1, and U373X2, which showed increased tumor growth, colony-forming potential, sphere-forming potential, and invasion ability. We further investigated using microarray analysis comparing malignant cells with their parental cells and mRNA expression analysis in grades II to IV glioma samples. Adipocyte enhancer binding protein 1, epiregulin (EREG), and microfibrillar associated protein 5 were identified as candidate genes associated with higher tumor grade and poor prognosis. Immunohistochemical analysis also indicated a correlation of a strong expression of EREG with short overall survival. Furthermore, both EREG stimulation and EREG introduction of GBM cell lines were found to increase phosphorylation of epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase and resulted in the promotion of colony formation, sphere formation, and in vivo tumor formation. Gefitinib treatment inhibited phosphorylation of EGFR and extracellular signal-regulated kinase and led to tumor regression in U373-overexpressed EREG. These results suggested that EREG is one of the molecules involved in glioma malignancy, and EGFR inhibitors may be a candidate therapeutic agent for EREG-overexpressing GBM patients.
• microRNA 31 functions as an endometrial cancer oncogene by suppressing Hippo tumor suppressor pathway (vol 13, 97, 2014)

  Takashi Mitamura, Hidemichi Watari, Lei Wang, Hiromi Kanno, Masaya Miyazaki, Makiko Kitagawa, Mohamed Kamel Hassan, Peixin Dong, Taichi Kimura, Mishie Tanino, Hiroshi Nishihara, Shinya Tanaka, Noriaki Sakuragi

  MOLECULAR CANCER 13 140  1476-4598 2014/06 [Refereed][Not invited]
  
• Immunohistochemical evaluation of O-6-methylguanine DNA methyltransferase (MGMT) expression in 117 cases of glioblastoma

  Masaya Miyazaki, Hiroshi Nishihara, Shunsuke Terasaka, Hiroyuki Kobayashi, Shigeru Yamaguchi, Tamio Ito, Yuuta Kamoshima, Shin Fujimoto, Sadao Kaneko, Masahito Katoh, Nobuaki Ishii, Hiromi Mohri, Mishie Tanino, Taichi Kimura, Shinya Tanaka

  NEUROPATHOLOGY 34 (3) 268 - 276 0919-6544 2014/06 [Refereed][Not invited]
   

  Temozolomide (TMZ) is an oral alkylating agent which is widely used in the treatment of glioblastoma (GBM) and is composed of astrocytic and/or oligodendroglial tumors, and the evaluation of O6-methylguanine DNA methyltransferase (MGMT) expression is important to predict the response to TMZ therapy. In this study, we conducted immunohistochemical analysis of 117 cases of Japanese GBM including 19 cases of GBM with oligodendroglioma component (GBMO), using a scoring system for quantitative evaluation of staining intensity and proportion of MGMT, and performed survival analysis of these patients. Immunohistochemically, 55 cases (47%) were positive for MGMT with various intensities and proportions (total score (TS)2), while 62 cases (53%) were negative (TS=0). The distribution of MGMT expression pattern was not affected by any clinicopathological parameters such as the histological subtype (GBM vs. GBMO), age and gender. The survival analysis of these patients revealed that the minimal expression of MGMT (TS2) was a significant unfavorable prognostic factor (P<0.001) as well as resectability (P=0.004). Moreover, multivariate analysis showed that minimal MGMT expression in GBM was the most potent independent predictor for progression free survival (P<0.001) and also overall patient survival (P<0.001). This is the first report employing the scoring system for both staining intensity and proportion to evaluate immunohistochemical MGMT expression in GBM. In addition, our results emphases the clinicopathological values of the immunohistochemical approach for MGMT expression in glioma patients as a routine laboratory examination.
• F-18-FDG PET/CT imaging for a gastrointestinal mantle cell lymphoma with multiple lymphomatous polyposis

  Makoto Saito, Masaya Miyazaki, Mishie Tanino, Shinya Tanaka, Kencho Miyashita, Koh Izumiyama, Akio Mori, Tatsuro Irie, Masanori Tanaka, Masanobu Morioka, Eriko Tsukamoto

  WORLD JOURNAL OF GASTROENTEROLOGY 20 (17) 5141 - 5146 1007-9327 2014/05 [Refereed][Not invited]
   

  Multiple lymphomatous polyposis (MLP) is an uncommon type of gastrointestinal lymphoma characterized by the presence of multiple polyps along the gastrointestinal tract. Most of this entity is in fact considered the counterpart of gastrointestinal tract involvement for mantle cell lymphoma (MCL). To our knowledge, there have been no reports on [fluorine-18]-fluorodeoxy-glucose (F-18-FDG)- positron emission tomography (PET)/computed tomography (CT) imaging for gastrointestinal MCL with MLP. We present the results of F-18-FDG PET/CT imaging in a patient with gastrointestinal tract involvement of MCL showing continuous MLP from the stomach to the rectum and intestinal intussusception. FDG-PET/CT findings were false negative in typical MLP spreading widely over the gastrointestinal tract, but uptake was noted in large lesions with deep infiltration considered atypical as MLP. On FDG-PET/CT imaging, the Ki-67 proliferative index, which is a cell proliferation marker, showed neither correlation with the presence of uptake nor the maximum standardized uptake value. (C) 2014 Baishideng Publishing Group Co., Limited. All rights reserved.
• Clinicopathologic Study of Pineal Parenchymal Tumors of Intermediate Differentiation

  Tamio Ito, Hiromi Kanno, Ken-ichi Sato, Mitsuteru Oikawa, Yoshimaru Ozaki, Hirohiko Nakamura, Shunsuke Terasaka, Hiroyuki Kobayashi, Kiyohiro Houkin, Kanako Hatanaka, Jyun-ichi Murata, Shinya Tanaka

  WORLD NEUROSURGERY 81 (5-6) 783 - 789 1878-8750 2014/05 [Refereed][Not invited]
   

  OBJECTIVE: Pineal parenchymal tumors of intermediate differentiation (PPTID) are extremely rare tumor entities, and only limited data are available regarding their pathologic features and biologic behaviors. Because grading criteria of pineal parenchymal tumors (PPTs) have yet to be established, the treatment strategy and prognosis of PPTIDs remain controversial. We describe the clinicopathologic study of six patients with PPTID and compare responses for the treatment and prognosis with cases of pineocytoma (PC) and pine-oblastoma (PB). From this analysis, we attempt to clarify the treatment strategy for PPTIDs. METHODS: This study included 15 patients with PPTs, consisting of 6 PCs, 6 PPTIDs, and 3 PBs. We focused on the 6 patients with PPTIDs. All PPTID cases were treated surgically, and radiotherapy and chemotherapy were administered as adjuvant therapies in some cases. We have earlier reported the histopathologic study (Neuropathology 32: 647-653, 2012). Briefly, we examined mitotic figures and necrosis by hematoxylin-eosin staining and immunohistochemical markers such as neuronal markers (synaptophysin, neurofilament (NF), and neuronal nuclear antigen), and an MIB-1 labeling index was determined. RESULTS: In the PPTID cases, the extent of resection was variable and the recurrence rates among patients varied according to stage and treatment. All PC patients underwent total resection with no recurrence. All PB patients underwent resection and adjuvant therapy with radiotherapy and chemotherapy. There were no recurrences in patients with PC or PB. The results of histopathologic findings have been already reported as mentioned above. Briefly, the results indicated no mitotic figure or necrosis in any of the six cases of PPTID, but those features were observed in PB cases. All cases even including PC and PB were immunopositive for neuronal markers. The MIB-1 labeling index of PPTID was 3.5%, whereas it was 0% in PC and 10.5% in PB. CONCLUSIONS: Good radiosensitivity of PPTIDs was observed in our series. Because there are cases with discrepancies between images and pathologic findings, it is very difficult to determine the proper treatment strategy for PPTIDs. Proliferative potential was correlated with World Health Organization grade, although the immunoreactivity of neuronal markers did not correlate with the histologic grade.
• Sustained elevation of Snail promotes glial-mesenchymal transition after irradiation in malignant glioma

  Roshan Mahabir, Mishie Tanino, Aiman Elmansuri, Lei Wang, Taichi Kimura, Tamio Itoh, Yusuke Ohba, Hiroshi Nishihara, Hiroki Shirato, Masumi Tsuda, Shinya Tanaka

  NEURO-ONCOLOGY 16 (5) 671 - 685 1522-8517 2014/05 [Refereed][Not invited]
   

  Ionizing irradiation is an effective treatment for malignant glioma (MG); however, a higher rate of recurrence with more aggressive phenotypes is a vital issue. Although epithelial-mesenchymal transition (EMT) is involved in irradiation-induced cancer progression, the role for such phenotypic transition in MG remains unknown. To investigate the mechanism of irradiation-dependent tumor progression in MG, we performed immunohistochemistry (IHC) and qRT-PCR using primary and recurrent MG specimens, MG cell lines, and primary culture cells of MG. siRNA technique was used for MG cell lines. In 22 cases of clinically recurrent MG, the expression of the mesenchymal markers vimentin and CD44 was found to be increased by IHC. In paired identical MG of 7 patients, the expression of collagen, MMPs, and YKL-40 were also elevated in the recurrent MGs, suggesting the The Cancer Genome Atlas-based mesenchymal subtype. Among EMT regulators, sustained elevation of Snail was observed in MG cells at 21 days after irradiation. Cells exhibited an upregulation of migration, invasion, numbers of focal adhesion, and MMP-2 production, and all of these mesenchymal features were abrogated by Snail knockdown. Intriguingly, phosphorylation of ERK1/2 and GSK-3 were increased after irradiation in a Snail-dependent manner, and TGF- was elevated in both fibroblasts and macrophages but not in MG cells after irradiation. It was noteworthy that irradiated cells also expressed stemness features such as SOX2 expression and tumor-forming potential in vivo. We here propose a novel concept of glial-mesenchymal transition after irradiation in which the sustained Snail expression plays an essential role.
• Metabolomic profiling of sodium fluoride-induced cytotoxicity in an oral squamous cell carcinoma cell line

  Sakagami, H, Sugimoto, M, Tanaka, S, Onuma, H, Ota, S, Kaneko, M, Soga, T, Tomita, M

  Metabolomics 10 (2) 270 - 279 1573-3882 2014/04 [Refereed][Not invited]
   

  Sodium fluoride (NaF) is used in dentistry as a preventive agent for dental caries because of its ability to remineralize the tooth surface and its antibacterial effect. Although one of its target molecules in bacteria is enolase, its site of action in human cells has not been identified. The aim of this study was to identify target metabolites that are coupled to NaF-induced cytotoxicity in the HSC-2 human oral squamous cell carcinoma cell line. Cell viability, membrane integrity and apoptosis induction were analyzed by MTT assay, trypan blue exclusion and caspase-3 activation, respectively. Cells were treated with a minimal cytotoxic concentration of NaF for various times and subjected to comprehensive metabolomics analysis using capillary electrophoresis-mass spectrometry. In the early stages, inhibition of the enolase reaction in glycolysis pathway was observed. This was coupled with rapid inhibition of the progression of TCA cycle. In the later stages, gradual increases in the AMP/ATP ratio (a putative marker of apoptosis) and oxidized products (e.g. GSSH, and methionine sulfoxide), and marginal changes in polyamine levels (putative marker of necrosis) were observed. This manuscript provides the new insight into the global impact of NaF on metabolic pathways in human oral squamous cell carcinoma cells.
• Differential diagnosis of small cell glioblastoma and anaplastic oligodendroglioma: a case report of an elderly man

  Kenta Takahashi, Masumi Tsuda, Hiromi Kanno, Junichi Murata, Roshan Mahabir, Yusuke Ishida, Taichi Kimura, Mishie Tanino, Hiroshi Nishihara, Kazuo Nagashima, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 31 (2) 118 - 123 1433-7398 2014/04 [Refereed][Not invited]
   

  Small cell glioblastoma is a histological subtype of glioblastoma with characteristic features of highly proliferative, monotonous small glial cells with high nuclear cytoplasm ratio. Morphologically, malignant lymphoma or small cell metastatic carcinoma should be carefully discriminated. Some cases are difficult to differentiate from anaplastic oligodendroglioma. In this report, we present a case of small cell glioblastoma of an elderly man. The lack of IDH1/2 mutation was confirmed by immunohistochemistry and direct sequencing. Fluorescence in situ hybridization revealed the lower rates of chromosome 1p and 19q deletion. Microsatellite analysis disclosed partial 10q alteration near the PTEN locus. Not only morphological and immunohistochemical examinations, but also cytogenetical investigations for IDH1/2 mutation, 1p/19q loss, and PTEN alteration, are strongly supportive methods for the differential diagnosis of small cell glioblastoma and anaplastic oligodendroglioma.
• microRNA 31 functions as an endometrial cancer oncogene by suppressing Hippo tumor suppressor pathway

  Takashi Mitamura, Hidemichi Watari, Lei Wang, Hiromi Kanno, Makiko Kitagawa, Mohamed Kamel Hassan, Taichi Kimura, Mishie Tanino, Hiroshi Nishihara, Shinya Tanaka, Noriaki Sakuragi

  MOLECULAR CANCER 13 97  1476-4598 2014/04 [Refereed][Not invited]
   

  Background: We aimed to investigate whether MIR31 is an oncogene in human endometrial cancer and identify the target molecules associated with the malignant phenotype.Methods: We investigated the growth potentials of MIR31-overexpressing HEC-50B cells in vitro and in vivo. In order to identify the target molecule of MIR31, a luciferase reporter assay was performed, and the corresponding downstream signaling pathway was examined using immunohistochemistry of human endometrial cancer tissues. We also investigated the MIR31 expression in 34 patients according to the postoperative risk of recurrence.Results: The overexpression of MIR31 significantly promoted anchorage-independent growth in vitro and significantly increased the tumor forming potential in vivo. MIR31 significantly suppressed the luciferase activity of mRNA combined with the LATS2 3'-UTR and consequently promoted the translocation of YAP1, a key molecule in the Hippo pathway, into the nucleus. Meanwhile, the nuclear localization of YAP1 increased the transcription of CCND1. Furthermore, the expression levels of MIR31 were significantly increased (10.7-fold) in the patients (n = 27) with a high risk of recurrence compared to that observed in the low-risk patients (n = 7), and this higher expression correlated with a poor survival.Conclusions: MIR31 functions as an oncogene in endometrial cancer by repressing the Hippo pathway. MIR31 is a potential new molecular marker for predicting the risk of recurrence and prognosis of endometrial cancer.
• 中枢神経系原発びまん性大細胞型B細胞性リンパ腫におけるDNAメチル化に関する解析

  畑中佳奈子, 畑中豊, 小林浩之, 菅野宏美, 西原広史, 田中伸哉, 三橋智子, 松野吉宏

  日本病理学会会誌 103 (1) 211  0300-9181 2014/03/26 [Not refereed][Not invited]
  
• A case of cervical juxtafacet cyst with extensive rim enhancement on Gd-DTPA MRI

  Toru Sasamori, Kazutoshi Hida, Kimio Anzai, Shunsuke Yano, Yasutaka Kato, Shinya Tanaka, Hisatoshi Saito, Kiyohiro Houkin

  CLINICAL IMAGING 38 (2) 199 - 201 0899-7071 2014/03 [Refereed][Not invited]
   

  The authors reported a case of cervical juxtafacet cyst with extensive rim enhancement on gadolinium-diethylenetriamine pentaacid magnetic resonance imaging. Operative finding revealed the epidural space around the mass filled with abundant venous plexus. Histological examination demonstrated that cyst wall was composed of the well-vascularized fibrous connective tissue with some inflammatory changes. We speculate that extensive rim enhancement of juxtafacet cyst may be attributed not only to the chronic inflammatory changes of cyst wall, but to engorged venous plexus within the widened epidural space. (c) 2014 Elsevier Inc. All rights reserved.
• Erratum to: Rapid immunohistochemistry based on alternating current electric field for intraoperative diagnosis of brain tumors [Brain Tumor Pathol, (2014), DOI:10.1007/s10014-014-0188-y]

  Mishie Tanino, Toshio Sasajima, Hiroshi Nanjo, Shiori Akesaka, Masami Kagaya, Taichi Kimura, Yusuke Ishida, Masaya Oda, Masataka Takahashi, Taku Sugawara, Toshiaki Yoshioka, Hiroshi Nishihara, Yoichi Akagami, Akiteru Goto, Yoshihiro Minamiya, Shinya Tanaka

  Brain Tumor Pathology 32 (1) 20 - 21 1861-387X 2014 [Refereed][Not invited]
  
• Erratum to: Clinicopathological evaluation of cyclooxygenase-2 expression in meningioma: immunohistochemical analysis of 76 cases of low and high-grade meningioma (Brain Tumor Pathology DOI:10.1007/s10014-012-0127-8)

  Yasutaka Kato, Hiroshi Nishihara, Hiromi Mohri, Hiromi Kanno, Hiroyuki Kobayashi, Taichi Kimura, Mishie Tanino, Shunsuke Terasaka, Shinya Tanaka

  Brain Tumor Pathology 31 (1) 31  1861-387X 2014 [Refereed][Not invited]
  
• C-ERC/mesothelin provokes lymphatic invasion of colorectal adenocarcinoma

  Futoshi Kawamata, Shigenori Homma, Hirofumi Kamachi, Takahiro Einama, Yasutaka Kato, Masumi Tsuda, Shinya Tanaka, Masahiro Maeda, Kazunori Kajino, Okio Hino, Norihiko Takahashi, Toshiya Kamiyama, Hiroshi Nishihara, Akinobu Taketomi, Satoru Todo

  JOURNAL OF GASTROENTEROLOGY 49 (1) 81 - 92 0944-1174 2014/01 [Not refereed][Not invited]
   

  Background Lymph node metastasis is a key event of colorectal cancer (CRC) progression. Mesothelin is expressed in various types of malignant tumor and associated with an unfavorable prognosis. The full-length mesothelin (Full-ERC) is cleaved by protease into membrane-bound C-ERC/mesothelin and N-ERC/mesothelin which is secreted into the blood. The aim of this study was to examine the biological role of mesothelin in CRC by clinicopathological analysis and in vitro lymphatic invasion assay. Methods Ninety-one cases of CRC specimens were immunohistochemically examined and the localization of mesothelin in luminal membrane and/or cytoplasm was also evaluated. Lymphatic invasion assay was also performed using the human CRC cell line, WiDr, which was transfected with Full-, N- and C-ERC/mesothelin expression plasmids (Full-WiDr, N-WiDr and C-WiDr). Results Immunohistochemically, "luminal membrane positive" of mesothelin was identified in 37.4 %, and correlated with lymphatic permeation and lymph node metastasis, but not with patients' prognosis. Interestingly, among the patients with lymph node metastasis (N = 38), "luminal membrane positive" of mesothelin significantly correlated with unfavorable patients' outcome. In addition, lymphatic invasion assay revealed that Full-WiDr and C-WiDr more significantly invaded human lymphatic endothelial cells than the Mock-WiDr (P < 0.01). Conclusion The luminal membrane expression of mesothelin was associated with unfavorable prognosis of CRC patients with lymph nodemetastasis. Moreover, this is the first report to prove the biological function of C-ERC/mesothelin associated with lymphatic invasion of cancer in vitro.
• Clinicopathological evaluation of cyclooxygenase-2 expression in meningioma: immunohistochemical analysis of 76 cases of low and high-grade meningioma

  Yasutaka Kato, Hiroshi Nishihara, Hiromi Mohri, Hiromi Kanno, Hiroyuki Kobayashi, Taichi Kimura, Mishie Tanino, Shunsuke Terasaka, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 31 (1) 23 - 30 1433-7398 2014/01 [Refereed][Not invited]
   

  Tumorigenic activity of cyclooxygenase-2 (COX-2), a rate-limiting enzyme in the production of prostaglandins (PGs), has been proved for some types of cancer, including brain tumors. We evaluated expression of COX-2 in meningioma, one of the most common intracranial tumors in adults which accounts for 24-30 % of intracranial tumors. We performed immunostaining for COX-2 in 76 cases of meningioma consisting of 44 cases of low-grade (WHO Grade I) and 32 cases of high-grade (29 cases of Grade II and 3 cases of Grade III) meningioma, and evaluated COX-2 expression levels on the basis of staining intensity and proportion in tumor cells. The expression level of COX-2 in meningioma cells was significantly correlated with WHO grade (P = 0.0153). In addition, COX-2 expression was significantly correlated with MIB-1 labeling index for all 76 cases of meningioma (P = 0.0075), suggesting tumor promotion by COX-2 in meningioma progression. Our results may indicate the therapeutic value of non-steroidal anti-inflammatory drugs against meningioma, especially for patients with elevated proliferation, to regulate the tumorigenic activity of COX-2 in meningioma cells.
• Combination of Exercise Training and Diet Restriction Normalizes Limited Exercise Capacity and Impaired Skeletal Muscle Function in Diet-Induced Diabetic Mice

  Tadashi Suga, Shintaro Kinugawa, Shingo Takada, Tomoyasu Kadoguchi, Arata Fukushima, Tsuneaki Homma, Yoshihiro Masaki, Takaaki Furihata, Masashige Takahashi, Mochamad A. Sobirin, Taisuke Ono, Kagami Hirabayashi, Takashi Yokota, Shinya Tanaka, Koichi Okita, Hiroyuki Tsutsui

  ENDOCRINOLOGY 155 (1) 68 - 80 0013-7227 2014/01 [Refereed][Not invited]
   

  Exercise training (EX) and diet restriction (DR) are essential for effective management of obesity and insulin resistance in diabetes mellitus. However, whether these interventions ameliorate the limited exercise capacity and impaired skeletal muscle function in diabetes patients remains unexplored. Therefore, we investigated the effects of EX and/or DR on exercise capacity and skeletal muscle function in diet-induced diabetic mice. Male C57BL/6J mice that were fed a high-fat diet (HFD) for 8 weeks were randomly assigned for an additional 4 weeks to 4 groups: control, EX, DR, and EX+DR. A lean group fed with a normal diet was also studied. Obesity and insulin resistance induced by a HFD were significantly but partially improved by EX or DR and completely reversed by EX+DR. Although exercise capacity decreased significantly with HFD compared with normal diet, it partially improved with EX and DR and completely reversed with EX+DR. In parallel, the impaired mitochondrial functionandenhanced oxidative stress in the skeletal muscle caused by the HFD were normalized only by EX+DR. Although obesity and insulin resistance were completely reversed by DR with an insulin-sensitizing drug or a long-term intervention, the exercise capacity and skeletal muscle function could not be normalized. Therefore, improvement in impaired skeletal muscle function, rather than obesity and insulin resistance, may be an important therapeutic target for normalization of the limited exercise capacity in diabetes. In conclusion, a comprehensive lifestyle therapy of exercise and diet normalizes the limited exercise capacity and impaired muscle function in diabetes mellitus.
• TRIM29 as a novel prostate basal cell marker for diagnosis of prostate cancer

  Yukiko Kanno, Masashi Watanabe, Taichi Kimura, Katsuya Nonomura, Shinya Tanaka, Shigetsugu Hatakeyama

  ACTA HISTOCHEMICA 116 (5) 708 - 712 0065-1281 2014 [Refereed][Not invited]
   

  Tripartite motif protein 29 (TRIM29) is one of the TRIM family proteins, some of which function as E3 ubiquitin ligases. In this study, we investigated the usefulness of TRIM29 for diagnosis of prostate cancer Prostate tissues including carcinoma and non-carcinoma tissues obtained by needle biopsy and radical prostatectomy were used. Immunohistochemistry was performed according to standard procedures using an antibody against TRIM29. Immunohistochemical staining with an antibody against 34 beta E12, which recognizes cytokeratins 1, 5, 10 and 14, was performed as a control. Basal cells of normal prostatic glands were stained with anti-TRIM29 antibody in all cases, whereas prostate cancer tissues had no or little staining with anti-TRIM29 antibody. TRIM29 is selectively expressed in basal cells of the normal prostate gland, and immunohistochemical staining with anti-TRIM29 antibody showed the same expression pattern as that with 34 beta E12 in prostate cancer and its benign mimics. Our data indicate that TRIM29 may be useful for distinguishing prostate cancers from benign tissues. (C) 2013 Elsevier GmbH. All rights reserved.
• Anaplastic transformation of papillary thyroid carcinoma in multiple lung metastases presenting with a malignant pleural effusion: A case report

  Tomoe Abe, Masaru Suzuki, Kaoruko Shimizu, Naofumi Shinagawa, Satoshi Oizumi, Yoshihiro Matsuno, Masaya Miyazaki, Mishie Tanino, Shinya Tanaka, Masaharu Nishimura

  Journal of Medical Case Reports 8 (1) 460  1752-1947 2014 [Refereed][Not invited]
   

  Introduction: Anaplastic transformation of well-differentiated papillary thyroid carcinoma at distant metastasis sites is rare. To the best of our knowledge, this is the first report of an autopsy case of anaplastic transformation of papillary thyroid carcinoma in multiple lung metastases presenting with a malignant pleural effusion. Case presentation: We report an autopsy case of a 61-year-old Japanese man with anaplastic transformation of papillary thyroid carcinoma with multiple lung metastases presenting with a malignant pleural effusion, which was difficult to diagnose by cytological examination before the autopsy. He presented with a 1-month history of progressive dyspnea, and examination of the left pleural effusion revealed a bloody exudate with an increase in thyroglobulin however, malignant cells in the pleural fluid were negative for thyroglobulin. Conclusion: It is important to be aware that anaplastic transformation of differentiated thyroid carcinoma could develop in lung metastases and could be a cause of a malignant pleural effusion.
• NS1-binding protein abrogates the elevation of cell viability by the influenza A virus NS1 protein in association with CRKL

  Masaya Miyazaki, Hiroshi Nishihara, Hideki Hasegawa, Masato Tashiro, Lei Wang, Taichi Kimura, Mishie Tanino, Masumi Tsuda, Shinya Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 441 (4) 953 - 957 0006-291X 2013/11 [Refereed][Not invited]
   

  The influenza A virus non-structural protein 1 (NS1) is a multifunctional virulence factor consisting of an RNA binding domain and several Src-homology (SH) 2 and SH3 binding motifs, which promotes virus replication in the host cell and helps to evade antiviral immunity. NS1 modulates general host cell physiology in association with various cellular molecules including NS1-binding protein (NS1-BP) and signaling adapter protein CRK-like (CRKL), while the physiological role of NS1-BP during influenza A virus infection especially in association with NS1 remains unclear. In this study, we analyzed the intracellular association of NS1-BP, NS1 and CRKL to elucidate the physiological roles of these molecules in the host cell. In HEK293T cells, enforced expression of NS1 of A/Beijing (H1N1) and A/Indonesia (H5N1) significantly induced excessive phosphorylation of ERR and elevated cell viability, while the over-expression of NS1-BP and the abrogation of CRKL using siRNA abolished such survival effect of NS1. The pull-down assay using GST-fusion CRKL revealed the formation of intracellular complexes of NS1-BP, NS1 and CRKL. In addition, we identified that the N-terminus SH3 domain of CRKL was essential for binding to NS1-BP using GST-fusion CRKL-truncate mutants. This is the first report to elucidate the novel function of NS1-BP collaborating with viral protein NS1 in modulation of host cell physiology. In addition, an alternative role of adaptor protein CRKL in association with NS1 and NS1-BP during influenza A virus infection is demonstrated. (C) 2013 Elsevier Inc. All rights reserved.
• Co-Overexpression of GEP100 and AMAP1 Proteins Correlates with Rapid Local Recurrence after Breast Conservative Therapy

  Rumiko Kinoshita, Jin-Min Nam, Yoichi M. Ito, Kanako C. Hatanaka, Ari Hashimoto, Haruka Handa, Yutaro Otsuka, Shigeru Hashimoto, Yasuhito Onodera, Mitsuchika Hosoda, Shunsuke Onodera, Shinichi Shimizu, Shinya Tanaka, Hiroki Shirato, Mishie Tanino, Hisataka Sabe

  PLOS ONE 8 (10) e76791  1932-6203 2013/10 [Refereed][Not invited]
   

  A major problem of current cancer research and therapy is prediction of tumor recurrence after initial treatment, rather than the simple biological characterization of the malignancy and proliferative properties of tumors. Breast conservation therapy (BCT) is a well-approved, standard treatment for patients with early stages of breast cancer, which consists of lumpectomy and whole-breast irradiation. In spite of extensive studies, only 'age' and 'Ki-67 positivity' have been identified to be well correlated with local recurrence after BCT. An Arf6 pathway, activated by GEP100 under receptor tyrosine kinases (RTKs) and employs AMAP1 as its effector, is crucial for invasion and metastasis of some breast cancer cells. This pathway activates beta 1 integrins and perturbs E-cadherin-based adhesions, hence appears to be integral for epithelial-mesenchymal transdifferentiation (EMT). We here show that expression of the Arf6 pathway components statistically correlates with rapid local recurrence after BCT. We retrospectively analyzed four hundred seventy-nine patients who received BCT in Hokkaido University Hospital, and found 20 patients had local recurrence. We then analyzed pathological samples of patients who experienced local recurrence by use of Kaplan-Meier analysis, Stepwise regression analysis and the t-test, coupled with immunostaining, and found that co-overexpression of GEP100 and AMAP1 correlates with rapidity of the local recurrence. Their margin-status, node-positivity, and estrogen receptor (ER)-or progesterone receptor (PgR)positivity did not correlated with the rapidity. This study is the first to show that expression of a certain set of proteins correlates with the rapidity of local recurrence. Our results are useful not only for prediction, but highlight the possibility of developing novel strategies to block local recurrence. We also discuss why mRNAs encoding these proteins have not been identified to correlate with local recurrence by previous conventional gene expression profiling analyses.
• Immunohistochemical molecular expression profile of metastatic brain tumor for potent personalized medicine

  Yasutaka Kato, Hiroshi Nishihara, Sayaka Yuzawa, Hiromi Mohri, Hiromi Kanno, Yutaka Hatanaka, Taichi Kimura, Mishie Tanino, Shinya Tanaka

  Brain Tumor Pathology 30 (3) 167 - 174 1433-7398 2013/07 [Refereed][Not invited]
   

  Recent progress in molecule-targeting therapy may yield personalized therapeutic strategies for patients with metastatic brain tumors (MBT), the most frequently encountered intracranial tumors. For this purpose, we investigated the molecular expression profile of MBT to establish the pathological basis for personalized diagnosis. We studied 166 MBT specimens including 70 cases of lung cancer and 34 cases of breast cancer, and performed immunostaining for EGFR, COX-2, and O-6-methylguanine-DNA methyltransferase (MGMT), among others, which could be target molecules for therapeutic agents or enable prediction of drug efficacy. Loss of MGMT expression was observed in approximately 20-40 % of MBT derived from lung, breast, and gastrointestinal cancers, indicating the possibility of treatment of MBT patients with temozolomide. In addition, MBT expressed a variety of receptor tyrosine kinases, for example EGFR and HER2, and signal transduction molecules, for example phospho-mTOR and COX-2, irrespective of tumor origin, enabling individualized medication with molecule-targeting drugs. We also identified alteration of molecular expression profile in 4 MBT cases during recurrence. Our results not only reveal the molecular characteristics of MBT but also suggest the possibility of potent personalized medicine for MBT patients. © 2012 The Japan Society of Brain Tumor Pathology.
• Expression of CD163 prevents apoptosis through the production of granulocyte colony-stimulating factor in meningioma

  Hiromi Kanno, Hiroshi Nishihara, Lei Wang, Sayaka Yuzawa, Hiroyuki Kobayashi, Masumi Tsuda, Taichi Kimura, Mishie Tanino, Shunsuke Terasaka, Shinya Tanaka

  NEURO-ONCOLOGY 15 (7) 853 - 864 1522-8517 2013/07 [Refereed][Not invited]
   

  Background. CD163 is a 130-kDa transmembrane protein expressed in human monocytes and macrophages, and the aberrant expression of CD163 in breast and colorectal cancer associated with patients' poor prognosis was reported. Here, we analyzed the expression of CD163 in meningioma, a common intracranial tumor, and its molecular mechanism in association with meningioma progression. Methods. First, we performed immunohistochemical analysis using 50 human meningioma specimens. Next, we established CD163-overexpressing human meningioma cell lines and investigated its roles in tumor progression in vitro and in vivo. Results. Immunohistochemically, 26 of 50 human meningioma specimens (52.0%) were positive for CD163 in tumor cells, including benign grade I (48.5%) and grade II (71.4%) cases. Furthermore, CD163 expression was correlated with histological atypical parameters that directly predict the prognosis of meningioma. CD163-overexpressing meningioma cells showed significant suppression of apoptosis and accelerated tumor growth in nude mice. In addition, unexpected splenomegaly affiliated with the xenograft predicted tumor-derived granulocyte colony-stimulating factor (G-CSF) production, which was confirmed by reverse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Conclusions. To our knowledge, this is the first report that demonstrates CD163 expression in meningioma not only by immunohistochemistry but also by reverse-transcription polymerase chain reaction, using primary culture cells, and provides the novel molecular function of CD163 to prevent apoptosis through the production of G-CSF in meningioma.
• Primary rhabdoid tumor with low grade glioma component of the central nervous system in a young adult

  Shogo Endo, Shunsuke Terasaka, Shigeru Yamaguchi, Hitoshi Ikeda, Tsutomu Kato, Hiroyuki Kobayashi, Shinya Tanaka, Kiyohiro Houkin

  NEUROPATHOLOGY 33 (2) 185 - 191 0919-6544 2013/04 [Refereed][Not invited]
   

  In the CNS, primary tumors with rhabdoid components are classified as atypical teratoid/rhabdoid tumor, rhabdoid meningioma or rhabdoid glioblastoma. The authors present a young adult patient with supratentorial rhabdoid tumor incidentally found after head trauma as a small pre-existing lesion in the parahippocampal gyrus. MRI demonstrated an area of hypointensity on T1-weighted images and hyperintensity on T2-weighted and fluid attenuated inversion recovery images. A serial MR scan revealed no change 3 months after the initial examination but drastic changes at 6 months. As the tumor and accompanying intratumoral hemorrhage enlarged rapidly, resection of the tumor was performed. Histopathology revealed that the main component of the tumor was typical rhabdoid cells with some necrotic areas. There were also pathological features consistent with oligoastrocytoma. The specimen had neither vascular proliferation usually seen in high-grade glioma nor the meningothelial pattern that suggests meningioma. Immunohistochemical findings revealed that cells were strongly positive for vimentin, epithelial membrane antigen and INI-1 antibody throughout the specimen. Further, monosomy 22 was detected by fluorescence in situ hybridization. The tumor was finally thought to be an unclassifiable primitive rhabdoid tumor with oligoastrocytoma that arose in the CNS. The patient died within 5 months of detection of the tumor, regardless of surgical resection, radiotherapy and chemotherapy.
• Intracranial mass-forming lesion associated with dural thickening and hypophysitis

  Hiromi Kanno, Mishie Tanino, Kentaro Watanabe, Yoshimaru Ozaki, Tamio Itoh, Taichi Kimura, Hiroshi Nishihara, Tomoo Itoh, Takuhito Narita, Kazuo Nagashima, Shinya Tanaka

  Neuropathology 33 (2) 213 - 216 0919-6544 2013/04 [Refereed][Not invited]
  
• In Japanese patients with type A gastritis with pernicious anemia the condition is very poorly associated with Helicobacter pylori infection

  Makoto Saito, Masanobu Morioka, Kentaro Wakasa, Koh Izumiyama, Akio Mori, Tatsuro Irie, Masanori Tanaka, Shinya Tanaka

  JOURNAL OF INFECTION AND CHEMOTHERAPY 19 (2) 208 - 210 1341-321X 2013/04 [Refereed][Not invited]
   

  There are conflicting views about the association between type A gastritis with pernicious anemia (PA) and infection with Helicobacter pylori, and currently, no definite conclusion has been reached. In this study, we evaluated H. pylori infection in patients with type A gastritis who developed PA. The study included a total of 25 Japanese patients (13 males and 12 females) who had been diagnosed with PA at our department, with a mean age of 71.2 years. We diagnosed infection with H. pylori by measuring serum H. pylori-IgG antibodies in all 25 patients, and we performed gastric biopsy in 17 patients. They were all negative for H. pylori-IgG antibody (0/25) and H. pylori on gastric biopsy (0/17). Although the prevalence of H. pylori infection (70-80 %) in our age-matched controls in Japan is higher than the prevalence in similar populations in western countries, we believe that type A gastritis with PA is very poorly associated with H. pylori infection.
• 脳腫瘍の現代病理学 BRAF異常とグリオーマ

  田中 伸哉, 谷野 美智枝, 津田 真寿美, 石田 雄介, 木村 太一, 西原 広史, 長嶋 和郎

  日本病理学会会誌 (一社)日本病理学会 102 (1) 203 - 203 0300-9181 2013/04
• 術中迅速脳腫瘍診断における電解非接触撹拌技術を搭載した迅速免疫染色装置使用の有用性

  谷野 美智枝, 明坂 詩織, 石田 雄介, 木村 太一, 西原 広史, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 102 (1) 349 - 349 0300-9181 2013/04
• 肺気腫合併肺線維症二剖検例における血管病変の解析

  石田 雄介, 谷野 美智枝, 加藤 容崇, 高橋 健太, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 102 (1) 428 - 428 0300-9181 2013/04
• SRC Signaling Is Crucial in the Growth of Synovial Sarcoma Cells

  Sebastian Michels, Marcel Trautmann, Elisabeth Sievers, Dagmar Kindler, Sebastian Huss, Marcus Renner, Nicolaus Friedrichs, Jutta Kirfel, Susanne Steiner, Elmar Endl, Peter Wurst, Lukas Heukamp, Roland Penzel, Olle Larsson, Akira Kawai, Shinya Tanaka, Hiroshi Sonobe, Peter Schirmacher, Gunhild Mechtersheimer, Eva Wardelmann, Reinhard Buettner, Wolfgang Hartmann

  CANCER RESEARCH 73 (8) 2518 - 2528 0008-5472 2013/04 [Refereed][Not invited]
   

  Synovial sarcoma is a soft-tissue malignancy characterized by a reciprocal t(X; 18) translocation encoding a chimeric transcriptional modifier. Several receptor tyrosine kinases have been found activated in synovial sarcoma; however, no convincing therapeutic concept has emerged from these findings. On the basis of the results of phosphokinase screening arrays, we here investigate the functional and therapeutic relevance of the SRC kinase in synovial sarcoma. Immunohistochemistry of phosphorylated SRC and its regulators CSK and PTP1B (PTPN1) was conducted in 30 synovial sarcomas. Functional aspects of SRC, including dependence of SRC activation on the SS18/SSX fusion proteins, were analyzed in vitro. Eventually, synovial sarcoma xenografts were treated with the SRC inhibitor dasatinib in vivo. Activated phospho (p)-(Tyr416)-SRC was detected in the majority of tumors; dysregulation of CSK or PTP1B was excluded as the reason for the activation of the kinase. Expression of the SS18/SSX fusion proteins in T-REx-293 cells was associated with increased p-(Tyr416)-SRC levels, linked with an induction of the insulin-like growth factor pathway. Treatment of synovial sarcoma cells with dasatinib led to apoptosis and inhibition of cellular proliferation, associated with reduced phosphorylation of FAK (PTK2), STAT3, IGF-IR, and AKT. Concurrent exposure of cells to dasatinib and chemotherapeutic agents resulted in additive effects. Cellular migration and invasion were dependent on signals transmitted by SRC involving regulation of the Rho GTPases Rac and RhoA. Treatment of nude mice with SYO-1 xenografts with dasatinib significantly inhibited tumor growth in vivo. In summary, SRC is of crucial biologic importance and represents a promising therapeutic target in synovial sarcoma. Cancer Res; 73(8); 2518-28. (C) 2013 AACR.
• Inhibition of GSH synthesis potentiates temozolomide-induced bystander effect in glioblastoma

  Shinji Kohsaka, Kenta Takahashi, Lei Wang, Mishie Tanino, Taichi Kimura, Hiroshi Nishihara, Shinya Tanaka

  CANCER LETTERS 331 (1) 68 - 75 0304-3835 2013/04 [Refereed][Not invited]
   

  Glioblastoma multiforme (GBM) is one of the most aggressive human tumors with poor prognosis. Current standard treatment includes chemotherapy using DNA alkylating agent temozolomide (TMZ) concomitant with surgical resection and/or irradiation. However, GBM patients exhibit various levels of the elevated expression of DNA repair enzyme, due to MGMT causing resistance to TMZ. Determination of the MGMT-positive population of primary tumor is important to evaluate the therapeutic efficacy of TMZ. Here we generated TMZ-resistant GBM cells by introducing MGMT into TMZ-sensitive GBM cell line KMG4, and established a model to assess the TMZ-induced bystander effect on TMZ-resistant cells. By mixing TMZ-resistant and -sensitive cells, GBM tumors with MGMT positivity as 50%, 10%, and 1% were generated in vivo. We could not observe any bystander effect of TMZ-induced cell death in tumor with 50% MGMT positivity. Although the bystander effect was observed within 20 days in the case of tumor with 1% MGMT positivity, final tumor size at day 28 was the same as control without sensitive cells. This bystander effect was observed in vitro using conditioned medium of TMZ-damaged GBM cells, and PCR array analysis indicated that the conditioned medium stimulated stress and toxicity pathway and upregulated anti-oxidants genes expression such as catalase and SOD2 in TMZ-resistant cells. In addition, the reduction of the activity of anti-stress mechanism by using inhibitor of GSH synthesis potentiated TMZ-induced bystander effect. These results suggest that GSH inhibitor might be one of the candidates for combination therapy with TMZ for TMZ-resistant GBM patients. (C) 2012 Elsevier Ireland Ltd. All rights reserved.
• Down-Regulation of microRNA-31 Induces Taxane Resistance in Ovarian Cancer Cells through Increase of Receptor Tyrosine Kinase MET.

  Hidemichi Watari, Takashi Mitamura, Shinya Tanaka, Noriaki Sakuragi

  REPRODUCTIVE SCIENCES 20 (S3) 313A - 313A 1933-7191 2013/03 [Refereed][Not invited]
  
• Relationship between Methyl CpG Binding Protein 2 and JC Viral Proteins

  Kenta Takahashi, Yasuko Orba, Taichi Kimura, Lei Wang, Shinji Kohsaka, Masumi Tsuda, Mishie Tanino, Hiroshi Nishihara, Kazuo Nagashima, Hirofumi Sawa, Shinya Tanaka

  JAPANESE JOURNAL OF INFECTIOUS DISEASES 66 (2) 126 - 132 1344-6304 2013/03 [Refereed][Not invited]
   

  JC virus (JCV) is a causative agent of progressive multifocal leukoencephalopathy (PML). Methyl CpG binding protein 2 (MeCP2) is a transcriptional control nuclear protein that is abundantly expressed in neurons. We previously observed that the MeCP2 protein is expressed in JCV large T antigen (TAg)-expressing glial cells in PML brains. To investigate the relationship between MeCP2 and JCV TAg, we examined the promoter activity and mRNA and protein expression levels of MeCP2 in JCV TAg-expressing cells. We found that JCV TAg enhances the promoter activity of MeCP2, but does not enhance the mRNA and protein levels of MeCP2. These results suggest that post-transcriptional mechanisms may play a role in MeCP2 expression.
• DPP6 as a candidate gene for neuroleptic-induced tardive dyskinesia.

  Tanaka S, Syu A, Ishiguro H, Inada T, Horiuchi Y, Ishikawa M, Koga M, Noguchi E, Ozaki N, Someya T, Kakita A, Takahashi H, Nawa H, Arinami T

  Pharmacogenomics J 13 (1) 27 - 34 1473-1150 2013/02 [Refereed][Not invited]
   

  We implemented a two-step approach to detect potential predictor gene variants for neuroleptic-induced tardive dyskinesia (TD) in schizophrenic subjects. First, we screened associations by using a genome-wide (Illumina HumanHapCNV370) SNP array in 61 Japanese schizophrenia patients with treatment-resistant TD and 61 Japanese schizophrenia patients without TD. Next, we performed a replication analysis in 36 treatment-resistant TD and 138 non-TD subjects. An association of an SNP in the DPP6 (dipeptidyl peptidase-like protein-6) gene, rs6977820, the most promising association identified by the screen, was significant in the replication sample (allelic P=0.008 in the replication sample, allelic P=4.6 x 10(-6), odds ratio 2.32 in the combined sample). The SNP is located in intron-1 of the DPP6 gene and the risk allele was associated with decreased DPP6 gene expression in the human postmortem prefrontal cortex. Chronic administration of haloperidol increased Dpp6 expression in mouse brains. DPP6 is an auxiliary subunit of Kv4 and regulates the properties of Kv4, which regulates the activity of dopaminergic neurons. The findings of this study indicate that an altered response of Kv4/DP
• Gene Regulation of Prominin-1 (CD133) in Normal and Cancerous Tissues

  Kouichi Tabu, Norihisa Bizen, Tetsuya Taga, Shinya Tanaka

  PROMININ-1 (CD133): NEW INSIGHTS ON STEM & CANCER STEM CELL BIOLOGY 777 73 - 85 0065-2598 2013 [Refereed][Not invited]
   

  A pentaspan membrane glycoprotein prominin-1 (frequently called CD133 in human) is widely used as a surface marker to identify and isolate normal stem/progenitor cells from various organs, although it is also expressed in some types of differentiated cells. Since CD133 was identified as a universal marker to isolate cancer stem cells (CSCs) in tumors derived from multiple tissues, much attention has been directed toward the relationship between its gene regulation and identity of CSCs (i.e., cancer stemness). Prominin-1 (PROM1) gene possesses five alternative promoters yielding multiple first exons within the 5'-untranslated region (UTR) and also splicing variants affecting the open reading frame (ORF) sequence, implicating the complicated gene regulation in a context-dependent manner. This chapter aims to organize the accumulated findings on prominin-1 with a focus on its altered expression and regulation in normal and cancerous cells and to discuss potential regulatory networks underlying cancer stemness.
• Downregulation of miRNA-31 induces taxane resistance in ovarian cancer cells through increase of receptor tyrosine kinase MET

  T. Mitamura, H. Watari, L. Wang, H. Kanno, M. K. Hassan, M. Miyazaki, Y. Katoh, T. Kimura, M. Tanino, H. Nishihara, S. Tanaka, N. Sakuragi

  Oncogenesis 2 2157-9024 2013 [Refereed][Not invited]
   

  Ovarian cancer is one of the most aggressive female reproductive tract tumors. Paclitaxel (PTX) is widely used for the treatment of ovarian cancer. However, ovarian cancers often acquire chemotherapeutic resistance to this agent. We investigated the mechanism of chemoresistance by analysis of microRNAs using the ovarian cancer cell line KFr13 and its PTX-resistant derivative (KFr13Tx). We found that miR-31 was downregulated in KFr13Tx cells, and that re-introduction of miR31 re-sensitized them to PTX both in vitro and in vivo. miR-31 was found to bind to the 3′-UTR of mRNA of MET, and the decrease in MET correlated to higher sensitivity to PTX. Furthermore, co-treatment of KFr13Tx cells with MET inhibitors sensitized the tumor cells to PTX both in vitro and in vivo. In addition, lower levels of miR31 and higher expression of MET in human ovarian cancer specimens were significantly correlated with PTX chemoresistance and poor prognosis. This study demonstrated miR31-dependent regulation of MET for chemoresistance of ovarian cancer, raising the possibility that combination therapy with a MET inhibitor and PTX will increase PTX efficacy. © 2013 Macmillan Publishers Limited All rights reserved.
• Large Solid-pseudopapillary Neoplasm of the Pancreas with Aberrant Protein Expression and Mutation of beta-Catenin: A Case Report and Literature Review of the Distribution of beta-Catenin Mutation

  Takahiko Kobayashi, Mariko Ozasa, Kencho Miyashita, Akiyoshi Saga, Kimiaki Miwa, Makoto Saito, Masanobu Morioka, Motoya Takeuchi, Nobuo Takenouchi, Takashi Yabiku, Hiromi Kanno, Sayaka Yuzawa, Mishie Tanino, Shinya Tanaka, Hiroshi Kawakami, Masahiro Asaka, Naoya Sakamoto

  INTERNAL MEDICINE 52 (18) 2051 - 2056 0918-2918 2013 [Refereed][Not invited]
   

  Solid-pseudopapillary neoplasms (SPN) are rare pancreatic tumors. The etiology of SPN involves mutations in the gene that encodes beta-catenin (CTNNB1). We herein report the case of a 23-year-old woman with a large SPN with proliferating tumor cells that displayed both solid and pseudo-papillary patterns. The simultaneous nuclear accumulation and loss of membrane localization of beta-catenin and E-cadherin was specifically observed in the tumor cells. Further, the tumor cells were shown to harbor a missense mutation in exon 3 of CTNNB1. We also present a review of the literature describing the clustering of CTNNB1 mutations in patients with SPN.
• Expression of O6-methylguanine DNA methyltransferase (MGMT) and immunohistochemical analysis of 12 pineal parenchymal tumors

  Hiromi Kanno, Hiroshi Nishihara, Mitsuteru Oikawa, Yoshimaru Ozaki, Junichi Murata, Yutaka Sawamura, Masahito Kato, Kanako Kubota, Mishie Tanino, Taichi Kimura, Kazuo Nagashima, Tamio Itoh, Shinya Tanaka

  NEUROPATHOLOGY 32 (6) 647 - 653 0919-6544 2012/12 [Refereed][Not invited]
   

  Pineal parenchymal tumors (PPTs) are rare neoplasms which occupy less than 1% of primary CNS tumors. Because of their rare incidence, previous reports on PPTs are limited in number and the useful molecular markers for deciding histological grading and even selecting chemotherapy are undetermined. In this study, we conducted immunohistochemical analysis of 12 PPT specimens, especially for expression of O6-methylguanine DNA methyltransferase (MGMT) to assess whether temozolomide (TMZ) could serve as a possible alternative therapy for PPTs. We analyzed 12 PPTs, consisting of three pineocytomas, six PPTs of intermediate differentiation (PPTIDs), and three pineoblastomas. Immunohistochemical analysis was performed using antibodies against MGMT, synaptophysin, neurofilament protein (NF), p53, and neuronal nuclear antigen (NeuN). Immunohistochemically, 11 out of 12 cases were positive for MGMT. The mean MIB-1 labeling index was less than 1% in pineocytoma, 3.5% in PPTID, and 10.5% in pineoblastoma. All 12 cases were positive for synaptophysin and 11 cases, except one PPTID case, showed positive for NF. Nuclear staining of NeuN was negative in all cases although cytoplasmic staining of NeuN was observed in five cases. No case was positive for p53. Eleven out of 12 cases of PPTs demonstrated MGMT expression, suggesting chemoresistancy to TMZ treatment. This is the first report showing MGMT expression in PPTs. In addition, MIB-1 labeling index correlated with WHO grade, although the immunoreactivity of synaptophysin, NF, NeuN and p53 did not correlate with the histological grade.
• Intracellular localization of mesothelin predicts patient prognosis of extrahepatic bile duct cancer

  Futoshi Kawamata, Hirofumi Kamachi, Takahiro Einama, Shigenori Homma, Munenori Tahara, Masaya Miyazaki, Shinya Tanaka, Toshiya Kamiyama, Hiroshi Nishihara, Akinobu Taketomi, Satoru Todo

  INTERNATIONAL JOURNAL OF ONCOLOGY 41 (6) 2109 - 2118 1019-6439 2012/12 [Refereed][Not invited]
   

  Mesothelin is expressed in various types of malignant tumors, and we recently reported that the expression of mesothelin was related to unfavorable patient outcome in pancreatic ductal adenocarcinoma and gastric adenocarcinoma. In this study, we examined the clinicopathological significance of mesothelin expression in extrahepatic bile duct cancer (EHBDCA), especially in terms of its association with the staining pattern. Tissue samples from 61 EHBDCA (16 hilar cholangiocarcinoma, 17 upper bile duct adenocarcinoma, 20 middle bile duct adenocarcinoma and 8 distal bile duct adenocarcinoma) were immunohistochemically examined. The expression levels of mesothelin in tumor cells was classified into the localization of mesothelin in luminal membrane and/or cytoplasm, in addition to high and low according to the staining intensity and proportion as a conventional analysis. 'High-level expression' of mesothelin (47.5%) was statistically correlated with liver metastasis (P=0.013) and poorer patient outcome (P=0.022), while 'luminal' membrane positive' of mesothelin (52.5%) was more significantly correlated with liver metastasis (P=0.006), peritoneal metastasis (P=0.024) and unfavorable patient outcome (P=0.017). Moreover, we found that 'cytoplasmic expression' isolated from 'luminal membrane negative' of mesothelin represented the best patient prognosis throughout this study. We describe the expression pattern level of mesothelin, i.e., in luminal membrane or cytoplasm both high and low level, evidently indicate the patient prognosis of EH BDCA, suggesting the pivotal role of mesothelin in cancer promotion depending on its intracellular localization.
• Pilot Study of Changes in Salivary Metabolic Profiles Induced by Template Therapy

  Tanaka, S, Taga, H, Maehara, K, Kaneshima, A, Machino, M, Onuma, H, Kaneko, M, Sakagami, H, Sugimoto, M, Soga, T, Tomita, M

  In Vivo. 26 (6) 1015 - 1020 0258-851X 2012/11 [Refereed][Not invited]
   

  Background: Occlusal raising method (so-called 'Template therapy') has been reported to alleviate various diseases and symptoms, but the underlying mechanism is not clear. We searched the low-molecular weight metabolite(s) in the saliva, the concentration of which is significantly changed by the template therapy. Materials and methods: One female patient with headache underwent the template therapy for 12 days, and her total saliva was subjected to non-targeted analysis using capillary electrophoresis time-of-flight mass spectrometry (CE-TOF-MS). Results: One hundred and thirteen substances were identified in the saliva. Glycine was the most abundant amino acid in the saliva, followed by alanine, serine and proline. After the start of the template therapy, her headache was alleviated, accompanied by a significant (p=0.042) increase of salivary concentration of glycine, as compared with total amino acids whereas that of other amino acids was not significantly changed. In the metabolomics profile, salivary concentration of large number of metabolites as compared with total metabolite concentration decreased, including N-acetylneuraminate (p=0.025) and p-hydroxyphenylacetate (p=0.039). Conclusion: This pilot study demonstrated, to our knowledge for the first time, that only glycine exhibited unique changes among total metabolites, suggesting its significant role in template therapy.
• 【分子標的薬-がんから他疾患までの治癒をめざして-】 基礎研究 分子標的薬耐性化メカニズムの解明 グリオーマにおけるMGMTの発現調節と抗がん剤耐性解除を目指した治療への応用

  高阪 真路, 菅野 宏美, 田中 伸哉

  日本臨床 (株)日本臨床社 70 (増刊8 分子標的薬) 346 - 352 0047-1852 2012/11 [Not refereed][Not invited]
  
• [Regulation of MGMT and application for the therapy to attenuate the chemoresistance].

  Kohsaka S, Kanno H, Tanaka S

  Nihon rinsho. Japanese journal of clinical medicine 70 Suppl 8 346 - 352 0047-1852 2012/11 [Refereed][Invited]
  
• Radiation induced intraparenchymal meningioma occurring 6 years after CNS germinoma: Case report

  Yuuta Kamoshima, Shunsuke Terasaka, Hiroyuki Kobayashi, Sadahiro Kaneko, Kanako Kubota, Shinya Tanaka, Kiyohiro Houkin

  CLINICAL NEUROLOGY AND NEUROSURGERY 114 (7) 1077 - 1080 0303-8467 2012/09 [Refereed][Not invited]
  
• A case of pilocytic astrocytoma in an adult

  Takeo Murahashi, Kenichi Sato, Tamio Ito, Yoshimaru Ozaki, Hironori Sugio, Hirohiko Nakamura, Shinya Tanaka

  Neurological Surgery 40 (9) 793 - 797 0301-2603 2012/09 [Refereed][Not invited]
   

  Pilocytic astrocytoma is the most common glioma in children, in whom the majority arise in the cerebellum. In contrast, pilocytic astrocytomas are less common in adults. The most frequent locations involved are the basal ganglia, cerebellum, optic chiasm, and hypothalamus. Overall survival rates are good. The case presented involved a pilocytic astrocytoma of the right parietal lobe in a 36-year-old man. Cranial magnetic resonance imaging (MRI) revealed a small mural nodule in the wall of the cyst, with no edema around the tumor. This nodule showed a hyperintense signal on gadolinium-enhanced MRI. Computed tomography (CT) scanning revealed a hypodense right parietal lobe mass with calcification. At surgery, the cyst contents were aspirated, and the mural nodule was excised. Postoperative radiotherapy was not given. Neuropathological examination revealed a pilocytic astrocytoma (Grade I). The MIB-index was 3.3%. There has been no recurrence after 1 year of postoperative follow-up.
• Prognostic Implication of Histological Oligodendroglial Tumor Component: Clinicopathological Analysis of 111 Cases of Malignant Gliomas

  Hiromi Kanno, Hiroshi Nishihara, Takuhito Narita, Shigeru Yamaguchi, Hiroyuki Kobayashi, Mishie Tanino, Taichi Kimura, Shunsuke Terasaka, Shinya Tanaka

  PLOS ONE 7 (7) e41669  1932-6203 2012/07 [Refereed][Not invited]
   

  The favorable prognosis of high-grade oligodendroglial tumor such as glioblastoma (GBM) with oligodendroglioma component (GBMO) has been suggested; however, the studies which examine the prognostic significance of oligodendroglial tumor were limited. In this study, we performed a histopathology-based reevaluation of 111 cases of high grade gliomas according to the latest World Health Organization (WHO), and compared the clinical outcomes between oligodendroglial tumors and pure astrocytic tumors. The survival analysis revealed that the patients with high grade oligodendroglial tumor including GBMO significantly indicated better prognosis compared to the patients with high grade pure astrocytic tumors (GBM and AA, anaplastic astrocytoma) as expected, and the obtained survival curves were almost identical to those from the patients with conventional Grade III or Grade IV tumors, respectively. Moreover, if the cases of oligodendroglial tumor were histopathologically excluded, the patients with AA exhibited extremely poor prognosis which was similar to that of GBM, suggesting that the histological identification of oligodendroglial tumor component, even partially, prescribe the prognosis of high grade glioma patients. This is the prominent report of retrospective clinicopathological analysis for high-grade gliomas throughout Grade III and IV, especially referring to the prognostic value of histological oligodendroglial tumor component; in addition, our results might offer an alternative aspect for the grading of high-grade astrocytic/oligodendroglial tumors.
• The JAK inhibitor tofacitinib regulates synovitis through inhibition of interferon-gamma and interleukin-17 production by human CD4+ T cells

  Keisuke Maeshima, Kunihiro Yamaoka, Satoshi Kubo, Kazuhisa Nakano, Shigeru Iwata, Kazuyoshi Saito, Masanobu Ohishi, Hisaaki Miyahara, Shinya Tanaka, Koji Ishii, Hironobu Yoshimatsu, Yoshiya Tanaka

  ARTHRITIS AND RHEUMATISM 6 64 (6) 1790 - 1798 0004-3591 2012/06 [Refereed][Not invited]
   

  Objective Tofacitinib (CP-690,550) is a novel JAK inhibitor that is currently in clinical trials for the treatment of rheumatoid arthritis (RA). The aim of this study was to examine the effects of tofacitinib in vitro and in vivo in RA, in order to elucidate the role of JAK in the disease process. Methods CD4+ T cells, CD14+ monocytes, and synovial fibroblasts (SFs) were purified from the synovium and peripheral blood of patients with RA and were evaluated for the effect of tofacitinib on cytokine production and cell proliferation. For in vivo analysis, synovium and cartilage samples obtained from patients with RA were implanted in immunodeficient mice (SCID-HuRAg mice), and tofacitinib was administered via an osmotic minipump. Results Tofacitinib treatment of CD4+ T cells originating from synovium and peripheral blood inhibited the production of interleukin-17 (IL-17) and interferon-? (IFN?) in a dose-dependent manner, affecting both proliferation and transcription, but had no effect on IL-6 and IL-8 production. Tofacitinib did not affect IL-6 and IL-8 production by RASFs and CD14+ monocytes. However, conditioned medium from CD4+ T cells cultured with tofacitinib inhibited IL-6 production by RASFs and IL-8 production by CD14+ monocytes. Treatment of SCID-HuRAg mice with tofacitinib decreased serum levels of human IL-6 and IL-8 and markedly suppressed invasion of synovial tissue into cartilage. Conclusion Tofacitinib directly suppressed the production of IL-17 and IFN? and the proliferation of CD4+ T cells, resulting in inhibition of IL-6 production by RASFs and IL-8 production by CD14+ cells and decreased cartilage destruction. In CD4+ T cells, presumably Th1 and Th17 cells, JAK plays a crucial role in RA synovitis.
• STAT3 Inhibition Overcomes Temozolomide Resistance in Glioblastoma by Downregulating MGMT Expression

  Shinji Kohsaka, Lei Wang, Kazuhiro Yachi, Roshan Mahabir, Takuhito Narita, Tamio Itoh, Mishie Tanino, Taichi Kimura, Hiroshi Nishihara, Shinya Tanaka

  MOLECULAR CANCER THERAPEUTICS 11 (6) 1289 - 1299 1535-7163 2012/06 [Refereed][Not invited]
   

  Glioblastoma multiforme (GBM) is one of the most aggressive human tumors with a poor prognosis. Current standard treatment includes chemotherapy with the DNA-alkylating agent temozolomide concomitant with surgical resection and/or irradiation. However, a number of cases are resistant to temozolomide-induced DNA damage due to elevated expression of the DNA repair enzyme O-6-methylguanine-DNA methyltransferase (MGMT). Here, we show that upregulation of both MGMT and STAT3 was accompanied with acquisition of temozolomide resistance in the GBM cell line U87. Inactivation of STAT3 by inhibitor or short hairpin RNA (shRNA) downregulated MGMT expression in GBM cell lines. MGMT upregulation was not observed by the treatment of interleukin (IL)-6 which is a strong activator of STAT3. Contrarily, forced expressed MGMT could be downregulated by STAT3 inhibitor which was partially rescued by the proteasome inhibitor, MG132, suggesting the STAT3-mediated posttranscriptional regulation of the protein levels of MGMT. Immunohistochemical analysis of 44 malignant glioma specimens showed significant positive correlation between expression levels of MGMT and phosphorylated STAT3 (p-STAT3; P < 0.001, gamma = 0.58). Importantly, the levels of both MGMT and p-STAT3 were increased in the recurrence compared with the primary lesion in paired identical tumors of 12 cases. Finally, we showed that STAT3 inhibitor or STAT3 knockdown potentiated temozolomide efficacy in temozolomide-resistant GBM cell lines. Therefore, STAT3 inhibitor might be one of the candidate reagents for combination therapy with temozolomide for patients with temozolomide-resistant GBM. Mol Cancer Ther; 11(6); 1289-99. (C)2012 AACR.
• CD133 Negatively Regulates Tumorigenicity via AKT Pathway in Synovial Sarcoma

  Taichi Kimura, Lei Wang, Kouichi Tabu, Hiroshi Nishihara, Yuji Mashita, Naoyuki Kikuchi, Mishie Tanino, Hiroaki Hiraga, Shinya Tanaka

  CANCER INVESTIGATION 30 (5) 390 - 397 0735-7907 2012/06 [Refereed][Not invited]
   

  Synovial sarcoma (SS) is an aggressive tumor that accounts for almost 10% of all soft tissue sarcomas. In this study, we found the expression of CD133 in human SS specimens, thus, we focused on the function of CD133 in SS. Separation of the CD133-positive and -negative subpopulations in SS cell lines clarified that the CD133-negative subpopulation exhibited enhanced growth and hyperphosphorylation of AKT. Treatment of Akt inhibitor suppressed the cell growth of CD133-negative subpopulation to the levels of CD133-positive cells. These results suggest that CD133 has negative effect on the growth of cells through AKT-dependent signalling pathway.
• Predictive factors of cervical spondylotic myelopathy in patients with lumbar spinal stenosis

  Hideki Iizuka, Keisuke Takahashi, Shinya Tanaka, Kohei Kawamura, Yoshitomo Okano, Hiromi Oda

  ARCHIVES OF ORTHOPAEDIC AND TRAUMA SURGERY 132 (5) 607 - 611 0936-8051 2012/05 [Refereed][Not invited]
   

  To analyze cervical spondylotic myelopathy (CSM) predictive factors in patients with lumbar spinal stenosis (LSS). Two hundred thirty-seven patients who visited for low back pain, lower limb pain and/or lower limb numbness and who were diagnosed with LSS were enrolled in this study. The ratio of males to females was 117-120, and the mean age was 68.8 years (range 45-87 years). LSS and CSM were diagnosed by characteristic symptoms, physical findings and MRI. We examined gender, age, Torg-Pavlov ratio (TPR), spondylolisthesis or spondylosis, LSS symptom types and number of stenosis segments with LSS to clarify predictive factors for CSM. There were 21 (8.86%) patients with coexistent CSM among 237 LSS patients. CSM morbidity was significantly more common among males compared with females. TPR was 0.71 +/- A 0.09 in the CSM patients and 0.81 +/- A 0.10 in the non-CSM patients. TPR of the CSM patients was significantly smaller than that of the non-CSM patients. We analyzed to determine the predictive factors of CSM and TPR was identified. The predictive value of TPR for CSM was 0.78. Torg-Pavlov ratio was the most important predictive factor of CSM in patients with LSS.
• F-18-Fluoromisonidazole positron emission tomography may differentiate glioblastoma multiforme from less malignant gliomas

  Kenji Hirata, Shunsuke Terasaka, Tohru Shiga, Naoya Hattori, Keiichi Magota, Hiroyuki Kobayashi, Shigeru Yamaguchi, Kiyohiro Houkin, Shinya Tanaka, Yuji Kuge, Nagara Tamaki

  EUROPEAN JOURNAL OF NUCLEAR MEDICINE AND MOLECULAR IMAGING 5 39 (5) 760 - 770 1619-7070 2012/05 [Refereed][Not invited]
   

  Purpose Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor and its prognosis is significantly poorer than those of less malignant gliomas. Pathologically, necrosis is one of the most important characteristics that differentiate GBM from lower grade gliomas; therefore, we hypothesized that F-18 fluoromisonidazole (FMISO), a radiotracer for hypoxia imaging, accumulates in GBM but not in lower grade gliomas. We aimed to evaluate the diagnostic value of FMISO positron emission tomography (PET) for the differential diagnosis of GBM from lower grade gliomas. Methods This prospective study included 23 patients with pathologically confirmed gliomas. All of the patients underwent FMISO PET and 18F-fluorodeoxyglucose (FDG) PET within a week. FMISO images were acquired 4 h after intravenous administration of 400 MBq of FMISO. Tracer uptake in the tumor was visually assessed. Lesion to normal tissue ratios and FMISO uptake volume were calculated. Results Of the 23 glioma patients, 14 were diagnosed as having GBM (grade IV glioma in the 2007 WHO classification), and the others were diagnosed as having non-GBM (5 grade III and 4 grade II). In visual assessment, all GBM patients showed FMISO uptake in the tumor greater than that in the surrounding brain tissues, whereas all the non-GBM patients showed FMISO uptake in the tumor equal to that in the surrounding brain tissues (p <= 0.001). One GBM patient was excluded from FDG PET study because of hyperglycemia. All GBM patients and three of the nine (33%) non-GBM patients showed FDG uptake greater than or equal to that in the gray matter. The sensitivity and specificity for diagnosing GBM were 100 and 100% for FMISO, and 100 and 66% for FDG, respectively. The lesion to cerebellum ratio of FMISO uptake was higher in GBM patients (2.74 +/- 0.60, range 1.71-3.81) than in non-GBM patients (1.22 +/- 0.06, range 1.09-1.29, p <= 0.001) with no overlap between the groups. The lesion to gray matter ratio of FDG was also higher in GBM patients (1.46 +/- 0.75, range 0.91-3.79) than in non-GBM patients (1.07 +/- 0.62, range 0.66-2.95, p <= 0.05); however, overlap of the ranges did not allow clear differentiation between GBM and non-GBM. The uptake volume of FMISO was larger in GBM ( 27.18 +/- 10.46%, range 14.02- 46.67%) than in non- GBM ( 6.07 +/- 2.50%, range 2.12- 9.22%, p <= 0.001). Conclusion These preliminary data suggest that FMISO PET may distinguish GBM from lower grade gliomas.
• 中間型松果体実質腫瘍の臨床病理学的検討

  伊東 民雄, 佐藤 憲市, 及川 光照, 中村 博彦, 寺坂 俊介, 宝金 清博, 田中 伸哉, 菅野 宏美, 久保田 佳奈子, 村田 純一

  Brain Tumor Pathology 日本脳腫瘍病理学会 29 (Suppl.) 176 - 176 1433-7398 2012/05
• A case of clear cell variant of solid-pseudopapillary tumor of the pancreas in an adult male patient

  Mishie Tanino, Shinji Kohsaka, Taichi Kimura, Kouichi Tabu, Hiroshi Nishihara, Hiroki Sawa, Hiroyuki Kawami, Hajime Kamada, Michio Shimizu, Shiriya Tanaka

  ANNALS OF DIAGNOSTIC PATHOLOGY 16 (2) 134 - 140 1092-9134 2012/04 [Refereed][Not invited]
   

  A clear cell variant of solid-pseudopapillary tumor (SPT) of the pancreas was initially reported in 2006 as a tumor that arose in the pancreatic body and tail in young adults; to date, only 4 cases of this entity have been reported. Here, we present the case of a 58-year-old man with clear cell variant of SPT with distinctive clinicopathologic features. The tumor was well demarcated, was 2.6 cm in size and mostly composed of multivacuolated clear cells with solid growth, and exhibited the characteristic immunohistochemical positivity of beta-catenin in the cytoplasm and nuclei of the neoplastic cells. In contrast to classical SPT with nuclear positivity, this case was negative for E-cadherin. Direct DNA sequencing of exon 3 of beta-catenin gene demonstrated a single amino acid substitution (serine to phenylalanine) in codon 37, which is the phosphorylation site by GSK beta and frequently found in classical SPT. Electron microscopy demonstrated enlarged mitochondria and endoplasmic reticulum. Despite the fact that previous cases of clear cell variant of SPT arose mainly in the pancreatic body and tail in female young adults (age, 26-32 years), this case suggested that it is possible for a clear cell variant of SPT to arise in the pancreatic head in a middle-aged man. Because the recognition of the clear cell variant of SPT is important for the appropriate diagnosis of primary pancreatic tumor, the present case with its distinctive characteristics may provide new information for a more profound understanding of the pancreatic SPT. (C) 2012 Elsevier Inc. All rights reserved.
• Ymer Acts as a Multifunctional Regulator in Nuclear Factor-kappa B and Fas Signaling Pathways

  Tadasuke Tsukiyama, Mayuko Matsuda-Tsukiyama, Miyuki Bohgaki, Sayuri Terai, Shinya Tanaka, Shigetsugu Hatakeyama

  MOLECULAR MEDICINE 18 (4) 587 - 597 1076-1551 2012/04 [Refereed][Not invited]
   

  The nuclear factor (NF)-kappa B family of transcription factors regulates diverse cellular functions, including inflammation, oncogenesis and apoptosis. It was reported that A20 plays a critical role in the termination of NF-kappa B signaling after activation. Previously, we showed that Ymer interacts and collaborates with A20, followed by degradation of receptor-interacting protein (RIP) and attenuation of NF-kappa B signaling. Here we show the function of Ymer in regulation of several signaling pathways including NF-kappa B on the basis of results obtained by using Ymer transgenic (Ymer Tg) mice. Ymer Tg mice exhibited impaired immune responses, including NF-kappa B and mitogen-activated protein kinase (MAPK) activation, cell proliferation and cytokine production, to tumor necrosis factor (TNF)-alpha, polyl:C or lipopolysaccharide ([PS) stimulation. Ymer Tg mice were more resistant to LPS-induced septic shock than wild-type mice. Transgene of Ymer inhibited the onset of glomerulonephritis in Ipr/Ipr mice as an autoimmune disease model. In contrast to the inflammatory immune response to LPS, Fas-mediated cell death was strongly induced in liver cells of Ymer Tg mice in which Ymer is abundantly expressed. These findings suggest that Ymer acts as a regulator downstream of several receptors and that Ymer functions as a positive or negative regulator in a signaling pathway-dependent manner. Online address: hffp://www.molmed.org doi: 10.2119/molmed.2011.00435
• TRIM67 Protein Negatively Regulates Ras Activity through Degradation of 80K-H and Induces Neuritogenesis

  Hiroaki Yaguchi, Fumihiko Okumura, Hidehisa Takahashi, Takahiro Kano, Hiroyuki Kameda, Motokazu Uchigashima, Shinya Tanaka, Masahiko Watanabe, Hidenao Sasaki, Shigetsugu Hatakeyama

  JOURNAL OF BIOLOGICAL CHEMISTRY 287 (15) 12050 - 12059 0021-9258 2012/04 [Refereed][Not invited]
   

  Tripartite motif (TRIM)-containing proteins, which are defined by the presence of a common domain structure composed of a RING finger, one or two B-box motifs and a coiled-coil motif, are involved in many biological processes including innate immunity, viral infection, carcinogenesis, and development. Here we show that TRIM67, which has a TRIM motif, an FN3 domain and a SPRY domain, is highly expressed in the cerebellum and that TRIM67 interacts with PRG-1 and 80K-H, which is involved in the Ras-mediated signaling pathway. Ectopic expression of TRIM67 results in degradation of endogenous 80K-H and attenuation of cell proliferation and enhances neuritogenesis in the neuroblastoma cell line N1E-115. Furthermore, morphological and biological changes caused by knockdown of 80K-H are similar to those observed by overexpression of TRIM67. These findings suggest that TRIM67 regulates Ras signaling via degradation of 80K-H, leading to neural differentiation including neuritogenesis.
• The Impact of Extent of Resection and Histological Subtype on the Outcome of Adult Patients with High-grade Gliomas

  Shigeru Yamaguchi, Hiroyuki Kobayashi, Shunsuke Terasaka, Nobuaki Ishii, Jun Ikeda, Hiromi Kanno, Hiroshi Nishihara, Shinya Tanaka, Kiyohiro Houkin

  JAPANESE JOURNAL OF CLINICAL ONCOLOGY 42 (4) 270 - 277 0368-2811 2012/04 [Refereed][Not invited]
   

  We reviewed the relationship between extent of resection and survival of patients with high-grade gliomas with special consideration of an oligodendroglial component.A retrospective review was performed on 160 adult patients with histological diagnosis of high-grade gliomas since 2000. All histological slides were categorized as high-grade astrocytomas or oligodendroglial tumors. Extent of resection was assessed by early post-operative magnetic resonance imaging and classified as complete resection, incomplete resection and biopsy. Measured outcomes were overall survival and progression-free survival. The independent association of extent of resection and survival was analyzed by the multivariate proportional hazard model adjusting for prognostic factors.The lesions were classified as high-grade astrocytomas in 93 patients and high-grade oligodendroglial tumors in 67 patients. In high-grade astrocytomas, the median survival after complete resection (n 36), incomplete resection (n 36) and biopsy (n 21) was 23.4, 15.3 and 12.6 months, respectively. Complete resection was independently associated with increased overall survival (P 0.001) and progression-free survival (P 0.002) compared with incomplete resection, while incomplete resection was not associated with survival benefit compared with biopsy by multivariate analysis. On the other hand, in high-grade oligodendroglial tumors, the majority of patients were still alive and there is no significant difference in the survival between complete resection (n 24) and incomplete resection (n 33), while even incomplete resection had a significantly longer overall survival (P 0.001) and progression-free survival (P 0.006) compared with biopsy (n 10).Maximal cytoreduction improves the survival of high-grade gliomas, although our data indicated that the impact of extent of resection in high-grade astrocytomas is different from that in high-grade oligodendroglial tumors.
• Prognostic Significance of Clusterin Expression in Advanced-Stage Cervical Cancer Treated With Curative Intended Radiotherapy

  Hidemichi Watari, Rumiko Kinoshita, Yimin Han, Lei Wang, Masayoshi Hosaka, Hiroshi Taguchi, Kazuhiko Tsuchiya, Shinya Tanaka, Hiroki Shirato, Noriaki Sakuragi

  INTERNATIONAL JOURNAL OF GYNECOLOGICAL CANCER 22 (3) 465 - 470 1048-891X 2012/03 [Refereed][Not invited]
   

  Objective: Overexpression of clusterin (CLU), an antiapoptotic molecule, has been reported to induce resistance to radiotherapy (RT) in a variety of cancer cell types. The aim of this study was to evaluate the significance of CLU expression to predict survival of patients with advanced-stage cervical cancer who received curative intended RT. Methods: Biopsy tissue specimens of advanced-stage cervical cancer before curative intended RT were obtained from 34 patients who were treated at Hokkaido University Hospital between 1998 and 2008 and whose complete medical records were available. The expression of CLU protein was analyzed by immunohistochemistry. Findings were evaluated in relation to several clinicopathological factors. Survival analyses were performed using the Kaplan-Meier curves and the log-rank test. Independent prognostic factors were determined by multivariate Cox regression analysis. Results: Clusterin protein was mainly present in the cytoplasm of cervical cancer cells. The expression of CLU protein in cervical cancer tissues before curative intended RT was not significantly related to any clinicopathological factors analyzed, including age, clinical stage, histologic type, and response to RT. Univariate analysis on prognostic factors showed that histologic type (P = 0.001), and CLU expression (P = 0.02) were related to survival. Multivariate analysis revealed that both histologic type (P = 0.002), and CLU expression (P = 0.02) were independent prognostic factors for overall survival. Conclusion: We conclude that CLU could be a new molecular marker to predict overall survival of patients with advanced-stage cervical cancer treated with curative intended RT.
• MUM1(IRF4)陽性NK/T細胞リンパ腫の一剖検例

  石田 雄介, 前田 豪樹, 岩田 祐司, 木村 仁, 星川 剛, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 101 (1) 376 - 376 0300-9181 2012/03
• 腎細胞癌に対するネクサバール投与と放射線照射との相乗作用により消化管穿孔が誘発された一剖検例

  佐藤 行真, 石田 雄介, 市原 浩司, 松川 雅則, 田中 伸哉

  日本病理学会会誌 (一社)日本病理学会 101 (1) 442 - 442 0300-9181 2012/03
• 中間型松果体実質腫瘍(PPTID)の3例

  大竹 安史, 佐藤 憲市, 伊東 民雄, 安斉 公雄, 尾崎 義丸, 及川 光照, 福井 崇人, 中村 博彦, 菅野 宏美, 田中 伸哉

  北海道脳神経疾患研究所医誌 (公財)北海道脳神経疾患研究所 22 (1) 33 - 39 1340-5764 2012/03 [Not refereed][Not invited]
   

  症例1(71歳女性)。後頭部痛を主訴に、頭部MRIとCTにて松果体部腫瘍が認められた。内視鏡的第三脳室底開窓術(ETV)および生検術を施行し、病理組織学的に中間型松果体実質腫瘍(PPTID)と診断されたが、術後10ヵ月経過で無症状である。症例2(33歳男性)。頭痛を主訴にMRIとCTにて閉塞性水頭症を伴う松果体腫瘍、および鞍上部と脊髄に播種が認められた。ETVと生検術を行い、病理組織学的にPPTIDと診断された。術後は化学療法、全脳全脊髄照射を追加したが、現在までPRで経過している。症例3(69歳女性)。歩行障害を主訴とした。頭部CTとMRIにて閉塞性水頭症を伴う松果体部腫瘍が認められ、ETVと生検術にて病理組織学的にPPTIDと診断された。以後、残存腫瘍に対し放射線療法を追加したところ、現在までPRで経過している。
• Analyses of IDH1 mutation and MGMT promoter methylation status for 5 cases of long-term survivors with glioblastoma

  Yuuta Kamoshima, Hiroaki Motegi, Shunsuke Terasaka, Hiroyuki Kobayashi, Shigeru Yamaguchi, Junichi Murata, Shinya Tanaka, Kiyohiro Houkin

  Neurological Surgery 40 (2) 129 - 135 0301-2603 2012/02/10 [Refereed][Not invited]
   

  Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor with an extremely poor prognosis in spite of multimodal treatment approaches. The median survival time of patients with GBM is 15 months, and only 3-5% of patients survive longer than 36 months. Those patients who survive over 36 months after the initial diagnosis are defined as long-term survivors. In this study, we retrospectively performed clinical and molecular analyses of five long-term survivors of GBM (> 36 months) and twenty four GBM patients with poor survival time as control group (< 36 months) to identify any prognostic factors that potentially contribute to survival. The O 6- methylguanine-DNA methyltransferase (MGMT) gene methylation status was evaluated by performing methylation specific polymerase chain reaction assays. The mutation of isocitrate dehydrogenase 1 and 2 were evaluated by the direct sequencing method. All five cases were primary GBMs and the coexistence of the oligodendroglioma component was checked for each case as GBM with oligodendroglioma component. All five cases showed MGMT promoter methylation (5/5). IDH1 mutation was detected in two of the long-term survivors with oligodendroglioma component (2/5) while no IDH1 mutation was detected in the control group. All patients were treated by gross total removal followed by radiotherapy and various chemotherapies including temozolomide. MGMT promoter methylation and IDH1 mutation might be favorable factors for long-term survival in GBM patients.
• Reoxygenation of Glioblastoma Multiforme Treated with Fractionated Radiotherapy Concomitant with Temozolomide: Changes Defined by F-18-fluoromisonidazole Positron Emission Tomography: Two Case Reports

  Takuhito Narita, Hidefumi Aoyama, Kenji Hirata, Shunsuke Onodera, Tohru Shiga, Hiroyuki Kobayashi, Junichi Murata, Shunsuke Terasaka, Shinya Tanaka, Kiyohiro Houkin

  JAPANESE JOURNAL OF CLINICAL ONCOLOGY 42 (2) 120 - 123 0368-2811 2012/02 [Refereed][Not invited]
   

  Two glioblastoma multiforme patients underwent F-18-FMISO (fluoromisonidazole) positron emission tomography study to access the tumor oxygenation status before and immediately after fractionated radiotherapy concomitant with temozolomide chemotherapy. In both cases, a prominent F-18-FMISO tumor accumulation observed in the first study was notably decreased in the second study, which was supposed to be a reoxygenation of the tumor. As far as we investigated, this is the first report of the changes of oxygenation status in glioblastoma multiforme treated through radiation therapy with temozolomide.
• CRKL plays a pivotal role in tumorigenesis of head and neck squamous cell carcinoma through the regulation of cell adhesion

  Hiroko Yanagi, Lei Wang, Hiroshi Nishihara, Taichi Kimura, Mishie Tanino, Teruki Yanagi, Satoshi Fukuda, Shinya Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 418 (1) 104 - 109 0006-291X 2012/02 [Refereed][Not invited]
   

  The signaling adapter protein CRK is an indispensable molecule involved in regulating the malignant potential of human cancers. CRK-like (CRKL) is a hematopoietic cell-dominant homologue of CRK that is reported to be phosphorylated by BCR-ABL tyrosine kinase in chronic myelogenous leukemia patients, but its biological function in non-hematopoietic tumors remains unclear. In this study, we explored the tumorigenic role of CRKL in head and neck squamous cell carcinoma (HNSCC) in vitro and in vivo. Immunoprecipitation analysis of HNSCC cell line, HSC-3 cells, showed that the dominant binding partner for C3G was CRKL, not CRK. To clarify the molecular function of CRKL, we established lentiviral shRNA-mediated CRKL-knockdown HNSCC cell lines. In CRKL-knockdown HSC-3 and HSC-4 cells, cell growth and motility were diminished compared to control cells. Cell adhesion assays showed that cell attachment onto both fibronectin- and collagen-coated dishes was significantly suppressed in CRKL-knockdown HSC-3 cells, while no significant change was observed for poly-L-lysine-coated dishes. Immunofluorescence staining revealed that focal adhesion was reduced in CRKL-knockdown HSC-3 cells. With a pull-down assay, CRKL-knockdown HSC-3 cells showed decreased amounts of active Rap1 compared to control cells. Moreover, in an in vivo assay, tumor formation of CRKL-knockdown HSC-3 cells in nude mice was significantly abrogated. Our results indicate that CRKL regulates HNSCC-cell growth, motility, and integrin-dependent cell adhesion, suggesting that CRKL plays a principal role in HNSCC tumorigenicity. (C) 2012 Elsevier Inc. All rights reserved.
• Role of Mast Cells and Basophils in IgE Responses and in Allergic Airway Hyperresponsiveness

  Minoru Sawaguchi, Shinya Tanaka, Yuriko Nakatani, Yasuyo Harada, Kaori Mukai, Yuko Matsunaga, Kenji Ishiwata, Keisuke Oboki, Taku Kambayashi, Naohiro Watanabe, Hajime Karasuyama, Susumu Nakae, Hiromasa Inoue, Masato Kubo

  JOURNAL OF IMMUNOLOGY 188 (4) 1809 - 1818 0022-1767 2012/02 [Refereed][Not invited]
   

  We established a diphtheria toxin (DT)-based conditional deletion system using 114 enhancer elements previously shown to be specific for IL-4 production in mast cells (MCs) or basophils (Mas-TRECK and Bas-TRECK mice). DT treatment of Bas-TRECK mice resulted in specific deletion of basophils, whereas both MCs and basophils were deleted in Mas-TRECK mice. DT-treated Mas-TRECK mice had impaired passive cutaneous anaphylaxis, IgE-mediated passive systemic anaphylaxis, and IgE-mediated chronic allergic inflammation, whereas DT-treated Bas-TRECK mice had impaired IgE-mediated chronic allergic inflammation. Using these mice, we also sought to tease out the role of MCs and basophils in airway hyperresponsiveness (AHR). Although MC deletion resulted in a slight increase in basal Ag-specific IgE levels and significant increases in basal IgE levels, we found that this deletion markedly impaired the AHR effector phase and was accompanied by decreased histamine levels. By contrast, basophil deletion had no effect on the AHR effector phase or on IgE production induced by systemic OVA immunization. Our results, using these newly established Mas-TRECK and Bas-TRECK models, demonstrated an indispensable role for MCs as effector cells in AHR. The Journal of Immunology, 2012, 188: 1809-1818.
• A fatal case of cytomegalovirus ventriculoencephalitis in a mycosis fungoides patient who received multiple umbilical cord blood cell transplantations

  Toshihiro Matsukawa, Hideki Goto, Kenta Takahashi, Shinsuke Asanuma, Atsushi Yasumoto, Mutsumi Takahata, Akio Shigematsu, Tomoyuki Endo, Junji Tanaka, Satoshi Hashino, Shinya Tanaka, Masahiro Imamura

  INTERNATIONAL JOURNAL OF HEMATOLOGY 95 (2) 217 - 222 0925-5710 2012/02 [Refereed][Not invited]
   

  Cytomegalovirus (CMV) infection is latent in the majority of adult humans. The reactivation of CMV causes pneumonia and gastrointestinal disease in severely immunosuppressed patients, who consequently suffer very high mortality due to CMV central nervous system disease. We report here a case involving a 28-year-old female patient with mycosis fungoides who underwent umbilical cord blood transplantation three times and developed CMV ventriculoencephalitis. The patient's CMV viremia was successfully preempted with ganciclovir (GCV) as indicated by undetectable CMV antigenemia; despite this successful treatment, the patient developed CMV ventriculoencephalitis. Foscarnet (FCV) therapy led to a temporary recovery, after which CMV ventriculoencephalitis recurred, and the patient died after receiving combination GCV and FCV therapy. Autopsy samples revealed CMV ventriculoencephalitis, as indicated by numerous inclusion-bearing cells (Owl's eye). It is likely that this patient harbored a GCV-resistant CMV strain; however, it was not possible to obtain nucleic acids suitable for use in assessing this possibility.
• Toll-like receptor 3 signaling converts tumor-supporting myeloid cells to tumoricidal effectors

  Hiroaki Shime, Misako Matsumoto, Hiroyuki Oshiumi, Shinya Tanaka, Akio Nakane, Yoichiro Iwakura, Hideaki Tahara, Norimitsu Inoue, Tsukasa Seya

  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 109 (6) 2066 - 2071 0027-8424 2012/02 [Refereed][Not invited]
   

  Smoldering inflammation often increases the risk of progression for malignant tumors and simultaneously matures myeloid dendritic cells (mDCs) for cell-mediated immunity. PolyI:C, a dsRNA analog, is reported to induce inflammation and potent antitumor immune responses via the Toll-like receptor 3/Toll-IL-1 receptor domain-containing adaptor molecule 1 (TICAM-1) and melanoma differentiation-associated protein 5/IFN-beta promoter stimulator 1 (IPS-1) pathways in mDCs to drive activation of natural killer cells and cytotoxic T lymphocytes. Here, we found that i.p. or s.c. injection of polyI:C to Lewis lung carcinoma tumor-implant mice resulted in tumor regression by converting tumor-supporting macrophages (Mfs) to tumor suppressors. F4/80(+)/Gr1(-) Mfs infiltrating the tumor respond to polyI:C to rapidly produce inflammatory cytokines and thereafter accelerate M1 polarization. TNF-alpha was increased within 1 h in both tumor and serum upon polyI: C injection into tumor-bearing mice, followed by tumor hemorrhagic necrosis and growth suppression. These tumor responses were abolished in TNF-alpha-/mice. Furthermore, F4/80(+) Mfs in tumors extracted from polyI:C-injected mice sustained Lewis lung carcinoma cytotoxic activity, and this activity was partly abrogated by anti-TNF-alpha Ab. Genes for supporting M1 polarization were subsequently up-regulated in the tumor-infiltrating Mfs. These responses were completely abrogated in TICAM-1(-/-) mice, and unaffected in myeloid differentiation factor 88(-/-) and IPS-1(-/-) mice. Thus, the TICAM-1 pathway is not only important to mature mDCs for cross-priming and natural killer cell activation in the induction of tumor immunity, but also critically engaged in tumor suppression by converting tumor-supporting Mfs to those with tumoricidal properties.
• The 3 ' Enhancer CNS2 Is a Critical Regulator of Interleukin-4-Mediated Humoral Immunity in Follicular Helper T Cells

  Yohsuke Harada, Shinya Tanaka, Yasutaka Motomura, Yasuyo Harada, Shin-ichiro Ohno, Shinji Ohno, Yusuke Yanagi, Hiromasa Inoue, Masato Kubo

  IMMUNITY 36 (2) 188 - 200 1074-7613 2012/02 [Refereed][Not invited]
   

  A main role for interleukin-4 (IL-4) is in humoral immunity, and follicular helper CD4(+) T (Tfh) cells may be an intrinsic IL-4 source. Here we demonstrate that conserved noncoding sequence 2 (CNS2) is an essential enhancer element for IL-4 expression in Tfh cells but not in Th2 cells. Mice with a CNS2 deletion had a reduction in IgG1 and IgE production and in IL-4 expression in Tfh cells. Tracking of CNS2 activity via a GFP reporter mouse demonstrated that CNS2-active cells expressed several markers of Tfh cells: CXCR5, PD-1, and ICOS; the transcriptional master regulator Bcl6; and the cytokines IL-21 and IL-4. These CNS2-active cells were mainly localized in B cell follicles and germinal centers. The GFP(+) Tfh cells were derived from GFP(-) naive T cells after in vivo systemic immunization. These results indicate that CNS2 is an essential enhancer element required for IL-4 expression in Tfh cells controlling humoral immunity.
• Histopathological and immunohistochemical findings of 20 autopsy cases with 2009 H1N1 virus infection.

  Nakajima N, Sato Y, Katano H, Hasegawa H, Kumasaka T, Hata S, Tanaka S, Amano T, Kasai T, Chong JM, Iizuka T, Nakazato I, Hino Y, Hamamatsu A, Horiguchi H, Tanaka T, Hasegawa A, Kanaya Y, Oku R, Oya T, Sata T

  Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 1 25 1 - 13 0893-3952 2012/01 [Refereed][Not invited]
  
• Neurohypophyseal germinoma with abundant fibrous tissue

  Shunsuke Terasaka, Masahito Kawabori, Hiroyuki Kobayashi, Junichi Murata, Hiromi Kanno, Shinya Tanaka, Kiyohiro Houkin

  BRAIN TUMOR PATHOLOGY 29 (1) 58 - 62 1433-7398 2012/01 [Refereed][Not invited]
   

  We report an unusual case of neurohypophyseal germinoma with abundant fibrous tissue and clival invasion that was initially misdiagnosed as lymphocytic hypophysitis. A 40-year-old woman presented with diabetes insipidus and panhypopituitarism after delivering her second son and which lasted for 4 years. Magnetic resonance imaging showed the intrasellar mass extending to the suprasellar region with enlarged pituitary stalk. The mass was heterogeneously enhanced and invaded the clivus. Biopsy of the intrasellar mass was performed via the trans-sphenoidal route, and histological examination revealed marked fibrous tissue and infiltration of lymphocytes, with no evidence of tumor cells. Lymphocytic hypophysitis was the initial diagnosis, and corticosteroid therapy was begun. Despite intensive treatment, the lesion enlarged and clinical symptoms worsened 2 weeks after surgery. Subtotal removal of the mass was performed, and a second histological examination revealed typical findings of the germinoma. Subsequently, the patient underwent chemoradiotherapy, and complete remission was achieved. Histological diagnosis is sometimes incorrect in fibrous tumors at the sellar region, and biopsy from several points is strongly recommended for this entity.
• Acute Phlegmonous Gastritis with Neutropenia

  Makoto Saito, Masanobu Morioka, Hiromi Kanno, Shinya Tanaka

  INTERNAL MEDICINE 51 (20) 2987 - 2988 0918-2918 2012 [Refereed][Not invited]
  
• Roles for Crk in Cancer Metastasis and Invasion

  Masumi Tsuda, Shinya Tanaka

  Genes and Cancer 3 (5-6) 334 - 340 1947-6019 2012 [Refereed][Not invited]
   

  The Crk family of adaptors is implicated in regulating various biological and pathological processes such as cell proliferation, adhesion, migration, invasion, phagocytosis, and survival. A large number of studies have shown that Crk plays an important role in aggressive and malignant behaviors of human cancers. In immunohistochemical analyses and gene-expression profiles, enhanced expression of Crk has been identified in adenocarcinomas of lung, breast, and stomach and in sarcomas and glioma. Overexpression of Crk in tumor cells induces the prominent tyrosine phosphorylations of scaffolding molecules such as p130Cas and paxillin through Src family tyrosine kinases and stimulates the activation loop of intracellular signalling, ultimately contributing to the increased motility and aggressive potential of cancer cells. Crk proteins thus are not simply conduits for intracellular signal transduction but also can control the amplitude of signalling. This review summarizes the significance of Crk and its mediated signaling assemblies, particularly in regulating tumor metastasis and invasion, and discusses the possibilities that they are potential cancer therapeutic targets. © The Author(s) 2012.
• Clusterin is a potential molecular predictor for ovarian cancer patient's survival: targeting clusterin improves response to paclitaxel.

  Mohamed K Hassan, Hidemichi Watari, Yimin Han, Takashi Mitamura, Masayoshi Hosaka, Lei Wang, Shinya Tanaka, Noriaki Sakuragi

  Journal of experimental & clinical cancer research : CR 30 113 - 113 0392-9078 2011/12/20 [Refereed][Not invited]
   

  BACKGROUND: Clusterin is a cytoprotective chaperone protein involved in numerous physiological processes, carcinogenesis, tumor growth and tissue remodelling. The purpose of this study was to investigate whether clusterin (CLU), an antiapoptotic molecule, could be a potential predictor molecule for ovarian cancer and whether or not targeting this molecule can improve survival of ovarian cancer patients. METHODS: Clusterin expression was compared between ten primary and their recurrent tumors from same patients immunohistochemically. We analyzed prognostic significance of CLU expression in another 47 ovarian cancer tissue samples by immunohistochemistry. We used small interference RNA to knock down CLU in the chemo-resistant ovarian cancer cell lines. KF-TX and SKOV-3-TX, paclitaxel-resistant ovarian cancer cells, were established from parental KF and SKOV-3 chemo-sensitive cell lines, respectively. Either siRNA or second generation antisense oligodeoxynucleotide against CLU (OGX-011), which is currently evaluated in clinical phase II trials in other cancer s, was used to modulate sensitivity to paclitaxel (TX) in ovarian cancer cells in vitro. Cellular viability assay, FACS analysis and annexin V staining were used to evaluate the comparative effect of CLU knocking down in ovarian cancer cells. RESULTS: Immunohistochemical analysis of CLU expression in primary ovarian cancer tissue specimens and their recurrent counterparts from same patients demonstrated higher expression of CLU in the recurrent resistant tumors compared with their primary tumors. High expression of CLU by immunohistochemistry among 47 surgical tissue specimens of early-stage (stage I/II) ovarian cancer, who underwent complete cytoreduction as a primary surgery, significantly related to poor survival, while none of other clinicopathological factors analyzed were related to survival in this patient cohort. Secretory CLU (s-CLU; 60 KDa) expression was upregulated in TX-resistant ovarian cancer cells compared to parental cells. Transfection of siRNA or OGX-011 clearly reduced CLU expression. Cell viability assay, FACS analysis and annexin V staining demonstrated that targeting CLU expression by siRNA or OGX-011 sensitized ovarian cancer cells to TX. CONCLUSION: We conclude that CLU could be a potential molecular target to predict survival while targeting this s-CLU may improve survival of patients with ovarian cancer.
• Clusterin is a potential molecular predictor for ovarian cancer patient's survival: targeting Clusterin improves response to paclitaxel

  Mohamed K. Hassan, Hidemichi Watari, Yimin Han, Takashi Mitamura, Masayoshi Hosaka, Lei Wang, Shinya Tanaka, Noriaki Sakuragi

  JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH 30 1756-9966 2011/12 [Refereed][Not invited]
   

  Background: Clusterin is a cytoprotective chaperone protein involved in numerous physiological processes, carcinogenesis, tumor growth and tissue remodelling. The purpose of this study was to investigate whether clusterin (CLU), an antiapoptotic molecule, could be a potential predictor molecule for ovarian cancer and whether or not targeting this molecule can improve survival of ovarian cancer patients. Methods: Clusterin expression was compared between ten primary and their recurrent tumors from same patients immunohistochemically. We analyzed prognostic significance of CLU expression in another 47 ovarian cancer tissue samples by immunohistochemistry. We used small interference RNA to knock down CLU in the chemo-resistant ovarian cancer cell lines. KF-TX and SKOV-3-TX, paclitaxel-resistant ovarian cancer cells, were established from parental KF and SKOV-3 chemo-sensitive cell lines, respectively. Either siRNA or second generation antisense oligodeoxynucleotide against CLU (OGX-011), which is currently evaluated in clinical phase II trials in other cancer s, was used to modulate sensitivity to paclitaxel (TX) in ovarian cancer cells in vitro. Cellular viability assay, FACS analysis and annexin V staining were used to evaluate the comparative effect of CLU knocking down in ovarian cancer cells. Results: Immunohistochemical analysis of CLU expression in primary ovarian cancer tissue specimens and their recurrent counterparts from same patients demonstrated higher expression of CLU in the recurrent resistant tumors compared with their primary tumors. High expression of CLU by immunohistochemistry among 47 surgical tissue specimens of early-stage (stage I/II) ovarian cancer, who underwent complete cytoreduction as a primary surgery, significantly related to poor survival, while none of other clinicopathological factors analyzed were related to survival in this patient cohort. Secretory CLU (s-CLU; 60 KDa) expression was upregulated in TX-resistant ovarian cancer cells compared to parental cells. Transfection of siRNA or OGX-011 clearly reduced CLU expression. Cell viability assay, FACS analysis and annexin V staining demonstrated that targeting CLU expression by siRNA or OGX-011 sensitized ovarian cancer cells to TX. Conclusion: We conclude that CLU could be a potential molecular target to predict survival while targeting this s-CLU may improve survival of patients with ovarian cancer.
• Lysine-specific demethylase 1 is highly expressed in solitary fibrous tumors, synovial sarcomas, rhabdomyosarcomas, desmoplastic small round cell tumors, and malignant peripheral nerve sheath tumors

  Hans-Ulrich Schildhaus, Rajjael Riegel, Wolfgang Hartmann, Susanne Steiner, Eva Wardelmann, Sabine Merkelbach-Bruse, Shinya Tanaka, Hiroshi Sonobe, Roland Schuele, Reinhard Buettner, Jutta Kirfel

  HUMAN PATHOLOGY 42 (11) 1667 - 1675 0046-8177 2011/11 [Refereed][Not invited]
   

  Epigenetic changes including histone methylation, histone acetylation, and DNA methylation are thought to play important roles in the onset and progression of cancer in numerous tumor types. Recent evidence shows that dysregulated epigenetic modifications are as significant as genetic mutations and can act as oncogenic driver lesions causing autonomous growth of cancer cells. Here, we investigated the role of lysine-specific demethylase 1 in mesenchymal tumors. Lysine-specific demethylase 1 is the first discovered histone lysine demethylase and can demethylate both H3K4me2/1 and H3K9me2/1. By analyzing a total of 468 tumors, we describe for the first time high lysine-specific demethylase 1 expression in several highly malignant sarcomas, including synovial sarcomas, rhabdomyosarcomas, desmoplastic small round cell tumors and malignant peripheral nerve sheath tumors. Among the intermediate tumors only solitary fibrous tumors were found to be highly lysine-specific demethylase I positive, whereas lysine-specific demethylase 1 expression was low or absent in benign tumors. Lysine-specific demethylase 1 inhibition with small molecule inhibitors resulted in growth inhibition of synovial sarcoma cells in vitro and an increase in global H3K4me2 methylation. Sarcomas continue to remain a clinical challenge and therefore the identification of both diagnostic markers and novel drug targets for the development of new therapeutic options are needed. Our results suggest that dysregulation of lysinespecific demethylase 1 is associated with highly malignant sarcomas proposing them as molecular tumor markers as well as targets for the treatment of these tumor types. (C) 2011 Elsevier Inc. All rights reserved.
• Effects of vertebroplasty for delayed-onset paraplegia caused by vertebral pseudarthrosis

  Fuminori Saito, Keisuke Takahashi, Shinya Tanaka, Tetuya Torio, Hideki Iizuka, Cui Wei, Hiromi Oda

  JOURNAL OF ORTHOPAEDIC SCIENCE 16 (6) 673 - 681 0949-2658 2011/11 [Refereed][Not invited]
   

  The effectiveness of percutaneous vertebroplasty for osteoporotic vertebral pseudarthrosis with delayed-onset paraplegia has not been reported. We performed vertebroplasty for such patients and tried to investigate the effectiveness of this surgery. We studied 11 patients (2001-2007) treated with percutaneous vertebroplasty for osteoporotic vertebral pseudarthrosis with delayed-onset paraplegia. The mean age of the patients was 71.9 +/- A 5.2 years and the affected vertebrae were located in the thoracolumbar junction. The mean period between the onset of motor weakness and the day of the surgery was 9.5 +/- A 5.7 weeks. Vertebroplasty was performed by filling the intravertebral cleft with polymethylmethacrylate. The clinical course was estimated using the Denis pain scale, the Eastern Cooperative Oncology Group performance status scale and the modified Medical Research Council grade before the surgery, 0, 1, 3, 6 months and 1 year following the surgery, and at the latest follow-up visit. The instability angle and local kyphotic angle were evaluated with X-rays. Significant improvements were observed in the pain scale in all patients, as they did not experience severe pain, and the performance status following the surgery. These conditions continued until the final clinical examination. Most patients had motor weakness, with a preoperative manual motor test score of 0-3, which gradually improved to 4-5 over the examination period. The stabilities of the affected vertebrae were confirmed on imaging at the final examination. Kyphotic changes were initially realigned, but a correction loss occurred in 7 of the 11 patients as a result of adjacent vertebral fractures. Increase in kyphosis following the surgery did not affect the muscle strength recovery. Bridging callus formations were observed around the affected vertebrae within 6 months in all cases. Percutaneous vertebroplasty for vertebral pseudarthrosis with delayed-onset paraplegia is effective for recovering muscle strength, improving performance status and relief from pain, despite local kyphosis.
• Bone Marrow-Derived Endothelial Progenitor Cells Participate in the Initiation of Moyamoya Disease

  Taku Sugiyama, Satoshi Kuroda, Naoki Nakayama, Shinya Tanaka, Kiyohiro Houkin

  NEUROLOGIA MEDICO-CHIRURGICA 51 (11) 767 - 773 0470-8105 2011/11 [Refereed][Not invited]
   

  The mechanisms through which moyamoya disease occurs and progresses remain unknown. Recent studies have indicated the involvement of circulating endothelial progenitor cells (EPCs) in the development of moyamoya disease. This study directly investigated the participation of EPCs in moyamoya disease, using specimens of the supraclinoid internal carotid artery collected from two adult patients. The specimens were stained with primary antibodies against CD34, CD133, and vascular endothelial growth factor receptor-2 (VEGFR2) to localize the circulating EPCs in the thickened intima of occlusive arterial lesion. The CD34- and VEGFR2-positive cells were densely found in the thickened intima of occlusive arterial lesion, particularly clustered in the superficial layer of thickened intima. However, the number of CD34- and CD133-positive cells was very small. The CD34-positive cells also expressed von Willebrand factor on the surface of thickened intima and were also positive for a-smooth muscle actin in the deeper layer. These findings suggest that circulating EPCs may be involved in the development of occlusive arterial lesion in moyamoya disease.
• Reusable and sustainable nanostructured skeleton catalyst: Heck reaction with nanoporous metallic glass Pd (PdNPore) as a support, stabilizer and ligand-free catalyst

  Tetsuro Kaneko, Shinya Tanaka, Naoki Asao, Yoshinori Yamamoto, Mingwei Chen, Wei Zhang, Akihisa Inoue

  Advanced Synthesis and Catalysis 353 (16) 2927 - 2932 1615-4150 2011/11 [Refereed][Not invited]
   

  Nanoporous metallic glass palladium (PdNPore), which was fabricated by de-alloying of a glassy metallic alloy Pd30Ni50P 20, exhibited a remarkable catalytic activity for the Heck reaction of versatile aryl iodides and aryl bromides. Moreover, the PdNPore can be reused several times without a significant loss of catalytic activity, and the PdNPore has ahigher resistance to leaching than palladium black and palladium on carbon. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
• Phosphatidylinositol-3 '-kinase/AKT signaling is essential in synovial sarcoma

  Nicolaus Friedrichs, Marcel Trautmann, Elmar Endl, Elisabeth Sievers, Dagmar Kindler, Peter Wurst, Jacqueline Czerwitzki, Susanne Steiner, Marcus Renner, Roland Penzel, Arend Koch, Olle Larsson, Shinya Tanaka, Akira Kawai, Peter Schirmacher, Gunhild Mechtersheimer, Eva Wardelmann, Reinhard Buettner, Wolfgang Hartmann

  INTERNATIONAL JOURNAL OF CANCER 129 (7) 1564 - 1575 0020-7136 2011/10 [Refereed][Not invited]
   

  Synovial sarcomas account for 5-10% of all malignant soft tissue tumors. They have been shown to express different membranous growth factor receptors, many of them signaling via intracellular kinase cascades. In our study, the functional role of PI3K/AKT signals in synovial sarcoma is analyzed with regard to tumor biology and therapeutic applicability. Immunohistochemical stainings of (Ser473)-phosphorylated (p)-AKT, its targets p-(Ser9)-GSK-3 beta and p-(Ser2448)-mTOR and the cell cycle regulators Cyclin D1 and p27(KIP1) were performed in 36 synovial sarcomas. The PIK3CA gene was screened for mutations. In vitro, four synovial sarcoma cell lines were treated with the PI3K inhibitor LY294002. Phosphorylation of AKT, GSK-3 beta and mTOR was assessed, and cellular proliferation and apoptosis were analyzed to functionally characterize the effects of PI3K inhibition. Finally, coincubations of LY294002 with cytotoxic drugs were performed. Most tumors showed significant expression levels of p-AKT, p-GSK-3 beta and p-mTOR, indicating activation of the PI3K/AKT signaling cascade in synovial sarcomas; Cyclin D1 and p27(KIP1) were differentially expressed. Mutations in the PIK3CA gene could be excluded. In vitro, PI3K inhibition diminished synovial sarcoma cell growth accompanied by reduced phosphorylation of AKT, GSK-3 beta and mTOR. Mechanistically, PI3K pathway inhibition lead to enhanced apoptosis and decreased cellular proliferation linked to reduced Cyclin D1 and increased p27(KIP1) levels. Simultaneous treatment of synovial sarcoma cell lines with LY294002 and cytotoxic drugs resulted in additive effects. In summary, PI3K signaling plays an essential role in growth control of synovial sarcomas and might be successfully targeted in multimodal therapeutic strategies.
• A case of lymphoplasmacyte-rich meningioma of the jugular foramen

  Hiromi Kanno, Hiroshi Nishihara, Keiji Hara, Yoshimaru Ozaki, Tamio Itoh, Taichi Kimura, Mishie Tanino, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 28 (4) 341 - 345 1433-7398 2011/10 [Refereed][Not invited]
   

  Lymphoplasmacyte-rich meningioma (LPRM), the most rare variant of meningiomas, features extensive lymphoplasmacytic infiltrates. Although the jugular foramen (JF) is occasionally involved by several types of tumor, such as paragangliomas and schwannomas, meningioma in the JF is an infrequent disease. Here we present an extremely rare case of LPRM found in the JF. A 55-year-old woman complained of paresis in her right eyelid and palsy in the right side of her lip. Hoarseness and dysphagia also occurred in the following month. Radiologic examinations disclosed a mass lesion in the right JF, and the tumor was operatively removed. Microscopically, the tumor was composed of extensive lymphoplasmacytic infiltration with mild vascular proliferation and scattered sheets of epithelioid cells with plump cytoplasm. Although the obvious whorl formation or psammoma bodies were not observed, by immunochemistry the epithelioid cells were positive for epithelial membrane antigen and also progesterone receptor, indicating a meningothelial cell origin. Considering the histological and radiologic findings, we finally diagnosed the case as LPRM, making this the second reported case of LPRM in the JF.
• Enhancement of TGF-beta Signaling Responses by the E3 Ubiquitin Ligase Arkadia Provides Tumor Suppression in Colorectal Cancer

  Vikas Sharma, Anna G. Antonacopoulou, Shinya Tanaka, Alexios A. Panoutsopoulos, Vasiliki Bravou, Haralabos P. Kalofonos, Vasso Episkopou

  CANCER RESEARCH 20 71 (20) 6438 - 6449 0008-5472 2011/10 [Refereed][Not invited]
   

  TGF-beta signaling provides tumor protection against colorectal cancer (CRC). Mechanisms that support its tumor-suppressive properties remain unclear. The ubiquitin ligase Arkadia/RNF111 enhances TGF-beta signaling responses by targeting repressors of the pathway for degradation. The corepressors SnoN/Ski, critical substrates of Arkadia, complex with the activated TGF-beta signaling effectors Smad2/3 (pSmad2/3) on the promoters of target genes and block their transcription. Arkadia degrades this complex including pSmad2/3 and unblocks the promoter. Here, we report that Arkadia is expressed highly in the mouse colonic epithelium. Heterozygous Akd(+/-) mice are normal but express less Arkadia. This leads to reduced expression of several TGF-beta target genes, suggesting that normal levels of Arkadia are required for efficient signaling responses. Critically, Akd(+/-) mice exhibit increased susceptibility to azoxymethane/dextran sodium sulfate carcinogen-induced CRC, as they develop four-fold more tumors than wild-type mice. Akd(+/-) tumors also exhibit a more aggressive pathology, higher proliferation index, and reduced cytostasis. Therefore, Arkadia functions as a tumor suppressor whose peak expression is required to suppress CRC development and progression. The accumulation of nuclear SnoN and pSmad2, along with the downregulation of TGF-beta target genes observed in Akd(+/-) colon and tumors, suggest that tumor-suppressing properties of Arkadia are mediated by its ability to derepress TGF-beta signaling. Consistent with this likelihood, we identified mutations in primary colorectal tumors from human patients that reduce Arkadia function and are associated with the accumulation of nuclear SNON. Collectively, our findings reveal that Arkadia enhances TGF-beta signaling responses and supports its tumor-suppressing properties in CRC. Cancer Res; 71(20); 6438-49. (C) 2011 AACR.
• 膠芽腫の術中細胞診標本を用いたリン酸化STAT3の検討

  青柳 瑛子, 谷野 美智枝, 菅野 宏美, 高阪 真路, 野田頭 未歩, 木村 太一, 西原 広史, 藤本 真, 村田 純一, 田中 伸哉

  日本臨床細胞学会北海道支部会報 北海道臨床細胞学会 20 1 - 4 0918-1628 2011/10 [Not refereed][Not invited]
   

  過去1年間に術中迅速病理診断で膠芽腫と診断された12症例の術中圧挫細胞診検体と永久標本を用いてsignal transducer and activator of transcription(STAT3)の免疫染色を行い、STAT3のリン酸化状態を評価するとともに、間葉系マーカーであるビメンチン発現との関連を検討した。その結果、いずれの細胞診検体にもSTAT3のリン酸化が確認され、陽性細胞比率により3群に分類されたが、その染色結果は組織標本のそれとほぼ一致していた。また、STAT3のリン酸化が亢進している群ではビメンチンの発現がみられるものが多く、組織標本ではリン酸化STAT3とビメンチンの共局在が確認された。細胞診標本でSTAT3のリン酸化状態を評価することは、腫瘍の特性を知るための多数のモダリティの一つになる可能性がある。
• A case of meningeal melanocytoma in the cerebellopontine angle

  Masaaki Mikamoto, Yoshinobu Seo, Tamio Ito, Jyoji Nakagawara, Hirohiko Nakamura, Shinya Tanaka

  Neurological Surgery 39 (9) 859 - 864 0301-2603 2011/09/10 [Refereed][Not invited]
   

  Meningeal melanocytomas are uncommon intracranial tumors and extremely rare in the cerebellopontine angle (CPA). The tumors are generally considered to be benign because they lack malignant features in histological examination, but several literatures describe malignant behavior of the tumors such as high frequency of local recurrence, malignant transformation with leptomeningeal seeding. We describe a case of meningeal melanocytoma in the CPA and discuss the features of the tumor. The case was a 43-year-old woman with a right CPA exta-axial mass suffering from vertigo and nausea. Computed tomography (CT) and magnetic resonance imaging (MRI) showed a mass in the right CPA. The mass was hyperintense on T1-weighted images and hypointense on T2-weighted images. Surgical removal was done and pathological diagnosis was made as meningeal melanocytoma. Twenty months after the first surgery, MRI revealed local recurrence of the tumor and subtotal resection was performed.
• AKT Has an Anti-Apoptotic Role in ABCA12-Deficient Keratinocytes

  Teruki Yanagi, Masashi Akiyama, Hiroshi Nishihara, Yuki Miyamura, Kaori Sakai, Shinya Tanaka, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 131 (9) 1942 - 1945 0022-202X 2011/09 [Refereed][Not invited]
  
• Follicular regulatory T cells expressing Foxp3 and Bcl-6 suppress germinal center reactions

  Yeonseok Chung, Shinya Tanaka, Fuliang Chu, Roza I. Nurieva, Gustavo J. Martinez, Seema Rawal, Yi-Hong Wang, Hoyong Lim, Joseph M. Reynolds, Xiao-hui Zhou, Hui-min Fan, Zhong-ming Liu, Sattva S. Neelapu, Chen Dong

  NATURE MEDICINE 17 (8) 983 - U102 1078-8956 2011/08 [Refereed][Not invited]
   

  Foxp3(+) regulatory T (T(reg)) cells suppress different types of immune responses to help maintain homeostasis in the body. How T(reg) cells regulate humoral immunity, including germinal center reactions, is unclear. Here we identify a subset of Treg cells expressing CXCR5 and Bcl-6 that localize to the germinal centers in mice and humans. The expression of CXCR5 on T(reg) cells depends on Bcl-6. These CXCR5(+) Bcl-6(+) T(reg) cells are absent in the thymus but can be generated de novo from CXCR5-Foxp3(+) natural T(reg) precursors. A lack of CXCR5+ T(reg) cells leads to greater germinal center reactions including germinal center B cells, affinity maturation of antibodies and the differentiation of plasma cells. These results unveil a Bcl-6-CXCR5 axis in T(reg) cells that drives the development of follicular regulatory T (T(FR)) cells that function to inhibit the germinal center reactions.
• Unique Inclusion Properties of Crystalline Powder p-tert-Butylthiacalix[4]arene toward Alcohols and Carboxylic Acids

  Naoya Morohashi, Shintaro Noji, Hiroko Nakayama, Yasutaka Kudo, Shinya Tanaka, Chizuko Kabuto, Tetsutaro Hattori

  ORGANIC LETTERS 13 (13) 3292 - 3295 1523-7060 2011/07 [Refereed][Not invited]
   

  Powdery crystals of p-tert-butylthiacalix[4]arene (2) selectively include EtOH from 1:1 mixtures of MeOH-EtOH and EtOH-PrOH, and EtCO(2)H from HCO(2)H-EtCO(2)H. On the other hand, no acid is included from HCO(2)H-MeCO(2)H, even though MeCO(2)H is included from the neat acid. The origins of these phenomena are discussed based on X-ray analysis of inclusion crystals prepared separately by crystallization.
• A population of BJ fibroblasts escaped from Ras-induced senescence susceptible to transformation

  Shinji Kohsaka, Ken Sasai, Kenta Takahashi, Tsuyoshi Akagi, Mishie Tanino, Taichi Kimura, Hiroshi Nishihara, Shinya Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 410 (4) 878 - 884 0006-291X 2011/07 [Refereed][Not invited]
   

  Oncogenic stimuli such as H-Ras induce oncogene-induced senescence (OIS) in fibroblasts to protect against transformation. Here we found that a population of the human diploid fibroblasts can escape from OIS induced by H-RasV12. We designated these OIS-escaped cells as OISEC (OIS-escaped cells). OISEC lost the expression of p16 which plays an important role for cell cycle arrest for induction of senescence, but OISEC preserved the p16 expression machinery and exhibited senescence by the treatment with hydrogen peroxide (H(2)O(2)) as stress-induced premature senescence (SIPS). OISEC did not possess anchorage-independent growth potential, and functional disruption of p53 and Rb by SV40 early region encoding large T and small t antigens, induced the aneuploidy phenotype and colony-forming potential of OISEC together with the exhibition of in vivo tumor formation. Finally, we also found that the distinctive feature of OISEC is expression of transcription factors, Oct3/4, SOX2, and Nanog which is closely related to stem-like cell features. This study highlights the presence of a cell population which escaped from OIS, and this OISEC may transform into malignant cancer cells by the additional hits of several genes in vivo. (C) 2011 Elsevier Inc. All rights reserved.
• 中間型松果体実質腫瘍(PPTID)の2例

  大竹 安史, 佐藤 憲市, 伊東 民雄, 安斉 公雄, 尾崎 義丸, 及川 光照, 福井 崇人, 中村 博彦, 菅野 宏美, 田中 伸哉

  脳神経外科ジャーナル 日本脳神経外科コングレス 20 (6) 456 - 462 0917-950X 2011/06 [Not refereed][Not invited]
   

  症例1:71歳女。後頭部痛を主訴とした。CTで松果体部に腫瘍を認め、MRIではT1強調画像で等〜低信号を呈し、T2強調画像、FLAIRでやや高信号を呈した。T1強調ガドリニウム造影では均一に造影され、境界明瞭で辺縁整であった。無症状であったため経過観察とし、6ヵ月後に脳室拡大が出現し、MRIのconstructive interference in steady state(CISS)で視床内側と腫瘍の癒着、中脳水道の狭窄を認めたため、右側脳室前角経由による内視鏡的生検および第三脳室底開窓術を施行した。病理所見は小型の細胞がびまん性に増殖し、rossete様構造や無核血管周囲腔を認め、pineocytomaに比べ細胞密度が高く中間型松果体実質腫瘍(PPTID)と診断された。術後10ヵ月現在、無症状で腫瘍の増大はない。症例2:33歳男。頭痛を主訴とした。CTで松果体を中心に腫瘍を認め、MRIではT1強調画像で等信号、T2強調画像、FLAIRで高信号を呈し、T1強調ガドリニウム造影では不均一に造影され、境界不明瞭で辺縁不整であった。腫瘍は広範囲に及び閉塞性水頭症がみられ、鞍上部および脊髄に播種も伴っていた。内視鏡的生検術および第三脳室底開窓術を施行し、病理所見は小型の細胞が密に増殖し、明らかなrosette形成や壊死所見は認めず、pineoblastomaに比べ細胞密度が低くPPTIDと診断された。術後化学療法で腫瘍は縮小し、全脳全脊髄照射を行いpartial responseで経過している。
• Syndecan-4 Prevents Cardiac Rupture and Dysfunction After Myocardial Infarction

  Yutaka Matsui, Masahiro Ikesue, Keiko Danzaki, Junko Morimoto, Mami Sato, Shinya Tanaka, Tetsuhito Kojima, Hiroyuki Tsutsui, Toshimitsu Uede

  CIRCULATION RESEARCH 108 (11) 1328 - U95 0009-7330 2011/05 [Refereed][Not invited]
   

  Rationale: Syndecan-4 (Syn4), a cell-surface heparan sulfate proteoglycan, has been detected in the infarct region after myocardial infarction (MI), but its functional significance has not been elucidated. Objective: We examined whether and how Syn4 regulates the cardiac healing process after MI. Methods and Results: Although the heart in Syn4-deficient (Syn4(-/-)) mice was morphologically and functionally normal, Syn4(-/-) mice exhibited impaired heart function and increased mortality rate as a result of cardiac ruptures after MI. Cardiac ruptures in Syn4(-/-) mice were associated with reduced inflammatory reaction and impaired granulation tissue formation during the early phase of MI, as evidenced by reduced numbers of leukocytes, fibroblasts, myofibroblasts, macrophages, and capillary vessels, along with reduced extracellular matrix protein deposition in the infarct region after MI. Transforming growth factor-beta 1-dependent cell signaling was preserved, whereas cell migration, fibronectin-induced cell signaling, and differentiation into myofibroblasts were defective in Syn4(-/-) cardiac fibroblasts. We also found that Syn4 was involved in basic fibroblast growth factor-dependent endothelial cell signaling, cell proliferation, and tube formation. Finally, overexpression of the shed form of Syn4 before MI creation led to an increase in mortality due to cardiac rupture via its action as a dominant-negative inhibitor of endogenous Syn4 signaling, which suggested a protective role of Syn4 signaling in MI. Conclusions: These results suggest that Syn4 plays an important role in the inflammatory response and granulation tissue formation, thereby preventing cardiac rupture and dysfunction after MI. (Circ Res. 2011; 108: 1328-1339.)
• High grade meningiomaの治療成績とMib-1 indexの関係の検討

  成田 拓人, 菅野 宏美, 湯澤 明夏, 寺坂 俊介, 小林 浩之, 村田 純一, 山口 秀, 西原 広史, 宝金 清博, 田中 伸哉

  Brain Tumor Pathology 日本脳腫瘍病理学会 28 (Suppl.) 092 - 092 1433-7398 2011/05
• 画像と病理所見から多中心性神経膠腫と診断した一例

  茂木 洋晃, 寺坂 俊介, 小林 浩之, 鴨嶋 雄大, 山口 秀, 遠藤 将吾, 宝金 清博, 高阪 真路, 田中 伸哉, 村田 純一

  Brain Tumor Pathology 日本脳腫瘍病理学会 28 (Suppl.) 123 - 123 1433-7398 2011/05
• Synthesis of Mono- and 1,3-Diaminocalix[4]arenes via Ullmann-Type Amination and Amidation of 1,3-Bistriflate Esters of Calix[4]arenes

  Yuka Nakamura, Shinya Tanaka, Ryuichi Serizawa, Naoya Morohashi, Tetsutaro Hattori

  JOURNAL OF ORGANIC CHEMISTRY 76 (7) 2168 - 2179 0022-3263 2011/04 [Refereed][Not invited]
   

  Practical methods are described for the preparation of monoamines 4 and 1,3-diamines 5, bearing one or two amino group(s) instead of the hydroxy group(s) at the 28-position or at both the 26- and 28-positions of p-tert-butylcalix[4]arene (1a) and p-tert-butylthiacalix[4]arene (1b), via the Ullmann-type amination or amidation. Thus, the copper-catalyzed or mediated amination of the 1,3-bistriflate ester (2a) of la with benzylamine affords either mono (benzylamino) triflate 7a or 1,3-bis(benzylamine) 8 in a high yield, depending on the reaction conditions. On the other hand, the 1,3-bistriflate ester (2b) of 1b resists disubstitution and produces, under stoichiometric conditions, mono(benzylamino) triflate 7b. The disubstitution of 2b is achieved by amidation with tosylamide, giving 1,3-bis(tosylamide) 17b. The hydrogenolysis of the benzylamino moiety of 7a, followed by the hydrolysis of the Tf moiety, affords monoamine 4a, while the hydrogenolysis of 8 affords 1,3-diamine Sa. The amino moiety of 7b can be deprotected under acidic conditions to give, after hydrolysis, monoamine 4b. The hydrolysis of 17b affords 1,3-diamine 5b. The overall yields of compounds 4a, 4b, 5a, and 5b are 72%, 45%, 78%, and 24%, respectively, based on commercially available compounds 1 and are much higher than the ones previously reported in the literature.
• [Morphological analysis of bone dynamics and metabolic bone disease. Bone histomorphometory of Human beings].

  Tanaka S, Oda H

  Clinical calcium 4 21 541 - 550 0917-5857 2011/04 [Refereed][Not invited]
  
• Optimization of lentiviral vector transduction into peripheral blood mononuclear cells in combination with the fibronectin fragment CH-296 stimulation

  Hideto Chono, Yumi Goto, Satoko Yamakawa, Shinya Tanaka, Yasuhiro Tosaka, Ikuei Nukaya, Junichi Mineno

  JOURNAL OF BIOCHEMISTRY 149 (3) 285 - 292 0021-924X 2011/03 [Refereed][Not invited]
   

  Large scale T-cell expansion and efficient gene transduction are required for adoptive T-cell gene therapy. Based on our previous observations, human peripheral blood mononuclear cells (PBMCs) can be expanded efficiently while conserving a naive phenotype by stimulating with both recombinant human fibronectin fragment (CH-296) and anti-CD3 monoclonal antibodies. In this article, we explored the possibility of using this co-stimulation method to generate engineered T cells using lentiviral vector. Human PBMCs were stimulated with anti-CD3 together with immobilized CH-296 or anti-CD28 antibody as well as anti-CD3/anti-CD28 conjugated beads and transduced with lentiviral vector simultaneously. Co-stimulation with CH-296 gave superior transduction efficiency than with anti-CD28. Next, PBMCs were stimulated and transduced with anti-CD3/CH-296 or with anti-CD3/CD28 beads. T-cell expansion, gene transfer efficiencies and immunophenotypes were analysed. Stimulation with anti-CD3/CH-296 resulted in more than 10-times higher cell expansion and higher gene transfer efficiency with conservation of the naive phenotype compared with anti-CD3/CD28 stimulation method. Thus, lentiviral transduction with anti-CD3/CH-296 co-stimulation is an efficient way to generate large numbers of genetically modified T cells and may be suitable for many gene therapy protocols that use adoptive T-cell transfer therapy.
• Leptin controls ketone body utilization in hypothalamic neuron

  Ryota Narishima, Masahiro Yamasaki, Shinya Hasegawa, Saki Yoshida, Shinya Tanaka, Tetsuya Fukui

  NEUROSCIENCE LETTERS 490 (3) 185 - 190 0304-3940 2011/03 [Refereed][Not invited]
   

  Leptin is an appetite-controlling peptide secreted from adipose tissue. Previously, we showed that the gene expression of acetoacetyl-CoA synthetase (AACS), the ketone body-utilizing enzyme for lipid synthesis, was suppressed by leptin deficiency-induced obesity in white adipose tissue. In this study, to clarify the effects of leptin on ketone body utilization in the central nervous system, we examined the effects of leptin signaling on AACS expression. In situ hybridization analysis of ob/ob and db/db mice revealed that AACS mRNA level was reduced by leptin deficiency in the arcuate nucleus (Arc) and ventromedial hypothalamic nucleus (VMH) in hypothalamus but not in other brain regions. Moreover, AACS mRNA level was increased by leptin treatment both in primary cultured neural cells and in N41 neural-like cells. In N41 cells, AACS level was decreased by AMPK inducer but increased by AMPK inhibitor. These results suggest that the up-regulation of AACS expression by leptin is due to the suppression of AMPK activity via neural leptin signaling and that the deficiency of this regulation may be responsible for neurological disorders in central appetite control. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
• Dopamine Induces IL-6-Dependent IL-17 Production via D1-Like Receptor on CD4 Naive T Cells and D1-Like Receptor Antagonist SCH-23390 Inhibits Cartilage Destruction in a Human Rheumatoid Arthritis/SCID Mouse Chimera Model

  Kazuhisa Nakano, Kunihiro Yamaoka, Kentaro Hanami, Kazuyoshi Saito, Yasuyuki Sasaguri, Nobuyuki Yanagihara, Shinya Tanaka, Ichiro Katsuki, Sho Matsushita, Yoshiya Tanaka

  JOURNAL OF IMMUNOLOGY 186 (6) 3745 - 3752 0022-1767 2011/03 [Refereed][Not invited]
   

  A major neurotransmitter dopamine transmits signals via five different seven-transmembrane G protein-coupled receptors termed D1-D5. Several studies have shown that dopamine not only mediates interactions into the nervous system, but can contribute to the modulation of immunity via receptors expressed on immune cells. We have previously shown an autocrine/paracrine release of dopamine by dendritic cells (DCs) during Ag presentation to naive CD4(+) T cells and found efficacious results of a D1-like receptor antagonist SCH-23390 in the experimental autoimmune encephalomyelitis mouse model of multiple sclerosis and in the NOD mouse model of type I diabetes, with inhibition of Th17 response. This study aimed to assess the role of dopaminergic signaling in Th17-mediated immune responses and in the pathogenesis of rheumatoid arthritis (RA). In human naive CD4(+) T cells, dopamine increased IL-6-dependent IL-17 production via D1-like receptors, in response to anti-CD3 plus anti-CD28 mAb. Furthermore, dopamine was localized with DCs in the synovial tissue of RA patients and significantly increased in RA synovial fluid. In the RA synovial/SCID mouse chimera model, although a selective D2-like receptor antagonist haloperidol significantly induced accumulation of IL-6(+) and IL-17(+) T cells with exacerbated cartilage destruction, SCH-23390 strongly suppressed these responses. Taken together, these findings indicate that dopamine released by DCs induces IL-6-Th17 axis and causes aggravation of synovial inflammation of RA, which is the first time, to our knowledge, that actual evidence has shown the pathological relevance of dopaminergic signaling with RA. The Journal of Immunology, 2011, 186: 3745-3752.
• A case of primary diffuse leptomeningeal gliomatosis, clinically indistinguishable from metastatic meningeal carcinomatosis

  Fumihito Nakano, Ichiro Yabe, Sachiko Tsuji-Akimoto, Akihiro Ishizu, Shinya Tanaka, Masanori Kasahara, Hidenao Sasaki

  Clinical Neurology 51 (3) 197 - 202 0009-918X 2011/03 [Refereed][Not invited]
   

  A 65 year old woman presented with progressive gait disturbance. She complained of appetite loss for 3 months. Her gait gradually became unsteady, and she was admitted to our hospital. On admission, slow mentation, bathyhypesthesia in left upper and both lower extremites, positive Romberg sign and wide-based gait were observed. Gd-enhanced MRI revealed mass lesions in the left temporal fossa and the cervical spinal canal with focal meningeal enhancement. Besides lesions in the central nervous system (CNS), systematic examination detected no additional malignancy. Repeated cytology of the cerebrospinal fluid was negative. After admission, her consciousness became reduced gradually. At 2 months after admission, she died of central respiratory failure. On autopsy, diffuse extension of the tumor cells was observed on the surface of CNS, and the mass lesions observed by MRI were extra-parenchymal. On microscopic examination, the mass was consisted of GFAP positive malignant cells, and included perivascular pseudorosette, pseudopalisading necrosis and many mitotic cells. The diagnosis of the case was made as primary diffuse leptomeningeal gliomatosis (PDLG). PDLG is a rare disorder that is difficult to diagnose by CSF cytology. The progress of PDLG is rapid, and appropriate treatment is rarely taken. However, the combination of temozolomide and the radiotherapy performed for a glioblastoma has been reported as a possible treatment for PDLG. We emphasize that, in possible cases of PDLG, a biopsy should be performed in the early stages, especially in cases showing features similar to those of metastatic meningeal carcinomatosis and have no malignant tumors by whole body examination.
• High expression of MeCP2 in JC virus-infected cells of progressive multifocal leukoencephalopathy brains

  Saya Shirai, Kenta Takahashi, Shinji Kohsaka, Tetsu Tsukamoto, Hiroshi Isogai, Shinichi Kudo, Hirofumi Sawa, Kazuo Nagashima, Shinya Tanaka

  NEUROPATHOLOGY 31 (1) 38 - 41 0919-6544 2011/02 [Refereed][Not invited]
   

  Mutations of the methyl CpG binding protein 2 (MeCP2) gene are a major cause of Rett syndrome. To investigate whether the expression of this gene was related to JC virus (JCV) infection, we examined brains of four progressive multifocal leukoencephalopathy (PML) patients. JCV infection was confirmed by immunohistochemical labeling with antibodies against JCV VP1, agnoprotein and large T antigen. MeCP2 expression was examined by immunohistochemistry using a specific polyclonal antibody against MeCP2. In normal brains and uninfected cortices of PML brains, MeCP2 expression was observed in the nuclei of neurons, but not observed in glial and endothelial cell nuclei. However, in PML brains intense immunolabeling was observed in abnormally enlarged glial nuclei of JCV-infected cells. Double immunolabeling using antibodies against large T antigen (visualized as blue) and MeCP2 (visualised as red) revealed dark red JCV-infected nuclei, which confirmed that the JCV infected nuclei expressed MeCP2. We conclude that MeCP2 is highly expressed in the JCV-infected nuclei of PML brain and these results may provide a new insight into the mechanism which regulates the MeCP2 expression in glial cells by the infection of JCV.
• Prenatal diagnosis of short-rib polydactyly syndrome type 3 (Verma-Naumoff type) by three-dimensional helical computed tomography

  Takahiro Yamada, Gen Nishimura, Keiichiro Nishida, Hideaki Sawai, Tokuhiko Omatsu, Taichi Kimura, Hiroshi Nishihara, Rina Shono, Shigeki Shimada, Mamoru Morikawa, Masato Mizushima, Takashi Yamada, Kazutoshi Cho, Shinya Tanaka, Hiroki Shirato, Hisanori Minakami

  JOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH 37 (2) 151 - 155 1341-8076 2011/02 [Refereed][Not invited]
   

  We present a case of short-rib polydactyly syndrome (SRPs) type 3 in which accurate prenatal diagnosis was feasible using both ultrasonography and 3D-CT. SRP encompass a heterogeneous group of lethal skeletal dysplasias. However, the phenotypes overlap with those of nonlethal skeletal dysplasias (i.e. Ellis-van Creveld syndrome and Jeune syndrome). As accurate prenatal diagnosis of SRP is helpful for parents, we used 3D-CT in the early third trimester to examine a fetus suggested to have phenotypes of 'short-rib dysplasia group' on ultrasonography. 3D-CT showed mild modification of the vertebral bodies, small ilia with horizontal acetabula and triangular partial ossification defects, and subtle metaphyseal irregularities of the femora. These CT findings and an extensive literature search regarding the phenotypes of various diseases categorized as short-rib dysplasia group led to a correct prenatal diagnosis of SRP type 3. This case exemplified the usefulness of 3D-CT for the precise prenatal diagnosis of skeletal dysplasias.
• A case of atypical teratoid/rhabdoid tumor in an adult, with long survival

  Kenta Takahashi, Hiroshi Nishihara, Masahito Katoh, Tomoaki Yoshinaga, Roshan Mahabir, Hiromi Kanno, Taichi Kimura, Mishie Tanino, Jun Ikeda, Yutaka Sawamura, Kazuo Nagashima, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 28 (1) 71 - 76 1433-7398 2011/02 [Refereed][Not invited]
   

  Atypical teratoid/rhabdoid tumor (AT/RT) is a malignant tumor that mostly occurs in early childhood and has poor prognosis despite aggressive therapy. Adult cases are rare and, as far as we are aware, only 30 cases have been reported to date. Here we present the case of a 27-year-old female with left parietal AT/RT with the chief complaint of numbness of the right superior limb. First, the tumor was surgically removed and the diagnosis was grade II glioma. With additional radiotherapy, the clinical course after surgery was favorable. After 6 years, she had an operation for recurrence and the diagnosis was grade III glioma. Temozolomide was prescribed, and a disease-free period of 2 years followed. Surgery was performed for a third time for second recurrence with histology of diffuse growth of rhabdoid cells. Immunohistochemistry was partially positive for vimentin and epithelial membrane antigen. Ki-67 labeling index was extremely high and tumor cells showed no staining of INI1 suggestive of diagnosis of AT/RT. We re-evaluated past specimens and none had immunoreactivity of INI1. Ki-67 labeling index and O-6 methylguanine DNA methyltransferase (MGMT) staining were also re-examined and both increased gradually. She is still alive without recurrence for more than 1 year. As far as we are aware, this is the second longest survival of an adult with AT/RT.
• Detection of histological anaplasia in gliomas with oligodendroglial components using positron emission tomography with F-18-FDG and C-11-methionine: report of two cases

  Shigeru Yamaguchi, Hiroyuki Kobayashi, Kenji Hirata, Tohru Shiga, Shinya Tanaka, Junichi Murata, Shunsuke Terasaka

  JOURNAL OF NEURO-ONCOLOGY 101 (2) 335 - 341 0167-594X 2011/01 [Refereed][Not invited]
   

  Gliomas are regionally heterogeneous tumors. Positron emission tomography (PET) with F-18-fluorodeoxyglucose (FDG) and C-11-methionine (MET) evaluates the heterogeneity of histological malignancy within the tumor. We present two patients with oligodendrocytic tumors that showed discrepancies in the highest uptake areas with these two tracers. PET studies with MET and FDG were performed on the same day, 2 weeks before surgery. In both cases, biopsy specimens were separately obtained from the highest MET and FDG uptake areas guided by intraoperative neuronavigation. Histological examinations demonstrated that the specimens from the highest MET uptake area revealed low-grade oligoastrocytoma or oligodendroglioma, whereas histological anaplasias were contained in the specimens from the highest FDG uptake area. With gliomas with oligodendroglial components, the MET uptake ratio does not always correspond to histological anaplasia, which can be detected only by FDG PET. Sole application of MET PET for preoperative evaluation may lead to misunderstanding of histological heterogeneity in gliomas, especially those with oligodendroglial components. FDG and MET tracers play complementary roles in preoperative evaluation of gliomas.
• The enhancer HS2 critically regulates GATA-3-mediated Il4 transcription in T(H)2 cells

  Shinya Tanaka, Yasutaka Motomura, Yoshie Suzuki, Ryoji Yagi, Hiromasa Inoue, Shoichiro Miyatake, Masato Kubo

  NATURE IMMUNOLOGY 12 (1) 77 - U103 1529-2908 2011/01 [Refereed][Not invited]
   

  GATA-3 is a master regulator of T helper type 2 (T(H)2) differentiation. However, the molecular basis of GATA-3-mediated T(H)2 lineage commitment is poorly understood. Here we identify the DNase I-hypersensitive site 2 (HS2) element located in the second intron of the interleukin 4 locus (Il4) as a critical enhancer strictly controlled by GATA-3 binding. Mice lacking HS2 showed substantial impairment in their asthmatic responses and their production of IL-4 but not of other T(H)2 cytokines. Overexpression of Gata3 in HS2-deficient T cells failed to restore Il4 expression. HS2 deletion impaired the trimethylation of histone H3 at Lys4 and acetylation of histone H3 at Lys9 and Lys14 in the Il4 locus. Our results indicate that HS2 is the target of GATA-3 in regulating chromosomal modification of the Il4 locus and is independent of the Il5 and Il13 loci.
• A nanostructured skeleton catalyst: Suzuki-coupling with a reusable and sustainable nanoporous metallic glass Pd-catalyst

  Shinya Tanaka, Tetsuro Kaneko, Naoki Asao, Yoshinori Yamamoto, Mingwei Chen, Wei Zhang, Akihisa Inoue

  CHEMICAL COMMUNICATIONS 47 (21) 5985 - 5987 1359-7345 2011 [Refereed][Not invited]
   

  Nanoporous metallic glass Pd, which was fabricated by dealloying of a glassy metallic alloy Pd(30)Ni(50)P(20), exhibited a remarkable catalytic activity for the Suzuki-coupling reaction between iodoarenes and arylboronic acids under mild conditions. Moreover, the catalyst can be reused several times without a significant loss of catalytic activity.
• Clinicopathologic Application of Lectin Histochemistry Bisecting GlcNAc in Glioblastoma

  Eiko Aoyanagi, Ken Sasai, Miho Nodagashira, Lei Wang, Hiroshi Nishihara, Hideyuki Ihara, Yoshitaka Ikeda, Shinya Tanaka

  APPLIED IMMUNOHISTOCHEMISTRY & MOLECULAR MORPHOLOGY 18 (6) 518 - 525 1541-2016 2010/12 [Refereed][Not invited]
   

  Glycosylation is one of the most common posttranslational modifications and changes in oligosaccharide structures are associated with many human diseases including a number of cancers. Thus, discovering aberrant glycosylation patterns that serve as markers for brain tumor progression and metastasis represents an attractive strategy to improve clinicopathologic diagnosis and to provide aids to the development of novel therapies. To identify glioblastoma (GBM) cells expressing glycoproteins that contain high levels of the bisecting N-acetylglucosamine (GlcNAc) structures, lectin histochemistry was carried out using erythroagglutinating phytohemagglutinin. Although GBM frequently expressed the bisecting GlcNAc, the lectin reactivity varied among tumor regions within individual specimens. Since detailed histopathologic analysis revealed that oligosaccharides with bisecting GlcNAc structures were preferably expressed in tumor regions with low KI67 immunopositivity, immunodetection of the bisecting GlcNAc could be useful to indicate less proliferative regions in human GBM. Our study highlights the potential use of lectin histochemistry to develop new methods for diagnosis that would improve future antiglioma therapy.
• Deficiency of vitamin A delays bone healing process in association with reduced BMP2 expression after drill-hole injury in mice

  Kazuhiro Tanaka, Shinya Tanaka, Akinori Sakai, Tadashi Ninomiya, Yoshinori Arai, Toshitaka Nakamura

  BONE 47 (6) 1006 - 1012 8756-3282 2010/12 [Refereed][Not invited]
   

  Although it is predicted that vitamin A and its active form retinoic acid regulate osteoblast lineage this has not been elucidated in growing mammalians To clarify the direct effect of retinoic acid on bone we observed the process of filling up newly generating bone Into a drill hole of the bone which is understood as membranous ossification in vitamin A-deficient mice Mice were assigned to three groups a vitamin A-deficient group (VAD) which was fed a diet without vitamin A from the 10th day of gestation to the end of the experiments a vitamin A deficient-sufficient group (VADS) which was fed a diet without vitamin A from the 10th day of gestation to 4 weeks of age and a vitamin A-sufficient group (VAS) which was fed a standard diet to the end of the experiment In mice at 10 weeks of age (day 0) a drill-hole injury was made with a diameter of 1 mm at the anterior portion of the diaphysis of the bilateral femurs In VAD retardation in repairing the drill hole was demonstrated by in vivo micro-CT and histomorphometry from day7 and after surgery During repair of the bone defect increases of bmp2 dlx5 msx2 col1a1 and osteocalcin mRNA expression were suppressed and runx2 p2 mRNA expression was accelerated in VAD Implantation of BMP2 in the bone defect of VAD normalized the delayed bone healing and mRNA expressions We concluded that vitamin A regulates bmp2 mRNA expression and plays a crucial role in osteoblastogenesis and bone formation (C) 2010 Elsevier Inc All rights reserved
• Radiation-induced osteosarcomas after treatment for frontal gliomas: a report of two cases

  Tamio Ito, Yoshimaru Ozaki, Ken-ichi Sato, Mitsuteru Oikawa, Mishie Tanino, Hirohiko Nakamura, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 27 (2) 103 - 109 1433-7398 2010/10 [Refereed][Not invited]
   

  Most radiation-induced osteosarcomas of the skull are reported to arise in the facial bone or paranasal sinus after radiotherapy for retinoblastoma and/or pituitary adenoma. Here we report two cases of radiation-induced osteosarcoma in the paranasal sinus after treatment for frontal glioma. Case 1 was a 56-year-old woman who underwent surgical resection of a left frontal tumor in October 1990. The histological diagnosis was a low-grade glioma, and radiotherapy of 54 Gy was administered. Sixteen years later, in September 2006, the patient noted an enlarging subcutaneous mass in the right frontal region. CT showed an osteolytic mass in the right frontal sinus. An open biopsy established the histopathological diagnosis of osteosarcoma, and the patient subsequently died of rapid tumor regrowth. Case 2 was a 58-year-old man who underwent partial removal of a bifrontal tumor in May 1996. The histological diagnosis was anaplastic oligoastrocytoma, and radiotherapy of 56 Gy was administered. Twelve years later, in March 2008, the patient was readmitted to our hospital for reasons of marked deterioration in general physical condition. Tumor recurrence was suspected in the left frontal lobe, and CT demonstrated an osteolytic mass in the left frontal and ethmoid sinus. A secondary operation was performed, and the pathological specimens were diagnosed as osteosarcoma. Radiotherapy was readministered, but the subject died of rapid tumor regrowth. From these clinicopathological findings, both cases were diagnosed as radiation-induced osteosarcoma. Radiation-induced osteosarcomas appeared 16 and 12 years after radiotherapy in cases 1 and 2, respectively. As the prognosis of radiation-induced osteosarcoma is poorer than that of primary osteosarcoma, careful attention is required for consideration of the long-term survival of patients with glioma.
• Smad2 Positively Regulates the Generation of Th17 Cells

  Gustavo J. Martinez, Zhengmao Zhang, Joseph M. Reynolds, Shinya Tanaka, Yeonseok Chung, Ting Liu, Elizabeth Robertson, Xia Lin, Xin-Hua Feng, Chen Dong

  JOURNAL OF BIOLOGICAL CHEMISTRY 285 (38) 29039 - 29043 0021-9258 2010/09 [Refereed][Not invited]
   

  Development of Foxp3(+) regulatory T cells and pro-inflammatory Th17 cells from naive CD4(+) T cells requires transforming growth factor-beta (TGF-beta) signaling. Although Smad4 and Smad3 have been previously shown to regulate Treg cell induction by TGF-beta, they are not required in the development of Th17 cells. Thus, how TGF-beta regulates Th17 cell differentiation remains unclear. In this study, we found that TGF-beta-induced Foxp3 expression was significantly reduced in the absence of Smad2. More importantly, Smad2 deficiency led to reduced Th17 differentiation in vitro and in vivo. In the experimental autoimmune encephalomyelitis model, Smad2 deficiency in T cells significantly ameliorated disease severity and reduced generation of Th17 cells. Furthermore, we found that Smad2 associated with retinoid acid receptor-related orphan receptor-gamma t (ROR gamma t) and enhanced ROR gamma t-induced Th17 cell generation. These results demonstrate that Smad2 positively regulates the generation of inflammatory Th17 cells.
• A Novel FRET-Based Biosensor for the Measurement of BCR-ABL Activity and Its Response to Drugs in Living Cells

  Tatsuaki Mizutani, Takeshi Kondo, Stephanie Darmanin, Masumi Tsuda, Shinya Tanaka, Minoru Tobiume, Masahiro Asaka, Yusuke Ohba

  CLINICAL CANCER RESEARCH 16 (15) 3964 - 3975 1078-0432 2010/08 [Refereed][Not invited]
   

  Purpose: To develop a novel diagnostic method for the assessment of drug efficacy in chronic myeloid leukemia (CML) patients individually, we generated a biosensor that enables the evaluation of BCR-ABL kinase activity in living cells using the principle of fluorescence resonance energy transfer (FRET). Experimental Design: To develop FRET-based biosensors, we used CrkL, the most characteristic substrate of BCR-ABL, and designed a protein in which CrkL is sandwiched between Venus, a variant of YFP, and enhanced cyan fluorescent protein, so that CrkL intramolecular binding of the SH2 domain to phosphorylated tyrosine (Y207) increases FRET efficiency. After evaluation of the properties of this biosensor by comparison with established methods including Western blotting and flow cytometry, BCR-ABL activity and its response to drugs were examined in CML patient cells. Results: After optimization, we obtained a biosensor that possesses higher sensitivity than that of established techniques with respect to measuring BCR-ABL activity and its suppression by imatinib. Thanks to its high sensitivity, this biosensor accurately gauges BCR-ABL activity in relatively small cell numbers and can also detect <1% minor drug-resistant populations within heterogeneous ones. We also noticed that this method enabled us to predict future onset of drug resistance as well as to monitor the disease status during imatinib therapy, using patient cells. Conclusion: In consideration of its quick and practical nature, this method is potentially a promising tool for the prediction of both current and future therapeutic responses in individual CML patients, which will be surely beneficial for both patients and clinicians. Clin Cancer Res; 16(15); 3964-75. (C) 2010 AACR.
• バイオマーカーによる診断、その他 個別化病理診断の基盤作成 胃癌におけるシグナル伝達分子のImmunoprofilingの検討(Diagnosis by biomaker, others Immunoprofiling of signaling molecule for gastric cancer: Pathological basis for Taylor made medicine)

  西原 広史, 菅野 宏美, 石川 麻倫, 大場 彩音, 田中 伸哉

  日本癌学会総会記事 69回 232 - 233 0546-0476 2010/08
• MassBank: a public repository for sharing mass spectral data for life sciences

  Horai, H, Arita, M, Kanaya, S, Nihei, Y, Ikeda, T, Suwa, K, Ojima, Y, Tanaka, K, Tanaka, S, Aoshima, K, Oda, Y, Kakazu, Y, Kusano, M, Tohge, T, Matsuda, F, Sawada, Y, Hirai, Y. M, Nakanishi, H, Ikeda, K, Akimoto, N, Maoka, T, Takahashi, H, Ara, T, Sakurai, N, Suzuki, H, Shibata, D, Neumann, S, Iida, T, Tanaka, K, Funatsu, K, Matsuura, F, Soga, T, Taguchi, R, Saito, K, Nishioka, T

  J. Mass Spectro. 45 (7) 703 - 714 1076-5174 2010/07/08 [Refereed][Not invited]
   

  MassBank is the first public repository of mass spectra of small chemical compounds for life sciences (<3000 Da). The database contains 605 electron-ionization mass spectrometry (EI-MS), 137 fast atom bombardment MS and 9276 electrospray ionization (ESI)-MS(n) data of 2337 authentic compounds of metabolites, 11 545 EI-MS and 834 other-MS data of 10,286 volatile natural and synthetic compounds, and 3045 ESI-MS(2) data of 679 synthetic drugs contributed by 16 research groups (January 2010). ESI-MS(2) data were analyzed under nonstandardized, independent experimental conditions. MassBank is a distributed database. Each research group provides data from its own MassBank data servers distributed on the Internet. MassBank users can access either all of the MassBank data or a subset of the data by specifying one or more experimental conditions. In a spectral search to retrieve mass spectra similar to a query mass spectrum, the similarity score is calculated by a weighted cosine correlation in which weighting exponents on peak intensity and the mass-to-charge ratio are optimized to the ESI-MS(2) data. MassBank also provides a merged spectrum for each compound prepared by merging the analyzed ESI-MS(2) data on an identical compound under different collision-induced dissociation conditions. Data merging has significantly improved the precision of the identification of a chemical compound by 21-23% at a similarity score of 0.6. Thus, MassBank is useful for the identification of chemical compounds and the publication of experimental data.
• Self-Improvement of Keratinocyte Differentiation Defects During Skin Maturation in ABCAl2-Deficient Harlequin lchthyosis Model Mice

  Teruki Yanagi, Masashi Akiyama, Hiroshi Nishihara, Junko Ishikawa, Kaori Sakai, Yuki Miyamura, Ayano Naoe, Takashi Kitahara, Shinya Tanaka, Hiroshi Shimizu

  AMERICAN JOURNAL OF PATHOLOGY 177 (1) 106 - 118 0002-9440 2010/07 [Refereed][Not invited]
   

  Harlequin ichthyosis (HI) is caused by loss-of-function mutations in the keratinocyte lipid transporter ABCA12. The patients often die in the first 1 or 2 weeks of life, although HI survivors' phenotypes improve within several weeks after birth. In order to clarify the mechanisms of phenotypic recovery, we studied grafted skin and keratinocytes from Abca12-disrupted (Abca12(-/-)) mice showing abnormal lipid transport. Abca12(-1-) neonatal epidermis showed significantly reduced total ceramide amounts and aberrant ceramide composition. Immunofluorescence and immunoblotting of Abca12(-1-) neonatal epidermis revealed defective profilaggrin/filaggrin conversion and reduced protein expression of the differentiation-specific molecules, loricrin, kallikrein 5, and transglutaminase 1, although their mRNA expression was up-regulated. In contrast, Abca12(-1-) skin grafts kept in a dry environment exhibited dramatic improvements in all these abnormalities. Increased transepidermal water loss, a parameter representing barrier defect, was remarkably decreased in grafted Abca12(-1-) skin. Ten-passage sub-cultured Abca12(-/-)keratinocytes showed restoration of intact ceramide distribution, differentiation-specific protein expression and profilaggrin/filaggrin conversion, which were defective in primary-cultures. Using cDNA microarray analysis, lipid transporters including four ATP-binding cassette transporters were up-regulated after sub-culture of Abca12(-/-) keratinocytes compared with primary-culture. These results indicate that disrupted keratinocyte differentiation during the fetal development is involved in the pathomechanism of HI and, during maturation, Abca12(-/-) epidermal keratinocytes regain normal differentiation processes. This restoration may account for the skin phenotype improvement observed in HI survivors. (Am J Pathol 2010, 177:106-118; DOI: 10.2353/ajpath.2010.091120)
• Combined use of positron emission tomography with 18F- fluorodeoxyglucose and 11C-methionine for preoperative evaluation of gliomas

  Shigeru Yamaguchi, Shunsuke Terasaka, Hiroyuki Kobayashi, Takuhito Narita, Kenji Hirata, Satoshi Shiga, Reiko Usui, Shinya Tanaka, Kanako Kubota, Junichi Murata, Katsuyuki Asaoka

  Neurological Surgery 38 (7) 621 - 628 0301-2603 2010/07 [Refereed][Not invited]
   

  Object: The aim of this study was to evaluate the usefulness of combined use of positron emission tomography (PET) with 18F-fluorodeoxyglucose (FDG) and 11C-methionine (MET) for the preoperative evaluation of gliomas and to investigate the feasibility of PET in glioma surgery. Methods: Preoperative FDG (n = 25) and/or MET (n = 22) PET studies were performed in 26 patients with primary and recurrent adult gliomas. We qualitatively (visual analysis) and quantitatively evaluated the uptake of both tracers in the tumor location. For quantitative analysis, data were analyzed by a region of interest method using the standard uptake value (SUV) and a calculated uptake ratio. We investigated the correlation among the tracer uptake ratios, histological tumor grading and tumor proliferation activity. Results: On visual inspection, no patient (0/9) with high uptake of FDG had low grade gliomas and 94% (14/15) had high grade gliomas, while uptake of MET was present in all patients. On quantitative analysis, histological tumor grade was most reflected in FDG uptake ratio compared with contralateral white matter. The tumor/normal brain (T/N) uptake ratio of MET increased stepwise with increasing histological grade but was not significantly different from tumor grade. In comparison of FDG and MET uptake ratio with proliferation activity, a significant correlation was shown for FDG uptake ratio, but not for the T/N ratio of MET. Conclusions: MET is useful in detecting and delineating the extent of the tumor, but not in evaluating tumor grade and proliferative activity. The FDG uptake ratio correlates well with tumor grade and proliferative activity. Preoperative PET studies with FDG and MET play complementary roles in the planning of glioma surgery, and integrated information from both tracers helps us to plan the extent of tumor resection.
• DOCK2 regulates cell proliferation through Rac and ERK activation in B cell lymphoma

  Lei Wang, Hiroshi Nishihara, Taichi Kimura, Yasutaka Kato, Mishie Tanino, Mitsufumi Nishio, Masato Obara, Tomoyuki Endo, Takao Koike, Shinya Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 395 (1) 111 - 115 0006-291X 2010/04 [Refereed][Not invited]
   

  DOCK2; a member of the CDM protein family, regulates cell motility and cytokine production through the activation of Rac in mammalian hematopoietic cells and plays a pivotal role in the modulation of the immune system. Here we demonstrated the alternative function of DOCK2 in hematopoietic tumor cells, especially in terms of its association with the tumor progression. Immunostaining for DOCK2 in 20 cases of human B cell lymphoma tissue specimens including diffuse large B cell lymphoma and follicular lymphoma revealed the prominent expression of DOCK2 in all of the lymphoma cells. DOCK2-knockdown (KD) of the B cell lymphoma cell lines, Ramos and Raji, using the lentiviral shRNA system presented decreased cell proliferation compared to the control cells. Furthermore, the tumor formation of DOCK2-KD Ramos cell in nude mice was significantly abrogated. Western blotting analysis and pull-down assay using GST-PAK-RBD kimeric protein suggested the presence of DOCK2-Rac-ERK pathway regulating the cell proliferation of these lymphoma cells. This is the first report to clarify the prominent role of DOCK2 in hematopoietic malignancy. (C) 2010 Elsevier Inc. All rights reserved.
• The Human Polyoma JC Virus Agnoprotein Acts as a Viroporin

  Tadaki Suzuki, Yasuko Orba, Yuki Okada, Yuji Sunden, Takashi Kimura, Shinya Tanaka, Kazuo Nagashima, William W. Hall, Hirofumi Sawa

  PLOS PATHOGENS 6 (3) e1000801  1553-7366 2010/03 [Refereed][Not invited]
   

  Virus infections can result in a range of cellular injuries and commonly this involves both the plasma and intracellular membranes, resulting in enhanced permeability. Viroporins are a group of proteins that interact with plasma membranes modifying permeability and can promote the release of viral particles. While these proteins are not essential for virus replication, their activity certainly promotes virus growth. Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease resulting from lytic infection of oligodendrocytes by the polyomavirus JC virus (JCV). The genome of JCV encodes six major proteins including a small auxiliary protein known as agnoprotein. Studies on other polyomavirus agnoproteins have suggested that the protein may contribute to viral propagation at various stages in the replication cycle, including transcription, translation, processing of late viral proteins, assembly of virions, and viral propagation. Previous studies from our and other laboratories have indicated that JCV agnoprotein plays an important, although as yet incompletely understood role in the propagation of JCV. Here, we demonstrate that agnoprotein possesses properties commonly associated with viroporins. Our findings demonstrate that: (i) A deletion mutant of agnoprotein is defective in virion release and viral propagation; (ii) Agnoprotein localizes to the ER early in infection, but is also found at the plasma membrane late in infection; (iii) Agnoprotein is an integral membrane protein and forms homo-oligomers; (iv) Agnoprotein enhances permeability of cells to the translation inhibitor hygromycin B; (v) Agnoprotein induces the influx of extracellular Ca(2+); (vi) The basic residues at amino acid positions 8 and 9 of agnoprotein key are determinants of the viroporin activity. The viroporin-like properties of agnoprotein result in increased membrane permeability and alterations in intracellular Ca(2+) homeostasis leading to membrane dysfunction and enhancement of virus release.
• Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumor stem-like hallmarks

  Kouichi Tabu, Taichi Kimura, Ken Sasai, Lei Wang, Norihisa Bizen, Hiroshi Nishihara, Tetsuya Taga, Shinya Tanaka

  MOLECULAR CANCER 9 39  1476-4598 2010/02 [Refereed][Not invited]
   

  Background: An increasing number of studies support the presence of stem-like cells in human malignancies. These cells are primarily responsible for tumor initiation and thus considered as a potential target to eradicate tumors. CD133 has been identified as an important cell surface marker to enrich the stem-like population in various human tumors. To reveal the molecular machinery underlying the stem-like features in tumor cells, we analyzed a promoter of CD133 gene using human colon carcinoma Caco-2 and synovial sarcoma Fuji cells, which endogenously express CD133 gene. Results: A reporter analysis revealed that P5 promoter, located far upstream in a human CD133 gene locus, exhibits the highest activity among the five putative promoters (P1 to P5). Deletion and mutation analysis identified two ETS binding sites in the P5 region as being essential for its promoter activity. Electrophoretic mobility shift assays demonstrated the specific binding between nuclear factors and the ETS binding sequence. Overexpression of dominant-negative forms of Ets2 and Elk1 resulted in the significant decrease of P5 activity. Furthermore, treatment of Fuji cells with a specific MEK/ERK inhibitor, U0126, also markedly decreased CD133 expression, but there was no significant effect in Caco-2 cells, suggesting cell type-specific regulation of CD133 expression. Instead, the side population, another hallmark of TSLCs, was dramatically diminished in Caco-2 cells by U0126. Finally, Ras-mediated oncogenic transformation in normal human astrocytes conferred the stem-like capability to form neurosphere-like colonies with the increase of CD133 mRNA expression. Conclusions: In conclusion, the Ras/ERK pathway at least in part contributes to the maintenance and the acquisition of stem-like hallmarks, although the extent of its contribution is varied in a cell type-specific manner. These findings could help our comprehensive understanding of tumor stemness, and also improve the development of eradicative therapies against human malignancies.
• Large T Antigen Promotes JC Virus Replication in G(2)-arrested Cells by Inducing ATM- and ATR-mediated G(2) Checkpoint Signaling

  Yasuko Orba, Tadaki Suzuki, Yoshinori Makino, Kanako Kubota, Shinya Tanaka, Takashi Kimura, Hirofumi Sawa

  JOURNAL OF BIOLOGICAL CHEMISTRY 285 (2) 1544 - 1554 0021-9258 2010/01 [Refereed][Not invited]
   

  Large T antigen (TAg) of the human polyomavirus JC virus (JCV) possesses DNA binding and helicase activities, which, together with various cellular proteins, are required for replication of the viral genome. We now show that JCV-infected cells expressing TAg accumulate in the G(2) phase of the cell cycle as a result of the activation of ATM- and ATR-mediated G(2) checkpoint pathways. Transient transfection of cells with a TAg expression vector also induced G(2) checkpoint signaling and G(2) arrest. Analysis of TAg mutants with different subnuclear localizations suggested that the association of TAg with cellular DNA contributes to the induction of G(2) arrest. Abrogation of G(2) arrest by inhibition of ATM and ATR, Chk1, and Wee1 suppressed JCV genome replication. In addition, abrogation of the G(2)-M transition by Cdc2 depletion disabled Wee1 depletion induced suppression of JCV genome replication, suggesting that JCV replication is facilitated by G(2) arrest resulting from G(2) checkpoint signaling. Moreover, inhibition of ATM and ATR by caffeine suppressed JCV production. The observation that oligodendrocytes productively infected with JCV in vivo also undergo G(2) arrest suggests that G(2) checkpoint inhibitors such as caffeine are potential therapeutic agents for JCV infection.
• Sudden Death of a Patient with Pandemic Influenza (A/H1N1pdm) Virus Infection by Acute Respiratory Distress Syndrome

  Akihiro Takiyama, Lei Wang, Mishie Tanino, Taichi Kimura, Naoki Kawagishi, Yasuyuki Kunieda, Harutaka Katano, Noriko Nakajima, Hideki Hasegawa, Tomoyuki Takagi, Hiroshi Nishihara, Tetsutaro Sata, Shinya Tanaka

  JAPANESE JOURNAL OF INFECTIOUS DISEASES 63 (1) 72 - 74 1344-6304 2010/01 [Refereed][Not invited]
   

  We describe an autopsy case of a patient with pandemic influenza (A/H1N1pdm) virus infection in Japan, who developed rapidly progressive viral pneumonia exhibiting diffuse alveolar damage. A 41-year-old female visited our hospital with a fever of 38.7 degrees C. She was a public health nurse with no underlying disease and had had contact with a group of elementary school students who had been infected with the influenza (A/H1N1pdm) virus I week earlier. She was prescribed oseltamivir and returned to the hotel where she was staying alone. The next day, she was found dead in her hotel room. At autopsy, both lungs were voluminous and microscopic examination revealed acute-stage, severe diffuse alveolar damage with remarkable mononuclear cell infiltration and hyaline membrane formation in the lungs. CD8-positive T lymphocytes were dominantly observed. Immunohistochemically, influenza A viral protein was confirmed in the damaged type 11 pneumocytes and also in the infiltrated macrophages. Real-time RT-PCR analysis of both pre- and post-mortem pharyngeal swabs confirmed a novel influenza (A/H1N1pdm) virus infection. This is the second autopsy case of influenza (A/H1N1pdm) virus infection in Japan, and the findings indicated that the patient died due to an exceptionally rapid progression of viral pneumonia. This case indicates that patients with influenza (A/H1N1pdm) virus infection should be carefully monitor for acute respiratory distress syndrome.
• Endoscopic Follow-up of 3 Cases with Gastrointestinal Tract Involvement of Mantle Cell Lymphoma

  Makoto Saito, Akio Mori, Tatsuro Irie, Masanori Tanaka, Masanobu Morioka, Mariko Ozasa, Takahiko Kobayashi, Akiyoshi Saga, Kimiaki Miwa, Shinya Tanaka

  INTERNAL MEDICINE 49 (3) 231 - 235 0918-2918 2010 [Refereed][Not invited]
   

  Gastrointestinal (GI) tract involvement of mantle cell lymphoma (MCL) presents as a variety of forms, ranging from multiple lymphomatous polyposis (MLP) to a slight mucosal change. We report 3 cases with GI tract involvement of MCL who were followed-up by endoscopy. The present study shows three new informations. MLP of the esophagus is rare, but it was observed in two of 3 patients who were extensively involved by MCL. Endoscopic follow-up in one patient suggested that lymphoma cells of MCL had invaded the lamina propria to submucosal layer before MLP developed. Two of the 3 cases showed a favorable clinical course with single-agent rituximab therapy.
• Primary Mediastinal Liposarcoma, with 6 Years of Follow-up to Autopsy, Revealed Histopathological Features of Primary and Metastatic Lesions

  Satoshi Konno, Satoshi Oizumi, Naofumi Shinagawa, Eiki Kikuchi, Jun Konishi, Kenichiro Ito, Nobuyuki Hizawa, Akihiro Takiyama, Shinya Tanaka, Masaharu Nishimura

  INTERNAL MEDICINE 49 (8) 771 - 775 0918-2918 2010 [Refereed][Not invited]
   

  Primary mediastinal liposarcoma was observed in a 73-year-old man. Because of tight adhesions to adjacent tissues, neither complete resection nor surgical debulking of the tumor was possible. A T-tube was inserted into the patient's trachea for severe dyspnea, and he was treated with radiotherapy and an oral peroxisome proliferator-activated receptor-gamma agonist. The patient died 6 years after the initial diagnosis. Autopsy revealed liposarcoma composed of 3 subtypes in the primary tumor: well-differentiated, dedifferentiated, and round cell components. Round cell and dedifferentiated liposarcomas were predominantly observed in the metastatic nodules.
• Elcatonin Injections Suppress Systemic Bone Resorption without Affecting Cortical Bone Regeneration after Drill-Hole Injuries in Mice

  Yuji Katae, Shinya Tanaka, Akinori Sakai, Masato Nagashima, Hideyuki Hirasawa, Toshitaka Nakamura

  JOURNAL OF ORTHOPAEDIC RESEARCH 27 (12) 1652 - 1658 0736-0266 2009/12 [Refereed][Not invited]
   

  It is assumed that there are systemic changes in mineral metabolism during fracture healing that may cause a predisposition to sequential fractures in osteoporotic patients who suffered from previous fractures. Initial therapies for patients with osteoporotic fractures are important to prevent disabilities in daily life consequent to bone and muscle atrophies, and sequential fractures, although systemic and local bone metabolism during fracture healing have not been well understood. We evaluated the effects of bone injury and elcatonin injection as an initial therapy on systemic and local bone turnover and bone wound healing. Two drill holes were made in the diaphysis of the left femur and tibia of 12-week-old male C57BL/6J mice. They were treated with three doses of elcatonin or a vehicle thrice a week until the end of the 28-day experiment. Urinary crosslinked C-telopeptide of type I collagen (CTX) increased and the bone mineral densities (BMDs) in the lumbar vertebrae decreased in the vehicle-treated mice. Elcatonin injection prevented increases in urinary CTX and reduction of the BMDs. In the noninjured femoral metaphysis, osteoclast surface increased until day 28, whereas elcatonin suppressed it. In the fracture site, elcatonin facilitated osteoblast proliferation and did not delay the healing of the bone defect. Bone injuries accelerated bone turnover systemically and locally, and the elcatonin injections suppressed the systemic acceleration of bone resorption without a delay of filling regenerated cortical bone in the bone defect. (C) 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1652-1658, 2009
• Differential oncogenic potential of geographically distinct Helicobacter pylori CagA isoforms in mice

  Motohiro Miura, Naomi Ohnishi, Shinya Tanaka, Kohei Yanagiya, Masanori Hatakeyama

  INTERNATIONAL JOURNAL OF CANCER 125 (11) 2497 - 2504 0020-7136 2009/12 [Refereed][Not invited]
   

  Infection with cagA-positive Helicobacter pylori is associated with gastric carcinoma. The cagA-encoded CagA protein is delivered into gastric epithelial cells and, upon tyrosine phosphorylation at the C-terminal EPIYA segments, binds and deregulates SHP-2 oncoprotein. On the basis of the differential alignment of the EPIYA segments, CagA can be subdivided into Western CagA, which is produced by H. pylori isolated in Western countries, and East Asian CagA, which is produced by H. pylori circulating in East Asian countries. Western CagA contains EPIYA-A, EPIYA-B and variable numbers of EPIYA-C segments, whereas East Asian CagA contains EPIYA-A, EPIYA-B and variable numbers of EPIYA-D segments. Upon tyrosine phosphorylation, EPIYA-C and EPIYA-D, respectively, serve as low-affinity and high-affinity SHP-2-binding sites. We previously reported that systemic expression of East Asian CagA (CagA-ABDD) induces gastrointestinal and hematopoietic malignancies in mice. In this study, we generated transgenic mice that systemically express Western CagA (CagA-ABCCC), the levels of which are comparable to those in mice expressing East Asian CagA. The mice developed gastric epithelial hypertrophy and gastrointestinal tumors and also showed lymphoid abnormality but not myeloid abnormalities such as granulocytosis and myeloid leukemia found in mice carrying East Asian CagA. The incidence of tumors in mice expressing Western CagA was significantly lower than that in mice expressing East Asian CagA. Our results indicate that Western CagA is qualitatively less oncogenic than East Asian CagA. Differential oncogenic potential of geographically distinct CagA isoforms may contribute to the differential prevalence of gastric carcinoma between East Asian countries and Western countries. (c) 2009 UICC
• Natural Occurring IL-17 Producing T Cells Regulate the Initial Phase of Neutrophil Mediated Airway Responses

  Shinya Tanaka, Takayuki Yoshimoto, Tetsuji Naka, Susumu Nakae, Yo-ichi Iwakura, Daniel Cua, Masato Kubo

  JOURNAL OF IMMUNOLOGY 183 (11) 7523 - 7530 0022-1767 2009/12 [Refereed][Not invited]
   

  Effector Th17 cells are a major source of IL-17, a critical inflammatory cytokine in autoimmune diseases and in host defenses during bacterial infections. Recently, splenic lymphoid tissue inducer-like cells have been reported to be a source of T cell independent IL-17. In this study, we report that the immune system contains a unique set of natural occurring IL-17 producing cell, "natural" Th17 (nTh17), which area memory-like T cell subset. The nTh17 cells can develop in the absence of the IL-6/STAT3 signaling axis required by inducible Th17 cells. The nTh17 cell population is distinct from conventional inducible Th17 cells, since nTh17 cells express substantial amounts of IL-17A (IL-17), but not IL-7F, under the control of the master regulator, ROR gamma t The nTh17 cells simultaneously produce IFN-gamma. DO11.10 transgenic mice with a Rag(-/-) background (DO11.10 Rag(-/-)) lack nTh17 cells, and, following intranasal administration of OVA, IL-17-dependent neutrophil infiltration occurs in DO11.10 transgenic mice, but not in DO11.10 Rag(-/-) mice. The impaired neutrophil-dependent airway response is restored by adaptive transfer of nTh17 cells into DO11.10 Rag(-/-) mice. These results demonstrate that a novel T cell subset, nTh17, facilitates the early phase of Ag-induced airway responses and host defenses against pathogen invasion before the establishment of acquired immunity. The Journal of Immunology, 2009, 183: 7523-7530.
• Oxidative stress in skeletal muscle impairs mitochondrial respiration and limits exercise capacity in type 2 diabetic mice

  Takashi Yokota, Shintaro Kinugawa, Kagami Hirabayashi, Shouji Matsushima, Naoki Inoue, Yukihiro Ohta, Sanae Hamaguchi, Mochamad A. Sobirin, Taisuke Ono, Tadashi Suga, Satoshi Kuroda, Shinya Tanaka, Fumio Terasaki, Koichi Okita, Hiroyuki Tsutsui

  AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY 297 (3) H1069 - H1077 0363-6135 2009/09 [Refereed][Not invited]
   

  Yokota T, Kinugawa S, Hirabayashi K, Matsushima S, Inoue N, Ohta Y, Hamaguchi S, Sobirin MA, Ono T, Suga T, Kuroda S, Tanaka S, Terasaki F, Okita K, Tsutsui H. Oxidative stress in skeletal muscle impairs mitochondrial respiration and limits exercise capacity in type 2 diabetic mice. Am J Physiol Heart Circ Physiol 297: H1069-H1077, 2009. First published July 17, 2009; doi: 10.1152/ajpheart.00267.2009.-Insulin resistance or diabetes is associated with limited exercise capacity, which can be caused by the abnormal energy metabolism in skeletal muscle. Oxidative stress is involved in mitochondrial dysfunction in diabetes. We hypothesized that increased oxidative stress could cause mitochondrial dysfunction in skeletal muscle and make contribution to exercise intolerance in diabetes. C57/BL6J mice were fed on normal diet or high fat diet (HFD) for 8 wk to induce obesity with insulin resistance and diabetes. Treadmill tests with expired gas analysis were performed to determine the exercise capacity and whole body oxygen uptake ((V) over dotO(2))). The work (vertical distance x body weight) to exhaustion was reduced in the HFD mice by 36%, accompanied by a 16% decrease of peak (V) over dotO(2). Mitochondrial ADP-stimulated respiration, electron transport chain complex I and III activities, and mitochondrial content in skeletal muscle were decreased in the HFD mice. Furthermore, superoxide production and NAD(P)H oxidase activity in skeletal muscle were significantly increased in the HFD mice. Intriguingly, the treatment of HFD-fed mice with apocynin [10 mmol/l; an inhibitor of NAD(P)H oxidase activation] improved exercise intolerance and mitochondrial dysfunction in skeletal muscle without affecting glucose metabolism itself. The exercise capacity and mitochondrial function in skeletal muscle were impaired in type 2 diabetes, which might be due to enhanced oxidative stress. Therapies designed to regulate oxidative stress and maintain mitochondrial function could be beneficial to improve the exercise capacity in type 2 diabetes.
• Adaptor Protein Crk Induces Src-Dependent Activation of p38 MAPK in Regulation of Synovial Sarcoma Cell Proliferation

  Takuya Watanabe, Masumi Tsuda, Shinya Tanaka, Yusuke Ohba, Hideaki Kawaguchi, Tokifumi Majima, Hirofumi Sawa, Akio Minami

  MOLECULAR CANCER RESEARCH 7 (9) 1582 - 1592 1541-7786 2009/09 [Refereed][Not invited]
   

  The adaptor protein Crk mediates intracellular signaling related to cell motility and proliferation and is implicated in human tumorigenesis. The role of Crk in the growth of human sarcoma has remained unclear, however. The present study shows that Crk-induced activation of Src and subsequent signaling by p38 mitogen-activated protein kinase (MAPK) contribute to the enhanced proliferation of human synovial sarcoma cells. Depletion of Crk by RNA interference markedly inhibited proliferation of the synovial sarcoma cell lines HS-SYII, SYO-1, and Fuji as well as prevented anchorage-independent growth. Conversely, reconstitution with CrkII by authentic small interfering RNA-resistant Crk gene restored proliferation in Crk-silenced SYO-1 cells. Crk-depleted synovial sarcoma cells manifested enhanced transcriptional activity and expression of the p16(INK4A) gene, resulting in their accumulation in G, phase of the cell cycle. In response to hepatocyte growth factor stimulation, Crk prominently induced the tyrosine phosphorylation of Grb2-associated binder 1 through activation of Src and focal adhesion kinase, and the Src family kinase inhibitor PP2 almost completely inhibited the proliferation of SYO-1 cells. Crk also induced the phosphorylation of p38 MAPK, and SB203580, a p38 MAPK-specific inhibitor, increased expression of p16(INK4A) gene in SYO-1 cells. Furthermore, SB203580 or depletion of p38 MAPK by small interfering RNA suppressed both the phosphorylation of Akt triggered by hepatocyte growth factor and the proliferation of SYO-1 cells. These results suggest that Crk promotes proliferation of human synovial sarcoma cells through activation of Src and its downstream signaling by a novel p38 MAPK-Akt pathway, with these signaling molecules providing potent new targets for molecular therapeutics. (Mol Cancer Res 2009;7(9):1582-92)
• Bcl6 Mediates the Development of T Follicular Helper Cells

  Roza I. Nurieva, Yeonseok Chung, Gustavo J. Martinez, Xuexian O. Yang, Shinya Tanaka, Tatyana D. Matskevitch, Yi-Hong Wang, Chen Dong

  SCIENCE 325 (5943) 1001 - 1005 0036-8075 2009/08 [Refereed][Not invited]
   

  A fundamental function of CD4(+) helper T (T(H)) cells is the regulation of B cell-mediated humoral immunity. Development of T follicular helper (T(FH)) cells that provide help to B cells is mediated by the cytokines interleukin-6 and interleukin-21 but is independent of T(H)1, T(H)2, and T(H)17 effector cell lineages. Here, we characterize the function of Bcl6, a transcription factor selectively expressed in T(FH) cells. Bcl6 expression is regulated by interleukin-6 and interleukin-21. Bcl6 overexpression induced T(FH)-related gene expression and inhibited other T(H) lineage cell differentiation in a DNA binding-dependent manner. Moreover, Bcl6 deficiency in T cells resulted in impaired T(FH) cell development and germinal center reactions, and altered production of other effector T cell subsets. Our data thus illustrate that Bcl6 is required for programming of T(FH) cell generation.
• A case of central nervous system lymphomatoid granulomatosis; characteristics of PET imaging and pathological findings

  Hiroshi Nishihara, Matsuyoshi Nakasato, Hiroki Sawa, Hiromi Murakami, Daisuke Yamamoto, Kenji Moriyama, Norifumi Kato, Ikuo Hashimoto, Hajime Kamada, Shinya Tanaka

  JOURNAL OF NEURO-ONCOLOGY 93 (2) 275 - 278 0167-594X 2009/06 [Refereed][Not invited]
   

  Lymphomatoid granulomatosis (LYG) in the central nervous system (CNS) is an uncommon lymphoproliferative disorder with low grade malignant potential. Here we report a case of CNS-LYG, in particular, its characteristics of radioisotope imaging and pathological findings. A 65-year-old man complained of visual disturbance and homonymous hemianopsia was designated. CT and MRI revealed an edematous, enhanced irregular and nodular lesion in the right occipital and parietal lobes. Although 18F-fluorodeoxyglucose (FDG)-positron emission tomography (PET) scan showed low uptake in the lesion, Methionine (MET)-PET scan indicated high uptake. Proton magnetic resonance spectroscopy ((1)H-MRS) at 3T revealed a decrease of the peak of the N-acetylaspartate (NAA), suggesting a possible neoplastic lesion. The patient was diagnosed with CNS-LYG based on the surgically removed material showing perivascular infiltration of CD3-positive small T-lymphocytes with granulomatous lesions. The post-operative steroid therapy was effective and the recurrence or exacerbation has not been observed by radiological imaging until now.
• The anabolic action of intermittent PTH in combination with cathepsin K Inhibitor or alendronate differs depending on the remodeling status in bone in ovariectomized mice

  Hirotoshi Yamane, Akinori Sakai, Toshiharu Mori, Shinya Tanaka, Kuniaki Moridera, Toshitaka Nakamura

  BONE 44 (6) 1055 - 1062 8756-3282 2009/06 [Refereed][Not invited]
   

  We hypothesized that the anabolic action of parathyroid hormone (PTH) with the anti-catabolic agents cathepsin K inhibitor and alendronate differs depending on the remodeling status in the bone. C57/BL/6J mice, 8 weeks of age, were subjected to ovariectomized (OVX) or sham surgery. At 6 weeks after surgery the, mice were treated with cathepsin K inhibitor, alendronate, or a vehicle (daily, for 8 weeks), with or without PTH (1-34) (5 times/week, for the last 4 weeks). We assessed the bone chemical markets of the serum and urine, bone mineral density (BMD), histomorphomery in the primary and secondary spongiosa of the proximal tibia after fluorescence labeling, primary cell Culture, and mRNA expressions in bone marrow cells. Cathepsin K inhibitor and alendronate significantly increased the BMD and the bone volume of the primary and secondary spongiosa, with a reduction of the urinary C-telopeptide of type I collagen that was increased by OVX, respectively. Cathepsin K inhibitor augmented the anabolic action of PTH on the BMD and bone Volume at both the primary and secondary spongiosa, while alendronate had the same effect on the BMD and bone volume only at the primary spongiosa. Cathepsin K inhibitor did not decrease serum osteocalcin with or without PTH, while alendronate did decrease it. Cathepsin K inhibitor did not decrease the values of osteoclast number or bone formation rate with or without PTH, while alendronate decreased those values and increased osteoclast apoptosis. The combination of PTH and cathepsin K inhibitor increased alkaline phosphatase-positive CFU-f formation and c-fos, osterix, and osteocalcin mRNA expressions of bone marrow cells as well as PTH alone, while the combination of PTH and alendronate decreased those values. This study demonstrated that alendronate enhances the anabolic action of PTH at the primary spongiosa, but blunts it in the remodeling trabecular bone, while cathepsin K inhibitor enhances the action at both sites in OVX mice. In conclusion, the anabolic action of intermittent PTH in combination with cathepsin K inhibitor or alendronate differs depending on the remodeling status of bone in OVX mice. (C) 2008 Elsevier Inc. All rights reserved.
• Human synovial sarcoma proto-oncogene Syt is essential for early embryonic development through the regulation of cell migration

  Taichi Kimura, Mieko Sakai, Kouichi Tabu, Lei Wang, Ryosuke Tsunematsu, Masumi Tsuda, Hirofumi Sawa, Kazuo Nagashima, Hiroshi Nishihara, Shigetsugu Hatakeyama, Keiko Nakayama, Marc Ladanyi, Shinya Tanaka, Keiichi I. Nakayama

  LABORATORY INVESTIGATION 89 (6) 645 - 656 0023-6837 2009/06 [Refereed][Not invited]
   

  SYT-SSX protein, resulted from chromosomal translocation, causes synovial sarcoma, which is a malignant tumor accounting for 10% of soft tissue sarcoma. However, biological functions of SYT (synovial sarcoma translocation), also known as SS18, are largely unclear, whereas it has been proven that Syt-null mice die at early stages of embryonic development. Here, we generated Syt-deficient mice and confirmed the reported phenotypes, including growth retardation, open neural tube and haplo-insufficient lethality, and therefore, there is no doubt that Syt is essential for embryonic development. However, placental defects, described in the earlier report, were rarely seen in our mice and we frequently observed cardiac defect in Syt-deficient mice. As the mechanisms responsible for embryonic lethality seem to be complicate, we performed additional experiments. By using primary cultured embryonic fibroblasts, we showed that Syt(-/-) MEFs deregulate actin organization and suppressed cell migration. These observations suggest that Syt may contribute to the signaling pathway important for various cellular functions in vivo and in vitro, and we propose that Syt-deficient MEFs would be a powerful means to understand the biological roles of SYT in vitro. Laboratory Investigation (2009) 89, 645-656; doi:10.1038/labinvest.2009.25; published online 30 March 2009
• HuR is exported to the cytoplasm in oral cancer cells in a different manner from that of normal cells

  H. Hasegawa, W. Kakuguchi, T. Kuroshima, T. Kitamura, S. Tanaka, Y. Kitagawa, Y. Totsuka, M. Shindoh, F. Higashino

  BRITISH JOURNAL OF CANCER 100 (12) 1943 - 1948 0007-0920 2009/06 [Refereed][Not invited]
   

  HuR, a ubiquitously expressed member of the Hu protein family that binds and stabilizes an AU-rich element (ARE)-containing mRNAs, is known to shuttle between the nucleus and the cytoplasm via several export pathways. When normal cells were treated with heat shock, HuR was exported to the cytoplasm in a chromosome maintenance region 1 (CRM1)-dependent manner. However, in this study, we demonstrate that HuR is exported to the cytoplasm in oral cancer cells even if the cells were treated with the inhibitor of the CRM1-independent export pathway. Immunohistochemical and biochemical analyses showed that HuR existed in both the cytoplasm and the nucleus in oral cancer cells, such as HSC-3 and Ca9.22, but existed entirely inside the nucleus in normal cells. AU-rich element-mRNAs were also exported to the cytoplasm and stabilised in the oral cancer cells, which were inhibited by HuR knockdown. This export of HuR was not affected by at least 7 h of treatment of leptomycin B (LMB), which is an inhibitor of the CRM1-dependent export pathway. These findings suggest that HuR is exported to the cytoplasm in oral carcinoma cells in a different manner from that of normal cells, and is likely to occur through the perturbation of a normal export pathway. British Journal of Cancer (2009) 100, 1943-1948; doi: 10.1038/sj.bjc.6605084 www.bjcancer.com (C) 2009 Cancer Research UK
• Crk adaptor protein-induced phosphorylation of Gab1 on tyrosine 307 via Src is important for organization of focal adhesions and enhanced cell migration

  Takuya Watanabe, Masumi Tsuda, Yoshinori Makino, Tassos Konstantinou, Hiroshi Nishihara, Tokifumi Majima, Akio Minami, Stephan M. Feller, Shinya Tanaka

  CELL RESEARCH 19 (5) 638 - 650 1001-0602 2009/05 [Refereed][Not invited]
   

  Upon growth factor stimulation, the scaffold protein, Gab1, is tyrosine phosphorylated and subsequently the adaptor protein, Crk, transmits signals from Gab1. We have previously shown that Crk overexpression, which is detectable in various human cancers, induces tyrosine phosphorylation of Gab1 without extracellular stimuli. In the present study, the underlying mechanisms were further investigated. Mutational analyses of CrkII demonstrated that the SH2 domain, but not the SH3(N) or the regulatory Y221 residue of CrkII, is critical for the induction of Gab1-Y307 phosphorylation. SH2 mutation of CrkII also decreased the interaction with Gab1. In GST pull-down assay, Crk-SH2 bound to wild-type Gab1, whereas Crk-SH3(N) interacted with the Gab1 mutant, which lacks the clustered tyrosine region (residues 242-410). Tyrosine phosphorylation of Gab1 was induced by all Crk family proteins, but not other SH2-containing signalling adaptors. Src-family kinase inhibitor, PP2, abrogates Crk-induced tyrosine phosphorylations of Gab1. Y307 phosphorylation was undetectable in fibroblasts lacking Src, Yes, and Fyn, even upon overexpression of Crk, whereas cells lacking only Yes and Fyn still contained Gab1 with phosphorylated Y307. Furthermore, Crk induced the phosphorylation of Src-Y416; accordingly the interaction between Crk and Csk was increased. The Gab1-Y307F mutant failed to localize near the plasma membrane even upon HGF stimulation and decreased cell migration. Moreover, Gab1-Y307F disturbed the localization of Crk, FAK, and paxillin, which are the typical components of focal adhesions. Taken together, these results indicate that Crk facilitates tyrosine phosphorylation of Gab1-Y307 through Src, contributing to the organization of focal adhesions and enhanced cell migration, thereby possibly promoting human cancer development.
• Crk and CrkL adaptor proteins: networks for physiological and pathological signaling

  Raymond B. Birge, Charalampos Kalodimos, Fuyuhiko Inagaki, Shinya Tanaka

  CELL COMMUNICATION AND SIGNALING 7 13  1478-811X 2009/05 [Refereed][Not invited]
   

  The Crk adaptor proteins (Crk and CrkL) constitute an integral part of a network of essential signal transduction pathways in humans and other organisms that act as major convergence points in tyrosine kinase signaling. Crk proteins integrate signals from a wide variety of sources, including growth factors, extracellular matrix molecules, bacterial pathogens, and apoptotic cells. Mounting evidence indicates that dysregulation of Crk proteins is associated with human diseases, including cancer and susceptibility to pathogen infections. Recent structural work has identified new and unusual insights into the regulation of Crk proteins, providing a rationale for how Crk can sense diverse signals and produce a myriad of biological responses.
• Integral Role of Transcription Factor 8 in the Negative Regulation of Tumor Angiogenesis

  Takayuki Inuzuka, Masumi Tsuda, Shinya Tanaka, Hideaki Kawaguchi, Yujiro Higashi, Yusuke Ohba

  CANCER RESEARCH 69 (4) 1678 - 1684 0008-5472 2009/02 [Refereed][Not invited]
   

  Angiogenesis is involved in various physiologic and pathological conditions, including tumor growth, and is tightly regulated by the orchestration of proangiogenic and antiangiogenic factors. Inhibition of vascular endothelial growth factor (VEGF), the best-established antiangiogenic treatment in cancer, has shown some effectiveness; however, the identification of novel regulators, whose function is independent of VEGF, is required to achieve better outcomes. Here, we show that transcription factor 8 (TCF8) is up-regulated in endothelial cells during angiogenesis, acting as a negative regulator. Furthermore, TCF8 is specifically expressed in the endothelium of tumor vessels. 1cf8-heterozygous knockout mice are more permissive than wildtype mice to the formation of tumor blood vessels in s.c. implanted melanoma, which seems to contribute to the more aggressive growth and the lung metastases of the tumor in mutant mice. Suppression of TCF8 facilitates angiogenesis in both in vitro and ex vivo models, and displays comprehensive cellular phenotypes, including enhanced cell invasion, impaired cell adhesion, and increased cell monolayer permeability due to, at least partly, MMPI overexpression, attenuation of focal adhesion formation, and insufficient VE-cadherin recruitment, respectively. Taken together, our findings define a novel, integral role for TCF8 in the regulation of pathologic angiogenesis, and propose TCF8 as a target for therapeutic intervention in cancer. [Cancer Res 2009;69(4):1678-84]
• Biological roles for signaling adaptor protein CRK

  Shinya Tanaka

  Seikagaku 81 (5) 361 - 376 0037-1017 2009 [Refereed][Not invited]
  
• CNS lymphomatoid granulomatosis with lymph node and bone marrow involvements

  Akihiro Takiyama, Hiroshi Nishihara, Ukihide Tateishi, Taichi Kimura, Wang Lei, Katsuji Marukawa, Tomoo Itoh, Satoshi Hashino, Kazuo Nagashima, Shinya Tanaka

  NEUROPATHOLOGY 28 (6) 640 - 644 0919-6544 2008/12 [Refereed][Not invited]
   

  Lymphomatoid granulomatosis (LYG) in the CNS is an uncommon lymphoproliferative disease with characteristic angiocentric lymphoreticular proliferative and granulomatous lesions exhibiting low-grade malignant potential. Here we report a rare case of CNS-LYG, which disseminated to the lymph node and bone marrow. A 50-year-old man was diagnosed with CNS-LYG based on brain biopsy showing perivascular infiltration of CD3-positive small T-lymphocytes without overt nuclear atypism. Eight months after the initial neurological symptoms, inguinal lymph node swelling was found and histopathologically diagnosed as peripheral T-cell lymphoma. TCR gamma-gene rearrangement study using both paraffin-embedded specimens of brain and inguinal lymph node demonstrated an identical clonal band. Considering the clinical course, we concluded lymph node involvement of CNS-LYG, suggesting the malignant potential of CNS-LYG.
• Harlequin ichthyosis model mouse reveals alveolar collapse and severe fetal skin barrier defects

  Teruki Yanagi, Masashi Akiyama, Hiroshi Nishihara, Kaori Sakai, Wataru Nishie, Shinya Tanaka, Hiroshi Shimizu

  HUMAN MOLECULAR GENETICS 17 (19) 3075 - 3083 0964-6906 2008/10 [Refereed][Not invited]
   

  Harlequin ichthyosis (HI), which is the most severe genodermatosis, is caused by loss-of-function mutations in ABCA12, a member of the ATP-binding cassette transporter family. To investigate the pathomechanism of HI and the function of the ABCA12 protein, we generated ABCA12-deficient mice (Abca12(-/-)) by targeting Abca12. Abca12(-/-) mice closely reproduce the human HI phenotype, showing marked hyperkeratosis with eclabium and skin fissure. Lamellar granule abnormalities and defective ceramide distribution were remarkable in the epidermis. Skin permeability assay of Abca12(-/-) fetuses revealed severe skin barrier dysfunction after the initiation of keratinization. Surprisingly, the Abca12(-/-) mice also demonstrated lung alveolar collapse immediately after birth. Lamellar bodies in alveolar type II cells of the Abca12(-/-) mice lacked normal lamellar structures. The level of surfactant protein B, an essential component of alveolar surfactant, was reduced in the Abca12(-/-) mice. Fetal therapeutic trials with systemic administration of retinoid or dexamethasone, which are effective for HI and respiratory distress, respectively, to the pregnant mother mice neither improved the skin phenotype nor extended the survival period. Our HI model mice reproduce the human HI skin phenotype soon after the initiation of fetal skin keratinization and provide evidence that ABCA12 plays pivotal roles in lung and skin barrier functions.
• Promoter hypomethylation regulates CD133 expression in human gliomas

  Kouichi Tabu, Ken Sasai, Taichi Kimura, Lei Wang, Eiko Aoyanagi, Shinji Kohsaka, Mishie Tanino, Hiroshi Nishihara, Shinya Tanaka

  CELL RESEARCH 18 (10) 1037 - 1046 1001-0602 2008/10 [Refereed][Not invited]
   

  Brain tumor-initiating cells (BTICs) have been enriched using antibodies against the cell surface protein CD133; however, the biological relevance and the regulatory mechanism of CD133 expression in human gliomas are not yet understood. In this study, we initially demonstrated that CD133 was overexpressed in high-grade human glioblastomas where CD133-positive cells were focally observed as a micro-cluster. In addition, CD133 transcripts with exon 1A, 1B, or 1C were predominantly expressed in glioblastomas. To elucidate the mechanism regulating this aberrant expression of CD133, three proximal promoters (P1, P2, and P3) containing a CpG island were isolated. In U251MG and T98G glioblastoma cells, the P1 region flanking exon 1A exhibited the highest activity among the three promoters, and this activity was significantly inactivated by in vitro methylation. After treatment with the demethylating agent 5-azacytidine and/or the histone deacetylase inhibitor valproic acid, the expression level of CD133 mRNA was significantly restored in glioma cells. Importantly, hypomethylation of CpG sites within the P1, P2, and P3 regions was observed by bisulfite sequencing in human glioblastoma tissues with abundant CD133 mRNA. Taken together, our results indicate that DNA hypomethylation is an important determinant of CD133 expression in glioblastomas, and this epigenetic event may be associated with the development of BTICs expressing CD133.
• Hypopharyngeal squamous cell carcinoma producing both granulocyte colony-stimulating factor and parathyroid hormone-related protein

  Kanae Tamura, Tomoaki Yoshinaga, Mishie Tanino, Taichi Kimura, Noriyuki Yamada, Masaharu Nishimura, Satoshi Fukuda, Hiroshi Nishihara, Masanobu Shindoh, Shinya Tanaka

  PATHOLOGY INTERNATIONAL 58 (10) 652 - 656 1320-5463 2008/10 [Refereed][Not invited]
   

  A 57-year-old woman was admitted to Hokkaido University Hospital because of dysphagia. Laryngoscopy indicated hypopharyngeal tumor histologically diagnosed as squamous cell carcinoma (SCC). A combination of radiotherapy and chemotherapy was performed for 2 months, and the hypopharyngeal lesion completely regressed. After 4 months, fever, anorexia, and malaise appeared, and chest X-ray and CT indicated two large tumors in the right lung. Transbroncheal lung biopsy (TBLB) specimens were diagnosed as SCC. Laboratory data showed high levels of serum granulocyte colony-stimulating factor (G-CSF) and parathyroid hormone-related protein (PTHrP). Subsequently, positron emission tomography (PET) showed multiple metastases to several organs including the liver, spine, skull, and thigh. Two months after readmission, the patient died with no success of chemotherapy. At autopsy, the lung tumor was clearly positive for both G-CSF and PTHrP on immunohistostaining. Retrospectively, examination showed that the primary pharyngeal tumor was focally positive for these two cytokines. Thus, a certain population of hypopharyngeal cancer producing G-CSF and PTHrP, spread to various organs and contributed to the rapid progression and poor prognosis. This case is presented with a discussion of several other cases in the literature.
• Establishment of a luciferase assay-based screening system: Fumitremorgin C selectively inhibits cellular proliferation of immortalized astrocytes expressing an active form of AKT

  Lei Wang, Ken Sasai, Tsuyoshi Akagi, Shinya Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 373 (3) 392 - 396 0006-291X 2008/08 [Refereed][Not invited]
   

  The AKT pathway is frequently activated in glioblastoma, and as such, inhibitors of this pathway could prove very useful as anti-glioblastoma therapies. Here we established immortalized astrocytes expressing Renilla luciferase as well as those expressing both an active form of AICT and firefly luciferase. Since both luciferase activities represent the numbers of corresponding cell lines, novel inhibitors of the AKT pathway can be identified by treating co-cultures containing the two types of luciferase-expressing cells with individual compounds. Indeed, such a screening system succeeded in identifying fumitremorgin C as an efficient inhibitor of the AICT pathway, which was further confirmed by the ability of fumitremorgin C to selectively inhibit the growth of immortalized astrocytes expressing an active form of AKT. The present study proposes a broadly applicable approach for identifying therapeutic agents that target the pathways and/or molecules responsible for cancer development. (c) 2008 Elsevier Inc. All rights reserved.
• Careful exclusion of non-neoplastic brain components is required for an appropriate evaluation of O-6-methylguanine-DNA methyltransferase status in glioma - Relationship between immunohistochemistry and methylation analysis

  Ken Sasai, Miho Nodagashira, Hiroshi Nishihara, Eiko Aoyanagi, Lei Wang, Masahito Katoh, Junichi Murata, Yoshimaru Ozaki, Tamio Ito, Shin Fujimoto, Sadao Kaneko, Kazuo Nagashima, Shinya Tanaka

  AMERICAN JOURNAL OF SURGICAL PATHOLOGY 32 (8) 1220 - 1227 0147-5185 2008/08 [Refereed][Not invited]
   

  Evaluation of O-6-methylguanine-DNA methyltransferase (MGMT) expression is important for antiglioma therapy as many clinical trials have demonstrated that promoter hypermethylation and low level expression of MGMT are associated with an enhanced response to alkylating agents. However, here we report that the current strategies used to evaluate MGMT status in gliomas are unreliable. We observed discordance in the MGMT expression status when immunohistochemical evaluation and polymerase chain reaction-based methylation assessments were used: 73% of gliomas with methylated MGMT promoter had substantial numbers of MGMT-immunopositive tumor cells. Furthermore, when MGMT expression was tested in tumor homogenates using reverse transcription-polymerase chain reaction, 43% of tumors were found positive, in comparison to only 24%, when histologic samples were assayed immunohistochemically. To explain these inconsistencies we undertook a detailed immunohistochemical evaluation of tumor samples and found that some gliomas demonstrated remarkably high expression of MGMT in the entire tumor whereas others contained only a small immunopositive area. Additionally, we found that gliomas contained various types of non-neoplastic cells expressing MGMT, including lymphocytes, vascular endothelial cells, and macrophages/microglias, which contribute to overall MGMT expression detected in tumor homogenates, and thus result in overestimation of tumor NIGMT expression. Therefore, to correctly establish MGMT expression in the tumor, which could be informative of glioma sensitivity to alkylating agents, exclusion of non-neoplastic brain components from analysis is required.
• Synthesis of dinuclear boron complexes of sulfinylcalix[4]arenes: Syn/anti stereocontrol by the arrangement of the sulfinyl functions

  Naoya Morohashi, Tsuyoshi Kitahara, Tomoko Arima, Shinya Tanaka, Yoshihiro Ohba, Tetsutaro Hattori

  ORGANIC LETTERS 10 (13) 2845 - 2848 1523-7060 2008/07 [Refereed][Not invited]
   

  Stereocontrol in the synthesis of dinuclear metal complexes of sulfinylcalix[4]arenes 2 has been achieved by the arrangement of sulfinyl functionalities. Thus, the treatment of the rtct isomer of 2 (2(rtct)) with an excess of Et(3)B affords syn dinuclear boron complex 4, while a similar treatment of rctt and rcct isomers 2(rctt) and 2(rcct) yields anti dinuclear complexes 5 and 6, respectively.
• An siRNA against JC virus (JCV) agnoprotein inhibits JCV infection in JCV-producing cells inoculated in nude mice

  Tomoko Matoba, Yasuko Orba, Tadaki Suzuki, Yoshinori Makino, Hideo Shichinohe, Satoshi Kuroda, Takahiro Ochiya, Hiroshi Itoh, Shinya Tanaka, Kazuo Nagashima, Hirofumi Sawa

  NEUROPATHOLOGY 28 (3) 286 - 294 0919-6544 2008/06 [Refereed][Not invited]
   

  JC virus (JCV) is the etiological agent of the demyelinating disease progressive multifocal leukoencephalopathy (PML). Because JCV has a very narrow host range, it has been difficult to develop an animal model of JCV infection; as a result, no effective therapy for PML has been established. In this study, we have tried to create an animal model that replaces an in vivo JCV infection. As a result, we have obtained a stable persistence of JCV-infected human cells in the mouse brain by inoculating the virus-infected cells into the nude mice brains. In this model, the JCV-infected cells were well preserved in the nude mouse brains for 2 weeks. We then treated JCV-injected brains with an siRNA against the JCV agnoprotein that is known to be an effective inhibitor of JCV infection in vitro. A highly purified type I collagen, atelocollagen, was used as a carrier for the siRNA. The siRNA inhibited the expression of JCV protein in inoculated JCV-infected cells in the mouse brain, compared to the medium containing only atelocollagen used as a placebo. Thus, the combination of siRNA and atelocollagen might be a candidate therapeutic agent for the treatment of JCV infection.
• Adaptor protein Crk is implicated in mucus formation in mucinous epithelial ovarian cancer (mEOC) cells MCAS

  Xu Dong-mei, Linghu Hua, Masumi Tsuda, Shinya Tanaka, Kazuo Nagashima

  CHINESE JOURNAL OF CANCER RESEARCH 20 (2) 121 - 125 1000-9604 2008/06 [Refereed][Not invited]
   

  Objective: The mucus production is an indicator for the histological grade of mucinous epithelial ovarian cancer (mEOC). In our previous study, Crk expression was targeted in the human ovarian mucinous adenocarcinoma cell line MCAS through RNA interference, resulting in the establishment of Crk knock down cells. These cells exhibited decreased tumorigenic potential both in vitro and in vivo. The purpose of this study was to investigate if there is any change in the capability of forming mucus in these Crk knock down cells. Methods: Cytoplasmic periodic acid Schiff (PAS) staining and particle excluding assay were conducted to assess the mucus formation within and around cells, respectively. Additionally, the amount of mucus formed in tumor lumps from nude mice model was measured following HE and PAS staining. Results: The increased mucus production in Crk knockdown mEOC cells (MCAS) was manifested by increased number of enlarged cells filled with vacuoles-like mucus observed by phase-contrast microscope and cytoplasmic PAS staining; and enhanced mucus secretion was represented by the assembly of pericellular matrix in particle excluding assay and increased mucus area in tumor lumps from nude mice models. Conclusion: The course of carcinogenesis in mEOC is associated with the altered pattern of mucus production and secretion. The adaptor protein Crk is implicated in both pathways.
• Genetic disruption of all NO synthase isoforms enhances BMD and bone turnover in mice in vivo: Involvement of the renin-angiotensin system

  Ken Sabanai, Masato Tsutsui, Akinori Sakai, Hideyuki Hirasawa, Shinya Tanaka, Eiichiro Nakamura, Akihide Tanimoto, Yasuyuki Sasaguri, Masako Ito, Hiroaki Shimokawa, Toshitaka Nakamura, Nobuyuki Yanagihara

  JOURNAL OF BONE AND MINERAL RESEARCH 23 (5) 633 - 643 0884-0431 2008/05 [Refereed][Not invited]
   

  Introduction: NO is synthesized by three different NO synthase (NOS) isoforms, including neuronal (nNOS), inducible (iNOS) and endothelial NOS (eNOS). The roles of NO in bone metabolism have been extensively investigated in pharmacological studies and in studies with NOS isoform-deficient mice. However, because of the nonspecificity of agents and compensation among the NOS isoforms, the ultimate roles of endogenous NO are still poorly understood. To address this point, we successfully generated mice in which all three NOS genes are completely disrupted. In this study, we examined whether bone metabolism is abnormal in those mice. Materials and Methods: Experiments were performed in 12-wk-old male wildtype, singly nNOS(-/-), iNOS(-/-), and eNOS(-/-) and triply n/i/eNOS(-/-) mice. BMD was assessed by DXA. The kinetics of osteoblastic bone formation and those of osteoclastic bone resorption were evaluated by measurements of morphological and biochemical markers. Results: BMD was significantly higher only in the triply NOS-/- mice but not in any singly NOS-/- mice compared with the wildtype mice. Markers of osteoblastic bone formation, including bone formation rate, mineral apposition rate, and serum alkaline phosphatase concentration, were also significantly larger only in the triply NOS-/- mice compared with wildtype mice. Furthermore, markers of osteoclastic bone resorption, including osteoclast number, osteoclast surface, and urinary deoxypyridinoline excretion, were again significantly greater only in the triply NOS-/- mice. Importantly, the renin-angiotensin system in bone was significantly activated in the triply NOS-/- mice, and long-term oral treatment with an angiotensin II type I (AT,) receptor blocker normalized this pathological bone remodeling in those mice. Conclusions: These results provide the first direct evidence that genetic disruption of the whole NOS system enhances BMD and bone turnover in mice in vivo through the AT, receptor pathway, showing the critical role of the endogenous NO/NOS system in maintaining bone homeostasis.
• A case of cerebral ganglioneuronal tumor in the parietal lobe of an adult

  Hiroshi Nishihara, Yoshimaru Ozaki, Tamio Ito, Tomoaki Yoshinaga, Kouichi Tabu, Mishie Tanino, Kazuo Nagashima, Shinya Tanaka

  BRAIN TUMOR PATHOLOGY 25 (1) 45 - 49 1433-7398 2008/04 [Refereed][Not invited]
   

  Central nervous system (CNS) neuroblastoma/ganglioneuroblastoma is one of the embryonal tumors with neuronal differentiation found in young adults, but it is most common in children, especially in those below the age of 5 years, whereas extraventricular neurocytoma, a rare neuroepithelial tumor with neuronal differentiation, mostly affects young adults. Here we present a rare case of cerebral ganglioneuronal tumor that occurred in a 32-year-old woman. The patient suffered from tonic convulsion, and computed tomography demonstrated a well-demarcated, round tumor 3.3 cm in size with marked calcification in the right parietal lobe. Histological analysis revealed diffuse infiltration of small, round cells with scattered large ganglion-like cells. Immunohistochemically, the tumor cells did not react with any neuronal molecules, except for chromogranin A in ganglion-like large tumor cells, but electron microscopy demonstrated the presence of synapse-like nerve terminal structures without mature postsynaptic density, suggesting the presence of neoplastic tumor components with neuronal differentiation; thus, this tumor was diagnosed as CNS ganglioneuroblastoma with possible low-grade malignancy because the Mib-1 labeling index was less than 3%-4%. Here we discuss the histological entity of cerebral ganglioneuronal tumors, including extraventricular neurocytoma.
• Clusterin expression predicts survival of invasive cervical cancer patients treated with radical hysterectomy and systematic lymphadenectomy

  Hidemichi Watari, Yoko Ohta, Mohamed Kamel Hassan, Ying Xiong, Shinya Tanaka, Noriaki Sakuragi

  GYNECOLOGIC ONCOLOGY 108 (3) 527 - 532 0090-8258 2008/03 [Refereed][Not invited]
   

  Objectives. The aim of this study was to evaluate the prognostic significance of clusterin expression in invasive cervical cancer patients treated with radical hysterectomy and systematic lymphadenectomy. Methods. Invasive cervical cancer specimens were obtained from 52 patients who underwent radical hysterectomy and systematic lymphadenectomy at Hokkaido University Hospital from 1997 to 2004. The expression of clusterin protein was analyzed by inummohistochemical staining. Findings were evaluated in relation to several clinicopathological factors. Survival analyses were performed by the Kaplan-Meier curves and the log-rank test. Independent prognostic factors were determined by multivariate Cox regression analysis. Results. Clusterin protein was present in the cytoplasm of cervical cancer cells. The expression of clusterin protein in invasive cervical cancer tissues was not related to any clinicopathologic factors analyzed. Patients with positive clusterin expression showed significantly worse prognosis than those with negative clusterin expression (p=0.017). Multivariate analysis including clusterin expression revealed that clusterin expression (p=0.006) and the number of positive node groups (p=0.002) were independent prognostic factors for survival. Survival of patients with invasive cervical cancer could be stratified into three groups by combination of clusterin expression and number of positive node groups with an estimated 5-year survival rate of 100.0% for no or one positive node group irrespective of clusterin expression (group A), 78.7% for multiple node groups with negative clusterin expression (group B), and 14.3% for multiple node groups with positive clusterin expression (group C) (p=0.03 for group A vs. group B, p=0.004 for group B vs. group C, and p<0.0001 for group A vs. group C). Conclusions. Clusterin expression and the number of positive node groups were independent prognostic factors for invasive cervical cancer patients treated with radical hysterectomy and systematic lymphadenectomy. Clusterin might be a new molecular marker to predict the survival of cervical cancer patients with multiple positive node groups. (C) 2007 Elsevier Inc. All rights reserved.
• Selective cyclooxygenase-2 inhibitor prevents reduction of trabecular bone mass in collagen-induced arthritic mice in association with suppression of RANKL/OPG ratio and IL-6 mRNA expression in synovial tissues but not in bone marrow cells

  Tomonori Taketa, Akinori Sakai, Shinya Tanaka, Kenichiro Nakai, Kunitaka Menuki, Hirotoshi Yamane, Kazuhiro Tanaka, Toshitaka Nakamura

  JOURNAL OF BONE AND MINERAL METABOLISM 26 (2) 143 - 151 0914-8779 2008/03 [Refereed][Not invited]
   

  We performed this study to clarify whether celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, prevents trabecular bone mass reduction by suppressing arthritis-related increase of bone resorption, and to discriminate differences in actions on bone among celecoxib, SC-58560 (a selective COX-1 inhibitor), and indomethacin. Eight-week-old DBA/1J male mice were divided into six groups as follows. Control untreated (Normal) and collagen-induced arthritic (CIA) mice were compared with four treatment groups: celecoxib was orally administered to CIA mice at doses of 0 (Vehicle), 16 (COX2L), and 75 (COX2H) mg/kg, in addition to two groups of mice treated with SC-58560 (COX1) or indomethacin (IND). Histomorphometry showed a significant decrease in tibial trabecular bone volume in arthritic mice, which was corrected by COX2H. The increased osteoclast surface and number in the Vehicle group were suppressed by COX2L, COX2H, and IND. The decreased bone formation rate in Vehicle was elevated by COX2H without statistical significance. A high ratio of mRNA expression of receptor activator of NF-kappa B ligand (RANKL)/osteoprotegerin (OPG) in Vehicle synovial tissue was suppressed by COX2L and COX2H. The increased expression of interleukin (IL)-6 mRNA in Vehicle was suppressed by COX2L, COX2H, and IND, although no difference in this expression was observed in bone marrow cells among all groups. In conclusion, in CIA mice, celecoxib suppresses arthritis-related increase in bone resorption at low and high doses and prevents trabecular bone mass reduction at high doses in association with suppression of osteoclast development in bone marrow through inhibition of RANKL/OPG ratio and IL-6 mRNA expression in inflammatory synovial tissue.
• PILR alpha is a herpes simplex virus-1 entry coreceptor that associates with glycoprotein B

  Takeshi Satoh, Jun Arii, Tadahiro Suenaga, Jing Wang, Amane Kogure, Junji Uehori, Noriko Arase, Ikuo Shiratori, Shinya Tanaka, Yasushi Kawaguchi, Patricia G. Spear, Lewis L. Lanier, Hisashi Arase

  CELL 132 (6) 935 - 944 0092-8674 2008/03 [Refereed][Not invited]
   

  Glycoprotein B(gB) is one of the essential components for infection by herpes simplex virus-1 (HSV-1). Although several cellular receptors that associate with glycoprotein D(gD), such as herpes virus entry mediator (HVEM) and Nectin-1, have been identified, specific molecules that mediate HSV-1 infection by associating with gB have not been elucidated. Here, we found that paired immunoglobulin-like type 2 receptor (PILR) alpha associates with gB, and cells transduced with PILR alpha become susceptible to HSV-1 infection. Furthermore, HSV-1 infection of human primary cells expressing both HVEM and PILR alpha was blocked by either anti-PILR alpha or anti-HVEM antibody. Our results demonstrate that cellular receptors for both gB and gD are required for HSV-1 infection and that PILR alpha plays an important role in HSV-1 infection as a coreceptor that associates with gB. These findings uncover a crucial aspect of the mechanism underlying HSV-1 infection.
• [Structural basis for the transforming activity of human cancer-related signaling adaptor protein Crk].

  Kobashigawa Y, Tanaka S, Inagaki F

  Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 2 53 (2) 148 - 156 0039-9450 2008/02 [Refereed][Not invited]
  
• Pharmacological cdk inhibitor R-Roscovitine suppresses JC virus proliferation

  Yasuko Orba, Yuji Sunden, Tadaki Suzuki, Kazuo Nagashima, Takashi Kimura, Shinya Tanaka, Hirofumi Sawa

  VIROLOGY 370 (1) 173 - 183 0042-6822 2008/01 [Refereed][Not invited]
   

  The human Polyomavirus JC virus (JCV) utilizes cellular proteins for viral replication and transcription in the host cell nucleus. These cellular proteins represent potential targets for antiviral drugs against the JCV. In this study, we examined the antiviral effects of the pharmacological cyclin-dependent kinase (cdk) inhibitor R-Roscovitine, which has been shown to have antiviral activity against other viruses. We found that Roscovitine significantly inhibited the viral production and cytopathic effects of the JCV in a JCV-infected cell line. Roscovitine attenuated the transcriptional activity of JCV late genes, but not early genes, and also prevented viral replication via inhibiting phosphorylation of the viral early protein, large T antigen. These data suggest that the JCV requires cdks to transcribe late genes and to replicate its own DNA. That Roscovitine exhibited antiviral activity in JCV-infected cells suggests that Roscovitine might have therapeutic utility in the treatment of progressive multifocal leukoencephalopathy (PML). (c) 2007 Elsevier Inc. All rights reserved.
• Transgenic expression of Helicobacter pylori CagA induces gastrointestinal and hematopoietic neoplasms in mouse

  Naomi Ohnishi, Hitomi Yuasa, Shinya Tanaka, Hirofumi Sawa, Motohiro Miura, Atsushi Matsui, Hideaki Higashi, Manabu Musashi, Kazuya Lwabuchi, Misao Suzuki, Gen Yamada, Takeshi Azuma, Masanori Hatakeyama

  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 105 (3) 1003 - 1008 0027-8424 2008/01 [Refereed][Not invited]
   

  Infection with cagA-positive Helicobacter pylori is associated with gastric adenocarcinoma and gastric mucosa-associated lymphoid tissue (MALT) lymphoma of B cell origin. The cagA-encoded CagA protein is delivered into gastric epithelial cells via the bacterial type IV secretion system and, upon tyrosine phosphorylation by Src family kinases, specifically binds to and aberrantly activates SHIP-2 tyrosine phosphatase, a bona fide oncoprotein in human malignancies. CagA also elicits junctional and polarity defects in epithelial cells by interacting with and inhibiting partitioning-defective 1 (PAR1)/microtubule affinity-regulating kinase (MARK) independently of CagA tyrosine phosphorylation. Despite these CagA activities that contribute to neoplastic transformation, a causal link between CagA and in vivo oncogenesis remains unknown. Here, we generated transgenic mice expressing wild-type or phosphorylation-resistant CagA throughout the body or predominantly in the stomach. Wild-type CagA transgenic mice showed gastric epithelial hyperplasia and some of the mice developed gastric polyps and adenocarcinomas of the stomach and small intestine. Systemic expression of wild-type CagA further induced leukocytosis with IL-3/GM-CSF hypersensitivity and some mice developed myeloid leukemias and B cell lymphomas, the hematological malignancies also caused by gain-of-function SHIP-2 mutations. Such pathological abnormalities were not observed in transgenic mice expressing phosphorylation-resistant CagA. These results provide first direct evidence for the role of CagA as a bacterium-derived oncoprotein (bacterial oncoprotein) that acts in mammals and further indicate the importance of CagA tyrosine phosphorylation, which enables CagA to deregulate SHIP-2, in the development of H. pylori-associated neoplasms.
• Regulation of the Il4 gene is independently controlled by proximal and distal 3' enhancers in mast cells and basophils

  Ryouji Yagi, Shinya Tanaka, Yasutaka Motomura, Masato Kubo

  MOLECULAR AND CELLULAR BIOLOGY 27 (23) 8087 - 8097 0270-7306 2007/12 [Refereed][Not invited]
   

  Mast cells and basophills are known to be a critical interleukin 4 (IL-4) source for establishing Th2 protective responses to parasitic infections. Chromatin structure and histone modification patterns in the Il13/Il4 locus of mast cells were similar to those of IL-4-producing type 2 helper T cells. However, using a transgenic approach, we found that Il4 gene expression was distinctly regulated by individual cis regulatory elements in cell types of different lineages. The distal 3' element contained conserved noncoding sequence 2 (CNS-2), which was a common enhancer for memory phenotype T cells, NKT cells, mast cells, and basophils. Targeted deletion of CNS-2 compromised production of IL-4 and several Th2 cytokines in connective-tissue-type and immature-type mast cells but not in basophills. Interestingly, the proximal 3' element containing DNase I-hypersensitive site 4 (HS4), which controls Il4 gene silencing in T-lineage cells, exhibited selective enhancer activity in basophils. These results indicate that CNS-2 is an essential enhancer for IN gene transcription in mast cell but not in basophils. The transcription of the Il4 gene in mast cells and basophils is independently regulated by CNS-2 and HS4 elements that may be critical for lineage-specific Il4 gene regulation in these cell types.
• Clinical usefulness of biliary scope for Pringle's maneuver in laparoscopic hepatectomy

  Shin-Ichiro Maehara, Eisuke Adachi, Mitsuo Shimada, Akinobu Taketomi, Ken Shirabe, Shinji Tanaka, Takashi Maeda, Keisuke Ikeda, Hidefurni Higashi, Yoshihiko Maehara

  JOURNAL OF THE AMERICAN COLLEGE OF SURGEONS 205 (6) 816 - 818 1072-7515 2007/12 [Refereed][Not invited]
  
• Signaling adaptor protein Crk is indispensable for malignant feature of glioblastoma cell line KMG4

  Lei Wang, Kouichi Tabu, Taichi Kimura, Masumi Tsuda, Hua Linghu, Mishie Tanino, Sadao Kaneko, Hiroshi Nishihara, Shinya Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 362 (4) 976 - 981 0006-291X 2007/11 [Refereed][Not invited]
   

  Signaling adaptor protein Crk has been shown to be involved in pathogenesis of human cancers including brain tumor where Crk was reported to be overexpressed. In this study, we addressed whether Crk is indispensable for malignant phenotype of brain tumor. In 20 surgical specimens of glioma, mRNA of both CrkI and CrkII was found to be elevated in malignant tumor. To define a precise role of Crk, we have established Crk-knockdown cell lines of glioblastoma KMG4 by siRNA, and early phase of cell adhesion to laminin was found to be suppressed. Wound healing assay revealed the decreased cell motility in Crk knockdown cells, and suppression of both anchorage-dependent and -independent growth were demonstrated in these cells. Furthermore, in vivo tumor forming potential was also markedly suppressed. These results suggest that Crk is required for early attachment to laminin, cell motility, and growth of glioblastoma cell line KMG4. (C) 2007 Elsevier Inc. All rights reserved.
• Oligodendrocyte lineage transcription factor 2 inhibits the motility of a human glial tumor cell line by activating RhoA

  Kouichi Tabu, Yusuke Ohba, Tadaki Suzuki, Yoshinori Makino, Taichi Kimura, Akiko Ohnishi, Mieko Sakai, Takuya Watanabe, Shinya Tanaka, Hirofurni Sawa

  MOLECULAR CANCER RESEARCH 5 (10) 1099 - 1109 1541-7786 2007/10 [Refereed][Not invited]
   

  The basic helix-loop-helix transcription factor, oligodendrocyte lineage transcription factor 2 (OLIG2), is specifically expressed in the developing and mature central nervous system and plays an important role in oligodendrogenesis from neural progenitors. It is also expressed in various types of glial tumors, but rarely in glioblastoma. Although we previously showed that OLIG2 expression inhibits glioma cell growth, its role in tumorigenesis remains incompletely understood. Here, we investigated the effect of OLIG2 expression on the migration of the human glioblastoma cell line U12-1. In these cells, OLIG2 expression is controlled by the Tet-off system. Induction of OLIG2 expression inhibited both the migration and invasiveness of U12-1 cells. OLIG2 expression also increased the activity of the GTPase RhoA as well as inducing the cells to form stress fibers and focal adhesions. Experiments using short interfering RNA against p27(KiP1) revealed that up-regulation of the p27(Kip1) protein was not essential for RhoA activation, rather it contributed independently to the decreased motility of OLIG2-expressing U12-1 cells. Alternatively, semiquantitative reverse transcription-PCR analysis revealed that mRNA expression of RhoGAP8, which regulates cell migration, was decreased by OLIG2 expression. Furthermore, expression of C3 transferase, which inhibits Rho via ADP ribosylation, attenuated the OLIG2-induced inhibition of cell motility. Imaging by fluorescence resonance energy transfer revealed that in U12-1 cells lacking OLIG2, the active form of RhoA was localized to protrusions of the cell membrane. In contrast, in OLIG2-expressing cells, it lined almost the entire plasma membrane. Thus, OLIG2 suppresses the motile phenotype of glioblastoma cells by activating RhoA.
• Immunohistochemical and gene rearrangement studies of central nervous system lymphomatoid granulomatosis

  Hiroshi Nishihara, Ukihide Tateishi, Tomoo Itoh, Kazuo Nagashima, Shinya Tanaka

  NEUROPATHOLOGY 27 (5) 413 - 418 0919-6544 2007/10 [Refereed][Not invited]
   

  Lymphomatoid granulomatosis (LYG) is a rare multisystem disorder with characteristic angiocentric lymphoproliferative features, most frequently involving the lung, skin, and rarely the CNS. LYG has been classified into three subtypes based on the relative proportions of atypical and inflammatory infiltrating cells. Most systemic LYGs have been shown to be EBV-associated, T-cell rich, B-cell proliferative disorders. Here, we present four cases of LYG arising from the CNS and have analyzed them by immunohistochemistry to assess the phenotype of the infiltrate, and by PCR-SSCP (single-strand conformation polymorphism) analysis for immunoglobulin heavy chain (IgH) and T-cell receptor (TcR) gamma gene rearrangements. Three cases revealed perivascular infiltration of T-cell dominant lymphoid cells, two cases showed monoclonal TcR gamma gene rearrangement, while the remaining case had a B-cell immunophenotype and monoclonal IgH gene rearrangement with EBV genome expression. This is the first report of a gene rearrangement study on CNS-LYG. We confirm that some cases of CNS-LYG are derived from T-cell monoclonal lymphoproliferative disease, although this disease should be classified as a borderline malignancy and should be separated from overt malignant lymphoma of CNS.
• Type I interferon prolongs cell cycle progression via p21(WAF1/CIP1) induction in human colon cancer cells

  Tomonari Katayama, Kazuaki Nakanishi, Hiroshi Nishihara, Naoya Kamiyama, Takahito Nakagawa, Toshiya Kamiyama, Ken Iseki, Shinya Tanaka, Satoru Todo

  INTERNATIONAL JOURNAL OF ONCOLOGY 31 (3) 613 - 620 1019-6439 2007/09 [Refereed][Not invited]
   

  Type I interferon (IFN) was originally identified as an immunomodulatory cytokine because of its antiviral activity. Further characterization of its biological effects revealed a prominent role in the direct control of cell growth and potent immunomodulatory and antiangiogenic actions. IFN-alpha and IFN-SS had both been classified as type I IFN, but differences in their antitumor activities were reported. We confirmed the difference in the antiproliferative activities of IFN-alpha 2b and IFN-SS toward HT29 and SW480 cells. IFN treatment was observed to prolong cell cycle progression; in particular, the accumulation of S-phase population was one of the most characteristic changes. The prolongation of S-phase progression and transition into G2/M-phase was suggested to be a crucial action of type I IFN on colon cancer. Additionally, IFN activated the p21 promoter gene and induced p21(WAFl/CIP1) expression. Furthermore, the cell cycle prolongation effect of IFN was suppressed when p21 expression was downregulated. Therefore, we confirmed that p21(WAFl/CIP1) was a crucial target molecule for the effects of IFN on the cell cycle. Additionally, the ability of p21 induction differed between IFN-alpha 2b and IFN-SS and correlated with their inhibitory activities toward cell growth. We conclude that type I IFN prolongs cell cycle progression by p21(WAFl/CIP1) induction in human colon cancer cells.
• Enteritis necroticans 'pigbel' in a Japanese diabetic adult

  Tomomichi Matsuda, Yuji Okada, Eiji Inagi, Yasushi Tanabe, Yozo Shimizu, Kazuo Nagashima, Jun Sakurai, Masahiro Nagahama, Shinya Tanaka

  PATHOLOGY INTERNATIONAL 57 (9) 622 - 626 1320-5463 2007/09 [Refereed][Not invited]
   

  Enteritis necroticans 'pigbel' is caused by Clostridium perfringens type C but has rarely been reported in developed countries. A 50-year-old Japanese man with untreated diabetes mellitus (DM) presented with diarrhea and abdominal pain. Intraoperative endoscopic and macroscopic examination disclosed segmental annular mucosal lesions characteristic of clostridial enteritis. Clostridial infection type C was verified on pathological, and immunohistochemical analysis. Although rare, the disease is likely to be underdiagnosed. Hence, the pathology and immunohistochemistry of segmental enteritis with annular mucosal lesions should be examined to establish a diagnosis of enteritis necroticans even in mildly affected patients, and especially those with DM.
• ApoE gene deficiency enhances the reduction of bone formation induced by a high-fat diet through the stimulation of p53-mediated apoptosis in osteoblastic cells

  Hideyuki Hirasawa, Shinya Tanaka, Akinori Sakai, Masato Tsutsui, Hiroaki Shimokawa, Hironori Miyata, Sawako Moriwaki, Shumpei Niida, Masako Ito, Toshitaka Nakamura

  JOURNAL OF BONE AND MINERAL RESEARCH 22 (7) 1020 - 1030 0884-0431 2007/07 [Refereed][Not invited]
   

  Osteoblast apoptosis increased in the tibias of apoE(-/-) mice fed with a high-fat diet, decreasing bone formation. The expression of p53 mRNA in marrow adherent cells increased. LDL or oxidized LDL increased apoptosis in the calvarial cells of apoE(-/-) mice. The increase in p53-mediated apoptosis is apparently related to a high-fat diet-induced osteopenia in apoE(-/-) mice. Introduction: The effects of high-fat loading and the apolipoprotein E (apoE) gene on bones have not been elucidated. We hypothesized that apoE gene deficiency (apoE(-/-)) modulates the effects of high-fat loading on bones. Materials and Methods: We assessed this hypothesis using wildtype (WT) and apoE(-/-) mice fed a standard (WTS and ApoES groups) or a high-fat diet (WTHf and ApoEHf groups). The concentration of serum lipid levels and bone chemical markers were measured. Histomorphometry of the femurs was performed using mu CT and a microscope. Bone marrow adherent cells from the femurs were used for colony-forming unit (CFU)-fibroblastic (CFU-f) assay and mRNA expressions analysis. The apoptotic cells in the tibias were counted. TUNEL fluorescein assay and Western analysis were performed in cultures of calvarial cells by the addition of low-density lipoprotein (LDL) or oxidized LDL. Results: In the ApoEHf group, the values of cortical bone volume and trabecular and endocortical bone formation of the femurs decreased, and urinary deoxypyridinoline increased. Subsequent analysis revealed that the number of apoptotic cells in the tibias of the ApoES group increased, and more so in the ApoEHf group. The ratio of alkaline phosphatase-positive CFU-f to total CFU-f was decreased in the ApoEHf group. p53 mRNA expression in adherent cells of the apoE(-/-) mice increased and had a significantly strong positive correlation with serum LDL. TUNEL fluorescein assay of osteoblastic cells revealed an increase of apoptotic cells in the apoE(-/-) mice. The number of apoptotic cells in the apoE(-/-) mice increased with the addition of 100 mu g/ml LDL or oxidized LDL. The p53 protein expression in apoE(-/-) cells exposed to 100 mu g/ml LDL or oxidized LDL increased. Conclusions: We concluded that apoE gene deficiency enhances the reduction of bone formation induced by a high-fat diet through the stimulation of p53-mediated apoptosis in osteoblastic cells.
• Role of growth factor receptor-bound protein 7 in hepatocellular carcinoma

  Shinji Itoh, Akinobu Taketomi, Shinji Tanaka, Norifumi Harimoto, Yo-ichi Yamashita, Shin-ichi Aishima, Takashi Maeda, Ken Shirabe, Mitsuo Shimada, Yoshihiko Maehara

  MOLECULAR CANCER RESEARCH 5 (7) 667 - 673 1541-7786 2007/07 [Refereed][Not invited]
   

  The human growth factor receptor-bound protein 7 (Grb7) is an adaptor molecule and is related to cell invasion. In this present study, we investigated the clinical and biological significance of Grb7 expression in human hepatocellular carcinoma (HCC). We reviewed 64 consecutive patients who had undergone liver resection for HCC, and we investigated the correlation between Grb7 expression and clinical outcome. To analyze the biological behavior of Grb7 in vitro and in vivo, we established Grb7 stable knockdown HCC cells using RNA interference technology. The positive staining of Grb7 protein was correlated with portal venous invasion (P < 0.01), hepatic venous invasion (P < 0.01), and intrahepatic metastasis (P < 0.05). Positive expression of Grb7 was significantly correlated with focal adhesion kinase (FAK) protein levels in HCC (P < 0.01). The Grb7- and FAK-positive group showed a significantly poorer prognosis as compared with the Grb7- and FAK-negative group (P < 0.05). Grb7 knockdown HCC cells exhibited significantly lower levels of invasion potential (P < 0.05) and motility (P < 0.05) than the control cells in vitro; moreover, Grb7 knockdown HCC cells showed delayed onset of the tumors compared with the control cells in vivo. Grb7 expression can modulate the invasive phenotype of HCC. Grb7 plays an important role in HCC progression and is strongly associated with expression of FAK. Grb7 could be a therapeutic target in HCC.
• Flt-1 tyrosine kinase-deficient homozygous mice result in decreased trabecular bone volume with reduced osteogenic potential

  Hajime Otomo, Akinori Sakai, Soshi Uchida, Shinya Tanaka, Makoto Watanuki, Sawako Moriwaki, Shumpei Niida, Toshitaka Nakamura

  BONE 40 (6) 1494 - 1501 8756-3282 2007/06 [Refereed][Not invited]
   

  To clarify the role of Fms-like tyrosine kinase-1 (Flt-1) signaling in bone dynamics, we examined C57BL/6J mice, aged 6, 9 and 16 weeks, with disruption of the flt1 tyrosine kinase domain gene (flt1(TK-/-)) and compared with age-matched wild-type (flt1(TK+/+)) mice. Dynamic histomorphometric analysis confirmed a significant decrease in the values of mineralizing surface (MS/BS), mineral apposition rate (MAR), and bone formation rate (BFR/BS) in the trabecular bone of the proximal tibiae of flt1(TK-/-) mice compared with those inf1(TK+/+) mice. The value of trabecular bone volume (BV/TV) was also significantly reduced in flt1(-/-) mice compared with that in,flt1(TK+/+) mice. The values of osteoclast surface (Oc.S/BS) and osteoclast number (Oc.N/BS) in flt1(TK-/-) mice were somewhat lower than those in flt1(TK+/+) mice. The values of bending load of the femur significantly decreased in flt1(TK-/-) mice. In addition, serum osteocalcin significantly decreased in flt1(TK-/-) mice compared with those in flt1(TK+/+) mice. Furthermore, there was a significant decreased mineralization of bone marrow stromal cultures from flt1(TK-/-) mice. These findings demonstrate that flt1(TK-/-) mice show lower trabecular bone volume than flt1(TK+/+) mice, providing powerful evidence that vascular endothelial growth factor signal pathway through the Flt-1 tyrosine kinase domain could be implicated in osteoblast development. (c) 2007 Elsevier Inc. All rights reserved.
• Structural basis for the transforming activity of human cancer-related signaling adaptor protein CRK

  Yoshihiro Kobashigawa, Mieko Sakai, Masato Naito, Masashi Yokochi, Hiroyuki Kumeta, Yoshinori Makino, Kenji Ogura, Shinya Tanaka, Fuyuhiko Inagaki

  Nature Structural and Molecular Biology 14 (6) 503 - 510 1545-9993 2007/06 [Refereed][Not invited]
   

  CRKI (SH2-SH3) and CRKII (SH2-SH3-SH3) are splicing isoforms of the oncoprotein CRK that regulate transcription and cytoskeletal reorganization for cell growth and motility by linking tyrosine kinases to small G proteins. CRKI shows substantial transforming activity, whereas the activity of CRKII is low, and phosphorylated CRKII has no biological activity whatsoever. The molecular mechanisms underlying the distinct biological activities of the CRK proteins remain elusive. We determined the solution structures of CRKI, CRKII and phosphorylated CRKII by NMR and identified the molecular mechanism that gives rise to their activities. Results from mutational analysis using rodent 3Y1 fibroblasts were consistent with those from the structural studies. Together, these data suggest that the linker region modulates the binding of CRKII to its targets, thus regulating cell growth and motility. © 2007 Nature Publishing Group.
• O-6-methylguanine-DNA methyltransferase is downregulated in transformed astrocyte cells: implications for anti-glioma therapies

  Ken Sasai, Tsuyoshi Akagi, Eiko Aoyanagi, Kouichi Tabu, Sadao Kaneko, Shinya Tanaka

  MOLECULAR CANCER 6 36  1476-4598 2007/06 [Refereed][Not invited]
   

  Background: A novel alkylating agent, temozolomide, has proven efficacious in the treatment of malignant gliomas. However, expression of O-6-methylguanine-DNA methyltransferase (MGMT) renders glioma cells resistant to the treatment, indicating that identification of mechanisms underlying the gene regulation of MGMT is highly required. Although glioma-derived cell lines have been widely employed to understand such mechanisms, those models harbor numerous unidentified genetic lesions specific for individual cell lines, which complicates the study of specific molecules and pathways. Results: We established glioma models by transforming normal human astrocyte cells via retroviral-mediated gene transfer of defined genetic elements and found that MGMT was downregulated in the transformed cells. Interestingly, inhibitors of DNA methylation and histone deacetylation failed to increase MGMT protein levels in the transformed astrocyte cells as well as cultured glioblastoma cell lines, whereas the treatment partially restored mRNA levels. These observations suggest that downregulation of MGMT may depend largely on cellular factors other than promoter-hypermethylation of MGMT genes, which is being used in the clinic to nominate patients for temozolomide treatment. Furthermore, we discovered that Valproic acid, one of histone deacetylase inhibitors, suppressed growth of the transformed astrocyte cells without increasing MGMT protein, suggesting that such epigenetic compounds may be used to some types of gliomas in combination with alkylating agents. Conclusion: Normal human astrocyte cells allow us to generate experimental models of human gliomas by direct manipulation with defined genetic elements, in contrast to tumor-derived cell lines which harbor numerous unknown genetic abnormalities. Thus, we propose that the study using the transformed astrocyte cells would be useful for identifying the mechanisms underlying MGMT regulation in tumor and for the development of rational drug combination in glioma therapies.
• Primary gastric Hodgkin's lymphoma expressing a B-cell profile including Oct-2 and Bob-1 proteins

  Makoto Saito, Shinya Tanaka, Akio Mori, Nobuyasu Toyoshima, Tatsuro Irie, Masanobu Morioka

  INTERNATIONAL JOURNAL OF HEMATOLOGY 85 (5) 421 - 425 0925-5710 2007/06 [Refereed][Not invited]
   

  Classic Hodgkin's lymphoma (cHL) most often involves lymph nodes, and gastric involvement is rare. Hodgkin's and Reed-Sternberg (H-RS) cells in cHL are known to often lack expression of several B-lineage markers, such as CD20, CD79a, Oct-2, and Bob-1. We present an extremely rare case of mixed-cellularity cHL in the stomach in which expression of these B-cells was detected immunohistochemically. The patient was an 83-year-old Japanese woman who developed a sensation of abdominal fullness and appetite loss. Endoscopic and abdominal computed tomography examinations revealed a gastric ulcer lesion and swelling of para-aortic lymph nodes, respectively. A subtotal gastrectomy was performed, and the histopathologic diagnosis was established as a typical cHL compatible with stomach origin. The patient underwent postoperative chemotherapy of 3 cycles of ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine) and has since been in complete remission. Immunohistochemically, the H-RS cells in the cHL were positive not only for CD30 but also for CD20, CD79a, Oct-2, and Bob-1, whereas they were negative for CD3, CD15, CD45, EMA, and ALK1. Our patient may have had an intermediate cHL disease overlapping that of non-Hodgkin's peripheral B-cell lymphoma, possibly reflecting derivation from germinal-center B-cells.
• The corrected blood urea nitrogen predicts the developmental quotient of extremely low-birth-weight infants at the corrected age of 36 months

  Ken Nagaya, Shinya Tanaka, Hiroyuki Kitajima, Masanori Fujimura

  EARLY HUMAN DEVELOPMENT 83 (5) 285 - 291 0378-3782 2007/05 [Refereed][Not invited]
   

  Background: Currently, there are no nutritional indices to predict cognitive function in extremely low-birth-weight (ELBW) infants. Objective: To assess the neonatal blood urea nitrogen (BUN) values in ELBW infants according to their cognitive function at the corrected age of 36 months. Methods: This was a retrospective study that assessed the neonatal factors affecting the developmental outcome in two groups "developmental quotient (DQ)>= 80" and "DQ<80", the groups were divided based on the DQ at the corrected age of 36 months. Between 1996 and 1999, 178 ELBW infants born at <28 weeks of gestation were admitted to our neonatal intensive care unit (NICU), of these, 32 died. Of the surviving 146 infants, 37 infants without any exclusion criteria (that would affect the cognitive function and BUN) except the nutritional factor, were assessed. Area under the curve (AUC) of corrected BUN (CBUN: BUN x 0.5/serum creatinine) from 28 to 84 days of life was used as an index of protein intake. Results: No significant differences were observed between the two groups with regard to the gestational age, birth weight, Z score of birth weight, and sex. However, compared to 15 infants with DQ<80, 22 infants with DQ >= 80 had significantly shorter duration of artificial ventilation and O-2 supplementation, a higher Apgar score at 5 min, and a higher AUC of CBUN. On multiple regression analysis, DQ >= 80 was observed to be significantly correlated with the AUC of CBUN (Odd's ratio 1.03, 95% confidence interval: 1.002-1.06).
• Triggering neural differentiation of ES cells by subtype switching of importin-alpha

  Noriko Yasuhara, Noriko Shibazaki, Shinya Tanaka, Masahiro Nagai, Yasunao Kamikawa, Souichi Oe, Munehiro Asally, Yusuke Kamachi, Hisato Kondoh, Yoshihiro Yoneda

  NATURE CELL BIOLOGY 9 (1) 72 - U93 1465-7392 2007/01 [Refereed][Not invited]
   

  Nuclear proteins are selectively imported into the nucleus by transport factors such as importin-alpha and importin-beta(1,2). Here, we show that the expression of importin-alpha subtypes is strictly regulated during neural differentiation of mouse embryonic stem (ES) cells, and that the switching of importin-alpha subtype expression is critical for neural differentiation. Moreover, reproducing the switching of importin-alpha subtype expression in undifferentiated ES cells induced neural differentiation in the presence of leukaemia inhibitory factor (LIF) and serum, coordinated with the regulated expression of Oct3/4, Brn2 and SOX2, which are involved in ES-neural identity determination. These transcription factors were selectively imported into the nucleus by specific subtypes of importin-alpha. Thus, importin-alpha subtype switching has a major impact on cell differentiation through the regulated nuclear import of a specific set of transcription factors. This is the first study to propose that transport factors should be considered as major players in cell-fate determination.
• IL-7/STAT5 cytokine signaling pathway is essential but insufficient for maintenance of naive CD4 T cell survival in peripheral lymphoid organs

  Yoh-ichi Seki, Jianying Yang, Mariko Okamoto, Shinya Tanaka, Ryo Goitsuka, Michael A. Farrar, Masato Kubo

  JOURNAL OF IMMUNOLOGY 178 (1) 262 - 270 0022-1767 2007/01 [Refereed][Not invited]
   

  Constitutive expression of suppressors of cytokine signaling (SOCS)l in T lineage in vivo attenuated cytokine signaling and resulted in a dramatic reduction in the number of naive CD44(low)CD62L(high) CD4 T cells in the spleen. After adoptive transfer of thymocytes from SOCS1 transgenic mice into normal recipients, naive CD4 T cells rapidly disappeared from the spleen within 1 wk. Likewise, T cell-specific deletion of STAT5a/b in vivo resulted in a similar phenotype characterized by loss of naive CD4 T cells. Thus, STAT5-mediated signaling is crucial for promoting naive T cell survival. However, forced expression of constitutively active STAT5 failed to rescue CD4 T cells in SOCS1 transgenic mice, implying that STAT5 activation is necessary but not sufficient for naive CD4 T cell survival. Although blockade of the IL-7R, a SOCS1 target, resulted in clear inhibition of naive T cell survival, the effect occurred 3 wk after anti-IL-7R Ab treatment, but not at earlier time points. These results suggest that IL-7-mediated STAT5 activation is essential for long-term survival of naive CD4 cells after export from thymus, and that another SOCS1-sensitive cytokine is critical for short-term naive T cell survival.
• Clinical study of therapeutic angiogenesis by autologous peripheral blood stem cell (PBSC) transplantation in 92 patients with critically ischemic limbs

  Akio Kawamura, Takashi Horie, Ichiro Tsuda, Yoshihiro Abe, Masahiro Yamada, Hidetoshi Egawa, Jun-Ichi Iida, Hiromi Sakata, Kazuhiko Onodera, Tohru Tamaki, Hidenori Furui, Kazutaka Kukita, Jun-Ichi Meguro, Motoki Yonekawa, Shinya Tanaka

  Journal of Artificial Organs 9 (4) 226 - 233 1434-7229 2006/12 [Refereed][Not invited]
   

  Patients with critically ischemic limbs due to maintenance hemodialysis and diabetes are increasing in number markedly in Japan. The difficulty of treating critically ischemic limbs is well recognized. Despite active medication and surgical therapy, many critically ischemic limbs are amputated. Ninety-two patients with critically ischemic limbs were treated by transplantation of autologous peripheral blood stem cells (PBSCs). The stem cells were mobilized into the peripheral blood by administration of granulocyte colony stimulating factor (G-CSF). The mobilized mononuclear cells were separated by an apheresis technique using a centrifuge. The separated mononuclear cells contained approximately 4.0 × 107 CD34-positive cells. The collected cell suspension was divided into aliquots of 0.5-1.0 ml and transplanted into the muscle of ischemic limbs at 50-70 transplantation points. At 1.5 months after PBSC transplantation, a strong immunostaining of CD34-positive cells and factor VIII, as well as capillary formation, was observed in the muscles into which stems cells had been transplanted. In each patient tested, the serum vascular endothelial growth factor (VEGF) level increased after stem cell transplantation the mean VEGF level increased by 176%. Of 11 diabetic patients (DM) who were not receiving hemodialysis (HD), there were no amputees regardless of their Fontaine classification. Of 19 patients in the HD(+)DM(-) category, there were no amputations in Fontaine stage I, II, and III patients, whereas three limbs and one toe were amputated in Fontaine stage IV patients. Of 13 patients in the HD(-)DM(+) category, none of the Fontaine stage I, II, or III patients underwent amputation, but six Fontaine stage IV patients underwent amputation. Of 49 patients in the HD(+)DM(+) category, 38 (78%) were classified as Fontaine stage IV, 71% (27/38) of whom had a toe or a limb amputated. In nine patients over 80 years of age, one toe and one limb were amputated. Nondiabetic, nondialyzed patients with ischemic limbs are strongly indicated for stem cell transplantation regardless of Fontaine classification. Therapeutic angiogenesis is effective for critically ischemic limbs resulting from hemodialysis and diabetes until Fontaine stage III, but is of limited effectiveness for stage IV cases. © 2006 The Japanese Society for Artificial Organs.
• Short communication: expression of human endogenous retrovirus-R gene links to differentiation of squamous cells.

  Otsuka N, Miyatake Y, Ishizu A, Tanaka S, Yamamoto Y, Ikeda H, Yoshiki T

  AIDS research and human retroviruses 22 (11) 1148 - 1151 0889-2229 2006/11 [Refereed][Not invited]
  
• Role of suppressor of cytokine signaling in ocular allergy

  Masato Kubo, Akemi Ozaki, Shinya Tanaka, Mariko Okamoto, Atsuki Fukushima

  CURRENT OPINION IN ALLERGY AND CLINICAL IMMUNOLOGY 6 (5) 361 - 366 1528-4050 2006/10 [Refereed][Not invited]
   

  Purpose of review The goal of this article is to evaluate developments in the knowledge of suppressor of cytokine signaling (SOCS) protein ocular allergy and the potential of SOCS proteins as targets for therapeutic strategies. Recent findings The family of proteins designated SOCS proteins plays an important role in Th2-mediated allergic responses through the control of the balance between Th1 Th2 cells. SOCS3 and SOCS5 are perdominantly expressed in Th2 and Th1 cells, respectively, and they reciprocally inhibit the Th1 and Th2 differentiation processes. SOCS3 is highly expressed at the disease site of allergic conjunctivits, and T-cell-specific expression SOCS3 deteriorates clinical and pathological features of allergic conjunctivitis. Reduction of the expression level or inhibition of function clearly reduces the severity of allergic conjunctivitis. On the other hand, constiutive expression of SOCS5, a specific inhibitor of IL-4 signaling, results in reduced eosinophil infiltration. Moreover, negative regulation of the Th2-mediated response by dominat-negative SOCS3 and SOCS5 reduced the incidence of allergic conjunctivitis in a mouse model. Summary The present article summarizes recent findings in terms of a role SOCS protein as a negative regulator in ocular allergy and its clinical application.
• Pancreatic cancer initially presenting with a pseudocyst at the splenic flexure

  Eiji Inagi, Shun Shimodan, Hisato Amizuka, Sanshiro Kigawa, Yozo Shimizu, Kazuo Nagashima, Shinya Tanaka

  PATHOLOGY INTERNATIONAL 56 (9) 558 - 562 1320-5463 2006/09 [Refereed][Not invited]
   

  Pancreatic cancer and pancreatitis associated pseudocyst are not uncommon disorders. However, occurrence of the cancer with an initial manifestation of pseudocyst has been rarely reported. Surgery was performed on a 44-year-old male patient for an abscess-like cavity situated at the mesenteric side of the colon and extending from the splenic flexure to the descending colon. The lesion was verified as a pseudocyst with fat necrosis due to leakage of pancreatic fluid. When further surgery was carried out 1 month later in order to manage the drainage site of the pancreatic fluid, cancer of the pancreas body was detected proximal to the drainage site. The cancer was a moderately differentiated ductal adenocarcinoma with wide peripancreatic infiltration. It is thought that the cancer-associated duct obstruction caused a local pancreatitis resulting in a large communicating pseudocyst, although the exact mechanism remains unresolved. The present case may be instructive in showing physicians that a pseudocyst may obscure the presence of pancreatic cancer.
• Cyclooxygenase-2 selective inhibition suppresses restoration of tibial trabecular bone formation in association with restriction of osteoblast maturation in skeletal reloading after hindlimb elevation of mice

  K Nakai, S Tanaka, A Sakai, M Nagashima, M Tanaka, H Otomo, T Nakamura

  BONE 39 (1) 83 - 92 8756-3282 2006/07 [Refereed][Not invited]
   

  To clarify the role of cyclooxygenase-2 (COX-2) in acute recovery of trabecular bone in reloaded hindlimbs of tail-suspended mice, we administered a COX-2 selective inhibitor in the mice during the reloading period after unloading. Experiments were conducted on 140 male C57BL/6J mice (8 weeks old). They were divided into ground control (GC) and unloading by tail suspension (UL) groups. On day 7, Group GC was divided into Groups GC + Vehicle (Veh) and GC + Celecoxib (Cel), while Group UL mice were fed on the ground [reloading (RL)] after 7-day unloading and were then divided into Groups RL + Veh and RL + Cel. Bone histomorphometry, osteogenic cell development, and mRNA expression of osteogenic molecules were assessed. At 5 days after reloading, the increase of bone formation rate and the ratio of osteocalcin mRNA expression per CFU-f colony in Group RL + Cel were significantly decreased compared with those in Group RL + Veh, while alkaline phosphatase-positive (ALP(+)) CFU-f formation and the ratios of cbfa-1, osterix, and type 1 collagen mRNA expression per CFU-f colony increased to the same levels in both RL groups. At 14 days after reloading, decreased bone volume by unloading in RL + Veh recovered to the same level as that of GC + Veh, but that in RL + Cel did not recover completely. The increase of c-fos mRNA expression in bone marrow cells at 1, 24, and 48 h after reloading, osteocalcin mRNA at 6 h, and osterix mRNA at 24 h were suppressed by COX-2 inhibitor. These data indicate that the COX-2 selective inhibitor celecoxib suppresses the restoration of tibial trabecular bone formation and the acute recovery of trabecular bone. These actions are closely related to restriction of c-fos and osteocalcin mRNA expressions and osteoblast differentiation in bone marrow cells. (c) 2006 Elsevier Inc. All rights reserved.
• Adaptor molecule Crk is required for sustained phosphorylation of Grb2-associated binder 1 and hepatocyte growth factor-induced cell motility of human synovial sarcoma cell lines

  Takuya Watanabe, Masumi Tsuda, Yoshinori Makino, Shin Ichihara, Hirofumi Sawa, Akio Minami, Naoki Mochizuki, Kazuo Nagashima, Shinya Tanaka

  MOLECULAR CANCER RESEARCH 4 (7) 499 - 510 1541-7786 2006/07 [Refereed][Not invited]
   

  Activation of the c-Met receptor tyrosine kinase through its ligand, hepatocyte growth factor (HGF), promotes mitogenic, motogenic, and morphogenic cellular responses. Aberrant HGF/c-Met signaling has been strongly implicated in tumor cell invasion and metastasis. Both HGF and its receptor c-Met have been shown to be overexpressed in human synovial sarcoma, which often metastasizes to the lung; however, little is known about HGF-mediated biological effects in this sarcoma. Here, we provide evidence that Crk adaptor protein is required for the sustained phosphorylation of c-Met-docking protein Grb2-associated binder 1 (Gab1) in response to HGF, leading to the enhanced cell motility of human synovial sarcoma cell lines SY0-1, HS-SY-II, and Fuji. HGF stimulation induced the sustained phosphorylation on Y307 of Gab1 where Crk was recruited. Crk knockdown by RNA interference disturbed this HGF-induced tyrosine phosphorylation of Gab1. By mutational analysis, we identified that Src homology 2 domain of Crk is indispensable for the induction of the phosphorylation on multiple Tyr-X-X-Pro motifs containing Y307 in Gab1. HGF remarkably stimulated cell motility and scattering of synovial sarcoma cell lines, consistent with the prominent activation of Rac1, extreme filopodia formation, and membrane ruffling. Importantly, the elimination of Crk in these cells induced the disorganization of actin cytoskeleton and complete abolishment of HGF-mediated Rac1 activation and cell motility. Time-lapse microscopic analysis revealed the significant attenuation in scattering of Crk knockdown cells following HGF treatment. Furthermore, the depletion of Crk remarkably inhibited the tumor formation and its invasive growth in vivo. These results suggest that the sustained phosphorylation of Gab1 through Crk in response to HGF contributes to the prominent activation of Rac1 leading to enhanced cell motility, scattering, and cell invasion, which may support the crucial role of Crk in the aggressiveness of human synovial sarcoma.
• The interleukin-4 enhancer CNS-2 is regulated by Notch signals and controls initial expression in NKT cells and memory-type CD4 T cells

  S Tanaka, J Tsukada, W Suzuki, K Hayashi, K Tanigaki, M Tsuji, H Inoue, T Honjo, M Kubo

  IMMUNITY 24 (6) 689 - 701 1074-7613 2006/06 [Refereed][Not invited]
   

  Epigenetic changes in chromatin structure at the T helper (Th2) locus correlate with interukin-4 (IL-4) and IL-13 expression during Th2 differentiation. By using a transgenic green fluorescence protein (GFP) reporter system, we show that conserved noncoding sequence-2 (CNS-2), located downstream of the 114 locus, is a constitutively active enhancer in NKT cells as well as in a subset of CD44(hi) memory phenotype CD4(+) T cells. CNS-2 enhancer activity and initial IL-4 expression in CD44(hi) CD4(+) T cells were abolished in mice with a CD4-specific deletion of the transcriptional mediator of Notch signalling, Rbp-j. Depletion of CNS-2 active CD4(+) T cells markedly decreased Th2 differentiation from naive CD4(+) T cells and antigen-specific IgE production after in vivo priming. These findings indicate that Notch-regulated CNS-2 enhancer controls initial IL-4 expression In NKT and memory phenotype CD4(+) T cells and that CNS-2 active CD44(hi) memory phenotype T cells are important in facilitating Th2 differentiation of naive CD4(+) T cells in allergic responses.
• Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS

  H Linghu, M Tsuda, Y Makino, M Sakai, T Watanabe, S Ichihara, H Sawa, K Nagashima, N Mochizuki, S Tanaka

  ONCOGENE 25 (25) 3547 - 3556 0950-9232 2006/06 [Not refereed][Not invited]
   

  Signaling adaptor protein Crk regulates cell motility and growth through its targets Dock180 and C3G, those are the guanine-nucleotide exchange factors (GEFs) for small GTPases Rac and Rap, respectively. Recently, overexpression of Crk has been reported in various human cancers. To de. ne the role for Crk in human cancer cells, Crk expression was targeted in the human ovarian cancer cell line MCAS through RNA interference, resulting in the establishment of three Crk knockdown cell lines. These cell lines exhibited disorganized actin fibers, reduced number of focal adhesions, and abolishment of lamellipodia formation. Decreased Rac activity was demonstrated by pull-down assay and FRET-based time-lapse microscopy, in association with suppression of both motility and invasion by phagokinetic track assay and transwell assay in these cells. Furthermore, Crk knockdown cells exhibited slow growth rates in culture and suppressed anchorage-dependent growth in soft agar. Tumor forming potential in nude mice was attenuated, and intraperitoneal dissemination was not observed when Crk knockdown cells were injected into the peritoneal cavity. These results suggest that the Crk is a key component of focal adhesion and involved in cell growth, invasion, and dissemination of human ovarian cancer cell line MCAS.
• Characterization and application of polyclonal antibodies that specifically recognize JC virus large T antigen

  Y Sunden, T Suzuki, Y Orba, T Umemura, M Asamoto, K Nagashima, S Tanaka, H Sawa

  ACTA NEUROPATHOLOGICA 111 (4) 379 - 387 0001-6322 2006/04 [Refereed][Not invited]
   

  Polyomavirus JC virus (JCV) is the causative agent of progressive multifocal leukoencephalopathy, a fatal demyelinating disease of the central nervous system. Similar to other polyomaviruses, such as simian vacuolating virus 40 (SV40) and BK virus (BKV), JCV is also associated with human tumours. The Polyomavirus early protein large T antigen (TAg) plays a crucial role in tumour pathogenesis. An antibody to SV40 TAg (PAb416), which cross-reacts with TAgs of both JCV and BKV, has been used widely for the detection of JCV and BKV TAgs. As a consequence, it is difficult to discriminate between the TAgs of SV40, BKV and JCV by immunohistochemical analyses. In the present study, we generated JCV TAg-specific polyclonal antibodies (JCT629 and JCT652) by immunization of New Zealand white rabbits with synthetic peptides reproducing the JCV TAg carboxyl-terminal region as immunogens. Immunoblotting analyses indicated that the new antibodies bind specifically to JCV TAg, and not to those of SV40 or BKV. We also demonstrated that these antibodies can be used for immunoprecipitation, immunocytochemical analyses and immunohistochemical staining of routinely processed specimens. In conclusion, the newly generated JCV-specific TAg antibodies may be useful both in the investigation of the pathophysiological function of JCV TAg and in discriminating between Polyomavirus-related clinical samples.
• A novel function of OLIG2 to suppress human glial tumor cell growth via p27(Kip1) transactivation

  K Tabu, A Ohnishi, Y Sunden, T Suzuki, M Tsuda, S Tanaka, T Sakai, K Nagashima, H Sawa

  JOURNAL OF CELL SCIENCE 119 (7) 1433 - 1441 0021-9533 2006/04 [Refereed][Not invited]
   

  The basic helix-loop-helix transcription factor OLIG2 is specifically expressed in cells of the oligodendrocyte lineage. It is also expressed in various tumors originating from glial cells; however, the expression of OLIG2 is rare or weak in glioblastomas, the most malignant gliomas. The role of OLIG2 in glioma remains unclear. To investigate the function of OLIG2 in glial tumor cells, we have established a glioblastoma cell line, U12-1, in which the expression of OLIG2 is induced by the Tet-off system. Induction of OLIG2 resulted in suppression of both the proliferation and anchorage-independent growth of U12-1. It also resulted in an increase in the expression of p27(Kip1). A luciferase assay revealed that the CTF site of the p27(Kip1) gene promoter was essential for OLIG2-dependent activation of p27(Kip1) gene transcription. Electrophoretic mobility shift assays confirmed that a nuclear extract of OLIG2-expressing U12-1 cells contained a protein complex that binds to the CTF site of the p27(Kip1) gene promoter. Furthermore, siRNA against p27(Kip1) rescued the OLIG2-mediated growth and DNA synthesis inhibition of U12-1 cells. These results indicate that OLIG2 suppresses the proliferation of U12-1 and that this effect is mediated by transactivation of the p27(Kip1) gene, and low expression of OLIG2 may be related to the malignant behavior of human glioblastoma.
• Elmo1 inhibits ubiquitylation of Dock180

  Y Makino, M Tsuda, S Ichihara, T Watanabe, M Sakai, H Sawa, K Nagashima, S Hatakeyama, S Tanaka

  JOURNAL OF CELL SCIENCE 119 (5) 923 - 932 0021-9533 2006/03 [Refereed][Not invited]
   

  Dock180, a member of the CDM family of proteins, plays roles in biological processes such as phagocytosis and motility through its association with the signalling adaptor protein Crk. Recently, the complex formation between Dock180 and Elmo1 was reported to function as a bipartite guanine nucleotide exchange factor for Rac. In this study, we demonstrated that the amount of Dock180 increased when Elmo1 was co-expressed. Dock180 was found to be ubiquitylated and Dock180 protein levels could be augmented by treatment with proteasome inhibitor. The ubiquitylation of Dock180 was enhanced by epidermal growth factor (EGF), Crk and adhesion-dependent signals. Furthermore, Elmo1 inhibited labiquitylation of Dock180, resulting in the increase in Dock180 levels. The Elmo1 mutant Delta 531, which encompasses amino acids required for Dock180 binding, preserved the inhibitory effects on ubiquitylation of Dock180. Upon EGF stimulation, both Dock180 and ubiquitin were demonstrated to translocate to the cell periphery by immunofluorescence, and we found ubiquitylation of Dock180 and its inhibition by Elmo1 to occur in cellular membrane fractions by in vivo ubiquitylation assay. These data suggest that Dock180 is ubiquitylated on the plasma membrane, and also that Elmo1 functions as an inhibitor of ubiquitylation of Dock180. Therefore, an ubiquitin-proteasome-dependent protein degradation mechanism might contribute to the local activation of Rac on the plasma membrane.
• Azathioprine suppresses ezrin-radixin-moesin-dependent T cell-APC conjugation through inhibition of Vav guanosine exchange activity on Rac proteins.

  Poppe D, Tiede I, Fritz G, Becker C, Bartsch B, Wirtz S, Strand D, Tanaka S, Galle PR, Bustelo XR, Neurath MF

  Journal of immunology (Baltimore, Md. : 1950) 1 176 640 - 651 0022-1767 2006/01 [Refereed][Not invited]
  
• A case of tanycytic ependymoma arising from the cerebral hemisphere

  Tamio Ito, Yoshimaru Ozaki, Hirohiko Nakamura

  BRAIN TUMOR PATHOLOGY 23 (2) 91 - 95 1433-7398 2006 [Refereed][Not invited]
   

  We report a rare case of tanycytic ependymoma arising from the cerebral hemisphere. A 59-year-old man was admitted to our hospital because of the incidental detection by MRI of a tumor lesion in the right temporo-occipital paratrigonal region. The mass showed low-to isointensity on T-1-weighted images and high intensity on T-2/proton-weighted images. Partial resection was performed using a transsulcal approach to avoid compromising the visual field. Most of the tumor cells showed elongated spindle shapes arranged in dense fascicles. A few true ependymal rosettes and perivascular pseudorosettes, were visible. The tumor cells were positive for GFAP, S-100, and vimentin, but negative for synaptophysin, EMA, and keratin. The MIB-1 labeling index was approximately 1%. Ultrastructurally, the tumor cells had ependymal cell features, such as microvilli and cilia. From these findings, a pathological diagnosis of tanycytic ependymoma was made.
• Induction of p21(WAF1/CIP1) by human synovial sarcoma-associated chimeric oncoprotein SYT-SSX1

  M Tsuda, T Watanabe, T Seki, T Kimura, H Sawa, A Minami, T Akagi, K Isobe, K Nagashima, S Tanaka

  ONCOGENE 24 (54) 7984 - 7990 0950-9232 2005/12 [Refereed][Not invited]
   

  Oncogenic protein provokes cell cycle arrest termed premature senescence. In this process Ras has been known to induce cyclin-dependent kinase inhibitor (CKI) p16(INK4A) in primary fibroblasts. Here, we present a novel finding that human chimeric oncoprotein SYT-SSX1 induces CKI p21(WAF1/CIP1) (p21) for suppression of cell growth. In human synovial sarcoma cell lines, the expression levels of p21 were high and the transcriptional activity of the p21 gene promoter was significantly elevated. The transient expression of SYT- SSX1-induced activation of the p21 gene promoter in human diploid fibroblasts. The N-terminus deletion form of SYT-SSX1, which failed to bind to hBRM one of the chromatin remodeling factors, preserved the p21 induction ability. This effect of SYT-SSX1 was similar in extent in both wild-type and p53-deficient HCT116 cell lines. Furthermore, the introduction of mutation in Sp1/Sp3 binding sites of the p21 gene promoter abolished the SYT-SSX1-induced transcriptional activity of its promoter. In SW13 cells, the stable expression of SYT-SSX1 suppressed cell growth in culture. These results suggest that SYT-SSX1 is able to induce p21 in a manner independent on hBRM and p53 but dependent on Sp1/Sp3.
• Pituicytoma incidentally found at autopsy

  H Takei, JC Goodman, S Tanaka, MB Bhattacharjee, A Bahrami, SZ Powell

  PATHOLOGY INTERNATIONAL 55 (11) 745 - 749 1320-5463 2005/11 [Refereed][Not invited]
   

  Pituicytoma is a rare benign neoplasm, occurring in the sellar and suprasellar regions. Reported herein is a case of asymptomatic pituicytoma, discovered at autopsy, in a 54-year-old Japanese woman. This is the first case report of pituicytoma, found incidentally at autopsy (incidentaloma), in which whole-mounted sections are available for histological and immunohistochemical studies. Grossly, the bisected pituitary gland revealed a round, white to light tan, 7 mm-diameter nodule. Microscopically, whole-mounted sections revealed a well-circumscribed nodule with no fibrous capsule, located mainly in the neurohypophysis and partially compressing the adenohypophysis. The tumor was composed primarily of bipolar, occasionally unipolar, cells with syncytial fibrillary cytoplasm, arranged in short curvilinear fascicles and/or storiform patterns. Unusual histological features were seen, which included a few groups of large pleomorphic tumor cells with abundant, glassy, eosinophilic cytoplasm, occasionally associated with multinucleated giant tumor cells, and scattered Herring bodies within the tumor. Immunohistochemically, the tumor showed diffuse strong expression of glial fibrillary acidic protein, S-100 protein, and vimentin. Epithelial membrane antigen immunoreactivity was focally observed, mainly in the large tumor cells. Distinction from other intrasellar tumors (granular cell tumor and pilocytic astrocytoma) is important. Because the immunohistochemical profiles of these tumors are similar, histological findings are crucial for distinction.
• Segmental and full paternal isodisomy for chromosome 14 in three patients: Narrowing the critical region and implication for the clinical features

  M Kagami, G Nishimura, T Okuyama, M Hayashidani, T Takeuchi, S Tanaka, F Ishino, K Kurosawa, T Ogata

  AMERICAN JOURNAL OF MEDICAL GENETICS PART A 138A (2) 127 - 132 1552-4825 2005/10 [Refereed][Not invited]
   

  We report on segmental and full paternal isodisomy for chromosome 14 in three previously unreported Japanese patients. Patient 1 was a 5(6)/(12)-year-old girl, Patient 2 was a male neonate, and Patient 3 was a 6(7)/(12)-year-old girl. Physical examination at birth showed various somatic features characteristic of paternal uniparental disomy for chromosome 14 (upd(14)pat) such as hairy forehead, protruding philtrum, micrognathia, small thorax, and abdominal wall defects in Patients 1-3, and the constellation of somatic features was persistently observed in Patients 1 and 3. Radiological studies at birth delineated unique bell-shaped thorax with coat-hanger appearance of the ribs in Patients 1-3, but the thoracic deformity ameliorated in Patients 1 and 3 by mid childhood. Chromosome analysis showed a 46,XX karyotype in Patients I and 3 and was not performed in Patient 2. Microsatellite analysis indicated full paternal isodisomy for chromosome 14 in Patients 1 and 2 and segmental paternal isodisomy for chromosome 14 distal to D14S981 at 14q23.3 in Patient 3. Methylation specific PCR assay for the differentially methylated region (DMR) of GTL2 at l4q32 yielded positive products with methylated allele specific primers and no products with unmethylated allele specific primers in Patients 1-3. Since clinical phenotype was similar between Patient 3 with segmental upd(14)pat and Patients 1 and 2 with full upd(14)pat, the results are keeping with the 14q32 localized imprinted genes as the critical components of the phenotype observed in upd(14)pat and help narrow the search for additional genes to the similar to 40 Mb region distal to D14S981. Furthermore, it is likely that the characteristic thoracic deformity ameliorates with age. (c) 2005 Wiley-Liss, Inc.
• Identification of FEZ1 as a protein that interacts with JC virus agnoprotein and microtubules - Role of agnoprotein-induced dissociation of FEZ1 from microtubules in viral propagation

  T Suzuki, Y Okada, S Semba, Y Orba, S Yamanouchi, S Endo, S Tanaka, T Fujita, S Kuroda, K Nagashima, H Sawa

  JOURNAL OF BIOLOGICAL CHEMISTRY 280 (26) 24948 - 24956 0021-9258 2005/07 [Refereed][Not invited]
   

  The human polyomavirus JC virus (JCV) is the causative agent of a fatal demyelinating disease, progressive multifocal leukoencephalopathy, and encodes six major proteins, including agnoprotein. Agnoprotein colocalizes with microtubules in JCV-infected cells, but its function is not fully understood. We have now identified fasciculation and elongation protein zeta 1 (FEZ1) as a protein that interacted with JCV agnoprotein in a yeast two-hybrid screen of a human brain cDNA library. An in vitro binding assay showed that agnoprotein interacted directly with FEZ1 and microtubules. A microtubule co-sedimentation assay revealed that FEZ1 also associates with microtubules and that agnoprotein induces the dissociation of FEZ1 from microtubules. Agnoprotein inhibited the promotion by FEZ1 of neurite outgrowth in PC12 cells. Conversely, overexpression of FEZ1 suppressed JCV protein expression and intracellular trafficking in JCV-infected cells. These results suggest that FEZ1 promotes neurite extension through its interaction with microtubules, and that agnoprotein facilitates JCV propagation by inducing the dissociation of FEZ1 from microtubules.
• Reduced expression of platelet endothelial cell adhesion molecule-1 in bone marrow cells in mice after skeletal unloading

  M Sakuma-Zenke, A Sakai, S Nakayamada, N Kunugita, T Tabata, S Uchida, S Tanaka, T Mori, K Nakai, Y Tanaka, T Nakamura

  JOURNAL OF BONE AND MINERAL RESEARCH 20 (6) 1002 - 1010 0884-0431 2005/06 [Refereed][Not invited]
   

  One week of tail suspension significantly decreased the expression of PECAM-1 in mouse tibial bone marrow cells but not those of a number of other vascular factors. Anti-PECAM-1 antibody suppressed both ALP(+) CFU-f formation and ALP production under co-culture of the osteoblastic cell line and the PECAM-1(+) endothelial cell line. This study suggests that the reduced ALP activity after skeletal unloading is related to downregulation of PECAM-1 expression in bone marrow cells in mice. Introduction: Vascular factors play a role in bone development and regeneration. We tested the hypothesis that skeletal unloading reduces osteogenic potential by inhibiting the molecules related to angiogenesis and/or vasculogenesis in bone marrow cells. Materials and Methods: Eight-week-old male mice were assigned to three groups after acclimatization for 1 week: ground control (GC), tail suspension (TS), and reloading after 7-day TS (RL). Bilateral tibial and humeral samples were used for analyses. MC3T3-E1, a mouse osteoblastic cell line, and EOMA and ISOS-1, mouse endothelial cell lines, were also used. Results: Flow cytometric analysis revealed that 7-day TS significantly decreased the expression of platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) in tibial bone marrow cells, but not those of angiopoietin-1, angiopoietin-2, Flk-1 (vascular endothelial growth factor receptor-2), and vascular endothelial cadherin. The expression of PECAM-1 in tibial marrow cells was reduced at day 3 of TS to 80% and still showed significantly low levels at day 7 of TS to 72% of that at the respective days of GC. This decreased expression of PECAM-1 after 7-day TS showed the GC level at 5-day reloading after 7-day TS. However, the expression of PECAM-1 in humeral marrow cells (internal bone marrow control) after TS and RL remained unchanged and equivalent to that of GC. The expression level of PECAM-1 mRNA was significantly lower at day 7 of F TS to 62% of that in GC. Double labeling analyses revealed that PECAM-1(+) cells mostly consisted of endothelial cells and partially of granulocytes. In bone marrow cell cultures, the formation of alkaline phosphatase (ALP)(+) colony forming units-fibroblastic was significantly reduced in the presence of anti-PECAM-1 antibody in the medium compared with the presence of immunoglobulin G (0.025 times as much as ALP production with immunoglobulin G). ALP production by cultured MC3T3-E1 was enhanced in combination with PECAM-1(+) EOMA (1.8 times as much as ALP production by MC3T3-E1 alone), but not in combination with PECAM-1(-) ISOS-1. Anti-PECAM-1 antibody inhibited the increase in ALP production under co-culture with EOMA. Conclusions: Our data show that the reduced ALP activity after skeletal unloading is closely correlated with reduced expression of PECAM-1 in bone marrow cells. We speculate that the loss of osteogenic potential after skeletal unloading is caused by the suppression of PECAM-1 signaling on endothelial cellular surface.
• A case of cervicomedullary junction tanycytic ependymoma associated with marked cyst formation

  Tamio Ito, Yoshimaru Ozaki, Jyoji Nakagawara, Hirohiko Nakamura, Shinya Tanaka, Kazuo Nagashima

  Brain Tumor Pathology 22 (1) 29 - 33 1433-7398 2005/06 [Refereed][Not invited]
   

  Tanycytic ependymomas are a subtype of ependymomas that were formally recognized as a new pathological entity in the latest World Health Organization (WHO) classification of 2000. They occur mostly in the spinal cord. Only a few reports have analyzed the proliferative potentials of these tumors however, it has been reported that the MIB-1 labeling index of tanycytic ependymoma is lower than that of other subtypes of WHO grade II ependymomas. We report a rare case of cervicomedullary junction tanycytic ependymoma associated with marked cyst formation. A 62-year-old man had a history of progressive gait disturbance, diplopia, and swallowing disturbance over a one-month period prior to admission. Magnetic resonance imaging (MRI) showed a cystic mass with a mural nodule at the cervicomedullary junction with Gd-DTPA enhancement. Cyst-subarachnoid shunt was performed using a far lateral approach. After 6 years, however, the man was readmitted to the hospital because of reaccumulation of the cyst. Partial removal of a mural nodule and a cyst-subarachnoid shunt were performed simultaneously by a midline suboccipital approach. The pathological diagnosis was tanycytic ependymoma. Postoperatively, the patient recovered well and was discharged from the hospital without further treatment. Most of the tumor cells had small, round nuclei pleomorphism was minimal. The cytoplasm was dilated. The tumor cells were positive for EMA and s-100, and negative for CD-34. GFAP was not determined due to difficulty caused by background glial processes. The MIB-1 labeling index was less than 1%. Ultrastructurally, the tumor cells had ependymal cell features, such as desmosomes and microvilli. Based on these findings, the pathological diagnosis was tanycytic ependymoma. © 2005 The Japan Society of Brain Tumor Pathology.
• Dissociation of heterochromatin protein 1 from lamin B receptor induced by human polyomavirus agnoprotein: role in nuclear egress of viral particles

  Y Okada, T Suzuki, Y Sunden, Y Orba, S Kose, N Imamoto, H Takahashi, S Tanaka, WW Hall, K Nagashima, H Sawa

  EMBO REPORTS 6 (5) 452 - 457 1469-221X 2005/05 [Refereed][Not invited]
   

  The nuclear envelope is one of the chief obstacles to the translocation of macromolecules that are larger than the diameter of nuclear pores. Heterochromatin protein 1 (HP1) bound to the lamin B receptor (LBR) is thought to contribute to reassembly of the nuclear envelope after cell division. Human polyomavirus agnoprotein ( Agno) has been shown to bind to HP1 alpha and to induce its dissociation from LBR, resulting in destabilization of the nuclear envelope. Fluorescence recovery after photobleaching showed that Agno increased the lateral mobility of LBR in the inner nuclear membrane. Biochemical and immunofluorescence analyses showed that Agno is targeted to the nuclear envelope and facilitates the nuclear egress of polyomavirus-like particles. These results indicate that dissociation of HP1a from LBR and consequent perturbation of the nuclear envelope induced by polyomavirus Agno promote the translocation of virions out of the nucleus.
• The newly established human hepatocyte cell line: application for the bioartificial liver

  N Harimoto, A Taketomi, D Kitagawa, Y Kuroda, S Itoh, T Gion, S Tanaka, K Shirabe, M Shimada, Y Maehara

  JOURNAL OF HEPATOLOGY 42 (4) 557 - 564 0168-8278 2005/04 [Refereed][Not invited]
   

  Background/Aims: Human hepatocyte cell lines are reported to lose many of their biochemical functions in a hybrid artificial liver support system (HALSS). Differentiation therapy is useful to up-regulate liver function. Methods: The human hepatoblastoma cell line HepG2 was transfected with HSV/tk gene. Albumin synthesis and ammonia removal activity were evaluated when HepG2/tk was cultured with histone deacetylase inhibitor (FR228) and peroxisome proliferator activated receptor-gamma ligand (pioglitazone). To investigate the function of HepG2/tk in vivo, cell transplantation for 90% hepatectonized rats was conducted. Results: We established stable cell lines which expressed HSV/tk and were sensitive to gancyclovir in vitro and in vivo. Both albumin synthesis rate and ammonia removal rate improved for HepG2/tk incubated with FR228 and pioglitazone for 3 days, which induced nuclear transport of p21. Rats with intrasplenic injection of HepG2/tk precultured for 3 days with FR228 and pioglitazone survived significantly longer than the control rats. The ammonia and total bilirubin concentrations were significantly lower in the test group than in the control group. The injection of gancyclovir inhibited the prolonged survival of the rats with precultured HepG2/tk. Conclusions: HepG2/tk is safe as well as enhancing high levels of liver function. It will be a potential cell source for HALLS in the future. (C) 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
• Bisphosphonate (YM529) delays the repair of cortical bone defect after drill-hole injury by reducing terminal differentiation of osteoblasts in the mouse femur

  M Nagashima, A Sakai, S Uchida, S Tanaka, M Tanaka, T Nakamura

  BONE 36 (3) 502 - 511 8756-3282 2005/03 [Refereed][Not invited]
   

  We evaluated the effects of YM529, a nitrogen-containing bisphosphonate, on the repair of cortical bone after drill-hole injury at the tissue-, cell- and gene-levels in the femur of mice. Eight-week-old male C57BL/6N mice were treated with an intravenous injection of 0.01, 0.05 and 0.1 mg/kg body weight (BW) of YM529, or the vehicle (VC) once a week from 8 weeks of age until sacrifice. At 10 weeks of age (day 0), a drill-hole was made in the diaphysis of bilateral femurs. Femoral specimens were obtained at days 3, 5, 7, 10, 14, 21 and 28 after surgery. Histology and histomorphometry confirmed the early woven bone fort-nation in 7 days after injury in all four groups, but the following lamellar bone repair in the cortical tissue area delayed only in the YM529-treated groups. Since the findings were not dose-dependent, following evaluations were performed in VC and YM529 0.1 mg/kg BW dose groups. Calcein-labeled surface of regenerated bone decreased at day 21 in the YM529 group. At day 0, CFU-f number and mineralized nodule area that developed from marrow cells were significantly smaller in YM529 group than in VC group. At day 5, however, these values increased to levels similar to those in VC group. The mRNA expression levels of BMP-2, cbfa 1, osterix, type I collagen, and osteocalcin in the injured bone and marrow cells at days 3 and 5 were similar in the two groups, but were higher in YM529 group at day 7 compared with that in the VC group. At day 14, the levels of these mRNAs were still high, while that of osteocalcin was significantly reduced compared to the VC group. These data indicate that the action of YM529 on bone formation is bimodal, stimulatory on the developments of osteogenic cells for the woven bone regeneration and inhibitory on the terminal differentiation of osteoblasts for the later remodeling, consequently leading to a delay in the lamellar bone healing in the cortical tissue area. (c) 2004 Elsevier Inc. All rights reserved.
• Establishment of an immunoscreening system using recombinant VP1 protein for the isolation of a monoclonal antibody that blocks JC virus infection

  C Henmi, H Sawa, H Iwata, Y Orba, S Tanaka, K Nagashima

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 327 (1) 242 - 251 0006-291X 2005/02 [Refereed][Not invited]
   

  Polyomavirus JC (JCV) infection causes the fatal human demyelinating disease, progressive multifocal leukoencephalopathy. Although the initial interaction of JCV with host cells occurs through direct binding of the major viral capsid protein (VP1) with cell-surface molecules possessing sialic acid, these molecules have not yet been identified. In order to isolate monoclonal antibodies which inhibit attachment of JCV, we established an immunoscreening system using virus-like particles consisting of the VP1. Using this system, among monoclonal antibodies against the cell membrane fraction from JCV-permissive human neuroblastoma IMR-32 cells, we isolated a monoclonal antibody designated as 24132 that specifically inhibited attachment and infection of JCV to IMR-32 cells. The antibody 24132 recognized a single molecule of around 60 kDa in molecular weight in the IMR-32 membrane fraction. Immunohistochemical staining with 24132 demonstrated immunoreactivity in the cell membrane of JCV-permissive cell lines and glial cells of the human brain. These results suggested that the molecule recognized by 24132 plays a role in JCV infection, and that it might participate as a receptor or a co-receptor in JCV attachment and entry into the cells. (C) 2004 Elsevier Inc. All rights reserved.
• Investigation of Simian virus 40 large T antigen in 18 autopsied malignant mesothelioma patients in Japan

  M Jin, H Sawa, T Suzuki, K Shimizu, Y Makino, S Tanaka, T Nojima, Y Fujioka, M Asamoto, N Suko, M Fujita, K Nagashima

  JOURNAL OF MEDICAL VIROLOGY 74 (4) 668 - 676 0146-6615 2004/12 [Refereed][Not invited]
   

  It has been reported that Simian virus 40 (SV40) is linked to human beings by inoculation of contaminated poliovaccines and may have a role in the etiology of malignant mesothelioma. However, there have been no reports describing the relationship between SV40 and malignant mesothelioma in Japan. A study was undertaken to investigate whether SV40 was related to patients of malignant mesothelioma in Japan by the polymerase chain reaction (PCR) assay, DNA sequence analysis, and immunohistochemical methods. Paraffin-embedded samples of the 18 autopsied patients with pleural malignant mesothelioma were collected from five hospitals in Japan. After isolation of DNA from paraffin blocks, PCR analyses followed by sequencing were performed using three different sets of primers for detection of SV40 large T antigen (TAg) gene. All 18 malignant mesothelioma samples were also immunohistochemically evaluated for expression of SV40 TAg protein with two different anti-SV40 TAg antibodies. SV40 TAg genome was detected in eight malignant mesothelioma cases. Only one of three primer pairs successfully amplified SV40 genome in the samples, whereas all pairs yielded a PCR product in the controls, suggesting a low content of virus DNA. No immunopositive staining for SV40 TAg was found in any of the samples. This study shows that SV40 genome was present in a subset of Japanese malignant mesothelioma patients who were unlikely to have received a contaminated polio vaccine based on their age.
• Crk associates with ERM proteins and promotes cell motility toward hyaluronic acid

  M Tsuda, Y Makino, T Iwahara, H Nishihara, H Sawa, K Nagashima, H Hanafusa, S Tanaka

  JOURNAL OF BIOLOGICAL CHEMISTRY 279 (45) 46843 - 46850 0021-9258 2004/11 [Refereed][Not invited]
   

  Cell migration is a well organized process regulated by the extracellular matrix-mediated cytoskeletal reorganization. The signaling adaptor protein Crk has been shown to regulate cell motility, but its precise role is still under investigation. Herein, we report that Crk associates with ERM family proteins ( including ezrin, radixin, and moesin), activates RhoA, and promotes cell motility toward hyaluronic acid. The binding of Crk with ERMs was demonstrated both by transient and stable protein expression systems in 293T cells and 3Y1 cells, and it was shown that v-Crk translocated the phosphorylated form of ERMs to microvilli in 3Y1 cells by immunofluorescence and immunoelectron microscopy. This v-Crk-dependent formation of microvilli was suppressed by inhibitors of Rho-associated kinase, and the activity of RhoA was elevated by coexpression of c-Crk-II and ERMs in 3Y1 cells. In concert with the activation of RhoA by Crk, Crk was found to associate with Rho-GDI, which has been shown to bind to ERMs. Furthermore, upon hyaluronic acid treatment, coexpression of c-Crk-II and ERMs enhanced cell motility, whereas the sole expression of c-Crk-II or either of the ERMs decreased the motility of 3Y1 cells. These results suggest that Crk may be involved in regulation of cell motility by a hyaluronic acid-dependent mechanism through an association with ERMs.
• Skeletal unloading alleviates the anabolic action of intermittent PTH(1-34) in mouse tibia in association with inhibition of PTH-induced increase in c-fos mRNA in bone marrow cells

  S Tanaka, A Sakai, M Tanaka, H Otomo, N Okimoto, T Sakata, T Nakamura

  JOURNAL OF BONE AND MINERAL RESEARCH 19 (11) 1813 - 1820 0884-0431 2004/11 [Refereed][Not invited]
   

  We analyzed the effect of unloading by tail suspension on the anabolic action of intermittent PTH in the tibia of growing mice. Unloading alleviated the PTH-induced increase of bone formation and accelerated bone resorption, consequently reducing bone mass. Reduction of the PTH-induced anabolic actions on bone was associated with unloading, which was apparently related to suppression of c-fos mRNA expression in bone marrow.
• Natural killer T cells accelerate atherogenesis in mice

  Y Nakai, K Iwabuchi, S Fujii, N Ishimori, N Dashtsoodol, K Watano, T Mishima, C Iwabuchi, S Tanaka, JS Bezbradica, T Nakayama, M Taniguchi, S Miyake, T Yamamura, A Kitabatake, S Joyce, L Van Kaer, K Onoe

  BLOOD 104 (7) 2051 - 2059 0006-4971 2004/10 [Refereed][Not invited]
   

  We have investigated the potential role of CD1d-restricted natural killer T (NKT) cells in the development of atherosclerosis in mice. When fed an atherogenic diet (AD), NKT cell-deficient CD1d(-/-) mice had significantly smaller atherosclerotic lesions than AD-fed C57BL/6 (wild-type [WT]) mice. A significant reduction in atherosclerotic lesions was also demonstrated in AD-fed, low-density lipoprotein receptor-deficient (Ldlr(-/-)) mice reconstituted with CD1d(-/-) bone marrow cells compared with the lesions observed in Ldlr(-/-) mice reconstituted with WT marrow cells. In addition, repeated injections of a-GalCer or the related glycolipid OCH to apolipoprotein E knockout (apoE(-/-)) mice during the early phase of atherosclerosis significantly enlarged the lesion areas compared with mice injected with vehicle control. However, administering alpha-GalCer to apoE(-/-) mice with established lesions did not significantly increase the lesion area but considerably decreased the collagen content. Atherosclerosis development in either AD-fed WT or apoE(-/-) mice was associated with the presence of Valpha14Jalpha18 transcripts in the atherosclerotic arterial walls, indicating that NKT cells were recruited to these lesions. Thioglycolate-elicited macrophages pulsed with oxidized low-density lipoproteins expressed enhanced CD1d levels and induced NKT cells to produce interferon-gamma, a potentially proatherogenic T-helper 1 (T(H)1) cytokine. Collectively, we conclude that NKT cells are proatherogenic in mice. (C) 2004 by The American Society of Hematology.
• Interplay of SOX and POU factors in regulation of the Nestin gene in neural primordial cells

  S Tanaka, Y Kamachi, A Tanouchi, H Hamada, NH Jing, H Kondoh

  MOLECULAR AND CELLULAR BIOLOGY 24 (20) 8834 - 8846 0270-7306 2004/10 [Refereed][Not invited]
   

  Intermediate-filament Nestin and group B1 SOX transcription factors (SOX1/2/3) are often employed as markers for neural primordium, suggesting their regulatory link. We have identified adjacent and essential SOX and POU factor binding sites in the Nestin neural enhancer. The 30-bp sequence of the enhancer including these sites (Nes30) showed a nervous system-specific and SOX-POU-dependent enhancer activity in multimeric forms in transfection assays and was utilized in assessing the specificity of the synergism; combinations of either group B1 or group C SOX (SOX11) with class III POU proved effective. In embryonic day 13.5 mouse spinal cord, Nestin was expressed in the cells with nuclei in the ventricular and subventricular zones. SOX1/2/3 expression was confined to the nuclei of the ventricular zone; SOX11 localized to the nuclei of both subventricular (high-level expression) and intermediate (low-level expression) zones. Class III POU (Brn2) was expressed at high levels, localizing to the nucleus in the ventricular and subventricular zones; moderate expression was observed in the intermediate zone, distributed in the cytoplasm. These data support the model that synergic interactions between group B1/C SOX and class III POU within the nucleus determine Nestin expression. Evidence also suggests that such interactions are involved in the regulation of neural primordial cells.
• Regulation of mineral-to-matrix ratio of lumbar trabecular bone in ovariectomized rats treated with risedronate in combination with or without vitamin K-2

  H Otomo, A Sakai, S Ikeda, S Tanaka, M Ito, RJ Phipps, T Nakamura

  JOURNAL OF BONE AND MINERAL METABOLISM 22 (5) 404 - 414 0914-8779 2004/09 [Refereed][Not invited]
   

  The relationship between bone turnover and bone tissue and material properties was examined in ovariectomized (OVX) rats treated with risedronate in combination with or without vitamin K-2. Seventy female rats, 18 weeks of age, were assigned to 7 groups (n=10): sham-operated + vehicle control; OVX + vehicle control; OVX + risedronate 0.1, 0.5, or 2.5 mg/kg/day po; OVX + vitamin K-2 similar to30 mg/kg/day po; OVX + vitamin K-2 (similar to30 mg/kg/day) and risedronate (0.5 mg/kg/day). Treatments were given daily for 9 months. To assess bone turnover, we measured serum osteocalcin and urinary deoxypyridinoline at 0, 3, and 9 months. To assess vertebral and femoral tissue and material properties, bone mass, bone mineral density (BMD by DXA), trabecular bone structure (vertebra: 3D-muCT), cortical bone structure (femur: histomorphometry), biomechanical properties, and mineral properties (mineral-to-matrix and carbonate-to-phosphate ratios by Fourier transform infrared microspectroscopy) were measured ex vivo at 9 months. Ovariectomy increased bone turnover and induced significant loss of bone mass/density, structure, mineral properties (mineral-to-matrix ratio), and strength. Risedronate produced dose-dependent inhibition of the ovariectomy-induced increase in turnover and loss of bone mass/density, structure, mineral-to-matrix ratio, and strength, with a lowest effective dose of 0.1-0.5 mg/kg/day. High-dose risedronate (2.5 mg/kg/day) did not induce increases in any parameter above that of sham control. Vitamin K-2 had no effects. In the OVX groups, urinary deoxypyridinoline at 3 and 9 months correlated significantly with vertebral BMD, trabecular bone volume, ultimate load, stiffness, and mineral-to-matrix ratio, and with femoral BMD, cortical area, and ultimate load. These results support the concept that changes in bone tissue and material properties can result directly from changes in bone turnover. Different effects among different drugs on material properties, including mineral-to-matrix ratio, may reflect differences in the relative rate and magnitude of osteoclastic bone resorption and osteoblastic primary bone mineralization.
• Inhibition of virus production in JC virus-infected cells by postinfection RNA interference

  Y Orba, H Sawa, H Iwata, S Tanaka, K Nagashima

  JOURNAL OF VIROLOGY 78 (13) 7270 - 7273 0022-538X 2004/07 [Refereed][Not invited]
   

  RNA interference has been applied for the prevention of virus infections in mammalian cells but has not succeeded in eliminating infections from already infected cells. We now show that the transfection of JC virus-infected SVG-A human glial cells with small interfering RNAs that target late viral proteins, including agnoprotein and VP1, results in a marked inhibition both of viral protein expression and of virus production. RNA interference directed against JC virus genes may thus provide a basis for the development of new strategies to control infections with this polyomavirus.
• Nuclear entry mechanism of the human polyomavirus JC virus-like particle - Role of importins and the nuclear pore complex

  QM Qu, H Sawa, T Suzuki, S Semba, C Henmi, Y Okada, M Tsuda, S Tanaka, WJ Atwood, K Nagashima

  JOURNAL OF BIOLOGICAL CHEMISTRY 279 (26) 27735 - 27742 0021-9258 2004/06 [Refereed][Not invited]
   

  JC virus (JCV) belongs to the polyomavirus family of double-stranded DNA viruses and causes progressive multifocal leukoencephalopathy in humans. Although transport of virions to the nucleus is an important step in JCV infection, the mechanism of this process has remained unclear. The outer shell of the JCV virion comprises the major capsid protein VP1, which possesses a putative nuclear localization signal (NLS), and virus-like particles (VLPs) consisting of recombinant VP1 exhibit a virion-like structure and physiological functions ( cellular attachment and intracytoplasmic trafficking) similar to those of JCV virions. We have now investigated the mechanism of nuclear transport of JCV with the use of VLPs. Wild-type VLPs (wtVLPs) entered the nucleus of most HeLa or SVG cells. The virion structure of VLPs was preserved during transport to the nucleus as revealed by confocal microscopy of cells inoculated with fluorescein isothiocyanate-labeled wtVLPs containing packaged Cy3. The nuclear transport of wtVLPs in digitonin-permeabilized cells was dependent on the addition of importins alpha and beta and was prevented by wheat germ agglutinin or by antibodies to the nuclear pore complex. The nuclear entry of VLPs composed of VP1 with a mutated NLS was greatly inhibited, compared with that of wtVLPs, in both intact and permeabilized cells. Unlike wtVLPs, the mutant VLPs did not bind to importins alpha or beta. Limited proteolysis analysis revealed that the NLS of VP1 was exposed on the surface of wtVLPs. These results suggest that JCV VLPs bind to cellular importins via the NLS of VP1 and are transported into the nucleus through the nuclear pore complex.
• Prostaglandin E-2 receptor (EP4) selective agonist (ONO-4819.CD) accelerates bone repair of femoral cortex after drill-hole injury associated with local upregulation of bone turnover in mature rats

  M Tanaka, A Sakai, S Uchida, S Tanaka, M Nagashima, T Katayama, K Yamaguchi, T Nakamura

  BONE 34 (6) 940 - 948 8756-3282 2004/06 [Refereed][Not invited]
   

  Prostaglandin E-2 (PGE(2)) is essential for fracture healing. Systemic administration of EP4 ligands such as PGE(2) and other synthetic EP4 agonists appears to transduce anabolic signals by binding to receptor EP4. Therefore, the present study was designed to test whether administration of EP4 agonist accelerates the healing of drill-hole injury in the femoral diaphysis. After surgery, a total of 128 Wistar rats, at the age of 12 weeks, were assigned to basal control (n = 8), and three groups with respective doses of 0 (vehicle control), 10 (low-dose), and 30 (high-dose) mug/kg body weight of the agent were subcutaneously injected twice a day. Femoral specimens were obtained at 0, 5, 7, 14, 2 1, and 28 days. In EP4 agonist-treated groups, the total bone volume of the regenerating bone in the defect did not significantly differ, but the regenerated cortical bone volume measured by histomorphometry and cortical bone mineral content (Ct. BMC) by pQCT dose-dependently increased at 14 and 21 days compared to the control. In the high-dose group, the value of osteoclast surface significantly increased compared with that in the control at 14 days. Expression levels of osteocalcin and TRAP mRNAs in the injured tissue increased at 14 days. Expression levels of EP4, BMP-2, and RANKL mRNAs increased at 7 days in the high-dose group. The bone mineral values of the lumbar bone at 28 days, measured by DXA, did not differ in the three groups. These data indicated that systemic administration of EP4 agonist ONO-4819.CD accelerated cortical bone healing after drill-hole injury by upregulating the local turnover of the regenerating bone. (C) 2004 Elsevier Inc. All rights reserved.
• Transcriptional co-activator activity of SYT is negatively regulated by BRM and Brg1.

  Ishida M, Tanaka S, Ohki M, Ohta T

  Genes to cells : devoted to molecular & cellular mechanisms 5 9 (5) 419 - 428 1356-9597 2004/05 [Refereed][Not invited]
  
• Helicobacter pylori CagA induces Ras-independent morphogenetic response through SHP-2 recruitment and activation

  H Higashi, A Nakaya, R Tsutsumi, K Yokoyama, Y Fujii, S Ishikawa, M Higuchi, A Takahashi, Y Kurashima, Y Teishikata, S Tanaka, T Azuma, M Hatakeyama

  JOURNAL OF BIOLOGICAL CHEMISTRY 279 (17) 17205 - 17216 0021-9258 2004/04 [Refereed][Not invited]
   

  The CagA protein of Helicobacter pylori, which is injected from the bacteria into bacteria-attached gastric epithelial cells, is associated with gastric carcinoma. CagA is tyrosine-phosphorylated by Src family kinases, binds the SH2 domain-containing SHP-2 phosphatase in a tyrosine phosphorylation-dependent manner, and deregulates its enzymatic activity. We established AGS human gastric epithelial cells that inducibly express wildtype or a phosphorylation-resistant CagA, in which tyrosine residues constituting the EPIYA motifs were substituted with alanines. Upon induction, wild-type CagA, but not the mutant CagA, elicited strong elongation of cell shape, termed the "hummingbird" phenotype. Time-lapse video microscopic analysis revealed that the CagA-expressing cells exhibited a marked increase in cell motility with successive rounds of elongation-contraction processes. Inhibition of CagA phosphorylation by an Src kinase inhibitor, PP2, or knockdown of SHP-2 expression by small interference RNA ( siRNA) abolished the CagA-mediated hummingbird phenotype. The morphogenetic activity of CagA also required Erk MAPK but was independent of Ras or Grb2. In AGS cells, CagA prolonged duration of Erk activation in response to serum stimulation. Conversely, inhibition of SHP-2 expression by siRNA abolished the sustained Erk activation. Thus, SHP-2 acts as a positive regulator of Erk activity in AGS cells. These results indicate that SHP-2 is involved in the Ras-independent modification of Erk signals that is necessary for the morphogenetic activity of CagA. Our work therefore suggests a key role of SHP-2 in the pathological activity of H. pylori virulence factor CagA.
• Application of laser capture microdissection to cytologic specimens for the detection of immunoglobulin heavy chain gene rearrangement in patients with malignant lymphoma

  Y Orba, S Tanaka, H Nishihara, N Kawamura, T Itoh, M Shimizu, H Sawa, K Nagashima

  CANCER CYTOPATHOLOGY 99 (4) 198 - 204 0008-543X 2003/08 [Refereed][Not invited]
   

  BACKGROUND. The demonstration of the monoclonality of immunoglobulin heavy chain (IgH) gene rearrangement is an indispensable method for the diagnosis of B-cell lymphoma as well as histocytochemical analysis. For the detection of IgH gene rearrangement, the extraction of DNA from a homogenous cell population is necessary. Recently, the laser capture microdissection (LCM) technique was shown to isolate specific cells from histopathologic specimens for molecular analysis. However, to the authors' knowledge the applicability of LCM to cytologic specimens has not yet been well established. METHODS. Using LCM, a homogenous population of B-cell lymphoma cells as both histologic sections and cytologic specimens was captured, and genomic DNA was extracted from the captured cells. IgH gene rearrangement was analyzed by the polymerase chain reaction (PCR)-based single-strand conformational polymorphism (SSCP) method. RESULTS. Genomic DNAs were extracted successfully from ethanol-fixed cytologic specimens, but cells were not captured from air-dried specimens. Using PCR-SSCP analysis, the monoclonality of the IgH gene rearrangement was detected in five cases of tissue sections among nine analyzed cases of malignant lymphoma diagnosed immunohistochemically. However, analysis of the cytologic specimens with LCM demonstrated the monoclonality of the IgH gene rearrangement in seven cases of lymphoma. CONCLUSIONS. The results of the current study suggest that the novel application of LCM to cytologic specimens occasionally exhibits high sensitivity for the detection of IgH gene rearrangement monoclonality compared with the use of histologic sections.
• Dietary calcium and phosphorus ratio regulates bone mineralization and turnover in vitamin D receptor knockout mice by affecting intestinal calcium and phosphorus absorption

  R Masuyama, Y Nakaya, S Katsumata, Y Kajita, M Uehara, S Tanaka, A Sakai, S Kato, T Nakamura, K Suzuki

  JOURNAL OF BONE AND MINERAL RESEARCH 18 (7) 1217 - 1226 0884-0431 2003/07 [Refereed][Not invited]
   

  The effects of the dietary Ca and P ratio, independent of any vitamin D effects, on bone mineralization and turnover was examined in 60 VDRKO mice fed different Ca/P ratio diets. High dietary Ca/P ratio promoted bone mineralization and turnover with adequate intestinal Ca and P transports in VDRKO mice. Introduction: To clarify the effects of the dietary calcium (Ca) and phosphorus (P) ratio (Ca/P ratio) on bone mineralization and turnover in null-vitamin D signal condition, vitamin D receptor knockout (VDRKO) mice were given diets containing, different Ca/P ratios. Materials and Methods: Five groups of 4-week-old VDRKO mice, 10 animals each, were fed diets for 4 weeks. Group 1 was wild-type littermate mice, fed the diet containing 0.5% Ca and P (Ca/P = 1). Group 2 was the control and was fed a similar diet (Ca/P = 1). Groups 3, 4, 5, and 6 were fed the following diets:0.5% Ca and 1.0% P (Ca/P = 0.5) 1.0% Ca and 1.0% P (Ca/P = 1), 1.0% Ca and 0.5% P (Ca/P = 2), and 0.5% Ca and 0.25% P (Ca/P = 2). Results and Conclusions: Compared with group 2, serum calcium and phosphor-us levels in groups 4-6 significantly increased. Serum parathyroid hormone levels increased in group 3 and decreased in group 5. The amounts of intestinal calcium absorption decreased in groups 3 and 4. Phosphorus absorption increased in group 3 and decreased in groups 4-6. Bone mineral content (BMC) and bone mineral density (BMD) of the femur in group 3 significantly decreased and increased in group 5. In the primary spongiosa of the proximal tibia, the trabecular bone volume (BV/TV) and osteoid thickness (O.Th) in group 3 significantly increased, and decreased in group 6. In groups 5 and 6. the numbers of the trabecular osteoclasts increased. In groups 2 and 4, and the secondary spongiosa was identified in 5 of 10 mice. In group 3, there was no secondary spongiosa in either mouse. Osteoid maturation time (OMT) significantly decreased, and bone formation rate (BFR/BS) increased in groups 4-6. These data indicate that the dietary Ca/P ratio regulates bone mineralization and turnover by affecting the intestinal calcium and phosphorus transports in VDRKO mice. They may suggest the existence of Ca/P ratio-dependent, vitamin D-independent calcium and phosphorus transport system in the intestine.
• キメラ遺伝子と病理診断 滑膜肉腫の臨床病理学特徴とSYT‐SSXキメラ遺伝子産物による細胞癌化のメカニズム

  田中伸哉, 津田真寿美, 平賀博明, 三浪明男, 長嶋和郎

  病理と臨床 21 (6) 593 - 603 0287-3745 2003/06/01 [Not refereed][Not invited]
  
• Effects of 1,25(OH)(2)D-3 on turnover, mineralization, and strength of bone in growing rats with liver cirrhosis induced by administration of carbon tetrachloride

  S Tanaka, H Tsurukami, A Sakai, N Okimoto, S Ikeda, H Otomo, T Nakamura

  BONE 32 (3) 275 - 283 8756-3282 2003/03 [Refereed][Not invited]
   

  To clarify the effects of 1 alpha, 25-dihydroxyvitamin D-3(1,25(OH)(2)D-3) on bone growth, strength, and turnover in growing rats with liver cirrhosis induced by carbon tetrachloride (CCl4) injection, groups of 4-week-old male Wistar rats (n = 10, each) were injected intraperitoneally with CCl4 twice weekly for 7 weeks. One group was treated with the vehicle alone (Group 1). Three CCl4-injected groups were orally administered 1,25(OH)(2)D-3 at doses of 0, 0.05, and 0.1 mug/kg, respectively (Groups 2, 3, and 4). At the end, serum levels of 1,25(OH)(2)D-3, IGF-I, and osteocalcin were reduced in Group 2 compared to Group 1, and the corresponding values in Group 4 were larger than those in Group 2. Urinary deoxypyridinoline levels increased in Group 2 compared to Group 1, and did not significantly differ in Groups 2-4. The values for bone sizes, mineral content (BMC) in the lumbar vertebra and femur, and ultimate bending load in the femur were reduced in Group 2 compared to Group 1, and lumbar BMC in Group 3 and bone sizes in Group 4 were larger than those in Group 2. The values for lumbar trabecular bone volume in Group 2 were reduced compared to Group 1, and the corresponding values in Group 4 were larger than those in Group 2. Bone formation rates, reduced in Group 2 compared to Group 1, did not differ in Groups 2-4. Parameters for trabecular osteoclasts did not differ among all groups. In the proximal tibia, the value of activation frequency (Ac.f) in Group 2 significantly decreased compared to Group 1. Ac.f values in Groups 3 and 4 were larger than that in Group 2. These data demonstrated that retardation of bone growth in CCl4-injected rats was associated with reduced serum 1,25(OH)(2)D-3 and IGF-I levels. The trabecular bone in the rats exhibited low turnover osteopenia. 1,25(OH)(2)D-3 administration partially prevented the growth disturbance, but did not substantially affect bone turnover. Factors other than 1,25(OH)(2)D-3 and IGF-I appeared to be critical in the low turnover osteopenia evident in liver cirrhosis. (C) 2003 Elsevier Science (USA). All rights reserved.
• CrkL directs ASAP1 to peripheral focal adhesions

  A Oda, Wada, I, K Miura, K Okawa, T Kadoya, T Kato, H Nishihara, M Maeda, S Tanaka, K Nagashima, C Nishitani, K Matsuno, M Ishino, LM Machesky, H Fujita, P Randazzo

  JOURNAL OF BIOLOGICAL CHEMISTRY 278 (8) 6456 - 6460 0021-9258 2003/02 [Refereed][Not invited]
   

  Searching for proteins in platelets that can interact with the N-terminal SH3 domain of CrkL (using a combination of a pull-down assay followed by mass spectrometry), we have found that human platelets express an ADP-ribosylation factor (Arf)-specific GTPase-activating protein (GAP), ASAP1, as a CrkL-binding protein. In spreading platelets, most endogenous ASAP1. is localized at peripheral focal adhesions. To determine the physiologic significance of the CrkL-ASAP1 association, we overexpressed CrkL, ASAP1, or both in combination in COS7 cells. Unlike endogenous ASAP1 in platelets, overexpressed ASAP1 showed diffuse cytoplasmic distribution. However, when co-expressed with wild-type CrkL, both endogenous and expressed ASAP1 accumulated at CrkL-induced focal adhesions. An SH2-mutated CrkL, which cannot localize at focal adhesions, failed to recruit ASAP1 into focal adhesions. Thus, CrkL appears to be a lynchpin between ASAP1 and peripheral focal adhesions.
• JC virus agnoprotein colocalizes with tubulin

  S Endo, Y Okada, Y Orba, H Nishibara, S Tanaka, K Nagashima, H Sawa

  JOURNAL OF NEUROVIROLOGY 9 10 - 14 1355-0284 2003 [Refereed][Not invited]
   

  The human polyomavirus JC (JCV) encodes an agnoprotein that consists of 71 amino acid residues, with a molecular weight of approximately 8 kDa, from the late protein coding region. The agnoprotein of JCV shares 50% to 60% homology with those of simian virus 40 (SV40) and BK virus (BKV), and the carboxyl-terminal region of JCV agnoprotein is relatively unique. By using specific antibody to the carboxyl-terminal region of JCV agnoprotein, the authors have demonstrated that JCV agnoprotein expressed in the JCV-infected cells, where it localized predominantly in the perinuclear region of the cytoplasm, and colocalizes with the cellular cytoskeletal protein, tubulin. The results suggest that JCV agnoprotein may play a role in the stability of microtubules and the preservation of JCV infected cells via an interaction with tubulin.
• DOCK2 associates with CrkL and regulates Rac1 in human leukemia cell lines

  H Nishihara, M Maeda, A Oda, M Tsuda, H Sawa, K Nagashima, S Tanaka

  BLOOD 100 (12) 3968 - 3974 0006-4971 2002/12 [Refereed][Not invited]
   

  The CDM (ced-5 of Caenorhabditis elegans, DOCK180 [downstream of Crk with molecular weight of 180 kDa] of humans, and myoblast city of Drosophila melanogaster) family of proteins has been shown to play a pivotal role in the integrin-mediated signaling pathway under the regulation of an adaptor molecule c-CT10- related kinase II (c-Crk-II) in adherent cells. Recently, hematopoietic cell-specific CDM protein DOCK2 has been shown to be indispensable for lymphocyte migration. However, the regulatory mechanism for DOCK2 is still unknown because DOCK2 lacks a c-Crk-II binding consensus motif. In this study, we demonstrated that DOCK2 bound to CrkL, which is present exclusively in hematopoietic cells both in vivo and in vitro, and we also found that 2 separate regions of DOCK2 contributed to its binding to Src homology 3 (SH3) domain of CrkL. Colocalization of DOCK2 with Crk-like (CrkL) and F-actin was shown by immunocytochemical analysis with the use of Jurkat cells. We also found that CrkL-induced activation of small guanine triphosphatase (GTPase) Rac1 was significantly inhibited by the DOCK2-dCS mutant in 293T cells. Furthermore, the association of DOCK2 and Vav, the guanine-nucleotide exchanging factor (GEF) for Rac1, was demonstrated in Jurkat cells. Finally, the stable expression of DOCK2-dCS mutant in Jurkat cells was shown to reduce cell attachment. These data suggest the presence of a novel protein complex of CrkL, DOCK2, and Vav to regulate Rac1 in leukemia cell lines. (C) 2002 by The American Society of Hematology.
• Oligosaccharides as receptors for JC virus

  R Komagome, H Sawa, T Suzuki, Y Suzuki, S Tanaka, WJ Atwood, K Nagashima

  JOURNAL OF VIROLOGY 76 (24) 12992 - 13000 0022-538X 2002/12 [Refereed][Not invited]
   

  JC virus (JCV) belongs to the polyomavirus family of double-stranded DNA viruses and in humans causes a demyelinating disease of the central nervous system, progressive multifocal leukoencephalopathy. Its hemagglutination activity and entry into host cells have been reported to depend on an N-linked glycoprotein containing sialic acid. In order to identify the receptors of JCV, we generated virus-like particles (VLP) consisting of major viral capsid protein VP1. We then developed an indirect VLP overlay assay to detect VLP binding to glycoproteins and a panel of glycolipids. We found that VLP bound to sialoglycoproteins, including alpha1-acid glycoprotein, fetuin, and transferrin receptor, and that this binding depended on alpha2-3-linked sialic acids and N-linked sugar chains. Neoglycoproteins were synthesized by using ovalbumin and conjugation with oligosaccharides containing the terminal alpha2-3- or alpha2-6-linked sialic acid or the branched alpha2-6-linked sialic acid. We show that the neoglycoprotein containing the terminal alpha2-6-linked sialic acid had the highest affinity for VLP, inhibited the hemagglutination activity of VLP and JCV, and inhibited the attachment of VLP to cells. We also demonstrate that VLP bound to specific glycolipids, such as lactosylceramide, and gangliosides, including GM3, GD2, GD3, GD1b, GT1b, and GQ1b, and that VLP bound weakly to GD1a but did not bind to GM1a, GM2, or galactocerebroside. Furthermore, the neoglycoprotein containing the terminal alpha2-6-linked sialic acid and the ganglioside GT1b inhibited JCV infection in the susceptible cell line IMR-32. These results suggest that the oligosaccharides of glycoproteins and glycolipids work as JCV receptors and may be feasible as anti-JCV agents.
• High incidence of human herpesvirus 6 infection with a high viral load in cord blood stem cell transplant recipients

  J Sashihara, K Tanaka-Taya, S Tanaka, K Amo, H Miyagawa, G Hosoi, T Taniguchi, T Fukui, N Kasuga, T Aono, M Sako, J Hara, K Yamanishi, S Okada

  BLOOD 100 (6) 2005 - 2011 0006-4971 2002/09 [Refereed][Not invited]
   

  Human herpesvirus 6 (HHV-6) infection in recipients of cord blood stem cell trans plants (CBSCTs) was estimated by semiquantitative and real-time quantitative polymerase chain reaction (PCR) and reverse-transcription PCR. Of the CBSCT recipients, 7 (70%) of 10 had active HHV-6 infection after transplantation, and all 7 were inferred from their age to have already had a primary infection. Because HHV-6 DNA is seldom detected in cord blood, these cases were considered likely to represent reactivatiom In contrast; the 3 patients Without HHV-6 infection were all believed to be naive regarding HHV-6 primary infection because of their age and the results of PCR assays given before the transplantation procedure. The incidence of HHV-6 infection after transplantation wag significantly higher (P < .05) than after bone marrow (BM) transplantation and peripheral blood stem cell (PBSC) transplantation, when recipients without primary HHV-6 infection prior to transplantation were excluded (CB-SCT, 100%; BMT/PBSCT, 56.3%). Real-time PCR,revealed a higher level of viral DNA in the Peripheral blood mononuclear cells from CBSCT recipients than from BMT/PBSCT recipients or patients with exanthem subitum (P < .05). HHV-6 mRNA of the U79/80 gene was Also detected by reverse-transcription PCR in all analyzed patients with HHV-6 infection. Its detection Was correlated with the emergence of viral DNA in the plasma and symptoms such as fever and rash. Thus, HHV-6 infection was more frequent and the viral load was higher in CBSCT recipients with prior primary infection.
• Regulation of bone mass for skeletal loading

  Akinori Sakai, Shinya Tanaka, Takeshi Sakata, Makoto Watanuki, Miyuki Zenke, Toshitaka Nakamura

  Journal of UOEH 24 (3) 281 - 287 0387-821X 2002/09/01 [Refereed][Not invited]
   

  Skeletal loading plays an important role in the maintenance of bone mass, bone shape and bone strength. Skeletal unloading, such as during space flight and long-term bed rest, induces bone loss in loaded bones in humans. The mechanotransduction system in bone tissue is not fully elucidated. Our observations demonstrate that generation of nitric oxide through inducible nitric oxide synthase is essential for the stimulation of bone formation upon mechanical reloading, and that disruption of p53 gene in response to 1-week unloading does not result in reductions in bone volume and bone formation. It should be significantly helpful for our understanding of various skeletal disorders to clarify the mechanism underlying regulation of bone mass after skeletal loading and unloading.
• Expression of JC virus agnoprotein in progressive multifocal leukoencephalopathy brain

  Y Okada, H Sawa, S Endo, Y Orba, T Umemura, H Nishihara, AC Stan, S Tanaka, H Takahashi, K Nagashima

  ACTA NEUROPATHOLOGICA 104 (2) 130 - 136 0001-6322 2002/08 [Refereed][Not invited]
   

  To examine the function of JC virus (JCV) agnoprotein, we examined the brains of cases of progressive multifocal leukoencephalopathy (PML), which is caused by JCV infection, using a newly generated antibody. The antibody reacted with 8 kDa protein specific for JCV agnoprotein by Western blotting. In vitro analyses showed that JCV capsid protein VP1 and large T antigen (T-Ag) were localized in the nuclei, but that agnoprotein was mainly detected in the cytoplasm of JCV-infected cells with an occasional nuclear staining. In the PML brain, an immunoreactive signal for agnoprotein was distributed in the perinuclear areas and cytoplasmic processes with occasional punctate staining in demyelinating lesions as well as adjacent myelinated areas. Agnoprotein presented mostly in the infected oligodendrocytes and partly in the astrocytes. Using double immunostaining, agnoprotein was seen to be expressed in the cytoplasmic processes of the cells, the nuclei of which were labeled with VP1 and T-Ag, where virus particles existed. Thus, JCV agnoprotein was mostly expressed in the infected oligodendrocytes and mainly localized in the cytoplasmic processes apart from virus particles in the demyelinated lesions.
• DOCK2 mediates T cell receptor-induced activation of Rac2 and IL-2 transcription

  H Nishihara, M Maeda, M Tsuda, Y Makino, H Sawa, K Nagashima, S Tanaka

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 296 (3) 716 - 720 0006-291X 2002/08 [Refereed][Not invited]
   

  DOCK2, a CDM family protein exclusively found in hematopoietic cells, has been shown to play a role in lymphocyte migration by the regulation of actin cytoskeleton. Although DOCK2 has been shown to induce the activation of Rac1, the regulatory mechanism of Rac2, which is a hematopoietic cell-specific small GTPase, is still unknown. In this study, we examined the role of DOCK2 in the activation of Rac2 in hematopoietic cells. DOCK2 was found to associate with the zeta subunit of the CD3 complex of T cell receptors in Jurkat cells and to activate forced expressed Rac2 in 293T cells. In addition, the stable expression of DOCK2 in Jurkat cells exhibited the elevated activity of endogenous Rac2. Furthermore, the transcriptional activity of interleukin-2 (IL-2) was enhanced in DOCK2-expressing Jurkat cells and the dominant negative form of Rac2 suppressed its elevated IL-2 promoter activity. These results suggest that DOCK2 mediates TCR-dependent activation of Rac2, leading to the regulation of IL-2 promoter activity in T cells. (C) 2002 Elsevier Science (USA). All rights reserved.
• Molecular and immunohistochemical analysis of signaling adaptor protein Crk in human cancers

  H Nishihara, S Tanaka, M Tsuda, S Oikawa, M Maeda, M Shimizu, H Shinomiya, A Tanigami, H Sawa, K Nagashima

  CANCER LETTERS 180 (1) 55 - 61 0304-3835 2002/06 [Refereed][Not invited]
   

  Crk is a signaling adaptor protein which is mostly composed of SH2 and SH3 domains. and has been shown to play a pivotal role in cell proliferation. differentiation, and migration. Because Crk was originally isolated as an avian sarcoma virus CT10 encoding oncoprotein v-Crk. we examined a potential role for c-Crk in the carcinogenesis of human cancers. First, to analyze gene mutations of c-Crk, we isolated a human bacterial artificial chromosome clone containing Crk genome and exon/intron structures, However, polymerase chain reaction-single strand conformation polymorphism methods failed to show any genomic mutations in the Crk exon which could be related to carcinogenesis. Second, immunohistochemical analysis of c-Crk-II demonstrated that the levels of c-Crk-II were significantly elevated in most of the tumors, particularly in the colon and lung cancers. Furthermore, immunoblot analysis using human lung cancer cell lines revealed that the expression levels of c-Crk-II were correlated to growth rates of L cells. The elevated expression levels of c-Crk-II might be related to the development of human cancers. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
• Advanced glycation end products induce angiogenesis in vivo

  T Okamoto, S Tanaka, AC Stan, T Koike, M Kase, Z Makita, H Sawa, K Nagashima

  MICROVASCULAR RESEARCH 63 (2) 186 - 195 0026-2862 2002/03 [Refereed][Not invited]
   

  Advanced glycation end products (AGEs) have been thought to participate in diabetic microangiopathy. However, the effects of AGEs on angiogenesis have so far been mainly examined either in vitro or by using cultured cells. In the present study, we have analyzed whether AGES induce angiogenesis in vivo by using the chorioallantoic membrane (CAM) assay. The CAM assay was carried out in embryonated hen eggs to determine the effects of AGEs. Following generation of AGEs based on bovine serum albumin (BSA), either AGE-BSA or nonglycated BSA was administered to the CAM and their effects on angiogenesis were assessed, together with an inhibitory effect of an anti-AGE antibody against AGE-BSA-induced angiogenesis. The histological features of AGE-induced vascular lumens were examined by immunohistochemical analysis for Factor VIII and smooth muscle a-actin. AGE-BSA induced angiogenesis in CAM in a dose- and time-dependent manner. AGE-induced angiogenesis on CAM was neutralized by the anti-AGE antibody. Inummohistochemical analysis demonstrated that AGE-induced vascular lumens were devoid of pericytes. Our data demonstrated that AGES are an angiogenetic factor and that our system of AGE-induced abnormal vessels in CAMs is useful in further investigations of the mechanism of diabetic retinal angiogenesis and can also be used to provide a therapeutic model for diabetic angiopathy. (C) 2002 Elsevier Science (USA).
• Signaling adaptor protein v-Crk activates Rho and regulates cell motility in 3Y1 rat fibroblast cell line.

  Tsuda M, Tanaka S, Sawa H, Hanafusa H, Nagashima K

  Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research 3 13 131 - 139 1044-9523 2002/03 [Refereed][Not invited]
  
• Mixed allogeneic chimerism with wild-type strains ameliorates atherosclerosis in apolipoprotein E-deficient mice

  N Ishimori, K Iwabuchi, S Fujii, K Watano, C Iwabuchi, M Ato, H Chiba, S Tanaka, A Kitabatake, K Onoe

  JOURNAL OF LEUKOCYTE BIOLOGY 69 (5) 732 - 740 0741-5400 2001/05 [Refereed][Not invited]
   

  Atherosclerosis involves inflammatory processes between vascular tissues and hematocytes with a hyperlipidemic background. To examine whether variations of hematocytes constitute one of the genetic components in atherosclerosis, irradiated apolipoprotein E (apoE)-deficient (apoE(-/-)) mice with hypercholesterolemia and preexisting atherosclerotic lesions were reconstituted with mixed bone marrow cells (BMC) from syngeneic and wild-type (apoE(+/+); atheroselerosis-resistant SJL or -susceptible B10.S) mice. Stable mixed allogeneic chimeras with small amounts of serum apoE were established without any detrimental complications. Compared with untreated apoE(-/-) mice or apoE(-/-) mice transplanted with syngeneic BMC alone, significant reduction of the cholesterol level and significant lesion regression were observed in the mixed chimeras. Furthermore, mixed chimeras given SJL BMC showed marked reductions in umbers of lesions compared with those reconstituted with B10.S BMC, Cholesterol levels in the former SJL chimeras, however, were significantly higher than those in the latter B10.S chimeras. These findings indicate that the resistance of SJL to atherosclerosis resides in the bone marrow-derived cells.
• Analysis of transforming activity of human synovial sarcoma-associated chimeric protein SYT-SSX1 bound to chromatin remodeling factor hBRM/hSNF2

  Nagai, M, Tanaka, S, Tsuda, M, Endo, S, Kato, H, Sonobe, H, Minami , A, Hiraga, H, Yamawaki, S, Nishihara, H, Sawa, H, Nagashima, K

  Proc.Natl.Acad.Sci.USA 98 (7) 3843 - 3848 0027-8424 2001 [Not refereed][Not invited]
  
• Downstream of Crk: activation of JNK by v-Crk through the guanine nucleotide exchange protein C3G

  Tanaka, S, Ouchi, T, Hanafusa, H

  Proc.Natl.Acad.Sci.USA 94 2356 - 2361 1997 [Not refereed][Not invited]
  
• Molecular analysis of phosphorylated tyrosine-binding proteins in the transduction of proliferation and differentiation signals

  K Nagashima, S Tanaka, S Ota, H Hasegawa, M Matsuda

  BRAIN TUMOR 151 - 159 1996 [Refereed][Not invited]
  
• C3G, a Guanine Nucleotide Releasing Protein Expressed Ubiquitously, Binds to the SH3 domain of CRK and GRB2/ASH

  Tanaka, S, Morishita, T, Hashimoto, Y, Hattori, S, Nakamura, S, Shibuya, M, Matsuoka, K, Takenawa, T, Kurata, T, Nagashima, K, Matsuda, M

  Proc.Natl.Acad.Sci.USA 91 (8) 3443 - 3447 0027-8424 1994 [Not refereed][Not invited]
  
• BOTH THE SH2 AND SH3 DOMAINS OF HUMAN CRK PROTEIN ARE REQUIRED FOR NEURONAL DIFFERENTIATION OF PC12 CELLS

  S TANAKA, S HATTORI, T KURATA, K NAGASHIMA, Y FUKUI, S NAKAMURA, M MATSUDA

  MOLECULAR AND CELLULAR BIOLOGY 13 (7) 4409 - 4415 0270-7306 1993/07 [Not refereed][Not invited]
   

  Human CRK protein is a homolog of the chicken v-crk oncogene product and consists mostly of src homology region 2 (SH2) and SH3, which are shared by many proteins, in particular those involved in signal transduction. SH2 has been shown to bind specifically to phosphotyrosine-containing peptides. We report here that both SH2 and SH3 are required for signaling from CRK protein. Microinjection of the CRK protein induced neurite formation of rat pheochromocytoma cell line PC12. This activity was abolished by mutation of the CRK protein in either SH2 or SH3. The neuronal differentiation induced by the CRK protein was blocked by an excess amount of peptides containing CRK SH3. Moreover, we identified three proteins, of 118, 125, and 136 kDa, which bound specifically to CRK SH3. The CRK-induced neuronal differentiation was also suppressed by monoclonal antibodies against either CRK SH2 or p21ras. These results suggest that both SH2 and SH3 of the CRK protein mediate specific protein-protein binding and that the resulting multimolecular complex generates a signal for neurite differentiation through activation of p21ras.
• PATHOLOGICAL FEATURES OF VIRUS-INFECTIONS OF THE CENTRAL-NERVOUS-SYSTEM (CNS) IN THE ACQUIRED-IMMUNODEFICIENCY-SYNDROME (AIDS)

  WW HALL, PM FARMER, H TAKAHASHI, S TANAKA, Y FURUTA, K NAGASHIMA

  ACTA PATHOLOGICA JAPONICA 41 (3) 172 - 181 0001-6632 1991/03 [Refereed][Not invited]
   

  Neurological disorders are a common cause of morbidity and mortality in the acquired immunodeficiency syndrome (AIDS). In this report we describe the neuropathological changes associated with both human immunodeficiency virus (HIV) infection and with the major opportunistic virus infections, cytomegalovirus (CMV), JC papovavirus (JCV) and herpes simplex virus (HSV) seen in AIDS. In addition "in situ" hybridization studies have been employed for the detection of virus genomic material in each case and the usefulness of this method in supporting the pathological diagnosis is demonstrated. Mechanisms whereby HIV infection results in leukoencephalopathy and the possible contributing roles of the three opportunistic virus infections are discussed.
• Pathological Features of Virus Infections of the Central Nervous System (CNS) in the Acquired Immunodeficiency Syndrome (AIDS)

  William W. Hall, Peter M. Farmer, Hidehiro Takahashi, Shinya Tanaka, Yasushi Furuta, Kazuo Nagashima

  Pathology International 41 (3) 172 - 181 1440-1827 1991 [Refereed][Not invited]
   

  Neurological disorders are a common cause of morbidity and mortality in the acquired immunodeficiency syndrome (AIDS). In this report we describe the neuropathological changes associated with both human immunodeficiency virus (HIV) infection and with the major opportunistic virus infections, cytomegalovirus (CMV), JC papovavirus (JCV) and herpes simplex virus (HSV) seen in AIDS. In addition “in situ” hybridization studies have been employed for the detection of virus genomic material in each case and the usefulness of this method in supporting the pathological diagnosis is demonstrated. Mechanisms whereby HIV infection results in leukoencephaiopathy and the possible contributing roles of the three opportunistic virus infections are discussed. Copyright © 1991, Wiley Blackwell. All rights reserved

Books etc

• がんゲノム病理学

  田中, 伸哉, 西原, 広史 
  文光堂 2021/11 (ISBN: 9784830604850) vii, 225p
• はじめの一歩の病理学

  深山, 正久, 田中, 伸哉, 豐國, 伸哉, 伊藤, 彰彦, 宮崎, 龍彦, 宇於崎, 宏, 田中, 文彦, 林, 祥剛, 大澤, 佳代, 酒々井, 眞澄, 小田, 義直, 前田, 大地, 川村, 公一, 後藤, 明輝 
  羊土社 2017/12 (ISBN: 9784758120845) 278, 1p
• 癌の分子病理学 : 病理診断から治療標的探索まで

  深山, 正久, 金井, 弥栄, 田中, 伸哉 
  文光堂 2016/04 vii, 349p
• はじめの一歩のイラスト病理学

  深山 正久編, 深山 正久 (Contributor田中 伸哉 第1章、第９章)
  羊土社 2012/11 (ISBN: 4758120366) 262

  
• Molecular Targets of CNS Tumors

  Ed. by Miklos Garami (ContributorISBN: 978-953-307-736-9)
  INTECH 2011/08
• 医療系学生のための病理学 第4版

  中村 仁志夫, 佐藤 達資, 石津 明洋, 田中 伸哉 (Joint editor)
  講談社 2010/11 (ISBN: 4061536966) 232

  
• 医療系学生のための病理学

  中村, 仁志夫, 佐藤, 達資, 石津, 明洋, 田中, 伸哉, 池田, 仁 
  講談社 2010/11 (ISBN: 9784061536968) xi, 208p, 図版 [8] p
• 病理形態学キーワード

  病理と臨床, 常任編集委員会, 小川 郁子 (Contributor異型グリア 田中伸哉)
  文光堂 2010
• Adaptor Proteins and Cancer

  TANAKA Shinya (ContributorISBN 978-81-7895-344-1)
  Thansworld Research Network 2008
• 細胞内シグナル伝達 (Bio Science 新用語ライブラリー)

  山本 雅 編, 山本 雅 (ContributorC3G・Sos 田中 伸哉)
  羊土社 1999/06 (ISBN: 4897062624) 217

  

MISC

• 非HIV関連かつ免疫抑制剤を使用していない進行性多巣性白質脳症5例の臨床的検討

  穴田麻眞子, 矢口裕章, 布村菫, 石丸誠己, 水島慶一, 工藤彰彦, 佐藤翔紀, 阿部恵, 江口克紀, 長井梓, 脇田雅大, 白井慎一, 岩田育子, 松島理明, 南尚哉, 中道一生, 松野吉宏, 田中伸哉, 矢部一郎  日本神経学会学術大会プログラム・抄録集  63rd-  2022
• Brainstem/spinal cord gliomas with IDH mutation: report of four cases

  澤谷亮佑, 伊師雪友, 山口秀, 岡本迪成, 茂木洋晃, 高宮宗一朗, 関俊隆, 大塚拓也, 若林健人, 松野吉宏, 種井善一, 田中伸哉, 藤村幹  日本脳腫瘍学会学術集会プログラム・抄録集  39th-  2021
• Glioma病理診断における統合診断フローチャート

  園田 順彦, 横尾 英明, 田中 伸哉, 木下 学, 中田 光俊, 西原 広史  Brain Tumor Pathology  36-  (Suppl.)  061  -061  2019/05  [Not refereed][Not invited]
  
• Artificial Intelligence Predicts the Genetic Information of The Integrated Diagnosis of Brain Tumors

  Yusuke Ishida, Masumi Tsuda, Jun Suzuka, Lei Wang, Satoshi Tanikawa, Hirokazu Sugino, Shinya Tanaka  MODERN PATHOLOGY  32-  2019/03
• Artificial Intelligence Predicts the Genetic Information of The Integrated Diagnosis of Brain Tumors

  Yusuke Ishida, Masumi Tsuda, Jun Suzuka, Lei Wang, Satoshi Tanikawa, Hirokazu Sugino, Shinya Tanaka  LABORATORY INVESTIGATION  99-  2019/03
• 日本から発信されたサイエンス(No.14)IPMN関連膵がんの発生・進化に関する分子経路 : Omori Y, Ono Y, Tanino M et al : Pathways of progression from intraductal papillary mucinous neoplasm to pancreatic ductal adenocarcinoma based on molecular features. Gastroenterology 156 : 647-661. e2, 2019

  大森 優子, 小野 裕介, 谷野 美智枝, 唐崎 秀則, 山口 浩, 古川 徹, 真口 宏介, 田中 伸哉, 水上 裕輔  消化器病学サイエンス = Science of gastroenterology  3-  (1)  16,49  -53  2019/03
• IPMN関連膵癌の発生・進化に関する分子経路

  大森優子, 小野裕介, 谷野美智枝, 唐崎秀則, 山口浩, 古川徹, 真口宏介, 田中伸哉, 水上裕輔  消化器病学サイエンス  3-  (1)  49  -53  2019/03  [Not refereed][Invited]
  
• A case of central nervous system methotrexate-associated lymphoproliferative disorders (MTX-LPD)

  Yuuki Ishida, Kenichi Sato, Yoshimaru Ozaki, Taku Asanome, Hirohiko Nakamura, Hirokazu Sugino, Shinya Tanaka  BRAIN PATHOLOGY  29-  164  -164  2019/02
• A case of cauda equina primary extramedullary plasmacytoma

  Yoshitaka Oda, Hirokazu Sugino, Izumi Koyanagi, Tanikawa Satoshi, Yuusuke Ishida, Masumi Tsuda, Shinya Tanaka  BRAIN PATHOLOGY  29-  165  -165  2019/02
• Development of bipolar charged hydrogel for neuronal tissue engineering

  Satoshi Tanikawa, Shingo Semba, Lei Wang, Mishie Tanino, Yusuke Ishida, Hirokazu Sugino, Jun Suzuka, Masumi Tsuda, Shinya Tanaka  BRAIN PATHOLOGY  29-  109  -109  2019/02
• A case of spinal rosette-forming glioneuronal tumor

  Mishie Tanino, Shuji Hamauchi, Hirokazu Sugino, Masumi Tsuda, Hidehiro Takei, Shinya Tanaka  BRAIN PATHOLOGY  29-  160  -160  2019/02
• 低分化胃癌の個別化病理診断のための遺伝子プロファイリング

  石川 麻倫, 西原 広史, 林 秀幸, 坂本 直哉, 田中 伸哉  日本消化器病学会雑誌  115-  (臨増大会)  A696  -A696  2018/10  [Not refereed][Not invited]
  
• IPMN関連膵癌におけるMolecular subtypeに基づいたクローン進化モデル

  大森優子, 大森優子, 小野裕介, 谷野美智枝, 唐崎秀則, 山口浩, 古川徹, 篠原敏也, 真口宏介, 水上裕輔, 水上裕輔, 田中伸哉  日本病理学会会誌  107-  (1)  236  2018/04  [Not refereed][Not invited]
  
• IDH1 Mutation Enhances Radiation Sensitivity by Regulating EMT and Apoptotic Pathway in Malignant Glioma

  Mishie A. Tanino, Arisa Kitazaki, Mei Kuzasa, Hirokazu Sugino, Yusuke Ishida, Lei Wang, Masumi Tsuda, Shinya Tanaka  MODERN PATHOLOGY  31-  667  -667  2018/03
• IDH1 Mutation Enhances Radiation Sensitivity by Regulating EMT and Apoptotic Pathway in Malignant Glioma

  Mishie A. Tanino, Arisa Kitazaki, Mei Kuzasa, Hirokazu Sugino, Yusuke Ishida, Lei Wang, Masumi Tsuda, Shinya Tanaka  LABORATORY INVESTIGATION  98-  667  -667  2018/03
• 低分化胃癌におけるSox10発現の臨床病理学的検討

  石川 麻倫, 西原 広史, 林 秀幸, 木村 太一, 石田 雄介, 王 磊, 津田 真寿美, 谷野 美智枝, 坂本 直哉, 田中 伸哉  日本消化器病学会雑誌  115-  (臨増総会)  A375  -A375  2018/03  [Not refereed][Not invited]
  
• IDH1 mutation contributes to apoptosis after multi-fractionated irradiation in malignant glioma

  Arisa Kitazaki, Mishie Tanino, Mei Kuzasa, Hirokazu Sugino, Lei Wang, Yusuke Ishida, Shingo Semba, Masumi Tsuda, Kaori Igarashi, Tomoyoshi Soga, Shinya Tanaka  CANCER SCIENCE  109-  1130  -1130  2018/01
• Expression of OTUB1 in human malignant mesothelioma

  Mei Kuzasa, Mishie Tanino, Arisa Kitazaki, Hirokazu Sugino, Yusuke Ishida, Lei Wang, Masumi Tsuda, Akira Takasawa, Hiroshi Hirano, Shinya Tanaka  CANCER SCIENCE  109-  1109  -1109  2018/01
• Aanalysis of relationship between BRAF V600E mutation and expression for p16 in pleomorphic xanthoastrocytoma

  Mishie A. Tanino, Hiroshi Nanjo, Masumi Tsuda, Hirokazu Sugino, Rei Wang, Yusuke Ishida, Shinya Tanaka  CANCER SCIENCE  109-  1127  -1127  2018/01
• 骨形成に再利用される合成HAp系インプラントの同位体顕微鏡観察

  野々山貴行, 鈴木裕貴, 木山竜二, WANG Lei, 津田真寿美, 安田和則, 田中伸哉, 永田康祐, 藤田龍介, 坂本直哉, 圦本尚義, GONG Jian Ping  日本セラミックス協会秋季シンポジウム講演予稿集(CD-ROM)  31st-  2018
• 低分化胃癌におけるSox10発現の臨床病理学的検討

  石川麻倫, 石川麻倫, 西原広史, 林秀幸, 木村太一, 石田雄介, 王磊, 津田真寿美, 谷野美智枝, 坂本直哉, 田中伸哉  日本消化器病学会雑誌(Web)  115-  2018
• 低分化胃癌の個別化病理診断のための遺伝子プロファイリング

  石川麻倫, 石川麻倫, 石川麻倫, 西原広史, 林秀幸, 坂本直哉, 田中伸哉  日本消化器病学会雑誌(Web)  115-  2018
• TAFRO症候群のリンパ節に浸潤する形質細胞のKi-67標識率は高い

  大森優子, 大森優子, 野口寛子, 篠原敏也, 永井友基, 酒井基, 石川麻倫, 石川麻倫, 谷野美智枝, 田中伸哉  日本病理学会会誌  107-  (1)  2018
• Lynch症候群への合併が疑われた膠芽腫の一例

  伊師雪友, 伊師雪友, 小林智絵, 江藤和範, 能條建, 北川道生, 茂木洋晃, 山口秀, 小林浩之, 田中伸哉, 外丸詩野  Brain Tumor Pathology  35-  (Supplement)  2018
• アダプター分子CRKはエクソソームのErbB2を制御し、膀胱癌の増殖・転移を亢進する

  津田 真寿美, 吉田 一彦, 松本 隆児, 近藤 恒徳, 篠原 信雄, 田中 伸哉  日本癌学会総会記事  76回-  E  -1059  2017/09  [Not refereed][Not invited]
  
• miR-23aによる膠芽腫の浸潤能亢進分子メカニズムの解明

  津田 真寿美, 谷地 一博, 高阪 真路, 三浪 友輔, 王 磊, 木村 太一, 谷野 美智枝, 西原 広史, 田中 伸哉  Brain Tumor Pathology  34-  (Suppl.)  093  -093  2017/05  [Not refereed][Not invited]
  
• グリオーマの日常診断におけるintegrated diagnosisの現状

  谷野 美智枝, 谷川 聖, 石田 雄介, 木村 太一, 岡田 佳奈子, 佐藤 真実, 津田 真寿美, 西原 広史, 長嶋 和郎, 田中 伸哉  Brain Tumor Pathology  34-  (Suppl.)  102  -102  2017/05  [Not refereed][Not invited]
  
• RANKLによるインテグリンα2の発現亢進はインテグリンβ2の細胞内輸送を介して細胞接着を亢進する

  大場雄介, 山田珠希, 山田珠希, 山田珠希, 津田真寿美, 藤岡容一朗, 藤岡真理, 堀内浩水, 堀口美香, 佐藤絢, ネパール ブラバ, 王せい, 柏木彩花, 西出真也, 南保明日香, 芳賀永, 田中伸哉, 進藤正信  日本生理学雑誌(Web)  79-  (2)  49 (WEB ONLY)  -49  2017/05  [Not refereed][Not invited]
  
• 脳腫瘍術中迅速診断に苦慮した星芽腫の1例

  鈴鹿 淳, 森谷 純, 竹浪 智子, 漆戸 万紗那, 湯澤 明夏, 木村 太一, 石田 雄介, 谷野 美智枝, 西原 広史, 田中 伸哉  日本臨床細胞学会雑誌  56-  (Suppl.1)  275  -275  2017/04  [Not refereed][Not invited]
  
• 今IPMNをどう診るか IPMN遺伝子解析の進歩 遺伝子異常からみたIPMN関連膵癌の特徴

  大森優子, 小野裕介, 谷野美智枝, 唐崎秀則, 高橋邦幸, 篠原敏也, 田中伸哉, 真口宏介, 水上裕輔  肝胆膵  74-  (4)  541‐549  2017/04  [Not refereed][Not invited]
  
• 組織形態とゲノム変化 良性腫瘍を中心として 孤立性線維性腫瘍/血管周皮腫におけるNAB2-STAT6融合遺伝子

  湯澤 明夏, 西原 広史, 田中 伸哉  日本病理学会会誌  106-  (1)  201  -201  2017/03  [Not refereed][Not invited]
  
• 髄膜腫におけるPOLR2A遺伝子変異の検討

  鈴木 佑季, 津田 真寿美, 湯澤 明夏, 木村 太一, 石田 雄介, 谷野 美智枝, 西原 広史, 田中 伸哉  日本病理学会会誌  106-  (1)  508  -508  2017/03  [Not refereed][Not invited]
  
• 髄膜発生孤在性線維性腫瘍/血管周皮腫(SFT/HPC)におけるNAB2-STAT6融合遺伝子の解析

  四宮 万里絵, 津田 真寿美, 湯澤 明夏, 木村 太一, 石田 雄介, 谷野 美智枝, 西原 広史, 田中 伸哉  日本病理学会会誌  106-  (1)  508  -509  2017/03  [Not refereed][Not invited]
  
• 原発不明癌症例の臨床病理学的解析

  高田 莉央, 鈴木 喬之, 谷野 美智枝, 木村 太一, 石田 雄介, 王 磊, 西原 広史, 田中 伸哉, 篠原 敏也, 後藤田 裕子  日本病理学会会誌  106-  (1)  511  -511  2017/03  [Not refereed][Not invited]
  
• 浸潤性膀胱癌の転移および薬剤耐性獲得におけるAKR1C1の役割

  津田 真寿美, 松本 隆児, 吉田 一彦, 谷野 美智枝, 木村 太一, 西原 広史, 阿部 崇重, 篠原 信雄, 野々村 克也, 田中 伸哉  日本病理学会会誌  106-  (1)  340  -340  2017/03  [Not refereed][Not invited]
  
• 合成ハイドロゲルが誘導する軟骨分化促進機序の解明

  仙葉愼吾, 後藤佳子, 北村信人, 北村信人, 黒野定, 近江谷克裕, 津田真寿美, 津田真寿美, 黒川孝幸, 黒川孝幸, 田中伸哉, 田中伸哉, GONG Jian Ping, GONG Jian Ping, 安田和則  再生医療  16-  267  2017/02/01  [Not refereed][Not invited]
  
• Preclinical Evidence of Anti-Tumor Activity Induced by EZH2 Inhibition in Human Models of Synovial Sarcoma (vol 11, e0158888, 2016)

  Satoshi Kawano, Alexandra R. Grassian, Masumi Tsuda, Sarah K. Knutson, Natalie M. Warholic, Galina Kuznetsov, Shanqin Xu, Yonghong Xiao, Roy M. Pollock, Jesse J. Smith, Kevin W. Kuntz, Scott Ribich, Yukinori Minoshima, Junji Matsui, Robert A. Copeland, Shinya Tanaka, Heike Keilhack  PLOS ONE  12-  (1)  2017/01  [Not refereed][Not invited]
  
• 前立腺癌診断におけるTripartite motif‐containing protein29(TRIM29)染色の有用性の検討

  吉田一彦, 吉田一彦, 谷野美智枝, 堀井理絵, 木村太一, 津田真寿美, 近藤恒徳, 秋山太, 畠山鎮次, 田邉一成, 田中伸哉  日本泌尿器科学会東部総会プログラム・抄録集  82nd-  222  2017  [Not refereed][Not invited]
  
• 家族性進行性核上性麻痺の遺伝子解析に基づいた孤発性進行性核上性麻痺の遺伝子解析

  矢部 一郎, 加藤 容崇, 谷川 聖, 三木 康生, 白井 慎一, 高橋 育子, 矢口 裕章, 藤岡 伸助, 國枝 保幸, 西原 広史, 田中 伸哉, 坪井 義夫, 若林 孝一, 佐々木 秀直  臨床神経学  56-  (Suppl.)  S317  -S317  2016/12  [Not refereed][Not invited]
  
• GENOTYPING OF GLIOMA INCLUDING 1p19q CODELETION BY TARGETED SEQUENCING

  Hiroshi Nishihara, Shigeki Tanishima, Sayaka Yuzawa, Lei Wang, Shigeru Yamaguchi, Hiroyuki Kobayashi, Shunsuke Terasaka, Shinya Tanaka  NEURO-ONCOLOGY  18-  118  -118  2016/11  [Not refereed][Not invited]
  
• SFT/HPCの術中迅速免疫細胞化学染色(R-ICC)を用いたSTAT6の有用性についての検討

  鈴鹿 淳, 森谷 純, 竹浪 智子, 漆戸 万紗那, 湯澤 明夏, 木村 太一, 西原 広史, 谷野 美智枝, 田中 伸哉  北海道臨床細胞学会会報  25-  15  -18  2016/10  [Not refereed][Not invited]
   

  孤在性線維性腫瘍(Solitary fibrous tumor:SFT)/血管周皮腫(Hemangiopericytoma:HPC)は全身に発生するが,脳原発例は再発や転移をきたすことが多く,外科的切除による全摘出が必要となる.しかし,腫瘍の発生部位や細胞形態の類似性により形態診断のみでは髄膜腫との鑑別は難しく,さらに術中迅速診断における鑑別は困難である.近年SFT/HPCにおいてNAB 2-STAT 6融合遺伝子が同定され,免疫組織化学(Immunohistochemistry:IHC)法を用いたSTAT 6の核内陽性像による診断の有用性が報告された.また当教室では電界撹拌により抗原抗体反応を迅速化したrapid immunohistochemistry(R-IHC),rapidimmunocytochemistry(R-ICC)法を用いた免疫染色を実施しており,術中迅速診断での有用性が期待される.今回我々は,SFT/HPC 2症例,髄膜腫16症例の術中迅速検体を使用し,SFT/HPCにおけるR-IHC,R-ICC法を用いたSTAT 6の免疫染色の有用性を検討した.R-IHC,R-ICC法では従来の方法と同様,SFT/HPCにおけるSTAT 6の核内陽性像が認められ,髄膜腫では認められなかった.以上より,SFT/HPCにおいてR-IHC,R-ICC法によるSTAT 6の核内移行の証明は有用であり,脳腫瘍術中迅速診断時の正診率の向上に寄与すると考えられた.(著者抄録)
• 術中迅速圧挫細胞診による血管評価に基づいた神経膠腫の悪性度の検討

  漆戸 万紗那, 森谷 純, 竹浪 智子, 鈴鹿 淳, 木村 太一, 西原 広史, 谷野 美智枝, 田中 伸哉  北海道臨床細胞学会会報  25-  19  -23  2016/10  [Not refereed][Not invited]
   

  目的:術中迅速圧挫細胞診における微小血管増生の程度から悪性度分類を行うことを試みた.対象:細胞診圧挫標本作製が可能であった術中迅速病理検体のうち,神経膠腫64症例(高悪性度47症例,低悪性度17症例)のPapanicolaou染色標本を用いた.方法:層構造や分岐の有無については目視で行い,内皮細胞の核の数を算定し層数とした.血管径については顕微鏡ソフトウェアを用いて計測した.結果:grade III以上の神経膠腫で血管内皮細胞の多層化がより多く認められ,分岐数の増加,構造の複雑化もgradeの上昇に伴い多く見られることがわかった.また,血管径を比較したところ,高悪性度群で有意に増大傾向があることが認められた.結語:細胞診圧挫標本を用いた神経膠腫における血管増生は,悪性度と相関があり,gradeを含めた診断を行う上で有用であると考えられた.(著者抄録)
• 肺がんのTranslational Science クリニカルシークエンスの臨床実装による肺癌に対する治療標的分子・個別化治療の探索

  西原 広史, 木下 一郎, 畑中 豊, 田中 伸哉, 秋田 弘俊  日本癌学会総会記事  75回-  SST1  -3  2016/10  [Not refereed][Not invited]
  
• チロシンキナーゼ阻害剤耐性膠芽腫細胞における腫瘍幹細胞性獲得とSFRP1の関連性

  鈴鹿 淳, 津田 真寿美, 王 磊, 谷野 美智枝, 木村 太一, 西原 広史, 田中 伸哉  日本癌学会総会記事  75回-  J  -2036  2016/10  [Not refereed][Not invited]
  
• 肺癌においてアダプター蛋白CrkはTGF-βシグナルと協調してEMTを誘導する

  谷野 美智枝, Elimansuri Aiman, Mahabir Roshan, 王 磊, 木村 太一, 西原 広史, 津田 真寿美, 田中 伸哉  日本癌学会総会記事  75回-  P  -1092  2016/10  [Not refereed][Not invited]
  
• LATS1は浸潤性乳がんにおいてE-cadherinの転写制御因子をリン酸化することによってE-cadherinの発現を抑制する

  向井 智美, 藪田 紀一, 谷野 美智枝, 鳥形 康輔, 関戸 好孝, 田中 伸哉, 野島 博  日本癌学会総会記事  75回-  J  -3099  2016/10  [Not refereed][Not invited]
  
• AKR1C1は膀胱癌の浸潤・転移と薬剤耐性を制御する

  津田 真寿美, 松本 隆児, 吉田 一彦, 谷野 美智枝, 木村 太一, 西原 広史, 阿部 崇重, 篠原 信雄, 野々村 克也, 田中 伸哉  日本癌学会総会記事  75回-  E  -1057  2016/10  [Not refereed][Not invited]
  
• 多形黄色星細胞腫におけるBRAFV600Eの遺伝子変異とリン酸化ERK及びp16の発現の検討

  谷野 美智枝, 津田 真寿美, 石田 雄介, 木村 太一, 西原 広史, 長嶋 和郎, 田中 伸哉  日本病理学会会誌  105-  (2)  75  -75  2016/09  [Not refereed][Not invited]
  
• 脳腫瘍術中迅速病理診断における迅速免疫染色装置(ラピート)の使用経験

  森谷 純, 谷野 美智枝, 木村 太一, 石田 雄介, 津田 真寿美, 西原 広史, 田中 伸哉  日本病理学会会誌  105-  (2)  76  -76  2016/09  [Not refereed][Not invited]
  
• 形態・分子診断と個別化治療 脳腫瘍クリニカルシークエンスの臨床実装にむけて

  西原 広史, 湯澤 明夏, 山口 秀, 小林 浩之, 寺坂 俊介, 田中 伸哉  Brain Tumor Pathology  33-  (Suppl.)  071  -071  2016/05  [Not refereed][Not invited]
  
• 髄膜発生SFT/HPCのNAB2-STAT6融合遺伝子パターンと臨床病理学的検討

  湯澤 明夏, 西原 広史, 王 磊, 津田 真寿美, 木村 太一, 谷野 美智枝, 田中 伸哉  Brain Tumor Pathology  33-  (Suppl.)  102  -102  2016/05  [Not refereed][Not invited]
  
• IPMN関連浸潤癌におけるRNF43,βcatenin,SMAD4,p53の蛋白発現および遺伝子変異プロファイルの解析検討

  大森優子, 大森優子, 谷野美智枝, 水上裕輔, 小野裕介, 安保義恭, 真口宏介, 西原広史, 篠原敏也, 田中伸哉  日本病理学会会誌  105-  (1)  368  2016/04/12  [Not refereed][Not invited]
  
• 臨床的シークエンス分析による髄膜腫の予後的影響(Prognostic impact for meningioma by clinical sequence system)

  湯澤 明夏, 西原 広史, 山口 秀, 毛利 普美, 王 磊, 木村 太一, 津田 真寿美, 谷野 美智枝, 佐藤 典宏, 田中 伸哉  日本病理学会会誌  105-  (1)  356  -356  2016/04  [Not refereed][Not invited]
  
• 大動脈穿破により突然死をきたした3剖検例

  石川麻倫, 宮崎将也, 小西崇夫, 小西崇夫, 中川麗, 谷野美智枝, 西原広史, 藤岡保範, 長嶋和郎, 田中伸哉  日本病理学会会誌  105-  (1)  2016
• 腹腔内出血を発症し死亡した急性膵炎の一例

  曽々木昇, 佐藤龍, 太田智之, 唐崎秀則, 石川麻倫, 木村太一, 田中伸哉, 齋藤博哉  日本腹部救急医学会雑誌  36-  (2)  2016
• CRKアダプター蛋白質はHGF/c-Metフィードバックループを介して膀胱癌のEMTと転移を誘導する

  王 磊, 松本 隆児, 津田 真寿美, 間石 奈湖, 安部 崇重, 木村 太一, 谷野 美智枝, 西原 広史, 樋田 京子, 大場 雄介, 篠原 信雄, 田中 伸哉  日本癌学会総会記事  74回-  J  -1142  2015/10  [Not refereed][Not invited]
  
• 膀胱癌転移巣におけるAldo-keto reductase(AKR)1C1の発現亢進は浸潤能と抗癌剤耐性能を反映する

  松本 隆児, 津田 真寿美, 安部 崇重, 篠原 信雄, 田中 伸哉, 野々村 克也  日本泌尿器科学会総会  103回-  471  -471  2015/04  [Not refereed][Not invited]
  
• 大腸癌におけるp53とCOX‐2発現の臨床病理学的解析

  加藤容崇, 西原広史, 川俣太, 小丹枝裕二, 毛利普美, 木村太一, 谷野美智枝, 武冨紹信, 田中伸哉  日本病理学会会誌  104-  (1)  297  2015/03/23  [Not refereed][Not invited]
  
• Predictive Value of MGMT Gene Promoter and LINE-1 Methylation Status in Primary Central Nervous System Diffuse Large B-Cell Lymphoma Treated With High-Dose MTX and Radiotherapy

  Kanako Hatanaka, Yutaka Hatanaka, Kyoko Fujii, Hiroyuki Kobayashi, Hiromi Kanno, Tomoko Mitsuhashi, Hiroshi Nishihara, Shinya Tanaka, Yoshihiro Matsuno  LABORATORY INVESTIGATION  95-  431A  -431A  2015/02  [Not refereed][Not invited]
  
• Predictive Value of MGMT Gene Promoter and LINE-1 Methylation Status in Primary Central Nervous System Diffuse Large B-Cell Lymphoma Treated With High-Dose MTX and Radiotherapy

  Kanako Hatanaka, Yutaka Hatanaka, Kyoko Fujii, Hiroyuki Kobayashi, Hiromi Kanno, Tomoko Mitsuhashi, Hiroshi Nishihara, Shinya Tanaka, Yoshihiro Matsuno  MODERN PATHOLOGY  28-  431A  -431A  2015/02  [Not refereed][Not invited]
  
• 胸水細胞診にて肺腺癌および甲状腺乳頭癌の未分化転化が鑑別に挙がった1剖検症例

  漆戸 万紗那, 谷野 美智枝, 森谷 純, 木村 太一, 西原 広史, 丸川 活司, 松野 吉宏, 田中 伸哉  北海道臨床細胞学会会報  24-  43  -47  2015
• 胃低分化腺癌及び印環細胞癌におけるactionable mutationの網羅的検討

  石川麻倫, 石川麻倫, 西原広史, 毛利普美, 毛利普美, 加藤容崇, WANG Lei, 木村太一, 津田真寿美, 谷野美智枝, 田中伸哉, 田中伸哉  日本病理学会会誌  104-  (1)  2015
• 次世代シークエンサーによるクリニカルシークエンスの基盤作成

  西原広史, 西原広史, 毛利普美, 毛利普美, 湯澤明夏, 石川麻倫, 西上耕平, 赤羽俊章, 澤田貴宏, 横内浩, 田中伸哉, 田中伸哉  日本臨床腫瘍学会学術集会(CD-ROM)  13th-  2015
• 非小細胞肺癌におけるHER2及び関連分子異常の観察研究(HOT1303‐A)とHER2陽性非小細胞肺癌に対するトラスツズマブの第II相試験(HOT1303‐B)の進行状況報告

  合田智宏, 木下一郎, 秋田弘俊, 大泉聡史, 西村正治, 田中伸哉, 原田敏之, 西原広史, 畑中豊, 松野吉宏, 天野虎次, 佐藤典宏, 磯部宏  肺癌(Web)  54-  (7)  992(J‐STAGE)  -992  2014/12  [Not refereed][Not invited]
  
• Conventional GBM vs GBMO: Are these two diseases the same or not? - Update information

  Tamio Ito, Hirohiko Nakamura, Shinya Tanaka, Tadashi Hasegawa  Neurological Surgery  42-  (11)  997  -1008  2014/11/01  [Not refereed][Not invited]
  
• IMMUNOHISTOCHEMICAL MOLECULAR EXPRESSION PROFILING FOR GLIOBLASTOMA

  Hiroshi Nishihara, Masaya Miyazaki, Hiroyuki Kobayashi, Shunsuke Terasaka, Shinya Tanaka  NEURO-ONCOLOGY  16-  2014/11  [Not refereed][Not invited]
  
• 電界撹拌迅速免疫染色機を使用した脳腫瘍術中迅速免疫染色の有用性

  MORIYA JUN, TANINO MICHIE, TAKENAMI TOMOKO, URUSHIDO MASANA, ENDO AKIKO, TAKIYAMA AKIHIRO, KIMURA TAICHI, NISHIHARA HIROSHI, TANAKA SHIN'YA  日本臨床細胞学会雑誌  53-  615  2014/10/10  [Not refereed][Not invited]
  
• ATDC5細胞におけるPAMPSゲル誘導軟骨分化特異的シグナル伝達経路の解明

  後藤佳子, 北村信人, 木村太一, 仙葉慎吾, 黒川孝幸, GONG Ping, 田中伸哉, 安田和則  日本整形外科学会雑誌  88-  (8)  S1529  2014/08/29  [Not refereed][Not invited]
  
• 大腸癌浸潤・転移におけるchorionic gonadotropin-βの機能解析とその臨床応用

  川俣 太, 本間 重紀, 西原 広史, 長津 明久, 旭 よう, 蒲池 浩文, 高橋 典彦, 津田 真寿美, 田中 伸哉, 神山 俊哉, 武冨 紹信  日本大腸肛門病学会雑誌  67-  (3)  240  -240  2014/03  [Not refereed][Not invited]
  
• 若年発症胃癌における分子プロファイリングによる臨床病理学的検討

  石川麻倫, 毛利普美, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  103-  (2)  2014
• 中枢神経原発血管炎5例の臨床像の検討

  白井 慎一, 矢口 裕章, 上床 尚, 佐久嶋 研, 廣谷 真, 加納 崇裕, 矢部 一郎, 田中 伸哉, 佐々木 秀直  臨床神経学  53-  (12)  1575  -1575  2013/12  [Not refereed][Not invited]
  
• Involvement of EphA2-mediated tyrosine phosphorylation of Shp2 in Shp2-regulated activation of extracellular signal-regulated kinase

  K. Miura, Y. Wakayama, M. Tanino, Y. Orba, H. Sawa, M. Hatakeyama, S. Tanaka, H. Sabe, N. Mochizuki  Oncogene  32-  (45)  5292  -5301  2013/11/07  [Not refereed][Not invited]
   

  Shp2 is a positive regulator for Erk activation downstream of receptor tyrosine kinases for growth factors. It has been controversial how Shp2 induces Erk activation. We here demonstrate that EphA2 is responsible for Shp2-mediated Erk activation by phosphorylating Tyr542 and Tyr580 of Shp2 in the cells stimulated with growth factors. In NMuMG mammary epithelial cells stimulated with hepatocyte growth factor (HGF), HGF-dependent Erk phosphorylation was prolonged only in the presence of EphA2. This Erk activation paralleled the phosphorylation of Tyr542/580 of Shp2 and the association of Grb2 with Shp2, suggesting the positive signal involving Grb2 signal to activate Ras-Erk pathway. Immunohistochemical studies of mammary cancer specimens revealed that the cancer progression was associated with both Tyr580 phosphorylation of Shp2 and increased expression of EphA2, which were also correlated with increased Erk phosphorylation. Overexpression of either Shp2Thr468Met (a phosphatase- defective mutant found in Lentigines, Electrocardiographic abnormalities, Ocular hypertelorism, Pulmonary stenosis, Abnormal genitalia, Retardation of growth and sensorineural Deafness (LEOPARD) syndrome) or Shp2Asn308Asp (a phosphatase-active mutant found in Noonan syndrome) with EphA2 exhibited comparable activation of Erk and stronger activation than wild-type Shp2, suggesting the phosphatase-independent Erk activation. Expression of Shp2Thr468Met with Tyr542/580Phe mutations resulted in the suppression of Erk activation. Phosphatase-active and -inactive, and wild-type Shp2s bound equally to Grb2, suggesting that phosphorylation of Tyr542/580 of Shp2 was essential but not sufficient for Shp2-mediated Erk activation. We found that Gab1 (Grb2-associated binder 1) was involved in the mutant Shp2-mediated Erk activation. Zebrafish injected with Shp2Thr468Met mRNA showed cardiac edema, whereas those depleted of EphA2b showed less phenotype, suggesting that EphA2 might partly account for the phenotype of LEOPARD syndrome. Collectively, tyrosine phosphorylation of Shp2 by EphA2 contributes to the phosphatase-independent Shp2-mediated activation of Erk and might be involved in Shp2-associated diseases. © 2013 Macmillan Publishers Limited.
• THE CLINICOPATHOLOGICAL EVALUATION OFCD 163 AND COX-2 IN MENINGIOMA; IMMUNOHISTOCHEMICAL ANALYSIS OF 76 CASES OF LOW- AND HIGH-GRADE MENINGIOMA

  Hiroshi Nishihara, Hiromi Kanno, Yasutaka Kato, Shinya Tanaka  NEURO-ONCOLOGY  15-  160  -160  2013/11  [Not refereed][Not invited]
  
• THE CLINICOPATHOLOGICAL EVALUATION OF O6-MEHYLGUANINE-DNA METHYLTRANSFERASE IN GLIOBLASTOMA MULTIFORMES

  Masaya Miyazaki, Hiroshi Nishihara, Tamio Itoh, Masahito Kato, Shin Fujimoto, Taichi Kimura, Mishie Tanino, Shinya Tanaka  NEURO-ONCOLOGY  15-  159  -159  2013/11  [Not refereed][Not invited]
  
• The mesenchymal phenotype in recurrent glioblastoma is due to irradiation induced Snail expression and resultant EMT.

  Roshan Mahabir, Mishie Tanino, Aiman Elmansuri, Masumi Tsuda, Taichi Kimura, Lei Wang, Hiroshi Nishihara, Shinya Tanaka  MOLECULAR CANCER THERAPEUTICS  12-  (11)  2013/11  [Not refereed][Not invited]
  
• Erratum: Immunohistochemical molecular expression profile of metastatic brain tumor for potent personalized medicine (Brain Tumor Pathol DOI: 10.1007/s10014-012-0124-y)

  Kato Y, Nishihara H, Yuzawa S, Mohri H, Kanno H, Hatanaka Y, Kimura T, Tanino M, Tanaka S  Brain Tumor Pathology  30-  (4)  266  -267  2013/10  [Not refereed][Not invited]
  
• 消化器癌におけるERC/Mesothelinの分子病理学的検討(Molecular and clinicopathological analysis for ERC/Mesothelin in digestive cancers)

  西原 広史, 川俣 太, 永生 高広, 津田 真寿美, 王 磊, 樋野 興夫, 武冨 紹信, 田中 伸哉  日本癌学会総会記事  72回-  308  -308  2013/10  [Not refereed][Not invited]
  
• 大腸癌浸潤・転移におけるchorionic gonadotropin‐βの機能解析とその臨床応用

  川俣太, 本間重紀, 西原広史, 長津明久, 旭火華, 蒲池浩文, 高橋典彦, 津田真寿美, 田中伸哉, 神山俊哉, 武冨紹信  日本消化器癌発生学会総会プログラム・抄録集  68回-  WS  -8  2013/07  [Not refereed][Not invited]
  
• 大腸癌91例の癌間質におけるCOX‐2発現の臨床病理学的解析

  加藤容崇, 西原広史, 川俣太, 武冨紹信, 田中伸哉  日本病理学会会誌  102-  (1)  323  2013/04/26  [Not refereed][Not invited]
  
• 消化器癌におけるERC/mesothelin発現の分子病理学的解析

  西原広史, 川俣太, 永生高広, 加藤容崇, 樋野興夫, 武冨紹信, 田中伸哉  日本病理学会会誌  102-  (1)  301  2013/04/26  [Not refereed][Not invited]
  
• 中枢神経に限局したアレルギー性肉芽腫性血管炎の1例

  白井 慎一, 上床 尚, 廣谷 真, 佐久嶋 研, 加納 崇裕, 鴨嶋 雄大, 矢部 一郎, 田中 伸哉, 佐々木 秀直  臨床神経学  53-  (4)  320  -320  2013/04  [Not refereed][Not invited]
  
• 大腸癌浸潤・転移におけるchorionic gonadotropin‐βの機能解析とその臨床応用

  川俣太, 本間重紀, 西原広史, 長津明久, 旭火華, 蒲池浩文, 高橋典彦, 津田真寿美, 田中伸哉, 神山俊哉, 武冨紹信  大腸癌研究会プログラム・抄録集  79th-  93  2013  [Not refereed][Not invited]
  
• Isolated granulomatous angiitis with eosinophilia in the central nervous system

  Shirai, S, Yabe, I. (corresponding aut, Sakushima, K, Kanno, H, Uwatoko, H, Hirotani, M, Kano, T, Kamoshima, Y, Tanaka, S, Sasaki, H  Neurology Clin Neurosci  1-  (3)  119  -121  2013  [Refereed][Not invited]
   

  A 73-year-old man was admitted for epileptic seizures. Diffuse white matter lesions were observed in the bilateral occipital lobes, and the left occipital lobe biopsy showed eosinophils and giant cells in the walls of medium to small blood vessels, fibrinoid necrosis, occluded blood vessels, and glia degeneration. Allergic granulomatous angiitis was pathologically diagnosed. No eosinophilic granulomatosis with polyangiitis findings were observed in other organs, and isolated granulomatous angiitis with eosinophilia in the central nervous system was clinically diagnosed.Prednisolone combined with immunosuppressant therapies improved his symptoms markedly.
• Co-overexpression of GEP100 With EGFR Correlates With Early Recurrence After Breast Conservation Therapy (BCT)

  R. Kinoshita, J. Nam, M. Hosoda, C. Kubota K, M. Tanino, A. Hashimoto, Y. M. Ito, S. Tanaka, H. Sabe, H. Shirato  INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS  84-  (3)  S227  -S228  2012/11  [Not refereed][Not invited]
  
• CLINICOPATHOLOGICAL ANALYSIS OF 133 MALIGNANT GLIOMAS; THE PROGNOSTIC VALUE OF OLIGODENDROGLIOMA COMPONENT

  Hiromi Kanno, Hiroshi Nishihara, Shinya Tanaka  NEURO-ONCOLOGY  14-  109  -109  2012/10  [Not refereed][Not invited]
  
• CLINICOPATHOLOGICAL ANALYSIS OF ANGIOCENTRIC, ANGIODESTRUCTIVE LYMPHOPROLIFERATIVE DISORDER IN CENTRAL NERVOUS SYSTEM

  Hiroshi Nishihara, Teruki Yanagi, Hiromi Kanno, Shinya Tanaka  NEURO-ONCOLOGY  14-  109  -109  2012/10  [Not refereed][Not invited]
  
• TRIM67は80K-Hを介してRasを抑制し, 神経突起形成を誘導する

  矢口 裕章, 奥村 文彦, 高橋 秀尚, 加納 崇裕, 亀田 浩之, 内ヶ島 基政, 田中 伸哉, 渡辺 雅彦, 佐々木 秀直, 畠山 鎮次  北海道醫學雜誌 = Acta medica Hokkaidonensia  87-  (4)  175  -175  2012/08/01
• 間質細胞のPDGFRβの発現は、膵癌の予後を規定する(Prognostic impact of PDGFR-beta expression in tumor stroma of pancreatic adenocarcinoma)

  西原 広史, 湯澤 明夏, 狩野 光伸, 田中 伸哉  日本癌学会総会記事  71回-  355  -355  2012/08  [Not refereed][Not invited]
  
• グリオブラストーマにおいてSTAT3阻害はMGMTを発現低下させることで抗がん剤 Temozolomide 耐性を解除する

  高阪 真路, 王 磊, 谷地 一博, 成田 拓人, 木村 太一, 谷野 美智枝, 西原 広史, 田中 伸哉  北海道醫學雜誌 = Acta medica Hokkaidonensia  87-  (4)  2012/08/01  [Not refereed][Not invited]
  
• F-18標識フルオロミソニダゾールとポジトロン断層撮影法によって神経膠芽腫とより低悪性度の神経膠腫を鑑別できるだろう

  平田 健司, 寺坂 俊介, 志賀 哲, 服部 直也, 孫田 恵一, 小林 浩之, 山口 秀, 宝金 清博, 田中 伸哉, 久下 裕司, 玉木 長良  北海道醫學雜誌 = Acta medica Hokkaidonensia  87-  (4)  179  -179  2012/08/01  [Not refereed][Not invited]
  
• Translocation-dependent activation of WNT signaling is essential in synovial sarcoma cells

  Marcel Trautmann, Elisabeth Sievers, Dagmar Kindler, Nicolaus Friedrichs, Arend Koch, Olie Larsson, Shinya Tanaka, Akira Kawai, Hiroshi Sonobe, Eva Wardelmann, Reinhard Buettner, Wolfgang Hartmann  CANCER RESEARCH  72-  2012/04  [Not refereed][Not invited]
  
• 臍帯血細胞移植後にサイトメガロウイルス脳室脳炎を発症した菌状息肉症の一剖検例

  高橋健太, 松川敏大, 後藤秀樹, 遠藤知之, 橋野聡, 木村太一, 谷野美智枝, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  101-  (1)  300  2012/03/26  [Not refereed][Not invited]
  
• 悪性髄膜腫の分子発現プロファイルに基づく予後解析

  西原広史, 湯澤明夏, 菅野宏美, 小林浩之, 寺坂俊介, 田中伸哉, 田中伸哉  日本病理学会会誌  101-  (1)  246  2012/03/26  [Not refereed][Not invited]
  
• 子宮体癌化学療法後に心不全を呈し薬剤性心筋障害を疑われた1剖検症例

  鈴木なつめ, ヘールナンデス真子, 木村太一, 谷野美智枝, 西原広史, 筒井裕之, 田中伸哉  日本病理学会会誌  101-  (1)  438  2012/03/26  [Not refereed][Not invited]
  
• Flail arm syndromeの一剖検例

  加藤容崇, 西原広史, 金藤公人, 木村太一, 谷野美智枝, 佐和弘基, 鎌田一, 長嶋和郎, 田中伸哉, 田中伸哉  日本病理学会会誌  101-  (1)  299  2012/03/26  [Not refereed][Not invited]
  
• 髄膜腫におけるCD163の発現と機能解析

  菅野宏美, 西原広史, 王磊, 木村太一, 谷野美智枝, 田中伸哉, 田中伸哉  日本病理学会会誌  101-  (1)  246  2012/03/26  [Not refereed][Not invited]
  
• 道化師様魚鱗癬モデルマウスは,肺胞拡張不全と重篤な皮膚バリア障害を来す

  柳輝希, 秋山真志, 坂井香織, 西江渉, 清水宏, 西原広史, 田中伸哉  日本皮膚科学会雑誌  122-  (1)  49  2012/01/20  [Not refereed][Not invited]
  
• ドネペジル(アリセプト)によるQT延長が原因と考えられた多形性心室頻拍(torsade de pointes)の1症例

  會澤 佳昭, 鈴木 章彦, 上村 明, 吉村 治彦, 大平 浩司, 原 豊道, 白間 信行, 加藤 総介, 中園 綾乃, 川畑 修平, 木村 孔一, 渡辺 雅弘, 板倉 てい子, 宮川 康宏, 小木 正明, 小木 睦美, 田中 伸哉, 西原 広史, 菅野 宏美, 谷野 美智恵, Rosman Mahabir  岩見沢市立総合病院医誌  37-  (1)  60  -61  2011/06  [Not refereed][Not invited]
  
• 肺静脈閉塞症における細胞増殖因子の関与

  谷野美智枝, 中村紘子, 木村太一, 大塚紀幸, 深澤雄一郎, 西川祐司, 池田健, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  100-  (1)  376  2011/03/26  [Not refereed][Not invited]
  
• 癌テーラーメード治療のための統合的個別化病理診断の基盤作成

  西原広史, 菅野宏美, 石川麻倫, 湯澤彩夏, 木村太一, 谷野美智枝, 田中伸哉, 田中伸哉  日本病理学会会誌  100-  (1)  349  2011/03/26  [Not refereed][Not invited]
  
• 急速な経過で死に至ったneuromyelitis opticaの一例

  長井梓, 高瀬香奈, 菅野宏美, 木村太一, 竹内朗子, 阿部剛典, 尾崎義丸, 谷野美智枝, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  100-  (1)  493  2011/03/26  [Not refereed][Not invited]
  
• IgG4‐related sclerosing pachymeningitisの一例

  菅野宏美, 谷野美智枝, 松岡絵美衣, 伊藤智雄, 渡邉健太郎, 尾崎義丸, 伊東民雄, 木村太一, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  100-  (1)  394  2011/03/26  [Not refereed][Not invited]
  
• GliomaにおけるMGMT発現調節による抗癌剤耐性解除の可能性の検討

  高阪真路, 王磊, 谷地一博, 成田拓人, 木村太一, 谷野美智枝, 西原広史, 田中伸哉  日本病理学会会誌  100-  (1)  349  2011/03/26  [Not refereed][Not invited]
  
• 中皮腫の細胞診を科学する 中皮腫における遺伝子異常検索 細胞診診断応用の可能性

  丸川 活司, 谷野 美智枝, 森谷 純, 山谷 幸恵, 田畑 佑希子, 畑中 豊, 久保田 佳奈子, 田中 伸哉, 松野 吉宏  日本臨床細胞学会雑誌  50-  (Suppl.1)  105  -105  2011/03  [Not refereed][Not invited]
  
• 胎児三次元CT(3D‐CT)を用いた短肋骨骨異形成症の出生前診断

  山田崇弘, 澤井英明, 西村玄, 西田圭一郎, 尾松徳彦, 木村太一, 西原広史, 島田茂樹, 森川守, 山田俊, 長和俊, 田中伸哉, 白土博樹, 櫻木範明, 水上尚典  北海道産科婦人科学会会誌  55-  (1)  135  2011/03  [Not refereed][Not invited]
  
• 疾患形成における標的分子の役割 急激な経過で死亡した新型インフルエンザ肺炎症例の病理学的検討

  瀧山晃弘, Wang Lei, 谷野美智枝, 木村太一, 西原広史, 田中伸哉  分子呼吸器病  15-  (1)  132-136  2011/03/01  [Not refereed][Not invited]
  
• Lower osteocalcin and osteopontin contents of the femoral head in hip fracture patients than osteoarthritis patients

  S. Tanaka, K. Narusawa, H. Onishi, M. Miura, A. Hijioka, Y. Kanazawa, S. Nishida, S. Ikeda, T. Nakamura  OSTEOPOROSIS INTERNATIONAL  22-  (2)  587  -597  2011/02  [Not refereed][Not invited]
   

  In patients with femoral neck fracture, clinical factors, bone metabolism markers (in serum, urine, and bone), bone mineral density, radiographic parameters, and bone histomorphometric parameters were investigated to detect determinants of fragility fracture. The osteocalcin/deoxypyridinoline ratio and osteopontin/calcium ratio of cortical bone were selected as significant predictors. Measurement of bone mineral density is widely used to assess bone strength, but this also depends on other bone components and on bone structure. The objective of this study was to investigate risk factors for fracture related to bone quality, the patient's history, and the patient's lifestyle. Twenty-one patients with femoral neck fracture and 18 patients with osteoarthritis were enrolled. Blood and urine samples were collected on admission to hospital, and bone samples were obtained from femoral necks resected during surgery. Multivariate logistic regression analysis was performed using osteoarthritis and femoral neck fracture as combined variables to assess the influence of alcohol or coffee intake, eating natto (fermented soybeans), osteocalcin and calcium concentrations, the osteocalcin/deoxypyridinoline ratio and osteopontin/calcium ratios of cortical bone and cancellous bone, various bone histomorphometric parameters, the bone mineral density of the lumbar spine and the intact contralateral femoral neck, and various radiographic parameters of the spine By forward stepwise multivariate analysis, the osteocalcin/deoxypyridinoline and osteopontin/calcium ratios of cortical bone were selected as significant factors for fracture (the odds ratios were 0.493 and < 0.001, respectively; both P < 0.001). A decrease of osteopontin and osteocalcin in bone is important for promoting vulnerability to hip fracture.
• Tumor Stem Cells: CD133 Gene Regulation and Tumor Stemness

  Tabu K, Taga T, Tanaka S  Stem Cells and Cancer Stem Cells, Vol.2  145  -153  2011  [Not refereed][Invited]
  
• Glioma Stem Cells

  Tabu K, Taga T, Tanaka S  Molecular Targets of CNS Tumors  151  -176  2011  [Not refereed][Not invited]
  
• Clinicopathological study of secondary gliosarcoma

  Tamio Ito, Yoshimaru Ozaki, Ken-Ichi Sato, Mitsuteru Oikawa, Hirohiko Nakamura, Shinya Tanaka, Mishie Tanino, Kazuo Nagashima  Japanese Journal of Neurosurgery  20-  (4)  289  -298  2011  [Not refereed][Not invited]
   

  Introduction: The gliosarcoma (GS) is a rare variant of glioblastoma (GB) containing distinct gliomatous andsarcomatous components. While uncommon, gliosarcomas have been known to arise secondarily, following con-ventional adjuvant therapy of malignant gliomas. We report a clinicopathological study of three secondary GS(SGS) cases. Clinical findings: All three patients were women, with a mean age of 44.3 years. Two had previously beendiagnosised with GB, and the third had anaplastic astrocytoma (AA). As initial treatment, all patients underwentresection, followed by radiation and chemotherapy. GS was diagnosed at the first recurrence in one patient, and atthe second recurrence in the other two. The mean latency of SGS induction, calculated from the diagnosis of GB/AA to the appearance of SGS, was 13 months. The mean length of survival from the time of SGS diagnosis was6.7 months. The mean overall survival from the initial GB/AA diagnosis was 19.7 months. Clinically, in the finalstage, fibrosarcomatous components were seen to characteristically grow into the subgalea or protrude out of thescalp after several recurrences. Pathological findings: In the two cases diagnosed with SGS at the second recurrence, fibroblastic cells had previously been recognized around the vessels at the first recurrence. Both cases also later exhibited abundantglial fibrillary acidic protein (GFAP) in the glial areas, and reticulin and fibronectin. in the fibrosarcomatous area.In the one SGS case diagnosed at the 1st recurrence, tumor cells showed gliosarcoma composed of chondrosarco-matous elements. Conclusions: SGS is a rare clinical entity that occasionally occurs after conventional adjuvant therapy formalignant gliomas. While the scarcity of cases makes research into the pathogenetic mechanism of SGS very diffi-cult, the strikingly poor survival rates of patients who have undergone combined therapy raise questions concern-ing the value of such therapeutic radiation. A larger number of patients would be valuable in helping to elucidatethe role of radiotherapy, and patient response to treatment.
• Progress of pathological research of brain tumor

  田中 伸哉, 長嶋 和郎  Japanese journal of clinical medicine  68-  111  -118  2010/12
• TGFβ1はGEP100-Arf6-AMAP1経路の活性化によりEMTを誘導し、この活性化は癌幹細胞性と関連する(TGFβ1 activates GEP100-Arf6-AMAP1 pathway to induce EMT, and possible relationship of this activation to cancer stemness)

  橋本 あり, 平野 真理子, 谷野 美智枝, 梅本 勉, 小野寺 康仁, 佐藤 宏紀, 木下 留美子, 南 ジンミン, 大塚 勇太郎, 福田 諭, 白土 博樹, 相沢 慎一, 橋本 茂, 田中 伸哉, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  83回・33回-  2P  -0237  2010/12  [Not refereed][Not invited]
  
• Differential diagnosis of CNS lymphomatoid granulomatosis REPLY

  Hiroshi Nishihara, Akihiro Takiyama, Shinya Tanaka  NEUROPATHOLOGY  30-  (3)  302  -303  2010/06  [Not refereed][Not invited]
  
• 脊髄腫瘍として発見され,ユーイング肉腫との鑑別を要したmyeloid sarcomaの一例

  湯澤明夏, 柴田ひな, 菅野宏美, 谷野美智枝, 矢野俊介, 木村太一, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  99-  (1)  372  2010/03/26  [Not refereed][Not invited]
  
• 神経膠腫のパラフィン包埋切片を用いたFISH法による1番短腕の欠失の遺伝子解析と予後の検討

  佐藤真実, 佐藤真実, 谷野美智枝, 木村太一, 西原広史, 伊東民雄, 佐和広基, 金子貞男, 村田純一, 加藤正仁, 田中伸哉, 田中伸哉  日本病理学会会誌  99-  (1)  352  2010/03/26  [Not refereed][Not invited]
  
• 神経症状を初発とし診断に苦慮した血管内リンパ腫の一例

  泉真祐子, 長谷川祐太, 高阪真路, 谷野美智枝, 木村太一, 古山裕康, 千葉進, 及川光照, 西原広史, 田中伸哉  日本病理学会会誌  99-  (1)  372  2010/03/26  [Not refereed][Not invited]
  
• 新型インフルエンザ(A/H1N1pdm)肺炎によるびまん性肺胞傷害により急死した1剖検例

  瀧山晃弘, 王磊, 谷野美智枝, 木村太一, 西原広史, 川岸直樹, 國枝保幸, 片野晴隆, 長谷川秀樹, 高木知敬, 佐多徹太郎, 田中伸哉, 田中伸哉  日本病理学会会誌  99-  (1)  297  2010/03/26  [Not refereed][Not invited]
  
• 放射線治療後7年間生存し得た原発性悪性心膜中皮腫の一例

  谷野美智枝, マハビール ロシャン, 菅野宏美, 鈴木宏明, 山城勝重, 木村太一, 西原広史, 丸川活司, 松野吉宏, 田中伸哉, 田中伸哉  日本病理学会会誌  99-  (1)  300  2010/03/26  [Not refereed][Not invited]
  
• 中枢神経原発びまん性大細胞型B細胞リンパ腫に対するMGMTを含む免疫染色の検討

  久保田佳奈子, 羽賀博典, 菅野宏美, 小林浩之, 西原広史, 田中伸哉, 松野吉宏  日本病理学会会誌  99-  (1)  353  2010/03/26  [Not refereed][Not invited]
  
• シグナル伝達分子のImmunoprofiling;胃癌20例における臨床病理学的検討

  石川麻倫, 大場彩音, 西原広史, 菅野宏美, 王磊, 木村太一, 谷野美智枝, 田中伸哉, 田中伸哉  日本病理学会会誌  99-  (1)  369  2010/03/26  [Not refereed][Not invited]
  
• Gliomaに特徴的な血管構造の臨床病理学的解析

  菅野宏美, 西原広史, 谷野美智枝, 木村太一, 田中伸哉  日本病理学会会誌  99-  (1)  229  2010/03/26  [Not refereed][Not invited]
  
• 捺印標本を用いたグリオーマのMGMT免疫染色の検討

  青柳瑛子, 王磊, 笹井研, 谷野美智枝, 木村太一, 西原広史, 藤本真, 石井伸明, 伊東民雄, 田中伸哉, 田中伸哉  日本臨床細胞学会雑誌  49-  220  2010/03/22  [Not refereed][Not invited]
  
• Immunoprofiling of signaling molecule for gastric cancer; Pathological basis for Taylor made medicine

  NISHIHARA Hiroshi, KANNO Hiromi, ISHIKAWA Marin, OHBA Ayane, TANAKA Shinya  日本癌学会学術総会記事  69th-  2010
• CNS lymphomatoid granulomatosis with lymph node and bone marrow involvements. (vol 28, pg 640, 2008)

  A. Takiyama, H. Nishihara, U. Tateishi, T. Kimura, W. Lei, K. Marukawa, T. Itoh, S. Hashino, K. Nagashima, S. Tanaka  NEUROPATHOLOGY  29-  (4)  520  -520  2009/08  [Not refereed][Not invited]
  
• FRETバイオセンサーを用いたCML分子標的薬剤の薬効評価系の構築(A FRET-based biosensor for the evaluation of the efficacy of molecular targeted drugs for chronic myeloid leukemia)

  水谷 龍明, 近藤 健, ダルマニン・ステファニー, 津田 真寿美, 田中 伸哉, 浅香 正博, 大場 雄介  日本癌学会総会記事  68回-  457  -457  2009/08  [Not refereed][Not invited]
  
• MDS患者のリンパ節に発生したtrilineage extramedullary myeloid cell tumorの1例

  西原広史, 高阪真路, 久保田加奈子, 田中伸哉, 松野吉宏  日本リンパ網内系学会会誌  49-  130  2009/06/10  [Not refereed][Not invited]
  
• 悪性胸膜中皮腫におけるシグナル伝達アダプター分子CRKを介したRac活性の可視化

  谷野美智枝, 王磊, 津田真寿美, 西原広史, 大場雄介, 矢野聖二, 曽根三郎, 田中伸哉  日本呼吸器学会雑誌  47-  237  2009/05/10  [Not refereed][Not invited]
  
• 脳腫瘍血管壁の組織病理学的解析 Glomeruloid vesselは肥厚した幼若なPericyteで構成されている

  西原 広史, 狩野 光伸, 田中 伸哉  Brain Tumor Pathology  26-  (Suppl.)  62  -62  2009/05  [Not refereed][Not invited]
  
• 間質性肺炎の経過中,急性に呼吸不全が悪化し死亡された11剖検例の検討

  谷野美智枝, 高阪真路, 木村太一, 西原広史, 西原広史, 進藤正信, 田中伸哉, 田中伸哉  日本病理学会会誌  98-  (1)  330  2009/03/20  [Not refereed][Not invited]
  
• 転落事故後の腹腔内出血にて死亡した一剖検例;Autopsy Imagingの有用性と限界

  川田淑子, 藤枝迪子, 藤枝迪子, 瀧山晃弘, 金藤きみと, 谷野美智枝, 西原広史, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  98-  (1)  397  2009/03/20  [Not refereed][Not invited]
  
• 滑膜肉腫細胞株におけるCD133の機能解析

  木村太一, 王磊, 平賀博明, 西原広史, 田中伸哉  日本病理学会会誌  98-  (1)  391  2009/03/20  [Not refereed][Not invited]
  
• N型糖鎖のbisectin GlcNAc構造は膠芽腫においてMIB1インデックスが低い領域に高発現する

  青柳瑛子, 笹井研, WANG Lei, 野田頭未歩, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  98-  (1)  300  2009/03/20  [Not refereed][Not invited]
  
• hCG産生大腸癌の分子病理学的検討

  吉永智彰, 吉永智彰, 西原広史, 西原広史, 福島祐介, 佐和弘基, 村上普美, 木村太一, 谷野美智枝, 田中伸哉, 田中伸哉  日本病理学会会誌  98-  (1)  227  2009/03/20  [Not refereed][Not invited]
  
• DOCK180はHodgkin細胞に発現し,細胞の形態制御に重要な役割を果たしている

  王磊, 西原広史, 西原広史, 田中伸哉, 田中伸哉  日本病理学会会誌  98-  (1)  276  2009/03/20  [Not refereed][Not invited]
  
• Clostridium属によるガス壊疽により死亡した2剖検例

  石川麻倫, 柴田頌太, 谷野美智枝, 木村太一, 西原広史, 篠原敏也, 田中伸哉, 田中伸哉  日本病理学会会誌  98-  (1)  397  2009/03/20  [Not refereed][Not invited]
  
• Tumefactive Multiple Sclerosis: A Case Report

  鈴木 聡, 及川 光照, 伊東 民雄, 佐光 一也, 尾崎 義丸, 吉永 智彰, 西原 広史, 田中 伸哉, 中村 博彦  北海道脳神経疾患研究所医誌 = Journal of Hokkaido Brain Research Foundation  19-  (1)  61  -64  2009/03/01  

  Multiple sclerosis (MS) is an autoimmune condition in which the immune system attacks the central nervous system, leading to demyelination in time and space. Tumefactive demyelinating lesions are generally thought of as solitary lesions greater than 2 cm, with MR imaging characteristics mimicking neoplasms. Without a history of multiple sclerosis, surgical biopsy is often performed in suspected tumors. Herein we describe a case of a patient whom over a six-month period developed relapsing-remitting tumefactive MS. Case: A 32-year-old woman presented with progressive left hemiparesis. Magnetic resonance imaging revealed large solitary right periventricular lesions with ring-like enhancement. Open biopsy via craniotomy was performed under the suspicion of tumor. A pathological examination showed massive demyelination. Tumefactive demyelinating lesions will show a good response to corticosteroid therapy with disappearance of the lesions on follow-up imaging.
• 腫瘤様多発性硬化症の一例

  鈴木聡, 及川光照, 伊東民雄, 佐光一也, 尾崎義丸, 吉永智彰, 西原広史, 田中伸哉, 中村博彦  北海道脳神経疾患研究所医誌  19-  (1)  61-64  2009/03/01  [Not refereed][Not invited]
  
• Involvement of Adaptor Protein Crk in Malignant Features of Human Mesothelioma.

  M. A. Tanino, L. Wang, S. Kohsaka, T. Kimura, K. Sasai, H. Nishihara, M. Tsuda, S. Yano, S. Tanaka  AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE  179-  2009  [Not refereed][Not invited]
  
• Negative regulation of angiogenesis by zinc-finger transcription factor TCF8

  Takayuki Inuzuka, Masumi Tsuda, Shinya Tanaka, Yujiro Higashi, Hideaki Kawaguchi, Yusuke Ohba  JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY  45-  S11  -S12  2008/10  [Not refereed][Not invited]
  
• ヒト正常血液細胞分化に伴うDOCK180の発現量変化

  熊野弘毅, 西尾充史, 藤本勝也, 西原広史, 遠藤知之, 山本聡, 坂井俊哉, 小原雅人, 田中伸哉, 佐藤典宏, 小池隆夫  臨床血液  49-  (9)  1194  -1194  2008/09/30  [Not refereed][Not invited]
  
• 胸膜中皮腫におけるシグナル伝達アダプター分子CRKの役割の解析

  谷野美智枝, 王磊, 西原広史, 松野吉宏, 矢野聖二, 曽根三郎, 田中伸哉  日本呼吸器学会雑誌  46-  289  2008/05/10  [Not refereed][Not invited]
  
• 肺出血及び消化管穿孔を来たした血管型Ehlers‐Danlos症候群の一剖検例

  瀧山晃弘, 谷野美智枝, 篠原敏也, 西原広史, 田中伸哉  日本病理学会会誌  97-  (1)  372  2008/03/31  [Not refereed][Not invited]
  
• 神経系,軟部組織及び胸壁における血管周皮腫と孤立性線維性腫瘍の組織病理学的解析

  谷野美智枝, 西原広史, 高阪真路, 木村太一, 久保田佳奈子, 伊藤智雄, 松野吉宏, 山城勝重, 長嶋和郎, 田中伸哉  日本病理学会会誌  97-  (1)  239  2008/03/31  [Not refereed][Not invited]
  
• 悪性膠腫におけるMGMT発理の病理組織学的解析

  野田頭未歩, 青柳瑛子, 笹井研, 王磊, 西原広史, 長嶋和郎, 田中伸哉  日本病理学会会誌  97-  (1)  244  2008/03/31  [Not refereed][Not invited]
  
• 悪性膠腫におけるCD133発現の分子細胞病理学的解析

  椨康一, 笹井研, 木村太一, 谷野美智枝, 青柳瑛子, 王磊, 高阪真路, 西原広史, 田中伸哉  日本病理学会会誌  97-  (1)  244  2008/03/31  [Not refereed][Not invited]
  
• Cortical dysplasiaに合併したpilocytic astrocytomaの一例

  吉永智彰, 西原広史, 長嶋和郎, 田中伸哉  日本病理学会会誌  97-  (1)  389  2008/03/31  [Not refereed][Not invited]
  
• Genetic disruption of all nitric oxide synthase isoforms enhances bone mineral density and bone turnover in mice in vivo: Involvement of the renin-angiotensin system

  Ken Sabanai, Masato Tsutsui, Akinori Sakai, Hideyuki Hirasawa, Shinya Tanaka, Eiichiro Nakamura, Toshitaka Nakamura, Nobuyuki Yanagihara  JOURNAL OF PHARMACOLOGICAL SCIENCES  106-  156P  -156P  2008  [Not refereed][Not invited]
  
• Nonenzymatic collagen cross-links induced by glycoxidation (pentosidine) predicts vertebral fractures

  Masataka Shiraki, Tatsuhiko Kuroda, Shiro Tanaka, Mitsuru Saito, Masao Fukunaga, Toshitaka Nakamura  Journal of Bone and Mineral Metabolism  26-  (1)  93  -100  2008/01  [Not refereed][Not invited]
   

  Advanced glycation end products (AGE) in collagen have been reported to decrease the mechanical property of bone. However, there are no available data on the relation between fracture risk and levels of glycoxidative (nonenzymatic) cross-links of collagen in clinical samples. A total of 432 Japanese elderly women who were not receiving any drug treatment for osteoporosis were selected and followed for 5.2 ± 3.3 (mean ± SD) years for this observational study. Vertebral fractures and bone mineral density were assessed at baseline and then at 1- to 2-year intervals or at indication of any symptom. Two types of collagen metabolites were measured at baseline: urinary N-terminal telopeptide of type I collagen (NTX), a marker of pyridinium cross-link, and urinary pentosidine, a nonenzymatic collagen cross-link produced by AGEs. A total of 97 incident vertebral fractures on 72 subjects were observed. Simple regression analysis using Cox's hazards model showed that log-transformed urinary NTX and pentosidine are significant risk factors for time-dependent incidence of vertebral fractures, in addition to the traditional risk factors (age, lumbar bone mineral density, and number of prevalent vertebral fractures). However, urinary excretion of pentosidine (hazard ratio, 1.33 95% CI, 1.01-1.76, P = 0.04) was a significant predictor of incident vertebral fracture after adjustment for other traditional risk factors. The present data suggest that AGE-related collagen cross-link is a novel risk for vertebral fracture. © 2008 Springer-Verlag.
• Allograft inflammatory factor-1 augments macrophage phagocytosis activity and accelerates the progression of atherosclerosis in ApoE-/- mice

  Mishima, T, Iwabuchi, K, Fujii, S, Tanaka, S, Ogura, H, Watano-Miyata, K, Ishimori, N, Andoh, Y, Nakai, Y, Iwabuchi, C, Ato, M, Kitabatake, A, Tsutsui, H, Onoé, K  Int. J. Mol. Med.  21-  (2)  181  -187  2008  [Not refereed][Not invited]
  
• 癌関連シグナル伝達アダプター分子CRKの構造生物学的解析

  田中伸哉, 酒井美恵子, 牧野吉倫, 西原広史, 小橋川敬博, 稲垣冬彦  日本病理学会会誌  96-  (1)  168  2007/02/05  [Not refereed][Not invited]
  
• 大脳に発生したGanglioneuroblastomaの一例

  吉永智彰, 西原広史, 谷野美智枝, 田中伸哉  日本病理学会会誌  96-  (1)  355  2007/02/05  [Not refereed][Not invited]
  
• 咽頭癌術後に多発転移を来たしたホルモン産生腫瘍の一例

  田村佳奈恵, 西原広史, 酒井美恵子, 谷野美智枝, 木村太一, 山田範幸, 鈴木章之, 鈴木清語, 進藤正信, 田中伸哉  日本病理学会会誌  96-  (1)  355  2007/02/05  [Not refereed][Not invited]
  
• 中枢神経発症Lymphomatoid granulomatosis(LYG)の分子病理学的検討

  西原広史, 立石宇貴秀, 長嶋和郎, 田中伸哉  日本病理学会会誌  96-  (1)  174  2007/02/05  [Not refereed][Not invited]
  
• Pharmacological cdk inhibitor suppresses JC virus proliferation

  Yasuko Orba, Yuji Sunden, Tadaki Suzuki, Takashi Kimura, Shinya Tanaka, Kazuo Nagashima, Hirofumi Sawa  JOURNAL OF NEUROVIROLOGY  13-  109  -110  2007  [Not refereed][Not invited]
  
• Biological effects of Natalizurnab on JC virus infectivity in its permissive human neural cell lines in vitro

  Tadaki Suzuki, Satoko Yamanouchi, Yuji Sunden, Yasuko Orba, Shinya Tanaka, Takashi Kimura, Hirofumi Sawa  JOURNAL OF NEUROVIROLOGY  13-  127  -127  2007  [Not refereed][Not invited]
  
• Case report of spermatocytic seminoma

  ICHIHARA Shin, TOKUYAMA Mika, KIMURA Taichi, AOYANAGI Eiko, NAKATA Yasunobu, TANAKA Shinya  診断病理 : Japanese journal of diagnostic pathology  23-  (4)  281  -283  2006/10/31  [Not refereed][Not invited]
  
• 脳腫瘍(神経膠芽腫)細胞株における癌遺伝子Crkの機能解析

  WAN Lei, 西原広史, 酒井美恵子, 牧野吉倫, 市原真, 田中伸哉  日本癌学会学術総会記事  65th-  164  2006/08/28  [Not refereed][Not invited]
  
• 膿ようを形成し,壊死性筋膜炎を併発した十二指腸原発GISTの一剖検例

  徳山実佳, 西原広史, 田中伸哉  日本病理学会会誌  95-  (1)  382  2006/03/18  [Not refereed][Not invited]
  
• 神経こう芽腫細胞株における癌遺伝子Crkの機能解析

  王らい, 内野晴登, 酒井美恵子, 牧野吉倫, 西原広史, 田中伸哉  日本病理学会会誌  95-  (1)  311  2006/03/18  [Not refereed][Not invited]
  
• 心筋梗塞後の細菌性心筋炎の一例

  種井善一, 的場智子, 腰山博昭, 伊東知子, 大久保亮, 金沢剛志, 北川寛, 工藤京平, 高谷紗帆, 田中諭, 東山明日美, 帆士縫, 山崎和義, 横畠絵美, 西原広史, 長嶋和郎, 長嶋和郎, 田中伸哉  日本病理学会会誌  95-  (1)  381  2006/03/18  [Not refereed][Not invited]
  
• 5歳小児に発症したspindle cell sarcomaの剖検例

  大村瞳, う ひょう, 上野山敦士, 小松佳奈, 松村馨, 西原広史, 野島孝之, 田中伸哉, 進藤正信  日本病理学会会誌  95-  (1)  384  2006/03/18  [Not refereed][Not invited]
  
• Effects of bisphosphonate (YM529) on osteoblastic function and differentiation in bone regeneration after drill-hole injury in mice

  M Nagashima, A Sakai, S Uchida, S Tanaka, M Tanaka, T Nakamura  JOURNAL OF BONE AND MINERAL RESEARCH  19-  S266  -S266  2004/10  [Not refereed][Not invited]
  
• The reduced osteogenic potential after skeletal unloading relates to the decreased expression of PECAM-1 in bone marrow cells in mice.

  M Sakuma-Zenke, A Sakai, S Nakayamada, N Kunugita, S Uchida, S Tanaka, T Mori, Y Tanaka, T Nakamura  JOURNAL OF BONE AND MINERAL RESEARCH  19-  S255  -S255  2004/10  [Not refereed][Not invited]
  
• NSAIDsの抗腫よう機序の解明:cAMPはc‐IAP2及びCOX‐2の発現を誘導し,大腸癌細胞のアポトーシスを抑制する

  西原広史, 田中伸哉, 長嶋和郎  日本癌学会総会記事  63rd-  211  2004/08/25  [Not refereed][Not invited]
  
• Identification of cell surface molecule as a candidate receptor for JC virus

  C. Henmi, H. Sawa, Y. Orba, S. Tanaka, K. Nagashima  JOURNAL OF NEUROVIROLOGY  14-  42  -42  2004  [Not refereed][Not invited]
  
• Human polyomavirus agnoprotein disrupts the interaction between HP1 alpha and LBR

  H. Sawa, Y. Okada, T. Suzuki, Y. Orba, Y. Sunden, C. Henmi, S. Semba, H. Takahashi, S. Tanaka, K. Nagashima  JOURNAL OF NEUROVIROLOGY  14-  33  -33  2004  [Not refereed][Not invited]
  
• JCウイルス外郭蛋白質VP1は糖脂質と糖蛋白質の糖鎖に結合する

  澤 洋文, 駒込 理佳, 田中 伸哉, 長嶋 和郎  Neuropathology : official journal the Japanese Society of Neuropathology  23-  199  -199  2003/05/01
• 上皮性の成分を伴ったglioblastomaの一例

  中川 智子, 佐藤 司, 西谷 幹雄, 近井 佳奈子, 田中 伸哉, 大西 晶子, 長嶋 和郎  Neuropathology : official journal the Japanese Society of Neuropathology  23-  296  -296  2003/05/01
• Advanced glycation end products induce angiogenesis in vivo

  AC Stan, T Okamoto, S Tanaka, T Koike, M Kase, Z Makita, H Sawa, K Nagashima  ACTA NEUROPATHOLOGICA  104-  (5)  570  -570  2002/11  [Not refereed][Not invited]
  
• Expression of JC virus agnoprotein in progressive multifocal leukoencephalopathy brain

  AC Stan, Y Okada, H Sawa, S Endo, Y Orba, T Umemura, H Nishihara, S Tanaka, H Takahashi, K Nagashima  ACTA NEUROPATHOLOGICA  104-  (5)  570  -570  2002/11  [Not refereed][Not invited]
  
• シグナル伝達アダプター分子CrkによるERMの局在変化と細胞運動能及び接着能の解析

  津田真寿美, 田中伸哉, 牧野吉倫, 西原広史, 沢洋文, 長嶋和郎  日本癌学会総会記事  61st-  317  2002/08/25  [Not refereed][Not invited]
  
• 遺伝子導入ベクターとしてのJC virus(JCV)VP1の有用性

  逸見 千寿香, 岩田 博司, 駒込 理佳, 佐藤 真実, 田中 伸哉, 澤 洋文, 長嶋 和郎  Neuropathology : official journal the Japanese Society of Neuropathology  22-  52  -52  2002/05/01
• 副腎骨髄脂肪腫を合併した腎腺腫の1例

  瀧山晃弘, 西原広史, 市原真, 小林真也, 田中伸哉, 伊藤智雄, 長嶋和郎  診断病理  19-  (2)  134-137  2002/04/30  [Not refereed][Not invited]
  
• アダプター分子v‐CrkによるRho依存的細胞骨格制御と細胞運動能の解析

  津田真寿美, 田中伸哉, 西原広史, 沢洋文, 長嶋和郎  日本病理学会会誌  91-  (1)  159  2002/02/28  [Not refereed][Not invited]
  
• Laser capture microdissection法を用いた胆汁細胞診標本からの細胞採取とK‐ras遺伝子変異の検索

  大場靖子, 西原広史, 田中伸哉, 沢洋文, 伊藤智雄, 長嶋和郎  日本病理学会会誌  91-  (1)  298  2002/02/28  [Not refereed][Not invited]
  
• Broad distribution of the JC virus receptor contrasts with a marked cellular restriction of virus replication

  S Suzuki, H Sawa, R Komagome, Y Orba, M Yamada, Y Okada, Y Ishida, H Nishihara, S Tanaka, K Nagashima  VIROLOGY  286-  (1)  100  -112  2001/07  [Not refereed][Not invited]
   

  To investigate the early events of JC virus (JCV) infection, including attachment, penetration, transport to the nuclei, and replication of the virus, we analyzed the susceptibility of 15 different cell lines to infection using a semiquantitative polymerase chain reaction (PCR) assay, in situ hybridization, laser scanning confocal microscopy, and a viral replication assay. The cell lines examined were human permissive and nonpermissive cells as well as cells of monkey and mouse origin. JCV entry into the nuclei of the all cell lines was observed within 10 min after inoculation, demonstrating that the virus receptor is widely distributed among mammalian cells. inhibition of viral entry by an anti-JCV VP1 antibody and sialidase treatment to remove sialic acid residues, which are considered a candidate for the JCV receptor, suggested that VP1 may interact with the cellular surface sialic acid. In addition, chlorpromazine, a clathrin-dependent pathway inhibitor, significantly suppressed entry of JCV into nuclei. In spite of the broad spectrum of cells susceptible to JCV entry, replication of the virus occurred exclusively in human neuroblastoma cell lines. These results suggest that whereas JCV can enter a wide variety of cell types and localize to the nuclei, cell-specific intranuclear mechanisms are required for virus replication. (C) 2001 Academic Press.
• JC virus large T protein transforms rodent cells but is not involved in human medulloblastoma

  H Hayashi, S Endo, S Suzuki, S Tanaka, H Sawa, Y Ozaki, Y Sawamura, K Nagashima  NEUROPATHOLOGY  21-  (2)  129  -137  2001/06  [Not refereed][Not invited]
   

  JC virus (JCV) together with Simian virus 40 (SV40) and BK virus (BKV), belong to the polyomavirus group and these viruses are neuro-oncogenic to rodents by expression of large T antigen (LT), which binds to cellular p53 and pRB thus reducing the anticancer potential of the cell. The function of LT has not been clarified because small t antigen (st) is transcribed from the same start codon as the overlapping reading frame of LT, and is translated as a different protein with the same N-terminal residues (1-81 amino acids) by a splice-site variant of mRNA. To elucidate the function of LT without st, we constructed plasmids that express LT only by deleting the splicing region including the C-terminus of st, and consequently stable cell lines were established that express only JCLT, SV40LT and BKLT. The growth rates of these cells were examined in colonies on soft agar and it was found that LT alone has a transforming capacity; the order of efficiency being SV40LT, BKLT and JCLT. In addition, to verify the involvement of JCV in human medulloblastoma, eight cases of medulloblastoma, six cases of frozen material and five cases of paraffin-embedded tissues which included three cases of frozen tissues, were examined. PCR assay, genomic Southern blotting, and in situ hybridization were applied to detect the JCV genome, and LT and st were examined by immunohistochemistry; the results were compared with JCV-infected tissues as a positive control. All methods failed to detect not only JCV genome but also LT protein in medulloblastoma and it was concluded that JCV LT has transforming activities in rodent cells, but is not related to human medulloblastoma.
• HHV8-negative primary effusion lymphoma of the peritoneal cavity presenting with a distinct immunohistochemical phenotype

  S Tanaka, H Katano, K Tsukamoto, M Jin, S Oikawa, H Nishihara, H Sawa, K Sawada, M Shimizu, T Sata, Y Fujioka, K Nagashima  PATHOLOGY INTERNATIONAL  51-  (4)  293  -300  2001/04  [Not refereed][Not invited]
   

  Primary effusion lymphoma (PEL) has been recognized as a body-cavity-based lymphoma that was originally reported to be associated with human herpes virus 8 (HHV8) infection, and was frequently found in human immunodeficiency virus-positive (HIV) patients. Here we describe an autopsy case of PEL of the peritoneal cavity in an immunocompetent patient. Cytological analysis of tumor cells within ascites revealed immunocytochemical features of keratin positivity and CD45 negativity. At autopsy, the presence of a massive volume of ascites as well as diffuse tumor cell infiltrates within the serosa of the intestine and mesenterium were observed, Tumor cells were morphologically similar to anaplastic large-cell lymphoma, but were immunohistochemically positive for keratin and epithelial membrane antigen (EMA). They also showed no reactivity to representative lymphocyte surface markers including CD45, in addition to being negative for CD30 and p80(NPM/ALK). Molecular analysis of the tumor cells revealed monoclonality of the immunoglobulin heavy-chain gene rearrangement which demonstrated a lymphoma of the B-cell lineage. Furthermore, HHV8 was not detected by immunohistochemical analysis, PCR or nested PCR technique. Based on these results, we consider the present case to be an HHV8-negative PEL with keratin and EMA positivity.
• 滑膜肉腫の癌化機構におけるSYT‐SSX1キメラ蛋白の役割

  長井真人, 田中伸哉, 平賀博明, 津田真寿美, 遠藤秀一, 沢洋文, 西原広史, 長嶋和郎  日本病理学会会誌  90-  (1)  190  2001/03/01  [Not refereed][Not invited]
  
• すい癌におけるSykの発現の検討 浸潤・転移への関与について

  渡辺佳明, 西原広史, 田中伸哉, 沢洋文, 清水道生, 長嶋和郎  日本病理学会会誌  90-  (1)  292  2001/03/01  [Not refereed][Not invited]
  
• JC virus(JCV) agnoproteinの発現と機能に関する研究

  岡田由紀, 沢洋文, 遠藤秀一, 大場靖子, 西原広史, 田中伸哉, 長嶋和郎  日本病理学会会誌  90-  (1)  349  2001/03/01  [Not refereed][Not invited]
  
• Neural cell specific activation of the JC virus promoter in the presence of HTLV-1 Tax.

  Y Okada, H Sawa, S Tanaka, S Suzuki, H Hasegawa, WW Hall, K Nagashima  AIDS RESEARCH AND HUMAN RETROVIRUSES  17-  S56  -S56  2001  [Not refereed][Not invited]
  
• Analysis of transforming activity of human synovial sarcoma-associated chimeric protein SYT-SSX1 bound to chromatin remodeling factor hBRM/hSNF2α

  Proc. Natl. Acad. Sci. USA  98-  (7)  3843  -3848  2001  [Not refereed][Not invited]
  
• 浮遊細胞特異的に発現するCDM‐family蛋白 DOCK2の機能の解析

  前田才恵, 西原広史, 田中伸哉, 松田道行, 沢洋文, 長嶋和郎  日本分子生物学会年会プログラム・講演要旨集  23rd-  432  2000/11/25  [Not refereed][Not invited]
  
• 新規tumor suppressorとしてのc‐Myc結合タンパク質MM‐1の同定と作用機作

  藤岡優子, 平敬宏, 佐藤亜希子, 田中伸哉, 西原広史, 有賀(井口)早苗, 長嶋和郎, 有賀寛芳  日本分子生物学会年会プログラム・講演要旨集  23rd-  339  2000/11/25  [Not refereed][Not invited]
  
• ヒト滑膜肉腫関連蛋白SYT‐SSX1とクロマチンリモデリング因子hBRMの結合の機能解析

  長井真人, 田中伸哉, 津田真寿美, 園部宏, 加藤宏幸, 平賀博明, 西原広史, 沢洋文, 長嶋和郎  日本分子生物学会年会プログラム・講演要旨集  23rd-  338  2000/11/25  [Not refereed][Not invited]
  
• Expression of JC virus early and late proteins in various cell lines and PML brains

  H Sawa, Y Okada, S Tanaka, Y Orba, S Endo, M Sasada, Y Hara, M Shintaku, K Nagashima  BRAIN PATHOLOGY  10-  (4)  757  -757  2000/09  [Not refereed][Not invited]
  
• 腫よう組織における前癌遺伝子産物CRKの発現と細胞増殖能に関する分子病理学的解析

  西原広史, 田中伸哉, 清水道生, 沢洋文, 松田道行, 長嶋和郎  Jpn J Cancer Res  91-  (Supplement (Sept))  499  2000/09/01  [Not refereed][Not invited]
  
• ヒト滑膜肉腫関連SYT‐SSX1遺伝子の形質転換機構

  長井真人, 田中伸哉, 沢洋文, 西原広史, 津田真寿美, 長嶋和郎  Jpn J Cancer Res  91-  (Supplement (Sept))  53  2000/09/01  [Not refereed][Not invited]
  
• JC virus 構成蛋白の発現と機能に関する研究

  岡田 由紀, 遠藤 秀一, 原 由紀子, 大場 靖子, 駒込 理佳, 田中 伸哉, 澤 洋文, 長嶋 和郎  Neuropathology : official journal the Japanese Society of Neuropathology  20-  59  -59  2000/06/01
• Dendritic distribution of JC virus agnoprotein in PML brain.

  K Nagashima, H Sawa, Y Ohba, S Endo, Y Okada, S Tanaka  JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY  59-  (5)  427  -427  2000/05  [Not refereed][Not invited]
  
• The Ras-mitogen-activated protein kinase pathway is critical for the activation of matrix metalloproteinase secretion and the invasiveness in v-crk-transformed 3Y1

  EB Liu, AA Thant, F Kikkawa, H Kurata, S Tanaka, A Nawa, S Mizutani, S Matsuda, H Hanafusa, M Hamaguchi  CANCER RESEARCH  60-  (9)  2361  -2364  2000/05  [Not refereed][Not invited]
   

  To search for the intracellular signaling pathway critical for the secretion of matrix metalloproteinases (MMP), we studied the effects of dominant negative Ras (S17N Ras) and dominant negative MEK1 (MEK1AA) expression in v-crk-transformed 3Y1. Expression of either S17N Ras or MEK1AA dramatically suppressed the augmented secretion of MMP-2 and MMP-9 in v-crk-transfected 3Y1. Similarly, a Ras farnesyltransferase inhibitor, manumycin A, and a MEK1 inhibitor, U0126, suppressed MMP secretion in a dose-dependent manner, whereas a PI3 kinase inhibitor, wortmannin, could not. In addition, the suppression of MMP secretion by S17N Ras showed good correlation with the inhibition of in vitro invasiveness of the cells. In contrast, expression of dominant negative C3G did not suppress MMP secretion, although it substantially blocked the c-Jun N-terminal kinase activation. Taken together, the Ras-MEK1 pathway, but not the C3G-JNK pathway, seems to play a key role in the activation of MMP secretion and, hence, the invasiveness of v-crk-transformed cells.
• ヒト腫よう組織における前癌遺伝子産物Crkの発現解析 免疫組織学的検討及びPCR‐SSCP法を用いた遺伝子変異の解析

  西原広史, 田中伸哉, 松田道行, 清水道生, 長嶋和郎  日本病理学会会誌  89-  (1)  224  2000/03/10  [Not refereed][Not invited]
  
• Transcriptional Activation of JC Virus by Human T-lymphotropic Virus Type (]G0001[) Tax Protein in Human Neuronal Cell Lines.

  J. Biol. Chem.  275-  (22)  17016  -17023  2000  [Not refereed][Not invited]
  
• Crk Activation of JNK via C3G and R-Ras

  J. Biol. Chem.  275-  (17)  12667  -12671  2000  [Not refereed][Not invited]
  
• HTLV-I TaxによるJC virusの活性化に関する研究

  岡田 由紀, 澤 洋文, 田中 伸哉, 伊藤 智雄, 長谷川 秀樹, 長嶋 和郎  Neuropathology : official journal the Japanese Society of Neuropathology  19-  69  -69  1999/06/03
• Activation of C3G guanine nucleotide exchange factor for Rap 1 by phosphorylation of tyrosive 504

  J. Biol. Chem.  274-  (20)  14376  -14381  1999  [Not refereed][Not invited]
  
• Adult Leigh syndrome with mitochondrial DNA mutation

  Acta Neuropathologica  274-  416  -422  1999  [Not refereed][Not invited]
  
• Meningeal large granular lymphocyte lymphoma

  T Nagashima, M Mori, K Kazumata, M Fujimoto, B Kuroda, M Nunomura, T Shinohara, H Hasegawa, Y Watanabe, S Tanaka, K Nagashima  NEUROPATHOLOGY  18-  (3)  336  -342  1998/09  [Not refereed][Not invited]
   

  A 57-year-old man who presented with leptomeningeal lymphoma was reported. The lymphoma cells in the cerebrospinal fluid were large in size, had atypical nuclei and contained many azurophilic granules in their pale cytoplasm, all of which were consistent with malignant lymphoma of large granular lymphocytes (LGL), Immunohistochemically, tumor cells were positive for cytoplasmic CD3, UCHL-1, and CD56, markers of natural killer (NK) cells, Epstein-Barr virus encoded small RNA (EBER) was detected in tumor cells by in situ hybridization, No evidence of parenchymal central nervous system or systemic tumor was identified, although at autopsy microscopic lymphoma involvement was found in the pituitary gland and kidney. To our knowledge this is the first autopsy case report of primary meningeal LGL lymphoma.
• Involvement of cell membrane factors in JC virus infection

  S Itoh, S Nukuzuma, C Nukuzuma, S Tanaka, K Nagashima  NEUROPATHOLOGY  18-  (1)  67  -72  1998/03  [Not refereed][Not invited]
   

  At present it remains unknown if the JC virus (JCV), the causative agent of progressive multifocal leukoencephalopathy, employs specific cellular receptors or cell membrane factors for viral entry. To investigate this, me have examined the cell-specific replication of a chimeric JCV, which contains the control region (CR) cloned from JCI and the coding sequences of the prototype Mad-1 virus. We examined and compared the hemagglutination (HA) titers produced during viral multiplication following microinjection, transfection with chloramphenicol acetyltransferase (CAT) assays, and inoculation using a permissive cell line IMR-32 and three non-permissive cell lines COS-7, CV-1, and A431. Virus microinjected into cells proliferated well in both IMR-32 and COS-7, but not in the non-permissive CV-1 and A431 cell lines. When cells were infected with the chimeric JCV, the HA titers were high in IMR-32, low in COS-7, and negative in the other cell lines. Chloramphenicol acetyltransferase assays demonstrated that the CR was active in IMR-32, COS-7, and CV-1, and inactive in A431 cells, The results suggest that not only nuclear, but also cell membrane factors play an important role in the susceptibility of cells to JCV infection.
• "Guanine-nucleotide exchange prowein C3G activates JNK1 by a Ras-independent mechanism : JNK1 activation inhibited by kinse negative forms of MLK3 and DLK mixed lineage kinases"

  J. Biol. Chem  273-  (3)  1281  -1284  1998  [Not refereed][Not invited]
  
• Multiple hilar cysts of the liver in patients with alcoholic cirrhosis: Report of three cases

  Y Fujioka, N Kawamura, S Tanaka, M Fujita, H Suzuki, K Nagashima  JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY  12-  (2)  137  -143  1997/02  [Not refereed][Not invited]
   

  The present report concerns the clinicopathological study of three patients with alcoholic cirrhosis (a 40-year-old man, a 52-year-old woman and a 48-year-old man) who had multiple cysts along their intrahepatic bile ducts. The cysts were visible on gross examination of the liver and, in two cases, an enlargement of the cysts had been detected in abdominal computed tomography scans performed 1 year apart. Histologically, the cysts consisted of proliferating and dilated peribiliary glands. The cysts occasionally compressed the original bile ducts. The latter showed mucosal hyperplasia with antral-type gland metaplasia. Neoplastic changes and necroinflammation were not seen. Immunohistochemical assays revealed that the peribiliary glands and antral-type glands expressed not only cytokeratin and carbohydrate antigen 19-9, but also c-MET protein, the hepatocyte growth factor receptor which may be related, at least in part, to the cystic dilatation of the peribiliary glands.
• Downstream of CrK adaptor signaling pathway : Activation of Jun Kinase by v-CrK through the guanine-nucleotide exchange protein C3G

  Proc. Natl. Acad. Sci. USA  94-  (6)  2356  -2361  1997  [Not refereed][Not invited]
  
• DOCK180, a major CRK-binding protein, alters cell morphology upon translocation to the cell membrane

  H Hasegawa, E Kiyokawa, S Tanaka, K Nagashima, N Gotoh, M Shibuya, T Kurata, M Matsuda  MOLECULAR AND CELLULAR BIOLOGY  16-  (4)  1770  -1776  1996/04  [Not refereed][Not invited]
   

  CRK belongs to a family of adaptor proteins that consist mostly of SH2 and SH3 domains, Far Western blotting with CRK SH3 has demonstrated that it binds to 135- to 145-, 160-, and 180-kDa proteins. The 135- to 145-kDa protein is C3G, a CRK SH3-binding guanine nucleotide exchange protein, Here, we report on the molecular cloning of the 18O-kDa protein, which is designated DOCK180 (18O-kDa protein downstream of CRK), The isolated cDNA contains a 5,598-bp open reading frame encoding an 1,866-amino-acid protein. The deduced amino acid sequence did not reveal any significant homology to known proteins, except that an SH3 domain was identified at its amino terminus. To examine the function of DOCK180, a Ki-Ras farnesylation signal was fused to the carboxyl terminus of DOCK180, a strategy that has been employed successfully for activation of adaptor-binding proteins in vivo., Whereas wild-type DOCK180 accumulated diffusely in the cytoplasm and did not have any effect on cell morphology, farnesylated DOCK180 was localized on the cytoplasmic membrane and changed spindle 3T3 cells to flat, polygonal cells, These results suggest that DOCK180 is a new effector molecule which transduces signals from tyrosine kinases through the CRK adaptor protein.
• "Analysis of the hepatocyte growth factor receptor in regeneration and oncogenesis of the liver"

  General and Diagnostic Pathol.  141-  179  -186  1996  [Not refereed][Not invited]
  
• "CRK Protein Binds to Two Guanine Nucleotide-Releasing Proteins for the Ras Family and Modulates Nerve Growth Factor-Induced activataion of Ras in PC12 Cells. "

  Mol. Cell. Biol.  14-  5495  -5500  1994  [Not refereed][Not invited]
  
• C3G, a guanine nucleotide-releasing protein expressed ubiquitously, binds to the SH3 domains of CRK and GRB2/ASH proteins

  Proc. Natl. Acad. Sci. USA  91-  (8)  3443  -3447  1994  [Not refereed][Not invited]
  
• BOTH THE SH2 AND SH3 DOMAINS OF HUMAN CRK PROTEIN ARE REQUIRED FOR NEURONAL DIFFERENTIATION OF PC12 CELLS

  S TANAKA, S HATTORI, T KURATA, K NAGASHIMA, Y FUKUI, S NAKAMURA, M MATSUDA  MOLECULAR AND CELLULAR BIOLOGY  13-  (7)  4409  -4415  1993/07  [Not refereed][Not invited]
   

  Human CRK protein is a homolog of the chicken v-crk oncogene product and consists mostly of src homology region 2 (SH2) and SH3, which are shared by many proteins, in particular those involved in signal transduction. SH2 has been shown to bind specifically to phosphotyrosine-containing peptides. We report here that both SH2 and SH3 are required for signaling from CRK protein. Microinjection of the CRK protein induced neurite formation of rat pheochromocytoma cell line PC12. This activity was abolished by mutation of the CRK protein in either SH2 or SH3. The neuronal differentiation induced by the CRK protein was blocked by an excess amount of peptides containing CRK SH3. Moreover, we identified three proteins, of 118, 125, and 136 kDa, which bound specifically to CRK SH3. The CRK-induced neuronal differentiation was also suppressed by monoclonal antibodies against either CRK SH2 or p21ras. These results suggest that both SH2 and SH3 of the CRK protein mediate specific protein-protein binding and that the resulting multimolecular complex generates a signal for neurite differentiation through activation of p21ras.
• STRUCTURE OF 85-KDA SUBUNIT OF HUMAN PHOSPHATIDYLINOSITOL 3-KINASE ANALYZED BY USING MONOCLONAL-ANTIBODIES

  S TANAKA, M MATSUDA, S NAGATA, T KURATA, K NAGASHIMA, Y SHIZAWA, Y FUKUI  JAPANESE JOURNAL OF CANCER RESEARCH  84-  (3)  279  -289  1993/03  [Not refereed][Not invited]
   

  An 85 kDa subunit (p85alpha) of phosphatidylinositol 3-kinase (PI-3K) has one SH3 and two SH2 regions [SH2(N) and SH2(C)], which direct protein-protein interaction. We have established eighteen hybridomas producing monoclonal antibodies against p85alpha to study the structure-function relationship of this protein. Epitope mapping using a series of deletion mutants expressed in E. coli showed that the monoclonal antibodies bound to at least 5 distinct epitope regions, which were well dispersed on p85alpha except for its carboxyl-terminus. Monoclonal antibodies against amino-terminal regions and polyclonal antibodies against carboxyl-terminal regions immunoprecipitated p85alpha expressed in human cells and in E. coli. On the other hand, monoclonal antibodies against the central part of p85alpha failed to immunoprecipitate p85alpha efficiently; however, they could immunoprecipitate p85alpha mutants with deletion of either the amino- or the carboxyl-terminal region. Similar results were obtained by immunocytochemistry using confocal microscopy. These results suggested that steric hindrance prevents binding of monoclonal antibodies to the central part of p85alpha where SH2(N) is located. The SH2(N) may have a distinct function from SH2(C), which is located at the carboxyl-terminal region and has been shown to mediate the binding of PI-3K to activated growth factor receptors.
• STRUCTURAL REQUIREMENT OF CRK SH2 REGION FOR BINDING TO PHOSPHOTYROSINE-CONTAINING PROTEINS - EVIDENCE FROM REACTIVITY TO MONOCLONAL-ANTIBODIES

  M MATSUDA, S NAGATA, S TANAKA, K NAGASHIMA, T KURATA  JOURNAL OF BIOLOGICAL CHEMISTRY  268-  (6)  4441  -4446  1993/02  [Not refereed][Not invited]
   

  The SH2 region of signal-transducing proteins mediates binding to phosphotyrosine-containing proteins. We analyzed the structure-function relationship of the SH2 region of the human CRK protein by using a series of monoclonal antibodies (mAbs). Seventeen mAbs against the CRK SH2 region were classified into 5 groups according to the reactivity with mutant CRK proteins expressed in COS7 cells and in Escherichia coli and by epitope scanning with synthetic nonapeptides. Two groups of mAbs (groups A and B) were reactive only with intact SH2. Mutation(s) in either the amino-terminal B box or the carboxyl-terminal C box, which are the two subdomains of SH2, abolished the reactivity of the CRK mutants to the mAbs of groups A and B. Group A mAbs competed the binding of the CRK SH2 region to the phosphotyrosine-containing proteins. Moreover, the spectrum of the CRK mutants which were recognized by group A mAbs coincided with that of the CRK mutants which could bind phosphotyrosine-containing proteins, suggesting that group A mAbs were directed against the site of binding to phosphotyrosine-containing proteins. Group C mAbs, directed against the region between the B and C boxes, were reactive with both wild-type and mutant CRK proteins and did not affect the capacity of SH2 to bind phosphotyrosine-containing proteins. Contrary to group A and B mAbs, mAbs belonging to groups D and E, which were mapped onto the C box, did not bind well to the native CRK proteins, but bound to CRK mutants with mutation(s) in either the B or the C box. These results suggest that the B and C boxes, which are separated by a hinge region, coordinately form the functional SH2 domain that binds to the phosphotyrosine-containing proteins and to the group A mAbs.
• "Two Species of Human CRK cDNA Encode Proteins with Distinct Biological Activity. "

  Mol. Cell. Biol.  12-  3482  -3489  1992  [Not refereed][Not invited]
  

Awards & Honors

• 2023/04 日本病理学賞
  
• 2022/10 北海道知事賞・北海道医師会賞
  
• 2003 日本病理学会学術奨励賞
  
• 2002 日本癌学会奨励賞
  

Research Grants & Projects

• 高強度ポーラスゲルによる３次元がん組織モデル創出とがん幹細胞標的新規治療法の開発

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 2021/04 -2024/03 

  Author : 津田 真寿美, 野々山 貴行, 田中 伸哉

   

  がんの再発は、がん治療に耐性を示すがん幹細胞が生き残ることによって起きる。がんの根絶には、がん組織におけるがん幹細胞の誕生・生存機構の解明が必須であるが、がん幹細胞が微量なことに加えて、従来のポリスチレンdish上でのがん細胞単独培養法は生体内のがん組織環境と大きく乖離しており、真の解明に至っていない。研究代表者らは、近年、高強度ダブルネットワークゲル（DNゲル）上にがん細胞を播種すると、24時間以内に迅速にがん幹細胞が誘導されるリプログラミング（初期化）現象を見出した（Nat. Biomed. Eng., 2021）。当該技術を基盤として、本研究ではがん組織におけるがん幹細胞の誕生・生存機序の解明を目指す。 令和3年度は、高頻度に再発を引き起こす肉腫の生体内環境を模倣するために、高強度DNゲルの作製技術と多孔質ゲルを得る凍結重合法を組み合わせて、高強度多孔質（ポーラス）ゲルを作製した。ポーラスゲルは、1st モノマーにアニオン性高分子NaAMPSを用い、2ndモノマーに中性高分子AAmを用いた。筋組織を模倣するため、細胞外基質としてフィブロネクチンを吸着後、GFPラベル筋芽細胞を播種し、細胞の接着や長期生存を検討した。AAmの濃度を変えることでポーラスサイズを調整し、さらに細胞の播種方法を改善することにより、ポーラスゲルの深部まで細胞が進展するよう3次元組織モデルを最適化した。このポーラスゲルをマウスの筋肉内に埋植すると、１週間以内に宿主細胞がゲル内に侵入して組織を復元した。一方、氷の成長方向を制御するフリーズキャスティング法を用いた凍結重合法により、筋組織を模倣した高度な配向構造を有するポーラスゲルの創製に成功した。
• Reprogramming of cancer stem cells by high-function hydrogels creates basis to establish novel cancer therapy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2019/04 -2024/03 

  Author : 田中 伸哉, 津田 真寿美, 高阪 真路, 黒川 孝幸, 前仲 勝実

   

  （研究の骨格）本研究は、申請者が高機能ポリマーハイドロゲルを用いることで、極めて短時間にがん細胞のリプログラミングを誘導して、がん幹細胞を同定する方法見出したことにはじまる。これは従来のがん幹細胞分離同定法とは異なり、がんの種類を問わず24時間以内にがん幹細胞を同定することができる画期的な方法であり、本研究では、高機能ゲルのどのような物理学的因子ががん細胞の遺伝子発現変化を短時間で誘導するのかを検討し、高機能ゲルを基盤としたがん幹細胞診断法を開発し、さらにがん幹細胞標的治療薬を大規模スクリーニングにより創出するものである。（今年度の実績）「ゲル開発」プロジェクトでは、これまでの当該研究の結果から、陰性荷電を有するpoly-2-acrylamido- 2-methylpropanesulfonic acid (PAMPS) と中性荷電のpoly-N, N’- dimethylacrylamide (PDMAAm)にがん幹細胞誘導能が存在することが判明しており、とくにこれらのダブルネットワーク（DN）構造が重要であることが判明していた。さらに正と負の荷電両方有する両性荷電ゲルPCDME （poly-carboxymethyl -dimethyl-methacryloyloxy ethanaminium）もがん幹細胞誘導能を有することが判明していたが、当該年度の研究において負荷電を有するPNaSS（poly-sodium p-styrene sulfonate)のシングルネットワークゲルが、DNゲルより高いがん幹細胞誘導能を有することが明らかとなった。また、「臨床応用」のプロジェクトでは、従来検討されていなかった、ヒト肝臓がん（肝細胞がん）の細胞株を用いてDNゲルにより誘導した肝細胞がん幹細胞の解析から、ケモカイン受容体の1つであるCMKLR1が新規マーカーであることを発見した。
• 電荷による細胞応答制御法の創出ーチャージハイドロゲルでウイルス感染症・がんを治す

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 2021/07 -2023/03 

  Author : 田中 伸哉

   

  （研究の骨格）医学の領域で感染症・がんは2大疾患であり、これらを制御することは喫緊の課題である。我々は最近、バイオマテリアルの１つであるハイドロゲルがヒト細胞に急速なゲノム変化を誘導することを見出した（Nat. Biomed. Eng, 5, 914-925, 2021）。本研究はこの技術を格段に発展させるもので、ハイドロゲルを用いて細胞周囲のチャージ（荷電状態）を変化させることで細胞のゲノム状態を制御し、１）ウイルスへの感染を防御し、また、２）がん幹細胞を誘導することで、がんの早期診断法およびがん幹細胞特異的治療薬を開発する。 （今年度の実績）（１）チャージゲルによるウイルス増殖制御法の創出：陰性荷電を有するPAMPSゲルとPNaSSゲル、さらに正と負の荷電の両方を有する両性荷電ゲルPCDME上でウイルス感受性細胞VEROを培養し、COVID-19感染後にウイルス増殖を検討した。ウイルス核酸レベルで評価した場合、PAMPSとPNaSS上でウイルス増殖が亢進する一方、PCDMEゲル上では抑制される傾向が認められた。 （２）チャージゲルによるがん幹細胞誘導：陰性荷電を有するAMPSと陽性荷電を有するAPTMAモノマーを様々な割合で配合したチャージハイドロゲルを作製し、患者由来膵癌初代細胞を培養した所、陽性にシフトしたチャージゲルに癌細胞は接着し、癌幹細胞マーカーの発現が亢進することが明らかとなった。
• 癌の転移先臓器決定と覚醒におけるCrkおよびExosomeの機能解析

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2018/04 -2021/03 

  Author : 津田 真寿美, 田中 伸哉, 王 磊

   

  がんの転移についてはこれまで多数の研究が実施されてきたが、転移先臓器で長年潜伏していたがん細胞が何をきっかけに覚醒・再増殖するのか、また解剖学的位置関係からは説明をつけにくい臓器に転移・再発する理由はなぜか、その詳細な分子メカニズムは未だ解明されていない。申請者らはこれまで、シグナル伝達アダプター分子Crkががんの増殖・接着・ 浸潤・転移の全てのステップにおいて重要な役割を果たしていること、また最近、Crkががん組織の多様性形成に関与すると共に、エクソソームを介して転移の成立と促進に寄与する という新知見を見出した。これは、アダプター分子が転移先臓器の環境を整備しがん転移を成立させることを示唆する画期的なデータである。本研究では、Crkによるエクソソームの多様性形成メカニズムを明らかにし、がん転移の臓器選択性とがん細胞の覚醒との関連性を解明する。 当該年度は、ヒト浸潤性膀胱癌細胞株UM-UC-3の親株およびCrkノックダウン細胞から、超遠心法によりエクソソームを分離した。エクソソームに内包されている分子を、nano-LC MS/MS(蛋白質)で解析した所、受容体型チロシンキナーゼ、細胞内チロシンキナーゼ、細胞増殖因子、細胞外マトリックス、細胞接着分子、マトリックスメタロプロテアーゼなどが内包されていることが明らかとなった。これらの結果は、Western blottingによって確認された。各エクソソームを血管内皮細胞HUVECsに投与すると、UM-UC-3由来のエクソソームはHUVECsの細胞増殖能力、およびマトゲルへの浸潤能が行進することが明らかとなった。Crkノックダウン細胞由来のエクソソームはこれらの効果は認められなかった。
• Establishment of synovial sarcoma initiating cell model and exploration of new therapeutic targets

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2013/04 -2016/03 

  Author : KIMURA TAICHI, TANAKA SHINYA

   

  In this study, to elucidate the maintenance and regulation mechanisms of synovial sarcoma initiating cells that are closely involved in the recurrence and metastasis, we examined SS18-SSX binding proteins which were specific for sphere-forming subpopulation by mass spectrometry analysis. By proteomic analysis, we obtained specific or enhanced 26 binding protein candidates, including PARP1 and NPM, which were localized in nucleus and associated with transcriptional regulation. These results suggest that the induction of SS18-SSX and SS18-SSX binding proteins specific for sphere-forming subpopulation may play important roles of the transcriptional regulation specific for synovial sarcoma initiating cells.
• Early pathological evaluation for irradiation effectiveness in uterine cervical cancer

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2012/04 -2015/03 

  Author : TANINO Mishie, HIDEMICHI Watari, SHINYA Tanaka, RUMIKO Kinoshita

   

  Ionizing irradiation is an effective treatment for uterine cervical cancer, however, a higher rate of recurrence with more aggressive phenotypes is a vital issue. Although epithelial-mesenchymal transition (EMT) is involved in irradiation-induced cancer progression, the role for such phenotypic transition in uterine cervical cancer remains unknown. To investigate the mechanism of irradiation-dependent tumor progression, cervical cancer cell lines were irradiated multifractionated 20Gy in two weeks. Knocking down Snail and Yap cell lines were established. EMT regulators, Hippo pathway effector, mesenchymal and stemness proteins were increased after irradiation. In phenotypical analysis, cells exhibited an upregulation of migration, invasion, numbers of focal adhesion. Knocking down Snail and Yap inhibit increase in mesenchymal markers. In conclusion, we propose inhibiting both EMT and Hippo pathway is useful to prevent malignat transtion after irradiation.
• Development of a new rapid immunohistochemical staining method using AC electric field

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2011/04 -2015/03 

  Author : MINAMIYA Yoshihiro, AKAGAMI Yoichi, TANINO Mishie, NANJO Hiroshi, NAKAMURA Ryuta, TANAKA Shinya, ITO Tomoo, KAGAYA Masami, KONNO Hayato, SASAJIMA Toshio

   

  For the intraoperative diagnosis of lymph-node micro-metastases with immunohistochemical staining, the staining time should be shortened. We invented a new method that allows to reduce the immunohistochemical-staining time by applying an electric field to the tissue sections. We can complete immunohistochemical staining which usually requires 2 hours, within 12 minutes using this method. We demonstrated that this method was useful for the intraoperative detection of lymph node micrometastases of patients with lung cancer. We also demonstrated that this method is useful for the grading of the glioma during surgery. We tried this method to the decision making of the surgery for lung cancer and brain tumor. We also tried this method to the decision making of the surgery of gastroenterological, gynecologic, pediatric cancer and cancer of otorhinolaryngology. And then, we were able to demonstrate the usefulness of this method for the decision making of these surgeries
• Clarification of signal transmission mechanism in mesenchymal cells during spontaneous cartilage regeneration induced by multi-functional synthetic gel

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2011/04 -2014/03 

  Author : YASUDA Kazunori, OMIYA Yoshihiro, OHASHI Toshiro, KUROKAWA Takayuki, KITAMURA Nobuto, TANAKA Shinya, SEMBA Shingo

   

  This study discovered several important phenomena and their molecular mechanisms during chondrogenic differentiation and regeneration of mesenchymal cells induced by the PAMPS/PDMAAm gel or soluble proteins:(1) ATP oscillation, which depends on oscillation of both the mitochondrial respiration and the glycolysis, was induced by calcium ion oscillation. The ATP oscillation does not control cell differentiation but controls cell aggregation and tissue formation via controlling secretion of soluble proteins. (2) Young's modulus of a chondrogenic precursor cell increased to a significantly greater degree during differentiation induced by the gel than during differentiation induced by Insulin. (3) Both the BMP-Smad signaling pathway and the IGF-depending signaling pathway were specifically activated during the differentiation induced by DN gel in comparison with that induced by Insulin. (4) The proteomics analysis showed that the gel has a function as a reservoir of soluble factors.
• 癌治療薬探索を目指した新規スプライシングモニタリングシステム構築への挑戦

  文部科学省：科学研究費補助金(挑戦的萌芽研究)

  Date (from‐to) : 2012 -2012 

  Author : 田中 伸哉

   

  本研究はこれまで全く未知であったmRNAのスプライシングに基づく発癌メカニズムを明らかにし、その治療薬を探索するシステムを構築することを目的とする。 具体的には多くのヒト癌の悪性化に関与することが判明しているシグナル伝達アダプター分子CRKに焦点をあて転移能の高いCRK-IIへの転換機構を明らかにする。CRKのスプライシングモニタリングシステムを構築し、CRKのスプライシングを制御する機構を分子レベルmicroRNAをスクリーニンし、これまでに全く未知のメカニズムを明らかにする。遺伝子発現は従来転写因子を中心とするエピジェネティックな解析が主流であり近年はmicroRNAが注目されているが、もう1つの重要なメカニズム１つであるスプライシング機構は解明が進んでおらず、疾患との関連、癌との関連では全く深い検討はなされていないのが現状である。 今回我々はシグナル伝達アダプター分子CRKがスプライシングバリアントにより機能が異なる点に着目し新規のスプライシングモニタリングシステムを構築するものであり、これまでにない時間空間的なスプライシング情報を得ることができ、正常細胞と癌細胞のスプライシングの差を明確にする画期的なシステムとなることが期待される。 本年度の研究においてヒトCRKゲノムをBACクローンより単離してその遺伝子配列を決定した。またCRKゲノムにGFPをN末端とC末端の２つの部分に分けて挿入し、スプライシングが生じるときにGFPの野生型の蛋白が発現するベクターをデザインした。またスプライシングによる生理作用をあらかじめ検討するためにCRK-IとCRK-IIをそれぞれ組み込んだ細胞を作成したところ、CRK-IIはepithelial mesenchymal transitionに積極的に関与することが明らかとなった。今後スプライシングとEMTの関連についても着目して研究を進める。
• 滑膜肉腫幹細胞の同定と創薬基盤の確立

  文部科学省：科学研究費補助金(基盤研究(B))

  Date (from‐to) : 2011 -2012 

  Author : 田中 伸哉

   

  滑膜肉腫は若年成人の関節周辺に発生する悪性度の高い腫瘍であり有効な治療法は未だ確立されていない。 本研究では根源的な治療法を開発するため、滑膜肉腫幹細胞を同定し腫瘍発生機構・病態を解明し、発癌メカニズムに基づく治療薬開発の基盤技術を確立する。発癌機構の解析は、①肉腫幹細胞の分離とマーカー分子の同定、② 滑膜肉腫キメラ遺伝子SYT-SSXによるクロマチンリモデリング測定システムの開発、③滑膜肉腫の腫瘍化に必須のシグナル伝達分子Crkによる転移・浸潤メカニズムの解明、の3つを柱とする。治療薬剤のリード化合物の探索は独自に開発した1分子特異的シグナル抑制薬剤スクリーニング法、2分子結合FRETイメージング法、分子構造に基づくin silico分子設計法を用いて、スクリーニング法の基盤を確立する。候補となる化合物は申請者らが確立した滑膜肉腫モデルマウスを改良し、臨床応用の可能性を含めて有効性を評価する。 本研究では肉腫幹細胞、クロマチンリモデリング機構、HGF-Crkシグナルの3点に焦点をあて、SYT-SSXの癌化のメカニズムについて分子生物学的に包括的・総合的な理解を目指す。特に肉腫幹細胞はこれまで報告がほとんどなく、治療上極めて重要な発見となる。クロマチンリモデリングと発癌との関連については、近年急速に解析が発展している分野であるが、滑膜肉腫については癌化に必須であるSYT-SSXとhBRMとの結合は当研究室で発見されたものであり独創性が高い。
• ウイルス因子と宿主因子の細胞レベルの相互作用の解明によるPMLの治療法の開発

  文部科学省：科学研究費補助金(基盤研究(B))

  Date (from‐to) : 2009 -2011 

  Author : 澤 洋文, 田中 伸哉

   

  本年度はJCウイルス(JCV)がコードする早期タンパク質であるT抗原(TAg)、および後期タンパク質であるVP1に着眼して研究を推進した。JCV感染細胞において、TAgの非特異的なDNA結合性が細胞周期に与える影響を検討するため、DNA結合性が低下もしくは増強するTAgの変異体を作成した。次に、これらの変異体を細胞に導入・発現させて細胞周期を解析した。野生型TAgを発現した細胞はG2/M期の細胞が増加していたが、DNA結合性が低下したTAg変異体を発現した細胞はG2/M期への細胞の集積は認められなかった。また、DNA結合性が増強しているTAg変異体を発現した細胞ではG2/M期に集積する細胞数が野生型よりさらに増加していた。本研究により、TAg発現による細胞周期のG2期停止の機構において、TAgが宿主細胞のDNAへ結合することが関与していることが明らかになった。また、JCV感染細胞でTAgの発現によって誘導されるG2期停止は効率的なウイルスDNAの複製に必須であることが判明した。次に、外郭タンパク質であるVP1内に6個存在するシステイン残基の変異体を作成して、JCVの粒子形成能を比較・検討した。本実験により、VP1タンパク質が互いに結合し5量体を形成すること、5量体の形成過程においてシステイン残基が必須であること、5量体の形成に続いて粒子形成が起こることが明らかになった。これま...
• Involvement of adaptor protein in the pathogenesis of idiopathic pulmonary fibrosis

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2009 -2011 

  Author : TANINO Mishie, TANAKA Shinya, BETSUYAKU Tomoko

   

  Idiopathic pulmonary fibrosis (IPF) is chronic, progressive, and fibrosing lung disease and there is no advantageous drug for longer survival in IPF patients. Recently, involvement of epithelial mesenchymal transition (EMT) is suggested as one of the pathogenesis of IPF. In this study, we have demonstrated CRKI/II is highly expressed in alveolar type II (AT II) cells in IPF/UIP specimens and CRK II overexpressed A549 cell line could be induced stronger EMT phenomenon under TGF-β1 stimulation. These results suggested suppression of CRK II in ATIIcells might be one of the therapeutic strategy for regulating lung fibrosis in IPF/UIP by regulation of EMT.
• 癌浸潤におけるアダプター分子CRKの解析と構造に基づく治療薬開発への応用

  文部科学省：科学研究費補助金(特定領域研究)

  Date (from‐to) : 2008 -2009 

  Author : 田中 伸哉, 西原 広史

   

  これまでの研究で、アダプター分子Crkの癌化における役割を解析し、癌腫、肉腫、脳腫瘍など様々なヒト癌細胞株を用いてsiRNA法にてCrk knockdown細胞株を樹立した。何れの種類のCrk knockdown細胞においても、接着能、浸潤能、足場非依存性増殖能、in vivo造腫瘍能など、癌細胞の悪性化を示す指標に、著明な減少がみられ、Crkはヒト卵巣癌、軟部肉腫、脳腫瘍において、悪性化に必須の分子であることが明かとなった(Oncogene, 25,2006 : Mol.Cancer Res., 7,2006)。また、Crkの癌化におけるシグナル伝達メカニズムを詳細に解析するために、NMRを用いた構造解析を行い(Nature Struct.Mol.Biol., 2007)、昨年度は、Crkのシグナル伝達を抑制する薬剤開発する前段階として、単一の遺伝子変化に対応する薬剤スクリーニングの系を確立した(BBRC,373,2008)。この系を用いてCrkを発現させたアストロサイトに対して、dual luciferase assayを行い96wellプレートで薬剤をスクリーニングして、Crkシグナル阻害薬を同定した。以後の研究では、真にCRKシグナルを抑制する薬剤か否かを個別に判定していき臨床応用可能か否かをin vivoの系で解析していきたい。また、今年度の研究において癌細胞の浸潤...
• Analysis of synovial sarcoma oncogene SYT-SSX for development of new therapy

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2007 -2009 

  Author : Shinya TANAKA, 笹井 研, Hiroshi NISHIHARA, Akio MINAMI

   

  The purpose of this study is to analyze the transforming mechanism of human synovial sarcoma-associated oncogene SYT-SSX and to establish the basic system for the development of new therapy. We originally proved that SYT-SSX is responsible oncogene for human synovial sarcoma, and in this study, we found that signalling adaptor protein Crk is involved in malignant feature of synovial sarcoma and one of the map kinase family protein as p38 plays an important role. Furthermore, SYT-SSX transgenic mice which develop synovial sarcoma and also SYT knockout mice were established in this study. The...
• JCウイルス蛋白による脱髄のモデル作製と治療法の開発

  文部科学省：科学研究費補助金(萌芽研究)

  Date (from‐to) : 2007 -2008 

  Author : 澤 洋文, 荒井 勇二, 田中 伸哉

   

  本研究は細胞側因子と相互作用し、細胞内の物質輸送に影響を与えるJCウイルス(JCV)のagnoproteinがオリゴデンドロサイトの機能障害を惹起させる。」という作業仮説を立て、agnoproteinを中枢神経に発現させるトランスジェニックマウスを作製し、JCV感染によって生じる、多発性硬化症とは異なった機序で発症する非炎症性の脱髄のモデルを確立し、ウイルス感染後の脱髄病変の形成過程の機序を詳細に検討することを目的として以下の実験を行った。平成20年度はマウスのスクリーニングを行うにあたり、従来行われていたSouthern blottingによる解析ではなく、mouseの血液から直接、導入遺伝子を検出するFTA paperを用いたPCRの系を使ってsample採取当日に結果を得ることの出来る系を構築した。作製したスクリーニングの系を用いて、agnoproteinおよびlate proteinのトランスジェニックマウスの子孫およびタモキシフェン反応性のCreリコンビナーゼを発現しているトランスジェニックマウスを交配させ、タモキシフェン投与によりJCウイルスのagnoproteinもしくはlate proteinをオリゴデンドロサイトに発現させるダブルトランスジェニックマウスを作製した。今後はタモキシフェン投与法の条件を決定し、タモキシフェン投与により脳のオリゴデンドロサイトにa...
• ヒト癌浸潤におけるアダプター分子クラックの解析

  文部科学省：科学研究費補助金(特定領域研究)

  Date (from‐to) : 2006 -2007 

  Author : 田中 伸哉, 西原 広史

   

  本研究ではアダプター分子Crkの癌化における役割を解析し、癌腫、肉腫、脳腫瘍など様々なヒト癌細胞株を用いてsiRNA法にてCrk knockdown細胞株を樹立した。何れの種類のCrk knockdown細胞においても、接着能、浸潤能、足場非依存性増殖能、in vivo造腫瘍能など、癌細胞の悪性化を示す指標に、著明な減少がみられ、Crkはヒト卵巣癌、軟部肉腫、脳腫瘍において、悪性化に必須の分子であることが明かとなった(Oncogene,25, 2006:M01. Cancer Res.,7, 2006)。また、Crkの癌化におけるシグナル伝達メカニズムを詳細に解析するために、NMRを用いた構造解析を行ったところ、Crk-Iはflexibleな分子で恒常的にシグナル標的分子と結合しているが、Crk-IIは、Grb2などのアダプタ一分子とは異なり、3つのSH領域がコンパクトな構造をとっており、この構造が2つのSH3領域の間にある15アミノ酸によって制御されることが判明し、この領域をISC (inter SH3 core)と命名した。ISCの変異体は、標的分子への親和性が回復し、また、細胞増殖能もCrk-Iと同様のレベルまで増加した。したがって、Crk-IIのcompactなstructureがシグナルの制御に関与することが判明した(Nature Struct. Mol. Biol...
• Analysis of human synovial sarcoma oncoprotein SYT-SSX and its therapeutic application.

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2004 -2006 

  Author : Shinya TANAKA, 澤 洋文, 三浪 明男, 赤城 剛, 澤 洋文, 三浪 明男

   

  The cause of synovial sarcoma had been unknown for long time, and in 1994, Clark et al., identify the chimeric gene of t(X;18) and named it SYT-SSX. Then molecular analysis of SYT-SSX has been advanced and we have shown that SYT-SSX serves oncogenic potential on cells (Nagai, M., et al., PNAS, 2001).In this project, SYT-SSX was found to bind to not only one of the chromatin remodeling factors, hBRM, but to BRG (Gene Cells, 9, 2004), and also shown to induce cellular senescence by the induction of p21 (Oncogene, 24, 2005). Furthermore, IGF2 has been shown to play an important role for growth...
• 癌浸潤に関与するユビキチン化を生細胞で時間的空間的に解析する

  文部科学省：科学研究費補助金(特定領域研究)

  Date (from‐to) : 2005 -2005 

  Author : 田中 伸哉, 西原 広史

   

  【目的】本研究では、細胞癌化とユビキチン化に焦点をあててその詳細なメカニズムの解析を行った。具体的には、シグナル伝達アダプター分子CRKを介する癌化のシグナル伝達機構の中でユビキチン化の受ける可能性のある分子を検討し、ユビキチン化を可視化する事を試みた。【方法と結果】(i)CRK自体が人の癌化に必須の分子であることをsiRNAを用いた実験で証明し報告した(Linghu, H., et al., Oncogene,2006,in press)。ヒト卵巣癌細胞にてCRKをノックダウンすることで細胞癌化能が著明に低下し、特に卵巣癌の腹腔内播種は完全に抑制された。(ii)次にCRKを介する細胞癌化機構の中でSH3領域の標的分子である低分子量G蛋白質活性化因子DOCK180がユビキチン化を受けることを見出した(Makino, Y., et al., JCS,119,923-932,2006)。さらにこのDOCK180のユビキチン化はDOCK180のSH3領域の標的分子であるElmo1によって抑制される事で、細胞の局所においてDOCK180の量が制御されることでRacの活性が制御されることが示された。さらに、我々は現在ELMO1がチロシンリン酸化を受ける事を見出しており、その特異的なチロシン残基を1箇所同定しており、現在その機能解析を行っている。(iii)YFP-DOCK180とCFP-...
• クロマチン構造変換モニタリングシステムの開発

  文部科学省：科学研究費補助金(萌芽研究)

  Date (from‐to) : 2004 -2005 

  Author : 田中 伸哉

   

  【目的】本研究はクロマチンリモデリングに対する視覚的なモニター系の開発を試みるものである。具体的には、我々が見出した滑膜肉腫細胞の原因遺伝子SYT-SSXとSWI/SNF型クロマチンリモデリング因子hBRMとの結合(Nagai,et al.PNAS,2001)に基づき系の確立を試みる。【方法と結果】(i)予備実験として、SYT-SSXが原因であるHuman Synovial sarcoma細胞株SYO-1においてFRETプローブが機能することを確認した。SYO-1細胞にRac活性のモニタリングプローブを導入し、HGFで刺激することで、効率にFRET(蛍光共鳴エネルギー移行)反応が誘導されることを確認した。(ii)本研究においてはSYT-SSXとhBRMが結合することに基づいて、CFP-SYT-SSXとYFP-hBRM(140-180AA)を共発現させて、2分子FRETの検出を試みたが、ノイズが多く有意な差は得られなかった。現在、SYT-SSXの構造変化に基づく、2分子FRETのプローブを作成中であり、現在有意なシグナルの検出を試みている。(iii)また、本研究においてはSYT-SSXのクロマチンリモデリング機能を解析する過程において、SYT-SSXは細胞に導入するとp21の発現を誘導しsenescenceを見出した(Tsuda, M.,et al.,Oncogene,2005...
• クロマチンリモデリングと細胞の癌化・老化の制御機構の解析

  文部科学省：科学研究費補助金(特定領域研究)

  Date (from‐to) : 2002 -2002 

  Author : 田中 伸哉

   

  本研究は滑膜肉腫関連癌遺伝子SYT-SSXがクロマチンリモデリング因子hBRMと結合することから、SYT-SSX遺伝子を用いて細胞の癌化・老化能の解析を行うものであり、今年度は以下の成果を得た。(1)特許出願。これまでの研究成果に基づき、SYT-SSXとhBRMの結合を阻害する50アミノ酸領域の同定が、滑膜肉腫の遺伝子治療の基盤技術となることを示し、国内特許を出願した(発明者:田中伸哉、長嶋和郎 名称:ヒト滑膜肉腫に対する遺伝子治療法、特願2002-050894)。(2)SYT-SSX1によるp21発現誘導。細胞老化と関連してSYT-SSX1及び各種変異体をラット線維芽細胞及びhBRM欠損SW13細胞株に導入すると、p21の発現量が増加することを見出した。SYT-SSX1とhBRMの共発現によりp21の発現量は更に亢進した。変異体解析によりこのp21の誘導には、SYT-SSX1のQPGY domain及びC末側が必要であることが明らかとなった。(3)SYT-SSX1によるp53非依存性p21プロモーターの活性化。293T細胞においてSYT-SSX1の過剰発現によりp21プロモーターを活性化することをルシフェラーゼアッセイにより確認した。(p21プロモータープラスミドは国立長寿研磯部先生より分与)。さらにp53欠損HCT116細胞株(Bert Vogelstein教授より分与)...
• Analysis of transforming mechanism of human synovial sarcoma oncogene SYT-SSX

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2001 -2002 

  Author : Shinya TANAKA, 赤城 剛, 三浪 明男, 澤 洋文

   

  Human synovial sarcoma occurs primarily around the joint of extremities and more than 90% of cases have chromosomal translocation t(X;18) resulting the formation of chimeric gene SYT-SSX (synovial sarcoma translocation-synovial sarcoma X break point). As both of SYT and SSX did not share any homology region of known functional proteins, the transforming mechanism of SYT-SSX has not yet been elucidated. In this project, we established rat fibroblast cell line 3Y1 expressing SYT, SSX, and SYT-SSX, and found that SYT-SSX induced anchorage-independent growth and tumor formation in nude mice. In...
• アダプター分子CRKによる癌細胞浸潤能の解析

  文部科学省：科学研究費補助金(奨励研究(A), 若手研究(B))

  Date (from‐to) : 2001 -2002 

  Author : 田中 伸哉

   

  本研究はアダプター分子CRKを介する癌細胞の浸潤メカニズムの解析を行い以下の新しい知見を得て論文発表を行った(2)は現在投稿中である。尚、(1)から(4)は研究代表者がCorresponding authorである。(5)は共同研究での成果である。(1)vCrkによるRhoの活性化:活性化型Crkであるv-Crkの発現誘導細胞を樹立して細胞の運動能および浸潤能を検討したところvCrkはRhoを活性化しストレスファイバー形成を促進することが判明した(Tsuda, et al. Cell Growth ＆ Differ)。(2)Crkによるヒアルロン酸CD44細胞運動系の制御:CrkはRho-GDIと結合することでRhoを活性化しさらに、ERMファミリー蛋白質を制御することで、ヒアルロン酸とその受容体であるCD44依存性に細胞運動能を亢進することを明らかにした(Tsuda et al.投稿中)。(3)Crkのヒト遺伝子解析単離解析とヒト癌組織を用いた分子病理学的解析:Crkはヒト癌細胞で高い発現が認められたが、その遺伝子レベルでの変異は検索した限りでは認めなかった(Nishihara, et al. Cancer Letter)。(4)ヒト血球系腫瘍細胞株におけるCrkLの解析:Crkの浸潤能にはRacの活性化因子DOCK180が関与することが考えられていたが、今回ヒト白血病細胞...
• Analysis of mechanisms of demyelination in progressive multifocal leukoencephalopathy

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(A))

  Date (from‐to) : 1998 -2000 

  Author : Kazuo NAGASHIMA, 田中 伸哉, 澤 洋文

   

  JC virus (JCV) causes in humans a demyelinating disease, progressive multifocal leukoencephalopathy (PML), but mechanisms of demyelination by JCV remain unknown.To investigate early events of JCV infection including attachment, penetration, and transport to the nuclei where viral replication occurs, we have analyzed the susceptibility of 15 different cell lines to infection using a semi-quantitative PCR assay, ISH, laser scanning confocal microscopy and a viral replication assay. JCV entry was observed in all cell lines within 10 min after inoculation. Inhibition of viral entry both by an a...
• シグナル伝達アダプター分子CRKによるJunキナーゼ活性化のメカニズムの解析

  文部科学省：科学研究費補助金(奨励研究(A))

  Date (from‐to) : 1998 -1999 

  Author : 田中 伸哉

   

  研究代表者は本研究助成において、申請時の目的を達成した。すなわちR-RasがC3Gの基質として機能し、Junキナーゼ(JNK)を活性化しv-Crkの癌化に関与することを見出し報告した(Mochizuki,N.,et al.,J.Biol.Chem.,2000,in press)。研究代表者はこれまでニワトリ肉腫のウイルスのコードする癌遺伝子v-Crkの癌化のメカニズムの解明を目的に研究を行ってきており、本研究においてはR-Rasがv-Crk/C3G複合体からのシグナルを媒介し癌化に関与することを明らかにした。V-CrkはSH2/SH3領域からなるアダプター分子でありそれ自身は何ら酵素活性を持っていないにもかかわらずチロシンキナーゼ活性を上昇させて細胞癌化を誘導するが、そのメカニズムは不明であった。我々は独自にv-Crk結合分子C3Gを単離し、その解析を進めていたが1997年までの段階でv-CrkはC3Gを介してJNKを活性化し細胞癌化に関与することを報告していた。しかしながらC3GのJNK活性化に関与する基質は不明であった。本研究では一連のsmall G蛋白を調べることによって、R-RasがC3Gの基質として機能しJNKを活性化することが明らかとなった。具体的には、活性化型R-Ras-V38は一過性発現によって293T細胞において、JNKを活性化し、野生型R-Ras-WTのJ...
• Analysis of apoptotic mechanism of cerebellar granular cells

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 1997 -1999 

  Author : Kazuo NAGASHIMA, 鈴木 宏明, 田中 伸哉, 澤 洋文

   

  We have established a rat model of injury of cerebellar granule cells by intoxication of methylmercury chloride (MMC). To elucidate the molecular mechanisms of brain damage induced by MMC we initial ly examined the pathologic examination by H&E staining, electron microscope, TUNEL staining, and DNA fragmentation. These examination revealed that cerebellar granule cell death by organic mercury intoxication occurs via an apoptotic process and was restricted to the granule cells and that the distributional pattern of apoptotic cerebellar granule cells was specific in rat cerebellar vermis. Sec...
• Yeast One Hybrid法によるJC virus複製機構の検討

  文部科学省：科学研究費補助金(萌芽的研究)

  Date (from‐to) : 1996 -1997 

  Author : 長嶋 和郎, 篠原 敏也, 田中 伸哉, 澤 洋文

   

  1.Clontech(Palo Alto,CA)社のyeast one-hybrid system)を用いてJC virus NF-1領域に特異的に結合する蛋白を同定した。NF-1配列を用いたスクリーニングで30個の陽性クローンから4個はこれまでにdata baseに登録されていない未知遺伝子であったためにこのDNA配列を確認し、NF-1/Xをcloningし、その発現が脳に多いことを示した。2.ヒトに無症候性に感染し、常時尿に排泄されている型Archetype JC virusであるCY株(調節領域が原始型)がCOS-7細胞で増殖することを見いだし、この細胞を用いた解析系を樹立した。Mad-1(調節領域がPML型)と調節領域をCYに変えたキメラウイルスCY/Mad-1で感染性を調べ、cellular tropsimに調節領域が関与していることを示した。3.Microinjection法により非感受性細胞であるCOS7でも細胞膜を越えてvirusを注入するとvirusの増殖が見られることから、virus感染に細胞膜因子が存在することを示唆した。4.過去2年間に日本国内にて発症が確認されたPMLの5症例のregulatory regionを解析し、4例のPML型と1例のArchetype JCの症例を見い出し、報告した。またPMLの髄液を用いたPCRによる診断方法を確立した。
• Analysis of human synovial sarcoma associated SYT-SSX gene
• Analysis of the mechanism for signaling pathway mediated by CrK

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• 2021年4月1日 

  - 

  2023年3月31日 

  大学院医学研究院副研究院長
• 2021年4月1日 

  - 

  2023年3月31日 

  大学院医学院副学院長
• 2023年4月1日 

  - 

  2025年3月31日 

  大学院医学研究院副研究院長
• 2023年4月1日 

  - 

  2025年3月31日 

  教育研究評議会評議員

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• 札幌ロータリークラブ会員

  Role : Others
• 日本医療安全調査機構北海道地域統括調査・支援医師

  Role : Investigator