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Last Updated :2024/07/25

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Master

Affiliation (Master)

  • Faculty of Medicine　Institute for Animal Experimentation

Affiliation (Master)

  • Faculty of Medicine　Institute for Animal Experimentation

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Profile and Settings

Profile and Settings

  • Name (Japanese)

    TAKEI

  • Name (Kana)

    NORIO

  • Name

    201501030526063351

Achievement

Research Areas

  • Life sciences / Laboratory animal science
  • Life sciences / Experimental pathology
  • Life sciences / Hematology and oncology

Research Experience

  • 2019/01 - Today Hokkaido University
  • 2015/06 - 2018/12 北海道大学 産学・地域協働推進機構 助教
  • 2011/04 - 2015/05 Sapporo Medical University School of Medicine

Published Papers

  • Masahiro Chiba, Joji Shimono, Takashi Ishio, Norio Takei, Kohei Kasahara, Reiki Ogasawara, Takahide Ara, Hideki Goto, Koh Izumiyama, Satoko Otsuguro, Liyanage P Perera, Hiroo Hasegawa, Michiyuki Maeda, Satoshi Hashino, Katsumi Maenaka, Takanori Teshima, Thomas A Waldmann, Yibin Yang, Masao Nakagawa
    Blood 140 (18) 1951 - 1963 2022/08/03 
    Adult T-cell leukemia/lymphoma (ATLL) is one of the aggressive peripheral T-cell neoplasms with a poor prognosis. Accumulating evidence demonstrates that escape from adaptive immunity is a hallmark for ATLL pathogenesis. However, the mechanisms by which ATLL cells evade NK-cell-mediated immunity have been poorly understood. Here we show that CD48 expression in ATLL cells determines the sensitivity for NK-cell-mediated cytotoxicity against ATLL cells. We performed unbiased genome-wide clustered regularly interspaced short palindromic repeat (CRISPR) screening using two ATLL derived cell lines and discovered CD48 as one of the best enriched genes whose knockout conferred resistance to YT-1 NK cell line mediated cytotoxicity. The ability of CD48-knockout ATLL cells to evade NK cell effector function was confirmed using human primary NK cells with reduced IFNg induction and degranulation. We found that primary ATLL cells had reduced CD48 expression along with disease progression. Furthermore, other subgroups among aggressive peripheral T-cell lymphomas (PTCL) also expressed lower levels of CD48 than normal T-cells, suggesting that CD48 is a key molecule in malignant T-cell evasion of NK cell surveillance. Thus, this study demonstrates that CD48 expression is likely critical for malignant T-cell lymphoma cell regulation of NK cell mediated immunity and provides a rationale for future evaluation of CD48 as a molecular biomarker in NK cell-associated immunotherapies.
  • Akinobu Ota, Ichiro Hanamura, Sivasundaram Karnan, Shingo Inaguma, Norio Takei, Vu Quang Lam, Shohei Mizuno, Jo Kanasugi, Md Wahiduzzaman, Md Lutfur Rahman, Toshinori Hyodo, Hiroyuki Konishi, Shinobu Tsuzuki, Hiroshi Ikeda, Akiyoshi Takami, Yoshitaka Hosokawa
    Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research 40 (8) 389 - 405 2020/08 [Refereed]
     
    [Figure: see text] Multiple myeloma (MM) remains an intractable hematological malignancy, despite recent advances in anti-MM drugs. Here, we show that role of PDZ binding kinase (PBK) in MM tumor growth. We identified that interleukin-6 (IL-6) readily increases PBK expression. Kaplan-Meier analysis showed that the MM patients with higher expression of PBK have a significant shorter survival time compared with those with moderate/lower expression of PBK. Knockout of PBK dramatically suppressed in vivo tumor growth in MM cells, while genome editing of PBK changing from asparagine to serine substitution (rs3779620) slightly suppresses the tumor formation. Mechanistically, loss of PBK increased the number of apoptotic cells with concomitant decrease in the phosphorylation level of Stat3 as well as caspase activities. A novel PBK inhibitor OTS514 significantly decreased KMS-11-derived tumor growth. These findings highlight the novel oncogenic role of PBK in tumor growth of myeloma, and it might be a novel therapeutic target for the treatment of patients with MM.
  • Sakurai T, Kamiyoshi A, Takei N, Watanabe S, Sato M, Shindo T
    Scientific reports 9 (1) 9923  2019/07 [Refereed][Not invited]
  • Takei N, Yoneda A, Kosaka M, Sakai-Sawada K, Tamura Y
    BMC cancer 19 (1) 510 - 510 2019/05 [Refereed][Not invited]
     
    BACKGROUND: Hypoxia is an important factor that contributes to tumour aggressiveness and correlates with poor prognosis and resistance to conventional therapy. Therefore, identifying hypoxic environments within tumours is extremely useful for understanding cancer biology and developing novel therapeutic strategies. Several studies have suggested that carbonic anhydrase 9 (CA9) is a reliable biomarker of hypoxia and a potential therapeutic target, while pimonidazole has been identified as an exogenous hypoxia marker. However, other studies have suggested that CA9 expression is not directly induced by hypoxia and it is not expressed in all types of tumours. Thus, in this study, we focused on endoplasmic reticulum disulphide oxidase 1α (ERO1α), a protein that localises in the endoplasmic reticulum and is involved in the formation of disulphide bonds in proteins, to determine whether it could serve as a potential tumour-hypoxia biomarker. METHODS: Using quantitative real-time polymerase chain reaction, we analysed the mRNA expression of ERO1α and CA9 in different normal and cancer cell lines. We also determined the protein expression levels of ERO1α and CA9 in these cell lines by western blotting. We then investigated the hypoxia-inducible ERO1α and CA9 expression and localisation in HCT116 and HeLa cells, which express low (CA9-low) and high (CA9-high) levels of CA9, respectively. A comparative analysis was performed using pimonidazole, an exogenous hypoxic marker, as a positive control. The expression and localisation of ERO1α and CA9 in tumour spheres during hypoxia were analysed by a tumour sphere formation assay. Finally, we used a mouse model to investigate the localisation of ERO1α and CA9 in tumour xenografts using several cell lines. RESULTS: We found that ERO1α expression increased under chronic hypoxia. Our results show that ERO1α was hypoxia-induced in all the tested cancer cell lines. Furthermore, in the comparative analysis using CA9 and pimonidazole, ERO1α had a similar localisation to pimonidazole in both CA9-low and CA9-high cell lines. CONCLUSION: ERO1α can serve as a novel endogenous chronic hypoxia marker that is more reliable than CA9 and can be used as a diagnostic biomarker and therapeutic target for cancer.
  • Mizuki Fujii, Akihiro Yoneda, Norio Takei, Kaori Sakai-Sawada, Marina Kosaka, Kenjiro Minomi, Atsuro Yokoyama, Yasuaki Tamura
    JOURNAL OF BIOLOGICAL CHEMISTRY 292 (38) 15649 - 15660 0021-9258 2017/09 [Refereed][Not invited]
     
    Upon liver injury, excessive deposition of collagen from activated hepatic stellate cells (HSCs) is a leading cause of liver fibrosis. An understanding of the mechanism by which collagen biosynthesis is regulated in HSCs will provide important clues for practical anti-fibrotic therapy. Endoplasmic reticulum oxidase 1 alpha (ERO1 alpha) functions as an oxidative enzyme of protein disulfide isomerase, which forms intramolecular disulfide bonds of membrane and secreted proteins. However, the role of ERO1 alpha in HSCs remains unclear. Here, we show that ERO1 alpha is expressed and mainly localized in the endoplasmic reticulum in human HSCs. When HSCs were transfected with ERO1 alpha siRNA or an ERO1 alpha shRNA-expressing plasmid, expression of ERO1 alpha was completely silenced. Silencing of ERO1 alpha expression in HSCs markedly suppressed their proliferation but did not induce apoptosis, which was accompanied by impaired secretion of collagen type 1. Silencing of ERO1 alpha expression induced impaired disulfide bond formation and inhibited autophagy via activation of the Akt/mammalian target of rapamycin signaling pathway, resulting in intracellular accumulation of collagen type 1 in HSCs. Furthermore, silencing of ERO1 alpha expression also promoted proteasome-dependent degradation of membrane type 1-matrix metalloproteinase (MT1-MMP), which stimulates cell proliferation through cleavage of secreted collagens. The inhibition of HSC proliferation was reversed by treatment with MT1-MMP-cleaved collagen type 1. The results suggest that ERO1 alpha plays a crucial role in HSC proliferation via posttranslational modification of collagen and MT1-MMP and, therefore, may be a suitable therapeutic target for managing liver fibrosis.
  • Norio Takei, Akihiro Yoneda, Kaori Sakai-Sawada, Marina Kosaka, Kenjiro Minomi, Yasuaki Tamura
    SCIENTIFIC REPORTS 7 (1) 9389  2045-2322 2017/08 [Refereed][Not invited]
     
    Endoplasmic reticulum disulphide oxidase 1 alpha (ERO1 alpha) is an oxidase localized in the endoplasmic reticulum that plays a role in the formation of disulphide bonds of secreted and cell-surface proteins. We previously showed that ERO1a is overexpressed in various types of cancer and we further identified ERO1 alpha expression as a novel factor related to poor prognosis in cancer. However, the biological functions of ERO1 alpha in cancer remain unclear. Here, we investigated the cell biological roles of ERO1 alpha in the human colon-cancer cell line HCT116. ERO1 alpha knockout (KO) by using CRISPR/Cas9 resulted in decreased tumourigenicity in vivo and reduced cell proliferation only under hypoxia in vitro, which suggested that ERO1 alpha promotes cancer progression specifically in a low-oxygen environment. Thus, we evaluated the function of ERO1 alpha in cell proliferation under hypoxia, and found that under hypoxic conditions, ERO1 alpha KO resulted in a contact-inhibited morphology and diminished motility of cells. We further showed that ERO1 alpha KO induced a change in integrin-beta 1 glycosylation and thus an attenuation of cell-surface integrin-beta 1 expression, which resulted in the aforementioned phenotype. Our study has established a previously unrecognized link between ERO1 alpha expression and integrin activation, and thus provides new evidence for the effectiveness of ERO1 alpha-targeted therapy for colorectal carcinoma.
  • Yasuaki Tamura, Akihiro Yoneda, Norio Takei, Kaori Sawada
    FRONTIERS IN IMMUNOLOGY 7 201  1664-3224 2016/05 [Refereed][Not invited]
     
    Although heat shock proteins (HSPs) primarily play a pivotal role in the maintenance of cellular homeostasis while reducing extracellular as well as intracellular stresses, their role in immunologically relevant scenarios, including activation of innate immunity as danger signals, antitumor immunity, and autoimmune diseases, is now gaining much attention. The most prominent feature of HSPs is that they function both in their own and as an HSP-ligand complex. We here show as a unique feature of extracellular HSPs that they target chaperoned molecules into a particular endosomal compartment of dendritic cells, thereby inducing innate and adaptive immune responses via spatiotemporal regulation.

MISC

Association Memberships

  • 日本ゲノム編集学会   日本癌学会   

Research Projects

  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 2023/04 -2027/03 
    Author : 武井 則雄, 太田 明伸, 桜井 敬之
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2021/04 -2024/03 
    Author : 桂田 武彦, 山本 幸司, 大西 俊介, 武井 則雄
     
    本研究では、羊膜MSCから産生されるHSPB6陽性エクソソームを用いた抗炎症効果の詳細を細胞実験ならびに炎症性腸疾患誘導性動物モデルを用いて、未だ適切な治療法がないヒト炎症性腸疾患に対する臨床応用への基盤を整える。本研究では、以下の通り、HSPB6過剰発現ならびにHSPB6欠損羊膜MSCの作成およびそれらエクソソームの樹立することを明らかにした。羊膜MSCのHSPB6過剰発現株ならびに遺伝子欠損株を樹立する。HSPB6過剰発現株の作成には、CMVプロモーター下にHSPB6遺伝子を挿入したプラスミドをMSCに導入後、薬剤選択による選別を行い、得られたクローンはウエスタンブロット法による野生型およびmockコントロールとの比較解析から、有意に発現増加の認められるクローンを選別することで樹立した。HSPB6欠損株の作成にはCRISPR/Cas9システムを採用し、ヒトHSPB6エクソン領域に特異的なgRNAを設計し、CAGプロモーター下でCas9およびT3プロモーター下でgRNAを発現するプラスミドを構築、MSCに導入し、genotypingによる標的遺伝子座での塩基の欠損が認められ、ウエスタンブロット法で標的分子の翻訳消失が認められるクローンを選別した。結果、20個のクローン中4～5個程度の目的クローンが選別することができた。さらに、その細胞からエクソソームを精製し、HSPB6の発現について確認したところ、HSPB6過剰発現株ではMOCKに比べてHSPB6の発現が増加していた一方、HSPB6欠損株では、HSPB6の発現が消失していることを確認した。これらのことから、本研究において、HSPB6過剰発現株ならびにHSPB6欠損発現株が樹立され、さらに両株から産生される目的となるエクソソームも精製することができた。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2021/04 -2024/03 
    Author : 太田 明伸, 武井 則雄, 花村 一朗
     
    多発性骨髄腫（Multiple myeloma， 以下MM）は形質細胞由来の難治性血液悪性腫瘍である。MMの臨床症状は多彩であり、治療やその効果の予測は難しい。臨床症状の原因として、細胞増殖の制御機構に異常をきたしたMM細胞の異常増殖が考えられているが、その分子学的機序には不明な点が多い。代表者は、MMの新規予後不良因子としてPDZ-binding kinase（以下PBK）を発見し、PBKがMM細胞の造腫瘍性に密接に関連することを示した。そこで本研究では、PBKを標的とした分子標的薬ならびにPBK関連分子との相互作用阻害がMMの新しい治療薬として有効である可能性を検証する方針とした。 はじめに、PBK遺伝子の生理的作用を見出す目的で、PBK遺伝子破壊（PBK-KO）マウスを樹立した。PBK-KOマウスは、正常に出産・発育し外見上特別な表現型を示さなかった。そこで、PBKが豊富に存在する精巣からRNAを抽出して、cDNAマイクロアレイ法とGene Set Enrichment Analysisにより遺伝子発現に関わる解析に着手した。 代表者は、公共のデータベースと遺伝子発現解析を利用したサブ解析から、PBKの発現レベルと高い相関性を示す遺伝子X（仮称）を見出した。遺伝子Xはインターロイキン６によって遺伝子発現が誘導され、その高発現はMM患者の生存率を有意に低下させること、MM細胞株の増殖に関連することを明らかにした。また、ゲノム編集法による内在性タグの導入と、タグ抗体を用いた免疫沈降によるプロテオミクス解析を実施し、遺伝子Xと相互作用する分子の同定を試みている。 さらに、PBKを高発現するMM細胞に対して増殖抑制効果の高い薬剤や化合物を同定するために、1500個のカスタム化合物ライブラリーを入手した。この化合物とPBKの発現を欠失させたアイソジェニッククローンを用いて、各種化合物に対する感受性の変化を解析中である。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2019/04 -2022/03 
    Author : SAKURAI TAKAYUKI
     
    The purpose of this study was to analyze the functional differentiation of RAMPs by analyzing DSS-induced colitis models, colorectal organoids, and macrophage cells using mice with mutations in RAMP subisoforms produced by our systematic multiple simultaneous genome editing methods. We found that RAMP1 and 3 are mainly involved in colitis in mice, and colitis increases in RAMP1-deficient mice, while RAMP3-deficient mice increased their resistance. Furthermore, we showed that RAMP1/RAMP3 signals respond to inflammatory signals simultaneously and synergistically in the inflammatory-stimulated colorectal epithelium, while RAMP1/RAMP3 signals respond to inflammatory signals simultaneously and inversely in macrophage cells.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2018/04 -2021/03 
    Author : Takei Norio
     
    In glucose metabolism, cancer cells are known to have a predominant energy metabolism by glycolysis, even in an aerobic environment, as compared to normal cells. This property has been suggested to be associated with malignant tumors of cancer in various stress environments within the tumor. In this study, we focused on the characteristic metabolic pathways of cancer that differ from these normal cells and conducted a study about the role of a orphan receptor, which is thought to be involved in its metabolism, and the verification of therapeutic effect of its inhibition. This result suggested that this molecule may be involved in the regulation of major metabolic enzymes in cancer-specific energy metabolic pathways, and expected to be a novel target molecule in cancer treatment.
  • 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 : Ota Akinobu
     
    Multiple myeloma (MM) is a complex plasma cell neoplasm, accounting for approximately 10% of all hematological malignancies. Novel molecular-targeted therapy based on their genomic abnormalities in the patients with MM is really wanted to improve their clinical outcome. In this study, we identified that interleukin-6 readily increases PBK expression. Kaplan-Meier analysis showed that the MM patients with higher expression of PBK have a significant shorter survival time compared with those with moderate/lower expression of PBK. Knockout of PBK dramatically suppressed in vivo tumor growth in MM cells. Mechanistically, loss of PBK increased the number of apoptotic cells with concomitant decrease in the phosphorylation level of Stat3. A novel PBK inhibitor OTS514 significantly decreased KMS-11-derived tumor growth. These findings highlight the novel oncogenic role of PBK in tumor growth of myeloma, and it might be a novel therapeutic target for the treatment of patients with MM.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)
    Date (from‐to) : 2016/04 -2018/03 
    Author : Takei Norio
     
    OR7C1 is a G protein coupled receptor and expressed in cancer stem cell and suggested the possibility as a novel factor of poor prognosis. However, the function in cancer remain unclear. In this study, to uncover the importance of OR7C1 expression in cancer biology, we generated conventional OR7C1 knockout clones by using the CRISPR/Cas9 system. As a result, OR7C1 KO decreased tumorigenicity and metastasis in vivo and cell proliferation under specific medium conditions in vitro. In conclusion, our findings suggest that OR7C1 is predominantly activated under specific condition such as starvation and contributes to cancer progression. Collectively, OR7C1 could be used as both a diagnostic biomarker of cancer exacerbation and a therapeutic target.
  • 日本学術振興会：科学研究費助成事業 若手研究(B)
    Date (from‐to) : 2014/04 -2016/03 
    Author : 武井 則雄
     
    前年度の研究結果を踏まえ、KOマウスのGVHD発症機構と関連の予想される主要組織適合抗原複合体(MHC)の発現等を含めたphenotype解析の追加実験および、in vitroでのallo recognitionにおけるサイトカイン産生などをWTとKOマウスで比較検証することでKOマウスを用いたGVHDマウスモデル解析におけるGVHD病態変化の要因を探索した。前年度からの本研究成果をまとめ現在論文投稿準備中である。

Industrial Property Rights

  • 特願2019-151138:MYH9の阻害物質を用いた、がん転移抑制  
    米田明弘, 田村保明, 武井則雄
  • 特願2018-228284:がん処置用RNAi分子  
    武井則雄, 田中洋行, 味呑憲二郎, 高橋博一, 田村保明, 米田明弘
  • 特願2018-228283:がんを処置するための組み合わせ  
    武井則雄, 田中洋行, 味呑憲二郎, 高橋博一, 田村保明, 米田明弘
  • 特願2018-155149:ERO1‐αの阻害物質を用いた、がん幹細胞分化誘導およびがん用化学療法の増強  
    田村保明 、武井則雄 、米田明弘
  • 特願2018-155148:HSP47の阻害物質を用いた、がん転移抑制  
    米田明弘, 田村保明, 武井則雄, 澤田香織
  • 特願2018-155147:HSP47の阻害物質を用いた化学療法剤感受性の増強  
    米田明弘, 田村保明, 武井則雄, 澤田香織

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