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Last Updated :2024/08/29

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Master

Affiliation (Master)

  • Hokkaido University Hospital

Affiliation (Master)

  • Hokkaido University Hospital

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Profile and Settings

Affiliation

  • Hokkaido University, Hokkaido University Hospital
  • Hokkaido University, Hokkaido University Hospital, 副センター長

Degree

  • MD, PhD（Hokkaido University）

Profile and Settings

  • Name (Japanese)

    Kinoshita

  • Name (Kana)

    Ichiro

  • Name

    200901016079944744

Affiliation

  • Hokkaido University, Hokkaido University Hospital
  • Hokkaido University, Hokkaido University Hospital, 副センター長

Achievement

Research Interests

  • 腫瘍内科学   呼吸器内科学   

Research Areas

  • Life sciences / Respiratory medicine

Research Experience

  • 2021/06 - Today Hokkaido University Hokkaido University Hospital
  • 2019/07 - Today Hokkaido University Hospital Division of Clinical Cancer Genomics Professor
  • 2012/09 - 2019/06 Hokkaido University Department of Medical Oncology, Graduate School of Medicine Associate Professor
  • 2005 - 北海道大学講師（大学院医学研究科腫瘍内科学分野） 講師
  • 2005 - Lecturer
  • 2004 - 北海道大学助手（大学院医学研究科腫瘍内科学分野） 助手
  • 2004 - Research Associate
  • 2001 - 北海道大学助手（医学部附属病院内科学第一講座） 助手
  • 2001 - Research Associate

Education

  •        - 1988  Hokkaido University  School of Medicine
  •        - 1988  Hokkaido University  Faculty of Medicine

Awards

  • 2009 Travel Award

Published Papers

  • Takuya Watanabe, Yoshitaka Saito, Yoh Takekuma, Yasushi Shimizu, Ichiro Kinoshita, Yoshito Komatsu, Mitsuru Sugawara
    International journal of clinical pharmacology and therapeutics 2024/03/03 
    OBJECTIVE: Irinotecan (IRI) is an anticancer drug that is frequently used to treat colorectal, gastric, and pancreatic cancers. Its side effects include cholinergic symptoms, such as diarrhea, abdominal pain, nausea, and hyperhidrosis. Anticholinergic medicines are frequently used for treatment or prophylaxis; however, the risk factors for the failure of a single prophylactic anticholinergic administration remain unclear. Moreover, an appropriate anticholinergic drug for prophylaxis remains unknown. Thus, we aimed to identify the risk factors associated with the failure of a single prophylactic dose of anticholinergic drugs for IRI-induced cholinergic symptoms and to evaluate the usefulness of multiple prophylactic doses of anticholinergic drugs. MATERIALS AND METHODS: Patients who underwent IRI treatment for colorectal, gastric, or pancreatic cancer and received prophylactic anticholinergic drugs for IRI-induced cholinergic symptoms (n = 135) were retrospectively evaluated. Univariate and multivariate logistic regression analyses were performed to identify the risk factors for failure of a single prophylactic dose of anticholinergic drugs. We also evaluated the efficacy of multiple prophylactic anticholinergic drug administration. RESULTS: Based on univariate and multivariate analyses, colorectal cancer, female sex, and prophylactic use of scopolamine butyl bromide were identified as risk factors for failure of a single prophylactic dose of anticholinergic drugs. The efficacy of multiple prophylactic doses was confirmed to be 95% of the patients who had a single prophylactic failure due to temporary effect but symptom appearance after a certain period of time (wearing-off). CONCLUSION: We determined that colorectal cancer, female sex, and prophylactic use of scopolamine butyl bromide were risk factors associated with the failure of a single prophylactic dose of anticholinergic drugs, and that multiple prophylactic doses for wearing-off can be a promising method.
  • Tatsunori Shimoi, Kuniko Sunami, Makoto Tahara, Satoshi Nishiwaki, Shota Tanaka, Eishi Baba, Masashi Kanai, Ichiro Kinoshita, Hidekazu Shirota, Hideyuki Hayashi, Naohiro Nishida, Toshio Kubo, Nobuaki Mamesaya, Yayoi Ando, Natsuko Okita, Taro Shibata, Kenichi Nakamura, Noboru Yamamoto
    EClinicalMedicine 69 102447 - 102447 2024/03 
    BACKGROUND: BRAF V600 mutations are common in melanoma, thyroid, and non-small-cell lung cancers. Despite dabrafenib and trametinib being standard treatments for certain cancers, their efficacy across various solid tumours remains unelucidated. The BELIEVE trial assessed the efficacy of dabrafenib and trametinib in solid tumours with BRAF V600E/R or non-V600 BRAF mutations. METHODS: Between October 1, 2019, and June 2022, at least 50 patients with measurable and seven without measurable diseases examined were enrolled in a subcohort of the BELIEVE trial (NCCH1901, jRCTs031190104). BRAF mutated solid tumour cases other than BRAF V600E mutated colorectal cancer, melanoma, and non-small cell lung cancer cases were included. Patients with solid tumours received dabrafenib (150 mg) twice daily and trametinib (2 mg) once daily until disease progression or intolerable toxicity was observed. The primary endpoint was overall response rate (ORR), and secondary endpoints included progression-free survival (PFS), 6-month PFS, and overall survival (OS). Bayesian analysis was performed using a prior distribution with a 30% expected response rate [Beta (0.6, 1.4)]. FINDINGS: Fourty-seven patients with measurable disease, mainly with the BRAF V600E mutation (94%), and three others with non-V600E BRAF mutations (V600R, G466A, and N486_P490del) were enrolled. The primary sites included the thyroid gland, central nervous system, liver, bile ducts, colorectum, and pancreas. The confirmed ORR was 28.0%; the expected value of posterior distribution [Beta (14.6, 37.4)] was 28.1%, although the primary endpoint was achieved, not exceeding an unexpectedly high response rate of 60% obtained using Bayesian analysis. The disease control rate (DCR) was 84.0%. The median PFS was 6.5 months (95% confidence interval [CI]; 4.2-7.2 months, 87.8% at 6 months). Responses were observed across seven tumour types. Median OS was 9.7 months (95% CI, 7.5-12.2 months). Additional patients without measurable diseases had a median PFS of 4.5 months. Adverse events (AEs) were consistent with previous reports, with 45.6% of patients experiencing grade ≥3 AEs. INTERPRETATION: This study reported promising efficacy against BRAF V600-mutant tumours. Dabrafenib and trametinib would offer a new therapeutic option for rare cancers, such as high-grade gliomas, biliary tract cancer, and thyroid cancer. FUNDING: This study was funded by the Japan Agency for Medical Research and Development (22ck0106622h0003) and a Health and Labour Sciences Research Grant (19EA1008).
  • YOSHITAKA SAITO, TATSUHIKO SAKAMOTO, MASAKI KOBAYASHI, YOH TAKEKUMA, ISSEI HIGUCHI, KEISUKE OKAMOTO, JUN SAKAKIBARA-KONISHI, YASUSHI SHIMIZU, ICHIRO KINOSHITA, MITSURU SUGAWARA
    In Vivo 38 (2) 800 - 806 0258-851X 2024/02/28
  • Tatsuhiko Sakamoto, Moeko Kado, Yoshitaka Saito, Kazuki Uchiyama, Ryota Kanno, Osamu Taniguchi, Yoh Takekuma, Jun Sakakibara-Konishi, Yasushi Shimizu, Ichiro Kinoshita, Mitsuru Sugawara
    Biological & pharmaceutical bulletin 47 (6) 1189 - 1195 2024 
    Although carboplatin (CBDCA) is classified as a moderately emetogenic agent, the majority of guidelines recommend the use of a neurokinin-1 receptor antagonist in addition to a 5-hydroxytryptamine type 3 receptor antagonist with dexamethasone (DEX) for CBDCA-containing chemotherapy because of its higher emetogenic risk. However, the additional efficacy of aprepitant (APR) in CBDCA-containing treatment remains controversial, and data on multiple-day treatments are limited. Etoposide (ETP) was administered on days 1-3 in the CBDCA + ETP regimen, and it is important to evaluate suitable antiemetic therapy for the regimen. Therefore, we evaluated the efficacy of additional APR in CBDCA + ETP. Patients were divided into two groups and retrospectively evaluated. One was the control group, which was prophylactically administered palonosetron (PALO) and DEX, and the other was the APR group, which received APR orally with PALO and DEX. The primary endpoint was complete response (CR) between the groups. The overall CR rates were 75.0 and 76.4% in the control and APR groups, respectively, with no significant difference (p = 1.00). In the acute phase, it was 88.9 and 97.2%, respectively, and 86.1 and 79.2% in the delayed phase, respectively, without significant differences (p = 0.10 and 0.38, respectively). The incidence and severity of nausea, vomiting, and anorexia were not significantly different between the two groups in the acute and delayed phases. Our findings suggest that combining APR with PALO and DEX does not improve the CR rate in CBDCA + ETP therapy.
  • Yoshitaka Saito, Shinya Tamaki, Daisuke Hirate, Shinya Takada, Kenta Takahashi, Yoh Takekuma, Jun Sakakibara-Konishi, Yasushi Shimizu, Ichiro Kinoshita, Mitsuru Sugawara
    Scientific reports 13 (1) 19457 - 19457 2023/11/09 [Refereed]
     
    Treatment using docetaxel (DOC) and ramucirumab (RAM) is an effective regimen in second or later line advanced non-small cell lung carcinoma (NSCLC) treatment. However, it induces severe adverse effects, resulting in treatment reduction such as dose reduction and/or discontinuation. This study aimed to reveal the factor(s) associated with treatment reduction in DOC + RAM. We retrospectively evaluated patients with advanced NSCLC (n = 155). Treatment reduction of the second course due to severe adverse effects was conducted in 25.8% of the participants, and relative dose intensity at the second course was 95.7 ± 8.4% for DOC and 91.9 ± 24.8% for RAM. Multivariate logistic regression analyses identified that baseline anemia and prophylactic granulocyte colony-stimulating factor (G-CSF) administration are preventive factors for the reduction (adjusted odds ratio, 0.29; 95% confidence interval, 0.12-0.66; P = 0.004 for baseline anemia, 0.18; 0.08-0.42; P < 0.0001 for prophylactic G-CSF administration). The primary cause of the reduction was febrile neutropenia, and the same factors were identified. Our study revealed that patients with baseline anemia and prophylactic G-CSF administration have less risk for treatment reduction in DOC + RAM for NSCLC treatment.
  • Yoshitaka Saito, Masaki Kobayashi, Shinya Tamaki, Katsuyuki Nakamura, Daisuke Hirate, Kenta Takahashi, Yoh Takekuma, Jun Sakakibara-Konishi, Yasushi Shimizu, Ichiro Kinoshita, Mitsuru Sugawara
    Scientific reports 13 (1) 17126 - 17126 2023/10/10 
    The occurrence of cisplatin (CDDP)-induced nephrotoxicity (CIN) has decreased with advancements in supportive care. In contrast, we reported that baseline diabetes mellitus (DM) complications significantly worsen CIN. This study aimed to determine further risk factors associated with CIN development in DM patients. Patients with thoracic cancer requiring DM pharmacotherapy, who received CDDP (≥ 60 mg/m2)-containing regimens using the short hydration method (n = 140), were enrolled in this retrospective multicenter observational study. The primary endpoint of the present study was the elucidation of risk factors (patient factors, DM medication influence, and treatment-related factors) associated with CIN development in patients with DM. Cisplatin-induced nephrotoxicity occurred in 22.1% of patients with DM. The median worst variation of serum creatinine levels and creatinine clearance (worst level - baseline level) was 0.16 mg/dL (range: - 0.12-1.41 mg/dL) and - 15.9 mL/min (- 85.5-24.3 mL/min), respectively. Multivariate logistic regression analyses identified female sex as the singular risk factor for CIN development in the DM population (adjusted odds ratio; 2.87, 95% confidence interval; 1.08-7.67, P = 0.04). Diabetes mellitus medication and treatment-related factors did not affect CIN development. In conclusion, our study revealed that female sex is significantly associated with CIN development in patients with DM and thoracic cancer.
  • Satoshi Takeuchi, Kenji Hirata, Keiichi Magota, Shiro Watanabe, Rika Moku, Akihiko Shiiya, Jun Taguchi, Shin Ariga, Tomohiro Goda, Yoshihito Ohhara, Takurou Noguchi, Yasushi Shimizu, Ichiro Kinoshita, Rio Honma, Yasushi Tsuji, Akihiro Homma, Hirotoshi Dosaka-Akita
    EJNMMI research 13 (1) 69 - 69 2023/07/17 
    BACKGROUND: Lenvatinib is widely used to treat unresectable and advanced thyroid carcinomas. We aimed to determine whether 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) performed 1 week after lenvatinib treatment initiation could predict treatment outcomes. RESULTS: This was a prospective, nonrandomised, multicentre study. Patients with pathologically confirmed differentiated thyroid carcinoma (DTC) and lesions refractory to radioiodine treatment were eligible for inclusion. Patients were treated with 24 mg lenvatinib as the initial dose and underwent PET/CT examination 1 week after treatment initiation. Contrast-enhanced CT was scheduled at least 4 weeks later as the gold standard for evaluation. The primary endpoint was to evaluate the discrimination power of maximum standardised uptake value (SUVmax) obtained by PET/CT compared to that obtained by contrast-enhanced CT. Evaluation was performed using the area under the receiver operating characteristic (ROC-AUC) curve. Twenty-one patients were included in this analysis. Receiver operating characteristic (ROC) curve analysis yielded an AUC of 0.714 for SUVmax after 1 week of lenvatinib treatment. The best cut-off value for the treatment response for SUVmax was 15.211. The sensitivity and specificity of this cut-off value were 0.583 and 0.857, respectively. The median progression-free survival was 26.3 months in patients with an under-cut-off value and 19.7 months in patients with an over-cut-off value (P = 0.078). CONCLUSIONS: The therapeutic effects of lenvatinib were detected earlier than those of CT because of decreased FDG uptake on PET/CT. PET/CT examination 1 week after the initiation of lenvatinib treatment may predict treatment outcomes in patients with DTC. TRIAL REGISTRATION: This trial was registered in the University Hospital Medical Information Network (UMIN) Clinical Trials Registry (number UMIN000022592) on 6 June, 2016.
  • Kanako Hagio, Junko Kikuchi, Kohichi Takada, Hiroki Tanabe, Minako Sugiyama, Yoshihito Ohhara, Toraji Amano, Satoshi Yuki, Yoshito Komatsu, Takahiro Osawa, Kanako C Hatanaka, Yutaka Hatanaka, Takashi Mitamura, Ichiro Yabe, Yoshihiro Matsuno, Atsushi Manabe, Akihiro Sakurai, Atsushi Ishiguro, Masato Takahashi, Hiroshi Yokouchi, Hirohito Naruse, Yusuke Mizukami, Hirotoshi Dosaka-Akita, Ichiro Kinoshita
    Cancer science 2023/05/19 
    Comprehensive genomic profiling (CGP) tests have been covered by public insurance in Japan for patients with advanced solid tumors who have completed or are completing standard treatments or do not have them. Therefore, genotype-matched drug candidates are often unapproved or off-label, and improving clinical trial access is critical, involving the appropriate timing of CGP tests. To address this issue, we analyzed the previous treatment data for 441 patients from an observational study on CGP tests discussed by the expert panel at Hokkaido University Hospital between August 2019 and May 2021. The median number of previous treatment lines was two; three or more lines accounted for 49%. Information on genotype-matched therapies was provided to 277 (63%). Genotype-matched clinical trials were ineligible because of an excess number of previous treatment lines or use of specific agents were found in 66 (15%) patients, with the highest proportion in breast and prostate cancers. Many patients met the exclusion criteria of one to two or more treatment lines across cancer types. In addition, previous use of specific agents was a frequent exclusion criterion for breast, prostate, colorectal, and ovarian cancers. The patients with tumor types with a low median number (two or fewer) of previous treatment lines, including most rare cancers, primary unknown cancers, and pancreatic cancers, had significantly fewer ineligible clinical trials. The earlier timing of CGP tests may improve access to genotype-matched clinical trials, with their proportion varying by cancer type. Each relevant society needs to advocate the desirable timing of CGP testing nationwide.
  • Osamu Taniguchi, Yoshitaka Saito, Yoh Takekuma, Hirotoshi Akita, Ichiro Kinoshita, Yasushi Shimizu, Naofumi Shinagawa, Mitsuru Sugawara
    International journal of clinical pharmacology and therapeutics 61 (6) 246 - 254 2023/03/27 
    OBJECTIVE: Chemotherapy-induced nausea and vomiting (CINV) and chemotherapy-associated dyspepsia syndrome (CADS) are frequently appearing adverse effects of cisplatin (CDDP)-containing chemotherapy. Antiemetic guidelines suggest that the administration of antacids such as proton pump inhibitors (PPIs) or histamine type-2 receptor antagonists be considered for CADS, although their efficacy for treating these symptoms remains unknown. This study aimed to reveal whether antacids attenuate gastrointestinal symptoms in CDDP-containing chemotherapy. MATERIALS AND METHODS: In total, 138 patients with lung cancer who received ≥ 75 mg/m2 CDDP-containing regimens were enrolled in this retrospective study. Patients were divided into an antacid group including patients administered PPIs or vonoprazan during all chemotherapy periods and controls without antacid administration. The primary endpoint was the comparison of anorexia incidence during the first cycle of chemotherapy. Secondary endpoints were CINV evaluation and risk factor analysis for the incidence of anorexia using logistic regression analysis. RESULTS: The incidence of anorexia during the first cycle was 54.4% in the control group and 60.3% in the antacid group, without significant differences (p = 0.60). The incidence of nausea was also similar between the groups (p = 1.00). Multivariate analysis suggested that antacid administration was not associated with anorexia. CONCLUSION: Baseline antacid administration does not affect gastrointestinal symptoms associated with CDDP-containing treatment in lung cancer.
  • EGFR変異陽性肺癌におけるCD24発現の検討
    椎谷 研彦, 野口 卓郎, 外丸 詩野, 有賀 伸, 高島 雄太, 品川 尚文, 木下 一郎, 松野 吉宏, 榊原 純, 秋田 弘俊
    日本病理学会会誌 (一社)日本病理学会 112 (1) 247 - 247 0300-9181 2023/03
  • Motoo Katabami, Ichiro Kinoshita, Shin Ariga, Yasushi Shimizu, Hirotoshi Dosaka-Akita
    PloS one 18 (5) e0285354  2023 
    Crystalline silica-induced inflammation possibly facilitates carcinogenesis. Here, we investigated its effect on lung epithelium damage. We prepared conditioned media of immortalized human bronchial epithelial cell lines (hereinafter bronchial cell lines) NL20, BEAS-2B, and 16HBE14o- pre-exposed to crystalline silica (autocrine crystalline silica conditioned medium), a phorbol myristate acetate-differentiated THP-1 macrophage line, and VA13 fibroblast line pre-exposed to crystalline silica (paracrine crystalline silica conditioned medium). As cigarette smoking imposes a combined effect on crystalline silica-induced carcinogenesis, a conditioned medium was also prepared using the tobacco carcinogen benzo[a]pyrene diol epoxide. Crystalline silica-exposed and growth-suppressed bronchial cell lines exhibited enhanced anchorage-independent growth in autocrine crystalline silica and benzo[a]pyrene diol epoxide conditioned medium compared with that in unexposed control conditioned medium. Crystalline silica-exposed nonadherent bronchial cell lines in autocrine crystalline silica and benzo[a]pyrene diol epoxide conditioned medium showed increased expression of cyclin A2, cdc2, and c-Myc, and of epigenetic regulators and enhancers, BRD4 and EZH2. Paracrine crystalline silica and benzo[a]pyrene diol epoxide conditioned medium also accelerated the growth of crystalline silica-exposed nonadherent bronchial cell lines. Culture supernatants of nonadherent NL20 and BEAS-2B in crystalline silica and benzo[a]pyrene diol epoxide conditioned medium had higher EGF concentrations, whereas those of nonadherent 16HBE14o- had higher TNF-α levels. Recombinant human EGF and TNF-α promoted anchorage-independent growth in all lines. Treatment with EGF and TNF-α neutralizing antibodies inhibited cell growth in crystalline silica conditioned medium. Recombinant human TNF-α induced BRD4 and EZH2 expression in nonadherent 16HBE14o-. The expression of γH2AX occasionally increased despite PARP1 upregulation in crystalline silica-exposed nonadherent lines with crystalline silica and benzo[a]pyrene diol epoxide conditioned medium. Collectively, crystalline silica- and benzo[a]pyrene diol epoxide-induced inflammatory microenvironments comprising upregulated EGF or TNF-α expression may promote crystalline silica-damaged nonadherent bronchial cell proliferation and oncogenic protein expression despite occasional γH2AX upregulation. Thus, carcinogenesis may be cooperatively aggravated by crystalline silica-induced inflammation and genotoxicity.
  • Akihiko Shiiya, Takuro Noguchi, Utano Tomaru, Shin Ariga, Yuta Takashima, Yoshihito Ohhara, Jun Taguchi, Satoshi Takeuchi, Yasushi Shimizu, Ichiro Kinoshita, Tomonobu Koizumi, Yoshihiro Matsuno, Naofumi Shinagawa, Jun Sakakibara-Konishi, Hirotoshi Dosaka-Akita
    Cancer science 2022/12/18 
    Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) elicit potent cell cycle arrest in EGFR-mutant non-small-cell lung cancer (NSCLC) cells. However, little is known about the mechanisms through which these drugs alter the tumor phenotype that contributes to the immune escape of EGFR-mutant cells. Using EGFR-mutant NSCLC cell lines and tissue samples from patients, we investigated the changes in immune checkpoints expressed in tumor cells following EGFR inhibition. Subsequently, we also analyzed the role of soluble factors from the dying tumor cells in the activation of immune signaling pathways involved in therapy resistance. Upon EGFR-TKI treatment, we found that EGFR-mutant cells upregulated the expression of innate immune checkpoint CD24 in vitro. We then analyzed biopsy samples from six patients who developed resistance to a first-generation EGFR-TKI without the acquired T790M mutation. Immunohistochemistry revealed that levels of tumor CD24 expression were increased upon treatment compared with those from pre-treatment samples. Monocyte-derived macrophages facilitated antibody-dependent cellular phagocytosis when EGFR-TKI-treated EGFR-mutant cells were incubated with anti-CD24 antibodies in vitro, suggesting that CD24 may be a therapeutical target for EGFR-mutant lung cancer. Moreover, EGFR inhibition accelerated the release of cell-free DNA (cfDNA) from dying tumor cells, which activated the type I interferon signaling pathways in human THP-1 monocytes in a stimulator of interferon genes-dependent manner. Our study indicates that EGFR inhibition in EGFR-mutant NSCLC cells fosters a tumor microenvironment associated with immune escape. Thus, CD24 targeted therapy and cfDNA monitoring may contribute to improved treatment outcomes in patients with EGFR-mutant NSCLC.
  • Yoshitaka Saito, Tatsuhiko Sakamoto, Yoh Takekuma, Masaki Kobayashi, Keisuke Okamoto, Naofumi Shinagawa, Yasushi Shimizu, Ichiro Kinoshita, Mitsuru Sugawara
    Scientific Reports 12 (1) 2022/12/17 
    Abstract Cisplatin (CDDP)-induced nephrotoxicity (CIN) is dose-limiting. We revealed that co-administration of non-steroid anti-inflammatory drugs and baseline comorbidity of diabetes mellitus (DM) are associated with CIN development in the short hydration method; however, the results were accessorily obtained without appropriate power calculation. This study aimed to demonstrate the influence of DM complications on CIN incidence in a real-world setting. Lung cancer patients receiving CDDP (≥ 75 mg/m2)-containing regimens with a short hydration method (n = 227) were retrospectively evaluated. The patients were divided into control and baseline DM complication groups. The primary endpoint was the evaluation of CIN incidence between the groups. Propensity score-matching was performed to confirm the robustness of the primary analysis results. CIN occurred in 6.8% of control and 27.0% of DM patients, respectively, with a significant difference in all-patient populations (P = 0.001). In addition, variation of serum creatinine and creatinine clearance significantly worsened in DM patients. Similar results were obtained in a propensity-matched population. Multivariate logistic regression analysis found that DM complication is a singular risk factor for CIN development (adjusted odds ratio; 4.31, 95% confidence interval; 1.62–11.50, P = 0.003). In conclusion, our study revealed that baseline DM complications significantly worsen CIN.
  • Yoichi Naito, Kuniko Sunami, Hidenori Kage, Keigo Komine, Toraji Amano, Mitsuho Imai, Takafumi Koyama, Daisuke Ennishi, Masashi Kanai, Hirotsugu Kenmotsu, Takahiro Maeda, Sachi Morita, Daisuke Sakai, Kousuke Watanabe, Hidekazu Shirota, Ichiro Kinoshita, Masashiro Yoshioka, Nobuaki Mamesaya, Mamoru Ito, Shinji Kohsaka, Yusuke Saigusa, Kouji Yamamoto, Makoto Hirata, Katsuya Tsuchihara, Takayuki Yoshino
    JAMA Network Open 5 (12) e2245081 - e2245081 2022/12/05 [Refereed]
     
    Importance Quality assurance of molecular tumor boards (MTBs) is crucial in cancer genome medicine. Objective To evaluate the concordance of recommendations by MTBs and centrally developed consensus treatment recommendations at all 12 leading institutions for cancer genomic medicine in Japan using 50 simulated cases. Design, Setting, and Participants This was a prospective quality improvement study of 50 simulated cancer cases. Molecular tumor boards from 12 core hospitals independently recommended treatment for 50 cases blinded to the centrally developed consensus treatment recommendations. The study’s central committee consisted of representatives from all 12 core hospitals in Japan who selected the 50 simulated cases from The Cancer Genome Atlas database, including frequently observed genomic alterations. The central committee recommended centrally developed consensus treatment. The concordance rate for genomically matched treatments between MTBs and centrally developed consensus treatment recommendations was evaluated. Data analysis was conducted from January 22 to March 3, 2021. Exposures Simulated cases of cancer. Main Outcomes and Measures The primary outcome was concordance, defined as the proportion of recommendations by MTBs concordant with centrally developed consensus treatment recommendations. A mixed-effects logistic regression model, adjusted for institutes as a random intercept, was applied. High evidence levels were defined as established biomarkers for which the treatment was ready for routine use in clinical practice, and low evidence levels were defined as biomarkers for genomically matched treatment that were under investigation. Results The Clinical Practice Guidance for Next-Generation Sequencing in Cancer Diagnosis and Treatment (edition 2.1) was used for evidence-level definition. The mean concordance between MTBs and centrally developed consensus treatment recommendations was 62% (95% CI, 57%-65%). Each MTB concordance varied from 48% to 86%. The concordance rate was higher in the subset of patients with colorectal cancer (100%; 95% CI, 94.0%-100%), ROS1 fusion (100%; 95% CI, 85.5%-100%), and high evidence level A/R (A: 88%; 95% CI, 81.8%-93.0%; R:100%; 95% CI, 92.6%-100%). Conversely, the concordance rate was lower in cases of cervical cancer (11%; 95% CI, 3.1%-26.1%), TP53 mutation (16%; 95% CI, 12.5%-19.9%), and low evidence level C/D/E (C: 30%; 95% CI, 24.7%-35.9%; D: 25%; 95% CI, 5.5%-57.2%; and E: 18%; 95% CI, 13.8%-23.0%). Multivariate analysis showed that evidence level (high [A/R] vs low [C/D/E]: odds ratio, 4.4; 95% CI, 1.8-10.8) and TP53 alteration (yes vs no: odds ratio, 0.06; 95% CI, 0.03-0.10) were significantly associated with concordance. Conclusions and Relevance The findings of this study suggest that genomically matched treatment recommendations differ among MTBs, particularly in genomic alterations with low evidence levels wherein treatment is being investigated. Sharing information on matched therapy for low evidence levels may be needed to improve the quality of MTBs.
  • Yoshihito Ohhara, Ichiro Kinoshita, Akira Suzuki, Makoto Imagawa, Jun Taguchi, Takuro Noguchi, Satoshi Takeuchi, Yasushi Shimizu, Hideyuki Seki, Junichi Suzuki, Hirotoshi Dosaka-Akita
    Oncology 2022/10/21 
    INTRODUCTION: Karyopherin alfa 2 (KPNA2) and karyopherin beta 1 (KPNB1), constitute nuclear transport protein complexes involved in nuclear import, and are significant in tumor progression. Although high KPNA2 expression was associated with poor prognosis in solid tumors, the relationship between KPNA2 and KPNB1 expression and their prognostic role in gastric cancer (GC) remains unclear. METHODS: Immunohistochemistry was used to correlate the expression of KPNA2 and KPNB1 with various features, including clinicopathologic characteristics in 130 patients with GC and survival in 94 patients with invasive lesions extending to the submucosa or deeper. RESULTS: High expression of KPNA2 and KPNB1 was found in 25% and 36% of the patients, respectively. Both were significantly related to tumor depth, lymph node metastasis, lymphatic invasion, venous invasion, and Ki-67 expression. KPNA2 expression was significantly related to that of KPNB1 (P < 0.001). Patients with high KPNB1 expression had poorer prognosis than those with low expression (P = 0.027), as was also observed in case of KPNA2 (P < 0.001). Patients with high expression of both KPNA2 and KPNB1 accounted for 18% and had a poorer prognosis than those with high expression of either and those with low expression of both (P = 0.001). Multivariate analysis revealed that high expression of both KPNA2 and KPNB1 was an independent prognostic factor in patients with GC (HR 3.46, 95% CI 1.64-2.73, P = 0.001). CONCLUSION: KPNA2 expression was correlated with KPNB1 expression, and high co-expression of KPNA2 and KPNB1 may represent a strong prognostic biomarker in GC.
  • Kohichi Takada, Tomohiro Kubo, Junko Kikuchi, Makoto Yoshida, Ayako Murota, Yohei Arihara, Hajime Nakamura, Hiroyuki Nagashima, Hiroki Tanabe, Shintaro Sugita, Yumi Tanaka, Ayana Miura, Yoshihito Ohhara, Atsushi Ishiguro, Hiroshi Yokouchi, Yasuyuki Kawamoto, Yusuke Mizukami, Hirofumi Ohnishi, Ichiro Kinoshita, Akihiro Sakurai
    Frontiers in Oncology 12 2022/09/02 
    Characterization of the genomic landscape of biliary tract cancer (BTC) may lead to applying genotype-matched therapy for patients with this disease. Evidence that comprehensive cancer genomic profiling (CGP) guides genotype-matched therapy to improve clinical outcomes is building. However, the significance of CGP in patients with BTC remains unclarified in clinical practice. Therefore, the purposes of this study were to assess the utility of CGP and identify associations between clinical outcomes and genomic alterations in patients with BTC. In this prospective analysis, detection rates for actionable genomic alterations and access rates for genotype-matched therapy were analyzed in 72 patients with advanced BTC who had undergone commercial CGP. Cox regression analyses assessed relationships between overall survival and genomic alterations detected with CGP. The most common genomic alterations detected were TP53 (41, 56.9%), followed by CDKN2A/B (24, 33.3%/20, 27.8%), and KRAS (20, 27.8%). Actionable genomic alterations were identified in 58.3% (42/72) of patients. Detection rates for FGFR2 fusions, IDH1 mutations, and BRAF V600E were low in this cohort. Eight (11.1%) patients received genotype-matched therapy. For patients with intrahepatic cholangiocarcinoma (ICC), CDKN2A/B loss was associated with shorter overall survival. These real-world data demonstrate that CGP can identify therapeutic options in patients with advanced BTC. CDKN2A/B loss was identified as a poor prognostic factor in patients with ICC. Thus, this study provides a rationale for considering CGP in planning therapeutic strategies for advanced BTC.
  • Yoshitaka Saito, Yoh Takekuma, Takashi Takeshita, Takuro Noguchi, Satoshi Takeuchi, Yasushi Shimizu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Mitsuru Sugawara
    Anticancer research 42 (7) 3693 - 3700 2022/07 
    BACKGROUND/AIM: Eribulin is an effective chemotherapeutic agent for the treatment of metastatic breast cancer and advanced or metastatic soft-tissue sarcomas. However, severe adverse effects (SAEs) occur in 30-40% of the patients, and significantly reduce the patients' quality of life and disturb the recommended treatment schedules. Neutropenia is the main cause of treatment suspension, delay, and/or dose reductions, also leading to relative dose intensity reduction. This study aimed to examine the risk factors for SAE occurrence after eribulin treatment. PATIENTS AND METHODS: Eighty patients with metastatic breast cancer or advanced or metastatic soft tissue sarcoma who received eribulin were retrospectively evaluated. Risk factors for SAE occurrence in the first cycle were primarily assessed. In addition, factors associated with SAE occurrence during all treatment cycles were evaluated. RESULTS: SAEs in the first cycle occurred in 45% of patients. The primary SAE was neutropenia (91.7%). The incidence of SAEs during all treatment cycles was 61.3%. Multivariate analyses suggested that lower baseline neutrophil and hemoglobin levels were risk factors for SAE occurrence and severe neutropenia incidence in the first cycle. An independent factor associated with SAE occurrence during all cycles was age ≥65 years and a tendency was confirmed for baseline anemia. CONCLUSION: Baseline neutropenia and anemia were risk factors for SAE occurrence during the first eribulin treatment cycle. Age ≥65 years was also associated with SAE occurrence during all treatment cycles. Patients with these risk factors should be carefully monitored for assessment and prophylaxis.
  • Yoshitaka Saito, Yoh Takekuma, Masaki Kobayashi, Naofumi Shinagawa, Takuro Noguchi, Satoshi Takeuchi, Yasushi Shimizu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Mitsuru Sugawara
    Anticancer research 42 (6) 3185 - 3193 2022/06 
    BACKGROUND/AIM: Short hydration is a method to change partial intravenous hydration to oral to administer cisplatin (CDDP); however, the most suitable form of oral hydration is unknown. This study aimed to determine whether oral rehydration solution (ORS) affects CDDP-induced nephrotoxicity (CIN) and electrolyte imbalance. PATIENTS AND METHODS: Lung cancer patients (n=200) who had received CDDP-including regimens (CDDP dosage ≥75 mg/m2) were retrospectively evaluated. We used logistic analysis to evaluate whether ORS intake could be a preventive factor for CIN (≥grade 2 serum creatinine elevation). Moreover, incidence of CIN and electrolyte imbalance and the variation in serum creatinine and electrolyte levels were compared between ORS and non-ORS (control) patients. RESULTS: CIN occurred in 9.8% of ORS patients, and 7.5% of non-ORS patients (p=0.79). The variation in serum creatinine level was also similar in both groups. Multivariate analysis suggested that ORS intake does not affect CIN, although CIN was associated with the coadministration of non-steroidal anti-inflammatory drugs and the presence of diabetes mellitus. The variations in serum electrolyte levels did not differ, and incidence of hyponatremia, hypokalemia, and hypochloremia was also similar between the groups. Moreover, patients in ORS group experienced significantly more anorexia compared to controls, and approximately 40% of the patients were unable to continue ORS intake. CONCLUSION: ORS intake in CDDP short hydration regimens does not affect CIN and CDDP-induced electrolyte imbalance; however, its intake is associated with the incidence of anorexia suggesting that ORS should not be used for oral hydration.
  • 進行固形がん患者に対する保険診療による包括的がんゲノムプロファイリング検査の実施時期についての検討
    菊地 順子, 大原 克仁, 天野 虎次, 矢部 一郎, 小松 嘉人, 松野 吉宏, 石黒 敦, 水上 裕輔, 櫻井 晃洋, 木下 一郎
    日本内科学会雑誌 (一社)日本内科学会 111 (Suppl.) 190 - 190 0021-5384 2022/02
  • Minako Sugiyama, Shinsuke Hirabayashi, Yukitomo Ishi, Junko Kikuchi, Ayako Ishikura, Hiroaki Motegi, Yuki Ueda, Saori Sawai, Kazuya Hara, Yukayo Terashita, Yuko Cho, Emi Takakuwa, Shohei Honda, Shigeru Yamaguchi, Ichiro Kinoshita, Atsushi Manabe
    Pediatric blood & cancer 68 (11) e29227  2021/11 [Refereed]
  • Shunji Takahashi, Masato Karayama, Masato Takahashi, Junichiro Watanabe, Hironobu Minami, Noboru Yamamoto, Ichiro Kinoshita, Chia-Chi Lin, Young-Hyuck Im, Issei Achiwa, Emi Kamiyama, Yasuyuki Okuda, Caleb Lee, Yung-Jue Bang
    Clinical cancer research : an official journal of the American Association for Cancer Research 2021/08/23 [Refereed]
     
    PURPOSE: To evaluate drug-drug interactions between the human epidermal growth factor receptor 2 (HER2)-targeted antibody-drug conjugate trastuzumab deruxtecan (T-DXd; DS-8201a) and the OATP1B/CYP3A inhibitor ritonavir or the strong CYP3A inhibitor itraconazole. PATIENTS AND METHODS: Patients with HER2-expressing advanced solid tumors were enrolled in this phase I, open-label, single-sequence crossover study (NCT03383692) and received i.v. T-DXd 5.4 mg/kg every 3 weeks. Patients received ritonavir (cohort 1) or itraconazole (cohort 2) from day 17 of cycle 2 through the end of cycle 3. Primary endpoints were maximum serum concentration (C max) and partial area under the concentration-time curve from beginning of cycle through day 17 (AUC17d) for T-DXd and deruxtecan (DXd) with (cycle 3) and without (cycle 2) ritonavir or itraconazole treatment. RESULTS: Forty patients were enrolled (cohort 1, n = 17; cohort 2, n = 23). T-DXd C max was similar whether combined with ritonavir [cohort 1, cycle 3/cycle 2; 90% confidence interval (CI): 1.05 (0.98-1.13)] or itraconazole [cohort 2, 1.03 (0.96-1.09)]. T-DXd AUC17d increased from cycle 2 to 3; however, the cycle 3/cycle 2 ratio upper CI bound remained at ≤1.25 for both cohorts. For DXd (cycle 3/cycle 2), C max ratio was 0.99 (90% CI, 0.85-1.14) for cohort 1 and 1.04 (0.92-1.18) for cohort 2; AUC17d ratio was 1.22 (1.08-1.37) and 1.18 (1.11-1.25), respectively. The safety profile of T-DXd plus ritonavir or itraconazole was consistent with previous studies of T-DXd monotherapy. T-DXd demonstrated promising antitumor activity across HER2-expressing solid-tumor types. CONCLUSIONS: T-DXd was safely combined with ritonavir or itraconazole without clinically meaningful impact on T-DXd or DXd pharmacokinetics.
  • Ichiro Kinoshita, Junko Kikuchi, Yoshihito Ohhara, Toraji Amano, Hirotoshi Akita
    Gan to kagaku ryoho. Cancer & chemotherapy 48 (7) 882 - 886 0385-0684 2021/07 
    Hokkaido University Hospital has been designated as a Core Hospital for Cancer Genomic Medicine and developed a system to provide cancer genomic medicine in Hokkaido with its liaison hospitals. Since being reimbursed in June 2019, comprehensive cancer genome profiling (CGP) testing showed certain therapeutic efficacy in patients with no standard treatment options, but it also revealed some problems such as the small number of patients who can receive therapeutic drugs matched with gene abnormalities. Since candidate drugs are often unapproved or off-label, it is necessary to smoothly introduce clinical trials, advanced medical treatment system, and patient-proposed health care service. At our hospital, we are focusing on sharing information on clinical trials being conducted in Hokkaido, launching investigator-initiated clinical trials, promoting patient-proposed health care service, promoting a registry study of genetic profiling and targeted therapies in patients with rare cancers and accompanying clinical trials, and incorporating pediatric cancer patients. This paper describes Hokkaido's cancer genomic medicine provision system, including its exit strategy, and the human resource development that serve as its foundation.
  • Takayuki Kiuchi, Utano Tomaru, Akihiro Ishizu, Makoto Imagawa, Sari Iwasaki, Akira Suzuki, Noriyuki Otsuka, Yoshihito Ohhara, Ichiro Kinoshita, Yoshihiro Matsuno, Hirotoshi Dosaka-Akita, Masanori Kasahara
    Journal of clinical pathology 74 (5) 300 - 306 2021/05 [Refereed]
     
    AIM: The immunoproteasome is a specific proteasome isoform whose proteolytic activity enhances the generation of antigenic peptides to be presented by major histocompatibility complex class I molecules to CD8+ T cells. Physiologically, it is expressed abundantly in immune cells and is induced in somatic cells by cytokines, especially interferon-γ. Recently, variable expression of immunoproteasomes has been demonstrated in different types of cancers. However, the clinical significance of immunoproteasome expression in malignant tumours is poorly understood. In this study, we performed clinicopathological evaluation of immunoproteasome subunit β5i in non-small cell lung carcinomas (NSCLCs). METHODS: Tumour tissues were collected from 155 patients with NSCLCs, and immunohistochemical analysis for β5i was performed in relation to the prognosis of patients. RESULTS: High expression of β5i was found in about 20% of all NSCLCs and was found significantly more frequently (40%) in the adenocarcinoma subset. High expression of β5i was associated with a better 5-year relative survival rate in patients with pStage I to II adenocarcinoma and was also a significant and independent favourable prognostic factor in adenocarcinoma patients. In addition, when we performed in vitro analysis using NSCLC cell lines, combined treatment with the immunoproteasome-specific inhibitor ONX0914 and the proteasome inhibitor MG132 enhanced cell death in β5i-expressing NSCLC cell lines. CONCLUSION: The expression of immunoproteasome can be explored as both a prognostic factor and a potential therapeutic target in NSCLCs. Since immunoproteasomes have crucial role in the antigen presentation, further studies may help to provide essential knowledge for therapeutic strategies in anticancer immunotherapy.
  • Junko Kikuchi, Yoshihito Ohhara, Kohichi Takada, Hiroki Tanabe, Kazuteru Hatanaka, Toraji Amano, Kanako C Hatanaka, Yutaka Hatanaka, Takashi Mitamura, Momoko Kato, Yuka Shibata, Ichiro Yabe, Akira Endoh, Yoshito Komatsu, Yoshihiro Matsuno, Minako Sugiyama, Atsushi Manabe, Akihiro Sakurai, Masato Takahashi, Hirohito Naruse, Yoshihiro Torimoto, Hirotoshi Dosaka-Akita, Ichiro Kinoshita
    Japanese journal of clinical oncology 51 (5) 753 - 761 2021/04/30 [Refereed]
     
    BACKGROUND: Comprehensive cancer genomic profiling has been used recently for patients with advanced solid cancers. Two cancer genomic profiling tests for patients with no standard treatment are covered by Japanese public health insurance since June 2019. METHODS: We prospectively analyzed data of 189 patients with solid cancers who underwent either of the two-cancer genomic profiling tests at Hokkaido University Hospital and its liaison hospitals and whose results were discussed in molecular tumor board at Hokkaido University Hospital between August 2019 and July 2020. RESULTS: All 189 patients had appropriate results. Actionable gene alterations were identified in 93 patients (49%). Frequent mutations included PIK3CA (12%) mutation, BRCA1/2 alteration (7%), ERBB2 amplification (6%) and tumor mutation burden-High (4%). The median turnaround time from sample shipping to acquisition by the expert panel was 26 days. Although 115 patients (61%) were provided with information for genotype-matched therapies, only 21 (11%) received them. Notably, four of eight patients below the age of 20 years were provided information for genotype-matched therapies, and three received them. Their response rates and disease control rates were 29% and 67%, respectively. Most patients who did not undergo the genotype-matched therapies were provided information for only investigational drugs in phases I and II at distant clinical trial sites in central Japan. Twenty-six patients were informed of suspected germline findings, while 11 patients (42%) received genetic counseling. CONCLUSIONS: The publicly reimbursed cancer genomic profilings may lead to the modest but favorable therapeutic efficacy of genotype-matched therapy for solid cancer patients with no standard therapy. However, poor access to genotype-matched therapy needs to be resolved.
  • Kanako Hagio, Toraji Amano, Hideyuki Hayashi, Takashi Takeshita, Tomohiro Oshino, Junko Kikuchi, Yoshihito Ohhara, Ichiro Yabe, Ichiro Kinoshita, Hiroshi Nishihara, Hiroko Yamashita
    Scientific reports 11 (1) 8109 - 8109 2021/04/14 [Refereed]
     
    Clinical targeted sequencing allows for the selection of patients expected to have a better treatment response, and reveals mechanisms of resistance to molecular targeted therapies based on actionable gene mutations. We underwent comprehensive genomic testing with either our original in-house CLHURC system or with OncoPrime. Samples from 24 patients with estrogen receptor-positive, human epidermal growth factor receptor 2-negative metastatic breast cancer underwent targeted sequencing between 2016 and 2018. Germline and somatic gene alterations and patients' prognosis were retrospectively analyzed according to the response to endocrine therapy. All of the patients had one or more germline and/or somatic gene alterations. Four patients with primary or secondary endocrine-resistant breast cancer harbored germline pathogenic variants of BRCA1, BRCA2, or PTEN. Among somatic gene alterations, TP53, PIK3CA, AKT1, ESR1, and MYC were the most frequently mutated genes. TP53 gene mutation was more frequently observed in patients with primary endocrine resistance compared to those with secondary endocrine resistance or endocrine-responsive breast cancer. Recurrent breast cancer patients carrying TP53-mutant tumors had significantly worse overall survival compared to those with TP53-wild type tumors. Our 160-gene cancer panel will be useful to identify clinically actionable gene alterations in breast cancer in clinical practice.
  • Yoshitaka Saito, Yoh Takekuma, Masaki Kobayashi, Naofumi Shinagawa, Yasushi Shimizu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Ken Iseki, Mitsuru Sugawara
    European journal of clinical pharmacology 77 (3) 381 - 388 2021/03 [Refereed]
     
    PURPOSE: Gefitinib is one of the standard treatments for non-small cell lung cancer (NSCLC) with epidermal growth factor receptor mutations. It has been reported that acid suppressants (AS) decrease the anti-tumor effect of gefitinib by reducing its solubility. AS is sometimes necessary in cancer patients; however, previous reports have not shown the most compatible AS with gefitinib administration in cancer patients. This study was conducted to determine if histamine type 2 receptor antagonists (H2RAs) can affect the anti-tumor efficacy of gefitinib. METHODS: Eighty-seven patients with NSCLC who were administered gefitinib were retrospectively investigated. Patients who were co-administered H2RA were compared with non-AS control patients. H2RA was administered once a day at about 3-5 or 8-12 h after gefitinib intake. The primary endpoint of this study was progression-free survival (PFS), and secondary endpoints were overall survival (OS), overall response rate (ORR), and adverse effects. RESULTS: Median PFS in H2RA group and control group was 8.0 months and 9.0 months, respectively, with no significant difference (p = 0.82). The incidence of liver dysfunction was significantly less in patients administered H2RA, whereas there were no differences between the two groups with regard to skin toxicity and diarrhea. Multivariate analysis suggested that H2RA co-administration is not a risk factor for worse PFS and OS (hazard ratio of 0.95, 0.86; 95% confidence interval of 0.60-1.48, 0.52-1.43; p = 0.82 and 0.60, respectively). CONCLUSION: This study demonstrated that concomitant administration of H2RA with gefitinib does not affect the efficacy of gefitinib.
  • Yoichi Naito, Hiroyuki Aburatani, Toraji Amano, Eishi Baba, Toru Furukawa, Tetsu Hayashida, Eiso Hiyama, Sadakatsu Ikeda, Masashi Kanai, Motohiro Kato, Ichiro Kinoshita, Naomi Kiyota, Takashi Kohno, Shinji Kohsaka, Keigo Komine, Itaru Matsumura, Yuji Miura, Yoshiaki Nakamura, Atsushi Natsume, Kazuto Nishio, Katsutoshi Oda, Naoyuki Oda, Natsuko Okita, Kumiko Oseto, Kuniko Sunami, Hideaki Takahashi, Masayuki Takeda, Shimon Tashiro, Shinichi Toyooka, Hideki Ueno, Shinichi Yachida, Takayuki Yoshino, Katsuya Tsuchihara
    International journal of clinical oncology 26 (2) 233 - 283 2021/02 [Refereed]
     
    BACKGROUND: To promote precision oncology in clinical practice, the Japanese Society of Medical Oncology, the Japanese Society of Clinical Oncology, and the Japanese Cancer Association, jointly published "Clinical practice guidance for next-generation sequencing in cancer diagnosis and treatment" in 2017. Since new information on cancer genomic medicine has emerged since the 1st edition of the guidance was released, including reimbursement for NGS-based multiplex gene panel tests in 2019, the guidance revision was made. METHODS: A working group was organized with 33 researchers from cancer genomic medicine designated core hospitals and other academic institutions. For an impartial evaluation of the draft version, eight committee members from each society conducted an external evaluation. Public comments were also made on the draft. The finalized Japanese version was published on the websites of the three societies in March 2020. RESULTS: The revised edition consists of two parts: an explanation of the cancer genomic profiling test (General Discussion) and clinical questions (CQs) that are of concern in clinical practice. Particularly, patient selection should be based on the expectation that the patient's post-test general condition and organ function will be able to tolerate drug therapy, and the optimal timing of test should be considered in consideration of subsequent treatment plans, not limited to treatment lines. CONCLUSION: We expect that the revised version will be used by healthcare professionals and will also need to be continually reviewed in line with future developments in cancer genome medicine.
  • Yasushi Yatabe, Koichi Goto, Shingo Matsumoto, Yutaka Hatanaka, Naoko Arakane, Sadakatsu Ikeda, Akira Inoue, Ichiro Kinoshita, Hideharu Kimura, Tomohiro Sakamoto, Miyako Satouchi, Junichi Shimizu, Kuniko Sunami, Koji Tsuta, Shinichi Toyooka, Kazuto Nishio, Kazumi Nishino, Masashi Mikubo, Tomoyuki Yokose, Hirotoshi Dosaka-Akita
    Japanese Journal of Lung Cancer 61 (5) 361 - 370 0386-9628 2021 
    MET, a proto-oncogene located in 7q21-q31, encodes a receptor tyrosine kinase, of which mutations, amplification, fusions and overexpression are reported to be associated with oncogenesis. MET exon 14 (METex14) skipping is one of such MET alterations, and this abnormality is caused by genetic deletions or mutations in the intron/exon boundary sites as splice-site abnormalities, resulting in the generation of a deleted transcript in exon 14. This exon encodes juxtamembrane domain, which contains the binding site of c-Cbl E 3 ubiquitin ligase. Therefore, lack of METex14 suppresses ubiquitination and degradation, which lead to functional MET activation. In 2020, tepotinib and capmatinib were approved for the treatment of advanced recurrent lung cancer with this alteration. To implement the molecular testing to detect METex14 skipping in clinical practice, a practical guidance was released from the Biomarker Committee of the Japan Lung Cancer Society, and the content is introduced in this article.
  • Kuniko Sunami, Hideaki Bando, Yasushi Yatabe, Yoichi Naito, Hideaki Takahashi, Katsuya Tsuchihara, Shinichi Toyooka, Koshi Mimori, Shinji Kohsaka, Hiroyuki Uetake, Ichiro Kinoshita, Keigo Komine, Masayuki Takeda, Tetsu Hayashida, Kenji Tamura, Kazuto Nishio, Noboru Yamamoto, Hiroyuki Aburatani, Takashi Khono, Hiroyuki Mano, Tetsuo Noda, Daisuke Aoki, Yuko Kitagawa, Masaki Mori, Gaku Muto, Hirotoshi Akita, Chikashi Ishioka, Issei Imoto, Hidehiko Miyake, Tomosho Nakayama
    Cancer Science 112 (9) 3911 - 3917 1347-9032 2021 [Refereed]
     
    Comprehensive genomic profiling (CGP) is being increasingly used for the routine clinical management of solid cancers. In July 2018, the use of tumor tissue-based CGP assays became available for all solid cancers under the universal health insurance system in Japan. Several restrictions presently exist, such as patient eligibility and limitations on the opportunities to perform such assays. The clinical implementation of CGP based on plasma circulating tumor DNA (ctDNA) is also expected to raise issues regarding the selection and use of tissue DNA and ctDNA CGP. A Joint Task Force for the Promotion of Cancer Genome Medicine comprised of three Japanese cancer-related societies has formulated a policy proposal for the appropriate use of plasma CGP (in Japanese), available at https://www.jca.gr.jp/researcher/topics/2021/files/20210120.pdf, http://www.jsco.or.jp/jpn/user_data/upload/File/20210120.pdf, and https://www.jsmo.or.jp/file/dl/newsj/2765.pdf. Based on these recommendations, the working group has summarized the respective advantages and cautions regarding the use of tissue DNA CGP and ctDNA CGP with reference to the advice of a multidisciplinary expert panel, the preferred use of plasma specimens over tissue, and multiple ctDNA testing. These recommendations have been prepared to maximize the benefits of performing CGP assays and might be applicable in other countries and regions.
  • Yasushi Yatabe, Kuniko Sunami, Koichi Goto, Kazuto Nishio, Naoko Aragane, Sadakatsu Ikeda, Akira Inoue, Ichiro Kinoshita, Hideharu Kimura, Tomohiro Sakamoto, Miyako Satouchi, Junichi Shimizu, Koji Tsuta, Shinichi Toyooka, Kazumi Nishino, Yutaka Hatanaka, Shingo Matsumoto, Masashi Mikubo, Tomoyuki Yokose, Hirotoshi Dosaka-Akita
    Pathology international 70 (12) 921 - 931 2020/12 [Refereed]
     
    The year 2019 was considered to be the first year of cancer genome medicine in Japan, with three gene-panel tests using next-generation sequencing (NGS) techniques being introduced into clinical practice. Among the three tests, the Oncomine CDx Target test was approved under the category of regular molecular testing for lung cancer, which meant that this test could be used to select patients for molecularly targeted drugs. Conversely, the other two tests, NCC OncoPanel and FoundationOne CDx, were assigned to be used under the National Cancer Genome Medicine Network, and implementation was restricted to patients for whom standard treatment was completed or expected to be completed. These NGS tests can detect a series of genetic alterations in individual tumors, which further promotes the development of therapeutic agents and elucidates molecular pathways. The NGS tests require appropriate tissue size and tumor cell content, which can be accessed only by pathologists. In this report, we review the current reimbursement schema in our national healthcare policy and the requirements of the specimens for NGS testing based on the recently published 'Guidance of Gene-panel Testing Using Next-Generation Sequencers for Lung Cancer', by the Japanese Society of Lung Cancer.
  • Hideyuki Hayashi, Shigeki Tanishima, Kyoko Fujii, Ryo Mori, Chihiro Okada, Emmy Yanagita, Yuka Shibata, Ryosuke Matsuoka, Toraji Amano, Takahiro Yamada, Ichiro Yabe, Ichiro Kinoshita, Yoshito Komatsu, Hirotoshi Dosaka-Akita, Hiroshi Nishihara
    Cancer science 111 (10) 3926 - 3937 2020/10 [Refereed]
     
    Precision medicine is a promising strategy for cancer treatment. In this study, we developed an in-house clinical sequencing system to perform a comprehensive cancer genomic profiling test as a clinical examination and analyzed the utility of this system. Genomic DNA was extracted from tumor tissues and peripheral blood cells collected from 161 patients with different stages and types of cancer. A comprehensive targeted amplicon exome sequencing for 160 cancer-related genes was performed using next-generation sequencing (NGS). The sequencing data were analyzed using an original bioinformatics pipeline, and multiple cancer-specific gene alterations were identified. The success rate of our test was 99% (160/161), while re-biopsy was required for 24% (39/161) of the cases. Potentially actionable and actionable gene alterations were detected in 91% (145/160) and 46% (73/160) of the patients, respectively. The actionable gene alterations were frequently detected in PIK3CA (9%), ERBB2 (8%), and EGFR (4%). High tumor mutation burden (TMB) (≥10 mut/Mb) was observed in 12% (19/160) of the patients. The secondary findings in germline variants considered to be associated with hereditary tumors were detected in 9% (15/160) of the patients. Seventeen patients (11%, 17/160) were treated with genotype-matched therapeutic agents, and the response rate was 47% (8/17). The median turnaround time for physicians was 20 days, and the median survival time after the initial visit was 8.7 months. The results of the present study prove the feasibility of implementing in-house clinical sequencing as a promising laboratory examination technique for precision cancer medicine.
  • Jun Taguchi, Yasushi Shimizu, Shin Ariga, Tomohiro Goda, Yoshihito Ohhara, Rio Honma, Takuro Noguchi, Satoshi Takeuchi, Ichiro Kinoshita, Toraji Amano, Takatsugu Mizumachi, Satoshi Kano, Miki Takahara, Takahisa Abe, Akihiro Homma, Hirotoshi Dosaka-Akita
    INTERNATIONAL JOURNAL OF CLINICAL ONCOLOGY 26 (1) 51 - 58 1341-9625 2020/09 [Refereed]
     
    Background The standard of care for first-line treatment of recurrent and/or metastatic squamous cell carcinoma of the head and neck (R/M SCCHN) in patients who cannot tolerate platinum-based regimens has not been clarified. We aimed to develop a new treatment regimen for patients with R/M SCCHN who are ineligible for platinum-based therapy, by evaluating the effects and safety of tegafur/gimeracil/oteracil (S-1) and cetuximab. Methods Platinum-ineligibility was defined as: elderly (aged >= 75 years), poor PS, comorbidity, platinum resistance and refusal to undergo platinum-based therapy. Patients received S-1 (80 mg/m(2)/day for 14 days followed by a seven-day break) and cetuximab (initial dose, 400 mg/m(2), followed by 250 mg/m(2)weekly) until disease progression or unacceptable toxicity. The primary endpoint was overall response rate (ORR). Results Between September 2014 and September 2018, we enrolled 23 patients. Among the 21 patients who were evaluable, 20 were male [median age, 69 years (range 49-82)]. The ORR was 9 (43%) of 21 patients [95% confidence interval (CI) 22-66]. One and eight patients achieved complete response (CR) and partial response (PR), respectively. The median overall survival (OS) was 13.7 months (95% CI 9.0-18.3) and progression-free survival (PFS) was 5.7 months (95% CI 3.1-8.2). Grade 3/4 adverse events included acneiform rash and skin reactions (33%), hypomagnesemia (19%), hand-foot syndrome (14%), fatigue (14%), mucositis (10%), and anorexia (10%). Conclusions Combination treatment with S-1 and cetuximab was effective and tolerated well by patients with platinum-ineligible R/M SCCHN.
  • Shin-Ichi Fukumoto, Satoshi Oizumi, Masao Harada, Noriaki Sukoh, Kosuke Nakano, Satoshi Fuke, Jun Sakakibara-Konishi, Kei Takamura, Kenichiro Ito, Yuka Fujita, Yutaka Nishigaki, Toshiyuki Harada, Kenji Akie, Ichiro Kinoshita, Toraji Amano, Hiroshi Isobe, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    Cancer chemotherapy and pharmacology 86 (1) 117 - 127 2020/07 [Refereed]
     
    PURPOSE: This study evaluated the efficacy and safety of platinum plus gemcitabine (P/G) combinations as postoperative adjuvant chemotherapies for non-small cell lung cancer. METHODS: Patients with postoperative stage IB-IIIA non-small cell lung cancer were randomly assigned to receive either cisplatin plus gemcitabine (GP arm) or carboplatin plus gemcitabine (GC arm) every 3 weeks for four cycles. The primary endpoint was 2-year disease-free survival (DFS). Secondary endpoints were safety, feasibility, overall survival (OS), and biomarker analyses. RESULTS: A total of 102 patients were randomized (stage IB, 22%; II, 36%; IIIA, 42%; histology: 74% adenocarcinoma). Of the 51 patients in each arm, 37 (73%) completed 4 cycles. During follow-up (median 5.8 years; range 0.1-9.7 years), estimated DFS and OS rates at 2 years were 59.6% and 86.3% with GP and 68.0% and 86.3% with GC, respectively. No significant difference in DFS was noted between arms (P = 0.163), although 3-, 4-, and 5-year DFS rates were higher with GC. Hematological toxic effects were comparable and non-hematological toxic effects were infrequent. DFS was significantly higher in the excision repair cross-complementation group 1 (ERCC1)-low group than in the ERCC1-high group for the GP arm (P = 0.045). CONCLUSION: Both P/G combination regimens were feasible and well-tolerated, and thus may represent valid options for postoperative adjuvant treatment of non-small cell lung cancer. Although no significant differences in DFS were evident between regimens, the present data favor the adoption of GC for further evaluation. CLINICAL TRIAL REGISTRATION: UMIN-CTR ( https://www.umin.ac.jp/ctr/ ) identifier: UMIN000000913.
  • 吉岡 正博, 山田 崇弘, 柴田 有香, 近藤 知大, 林 秀幸, 西原 広史, 矢部 一郎, 秋田 弘俊, 武藤 学, 木下 一郎, 小杉 真司
    日本遺伝カウンセリング学会誌 日本遺伝カウンセリング学会 41 (2) 75 - 75 1347-9628 2020/06
  • Tetsuaki Shoji, Eiki Kikuchi, Junko Kikuchi, Yuta Takashima, Megumi Furuta, Hirofumi Takahashi, Kosuke Tsuji, Makie Maeda, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Jun Sakakibara-Konishi, Satoshi Konno
    Cancer chemotherapy and pharmacology 85 (5) 843 - 853 2020/03/30 [Refereed][Not invited]
     
    PURPOSE: We evaluated the expression of proteasome subunits to assess whether the proteasome could be a therapeutic target in cisplatin-resistant lung cancer cells. METHODS: Cisplatin-resistant (CR) variants were established from three non-small cell lung cancer (NSCLC) cell lines (A549, H1299, and H1975) and two small cell lung cancer (SCLC) cell lines (SBC3 and SBC5). The expression of proteasome subunits, the sensitivity to immunoproteasome inhibitors, and 20S proteasomal proteolytic activity were examined in the CR variants of the lung cancer cell lines. RESULTS: All five CR cell lines highly expressed one or both of the immunoproteasome subunit genes, PSMB8 and PSMB9, while no clear trend was observed in the expression of constitutive proteasome subunits. The CR cells expressed significantly higher levels of PSMB8 and PSMB9 proteins, as well. The CR variants of the H1299 and SBC3 cell lines were more sensitive to immunoproteasome inhibitors, and had significantly more proteasomal proteolytic activity than their parental counterparts. CONCLUSIONS: The immunoproteasome may be an effective therapeutic target in a subset of CR lung cancers. Proteasomal proteolytic activity may be a predictive marker for the efficacy of immunoproteasome inhibitors in cisplatin-resistant SCLC and NSCLC.
  • Yasuyo Adachi, Norikazu Iwata, Yasushi Adachi, Hiroko Nakamura, Takefumi Kikuchi, Masahiro Nakamura, Hiroaki Mita, Yukinari Yoshida, Ichiro Kinoshita, Yoshifumi Ishii, Takao Endo
    Nihon Shokakibyo Gakkai zasshi = The Japanese journal of gastro-enterology 117 (6) 532 - 541 2020 
    IgG4-related autoimmune hepatitis (IgG4-AIH) is characterized by hepatic inflammation and is considered an IgG4-related disease. Several inflammatory pseudotumors (IPTs) are also considered as IgG4-related diseases;however, there have been no reports of cases wherein both diseases occurred concurrently. An older adult with liver dysfunction was admitted to the hospital and was diagnosed with IgG4-AIH following a liver biopsy;IgG4-positive plasma cell infiltration in the portal tract and high serum IgG4 concentration were detected. A few months following biopsy, imaging studies revealed two IPTs in the liver. The patient was diagnosed with cryptogenic organized pneumonia several months after imaging and was treated with steroids in a different hospital. Her liver dysfunction improved, and one of the two IPTs disappeared in response to steroid treatment. The following is an account of a rare case of IgG4-AIH with IPTs of the liver.
  • Takashima Y, Kikuchi E, Kikuchi J, Suzuki M, Kikuchi H, Maeda M, Shoji T, Furuta M, Kinoshita I, Dosaka-Akita H, Sakakibara-Konishi J, Konno S
    International journal of cancer 146 (4) 1114 - 1124 0020-7136 2019/06 [Refereed][Not invited]
     
    Bromodomain and extraterminal domain (BET) inhibitors are broadly active against distinct types of cancer, including nonsmall cell lung cancer (NSCLC). Previous studies have addressed the effect of BET-inhibiting drugs on the expression of oncogenes such as c-Myc, but DNA damage repair pathways have also been reported to be involved in the efficacy of these drugs. AZD1775, an inhibitor of the G2-M cell cycle checkpoint kinase WEE1, induces DNA damage by promoting premature mitotic entry. Thus, we hypothesized that BET inhibition would increase AZD1775-induced cytotoxicity by impairing DNA damage repair. Here, we demonstrate that combined inhibition of BET and WEE1 synergistically suppresses NSCLC growth both in vitro and in vivo. Two BET inhibitors, JQ1 and AZD5153, increased and prolonged AZD1775-induced DNA double-strand breaks (DSBs) and concomitantly repressed genes related to nonhomologous end joining (NHEJ), including XRCC4 and SHLD1. Furthermore, pharmaceutical inhibition of BET or knockdown of the BET protein BRD4 markedly diminished NHEJ activity, and the BET-inhibitor treatment also repressed myelin transcription factor 1 (MYT1) expression and promoted mitotic entry with subsequent mitotic catastrophe when combined with WEE1 inhibition. Our findings reveal that BET proteins, predominantly BRD4, play an essential role in DSB repair through the NHEJ pathway, and further suggest that combined inhibition of BET and WEE1 could serve as a novel therapeutic strategy for NSCLC.
  • Saito Y, Kobayashi M, Yamada T, Sakakibara-Konishi J, Shinagawa N, Kinoshita I, Dosaka-Akita H, Iseki K
    Supportive care in cancer : official journal of the Multinational Association of Supportive Care in Cancer 28 (1) 221 - 227 0941-4355 2019/04 [Refereed][Not invited]
     
    PURPOSE: Paclitaxel-associated acute pain syndrome (P-APS) affects 80% of patients undergoing therapy. Although it has been shown that prednisone administration for 5 days relieves P-APS, detailed results have not been reported thus far. Therefore, in this study, we evaluated the preventive effect of dexamethasone (DEX) administration against P-APS. METHODS: A total of 60 patients who received carboplatin (area under the curve; AUC = 5-6) plus paclitaxel (200 mg/m2) (plus bevacizumab 15 mg/kg, if non-squamous carcinoma of lung) were enrolled. Eight milligrams of DEX was orally administered on days 2 and 3 to the DEX group patients, and the frequency, severity, duration of P-APS, and other adverse effects in the first cycle were retrospectively evaluated and compared to those observed in control group patients, who were not administered DEX on days 2 and 3. RESULTS: No difference in terms of patient characteristics, except for type of cancer, was observed between groups. The results showed that the frequency of all grade P-APS was approximately 70% and there was no difference between groups. Frequency of ≥ G2 P-APS was 40% in the control group and 14% in the DEX group, demonstrating a significant reduction. Duration of P-APS was 5.8 days in the control group and 4.3 days in the DEX group, which tended to become shorter following additional DEX administration, although this was not significant. Adverse effects other than P-APS induced by chemotherapy were similar between the two groups. CONCLUSION: Additional DEX administration is safe and useful for the attenuation of the severity of P-APS.
  • Furuta M, Kikuchi H, Shoji T, Takashima Y, Kikuchi E, Kikuchi J, Kinoshita I, Dosaka-Akita H, Sakakibara-Konishi J
    Cancer science 110 (5) 1599 - 1608 1347-9032 2019/03 [Refereed][Not invited]
     
    Delta-like protein 3 (DLL3) is a ligand of Notch signaling, which mediates cell-fate decisions and is tumor-suppressive or oncogenic depending on the cellular context. Previous studies show that DLL3 is highly expressed in small cell lung cancer (SCLC) but not in normal lung tissue, suggesting that DLL3 might be associated with neuroendocrine tumorigenesis. However, its role in SCLC remains unclear. To investigate the role of DLL3 in tumorigenesis in SCLC, we performed loss-of-function and gain-of-function assays using SCLC cell lines. In vitro analysis of cell migration and invasion by transwell assay showed that DLL3 knockdown reduced migration and invasion of SCLC cells, whereas DLL3 overexpression increased these activities. In addition, DLL3 positively regulated SNAI1 expression and knockdown of SNAI1 attenuated the migration and invasion ability of SCLC cells. Moreover, upregulated DLL3 expression induced subcutaneous tumor growth in mouse models. These results indicate that DLL3 promoted tumor growth, migration and invasion in an SCLC model by modulating SNAI1/Snail.
  • Saito Y, Yamada T, Kobayashi M, Sakakibara-Konishi J, Shinagawa N, Kinoshita I, Dosaka-Akita H, Iseki K
    Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan 139 (12) 1601 - 1608 0031-6903 2019 [Refereed][Not invited]
     
    Paclitaxel (PTX)-associated acute pain syndrome (P-APS) is characterized by disabling but transient arthralgia and myalgia in up to 80% of patients administered with PTX. Non-steroidal anti-inflammatory drugs (NSAIDs) are widely administered to patients with cancer who have pain or fever, and are mainly used to manage P-APS. In this study, we investigated how P-APS appear in the patients who were administered NSAIDs prior to PTX injection. The incidence or severity and duration of P-APS in patients previously administered NSAIDs were compared to those of patients who were not administered NSAIDs. The relationship between previously administered NSAIDs and rescue administration for the relief of P-APS was also evaluated. It was revealed that the incidence and duration of P-APS were 72% and 4.67±2.30 d, respectively, in the control group and 84% and 6.19±3.30 d, respectively, in the NSAIDs group. There was no significant difference in the incidence and duration and the severity of P-APS between the two groups. Patients who were previously administered NSAIDs tended to obtain less pain relief from NSAIDs administered as rescue medications, and needed other medication. Univariate and multivariate analysis revealed no correlation between previously administered NSAIDs or patient characteristics and the incidence of P-APS. In this study, it was found that clinical condition that needs NSAIDs and previously administered NSAIDs prior to PTX injection do not affect the incidence, severity, and duration of P-APS. These results will help in educating patients about their medications and will contribute to the management of P-APS.
  • 高島 雄太, 榊原 純, 畑中 豊, 畑中 佳奈子, 大原 克仁, 大泉 聡史, 樋田 泰浩, 加賀 基知三, 木下 一郎, 秋田 弘俊, 松野 吉宏, 品川 尚文
    肺癌 (NPO)日本肺癌学会 58 (6) 633 - 633 0386-9628 2018/10 [Refereed][Not invited]
  • Takeuchi S, Goda T, Taguchi J, Douhata Y, Honma R, Ariga S, Ohhara Y, Shimizu Y, Kinoshita I, Fukuda I, Nagashima Y, Akita H
    Internal medicine (Tokyo, Japan) 57 (16) 2431 - 2436 0918-2918 2018/08 [Refereed][Not invited]
  • Takeuchi S, Shiga T, Hirata K, Taguchi J, Magota K, Ariga S, Gouda T, Ohhara Y, Homma R, Shimizu Y, Kinoshita I, Tsuji Y, Homma A, Iijima H, Tamaki N, Dosaka-Akita H
    BMJ open 8 (8) e021001  2018/08 [Refereed][Not invited]
     
    INTRODUCTION: Lenvatinib, an oral molecular targeted drug, is used to treat patients with unresectable or advanced thyroid carcinoma that is refractory to radioiodine treatment. Effective methods for evaluating molecular targeted drugs are a critical unmet need owing to their expensive costs and unique adverse events. The aim of this study is to determine whether 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET)/CT at 1 week after commencing lenvatinib can predict treatment outcomes. DESIGN AND METHODS: This study is planned as a non-randomised single-arm multicentre study; patients with pathologically confirmed differentiated thyroid carcinoma (DTC) with lesions that are refractory to radioiodine treatment are eligible. The main exclusion criteria are medullary or anaplastic carcinoma, prior treatment with chemotherapy, poor general condition and thromboembolism-requiring treatment. Patients to be included in the study will be treated with lenvatinib and undergo FDG-PET/CT examination twice: before and 1 week after the initiation of treatment. Contrast-enhanced CT, the gold standard for evaluation, will be performed at least 4 weeks after the initiation of treatment. The primary objective is to evaluate the ability of the lesion maximum standard uptake value for FDG PET/CT performed 1 week after the initiation of treatment to predict outcomes compared with the response evaluation obtained via contrast-enhanced CT performed at least 4 weeks after the initiation of treatment. ETHICS AND DISSEMINATION: This study is conducted in accordance with the Declaration of Helsinki and has received ethical approval from the institutional review board of the Hokkaido University Hospital (approval number: 015-402). The results of this study will be disseminated through a presentation at a conference and the publication of the data in a peer-reviewed journal. The study will be implemented and reported in line with the SPIRIT statement. TRIAL REGISTRATION NUMBER: UMIN000022592.
  • Inoue T, Katoh N, Ito YM, Kimura T, Nagata Y, Kuriyama K, Onishi H, Yoshitake T, Shioyama Y, Iizuka Y, Inaba K, Konishi K, Kokubo M, Karasawa K, Kozuka T, Tanaka K, Sakakibara-Konishi J, Kinoshita I, Shirato H
    Lung cancer (Amsterdam, Netherlands) 122 107 - 112 0169-5002 2018/08 [Refereed][Not invited]
     
    OBJECTIVES: Even with advanced image guidance, biopsies occasionally fail to diagnose small lung lesions, which are highly suggestive of primary lung cancer by radiological examination. The aim of this study was to evaluate the outcome of stereotactic body radiotherapy (SBRT) to treat small lung lesions clinically diagnosed as primary lung cancer. MATERIALS AND METHODS: This is a prospective, multi-institutional observation study. Strict inclusion and exclusion criteria were determined in a nation-wide consensus meeting and used to include patients who were clinically diagnosed with primary lung cancer using precise imaging modalities, for whom further surgical intervention was not feasible, who refused watchful waiting, and who were highly tolerable of SBRT with informed consent. SBRT was performed with 48 Gy in 4 fractions at the tumor isocenter. RESULTS: From August 2009 to August 2014, 62 patients from 11 institutions were enrolled. Their median age was 80 years. The tumors ranged in size from 9 to 30 mm in diameter (median, 18 mm). The median follow-up interval was 55 months. The 3-year overall survival rate was 83.3% (95% confidence interval (CI) 71.1-90.7%) for all the patients and 94.7% (95% CI 68.1-99.2%) for the patients younger than 75 years. Local failure, regional lymph node metastases and distant metastases occurred in 4 (6.4%), 3 (4.8%) and 11 (17.7%) patients, respectively. Grades 3 and 4 toxicities were observed in 8 (12.9%) patients and 1 (1.6%) patient, respectively. No grade 5 toxicities were observed. CONCLUSIONS: SBRT is safe and effective for patients with small lung lesions clinically diagnosed as primary lung cancer that satisfied the proposed strict indication criteria as previously reported. A prospective interventional study is required to ascertain if SBRT is an alternative strategy for these patients.
  • Hayashi H, Tanishima S, Fujii K, Mori R, Okamura Y, Yanagita E, Matsuoka R, Amano T, Kinoshita I, Komatsu Y, Dosaka-Akita H, Nishihara H
    Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.] 18 (6) 647 - 654 1424-3903 2018/07 [Refereed][Not invited]
     
    BACKGROUND: Precision medicine guided by comprehensive genome sequencing represents a potential treatment strategy for pancreatic cancer. However, clinical sequencing for pancreatic cancer entails several practical difficulties. We have launched an in-house clinical sequencing system and started genomic testing for patients with cancer in clinical practice. We have analyzed the clinical utility of this system in pancreatic cancer. METHODS: We retrospectively reviewed 20 patients with pancreatic cancer who visited our division. Genomic DNA was extracted from both tumor tissue and peripheral blood mononuclear cells obtained from the patients. We performed a comprehensive genomic testing using targeted amplicon sequencing for 160 cancer-related genes. The primary endpoints were the detection rates of potential actionable and druggable gene alterations. The secondary endpoints were the detection rate of secondary germline findings, the rate of re-biopsy required for genome sequencing, survival time after the initial visit (post-sequencing survival time), and turnaround time. RESULTS: Although re-biopsy was required for 25% (5/20) of all patients, genomic testing was performed in all patients. Actionable and druggable gene alterations were detected in 100% (20/20) and 35% (7/20) of patients, respectively, whereas secondary germline findings were detected in 5% (1/20) of patients. The median turnaround times for physicians and patients were 20 and 26 days, respectively. The median post-sequencing survival time was 10.3 months. Only 10% (2/20) of all patients were treated with therapeutic agents based on the outcomes of genomic testing. CONCLUSIONS: The clinical application of comprehensive genomic testing for pancreatic cancer was feasible and promising in clinical practice.
  • Takashima Y, Sakakibara-Konishi J, Hatanaka Y, Hatanaka KC, Ohhara Y, Oizumi S, Hida Y, Kaga K, Kinoshita I, Dosaka-Akita H, Matsuno Y, Nishimura M
    Clinical lung cancer 19 (4) 352 - 359.e1 1525-7304 2018/07 [Refereed][Not invited]
  • Satoshi Oizumi, Shunichi Sugawara, Koichi Minato, Toshiyuki Harada, Akira Inoue, Yuka Fujita, Makoto Maemondo, Satoshi Watanabe, Kazuhiko Ito, Akihiko Gemma, Yoshiki Demura, Shinichi Fukumoto, Hiroshi Isobe, Ichiro Kinoshita, Satoshi Morita, Kunihiko Kobayashi, Koichi Hagiwara, Keisuke Aiba, Toshihiro Nukiwa
    ESMO open 3 (2) e000313  2018 
    Background: The North-East Japan Study Group (NEJ) 005/Tokyo Cooperative Oncology Group (TCOG) 0902 study has reported that first-line concurrent and sequential alternating combination therapies of an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (gefitinib) plus platinum-based doublet chemotherapy (carboplatin/pemetrexed) offer promising efficacy with predictable toxicities for patients with EGFR-mutant non-small cell lung cancer. However, overall survival (OS) data were insufficient in the primary report because of the lack of death events. Patients and methods: Progression-free survival (PFS) and OS were re-evaluated at the final data cut-off point (March 2017) for the entire population (n=80). Results: At the median follow-up time of 35.6 months, 88.8% of patients had progressive disease and 77.5% of patients had died. Median PFS was 17.5 months for the concurrent regimen and 15.3 months for the sequential alternating regimen (P=0.13). Median OS was 41.9 and 30.7 months, respectively (P=0.036). Updated response rates were similar in both groups (90.2% and 82.1%, respectively; P=0.34). Patients with Del19 tumours displayed relatively better OS (median: 45.3 vs 33.3 months, respectively) than those with L858R (31.4 vs 28.9 months, respectively). No severe adverse events, including interstitial lung disease, occurred in the period since the primary report. Conclusions: This updated analysis confirms that PFS is improved with first-line combination therapy compared with gefitinib monotherapy and that the concurrent regimen, in particular, offers an OS benefit of 42 months in the EGFR-mutated setting. Our ongoing NEJ009 study will clarify whether this combination strategy can be incorporated into routine clinical practice. Trial registration number: UMIN C000002789, Post-results.
  • A phase II study of carboplatin, pemetrexed, and bevacizumab followed by erlotinib and bevacizumab maintenance for nonsquamous non-small cell lung cancer with wild-type EGFR (HOT1101)
    Taichi Takashina, Hajime Asahina, Satoshi Oizumi, Noriyuki Yamada, Masao Harada, Kei Takamura, Hiroshi Yokouchi, Toshiyuki Harada, Osamu Honjo, Takahiro Ogi, Naoto Morikawa, Ichiro Kinoshita, Ryoichi Honda, Kosuke Nakano, Kenya Kanazawa, Toraji Amano, Hirotoshi Dosaka‐Akita, Hiroshi Isobe, Masaharu Nishimura on behalf of, Hokkaido Lung, Cancer Clinical, Study Group
    Int J Clin Oncol. 2018 [Refereed][Not invited]
  • Numb has distinct function in lung adenocarcinoma and squamous cell carcinoma.
    Kikuchi H, Sakakibara-Konishi J, Furuta M, Kikuchi E, Kikuchi J, Oizumi S, Hida Y, Kaga K, Kinoshita I, Dosaka-Akita H, Nishimura M
    Oncotarget. 9 29379 - 29391 2018 [Refereed][Not invited]
  • EGFR遺伝子変異とEGFRチロシンキナーゼ阻害薬(EGFR-TKI)に対する反応が異なる肺3重癌の1例
    合田 智宏, 木下 一郎, 堂畑 雄一, 有賀 伸, 田口 純, 本間 理央, 竹内 啓, 清水 康, 秋田 弘俊, 品川 尚文, 樋田 泰浩, 加賀 基知三
    肺癌 (NPO)日本肺癌学会 57 (7) 915 - 915 0386-9628 2017/12 [Refereed][Not invited]
  • Yasuyuki Ikezawa, Hajime Asahina, Satoshi Oizumi, Masahiro Watanabe, Kei Takamura, Yasutaka Kawai, Noriyuki Yamada, Toshiyuki Harada, Ichiro Kinoshita, Yuka Fujita, Eisaku Miyauchi, Takahiro Ogi, Toraji Amano, Megumi Furuta, Jun Sakakibara-Konishi, Hiroshi Nishihara, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Masaharu Nishimura
    CANCER CHEMOTHERAPY AND PHARMACOLOGY 80 (5) 955 - 963 0344-5704 2017/11 [Refereed][Not invited]
     
    A high proportion of patients with wild-type EGFR non-small cell lung cancer (NSCLC) receive third-line therapy and beyond, with no prospective randomized trials addressing the issue. This study aimed to select the most suitable regimen as a third- or fourth-line therapy for wild-type EGFR NSCLC. This multicenter, randomized phase II study in Japan included patients with recurrent or advanced NSCLC with wild-type or unknown EGFR, who progressed after two or three previous chemotherapies. The patients were randomly assigned to erlotinib (150 mg/day, days 1-21) or S-1 (80-120 mg/day, days 1-14) every 3 weeks until disease progression or unacceptable toxicity. The primary endpoint was disease control rate (DCR). The secondary endpoints included overall survival (OS), progression-free survival (PFS), objective response rate (ORR), toxicity, and quality of life (QOL). From 2011 to 2016, 37 patients were randomly assigned to receive erlotinib (E arm, n = 19) and S-1 (S arm, n = 18). This study was terminated prematurely because of poor patient accrual. DCR/ORR were 42.1%/15.8% in the E arm and 66.7%/16.7% in the S arm. Median PFS/OS were 1.6 months/8.0 months in the E arm and 3.3 months/12.2 months in the S arm. In both groups, the most commonly reported grade 3-4 toxicities were fatigue, anorexia, and nausea. One grade 5 pneumonitis occurred in the S arm. No significant difference was seen in QOL. S-1 as a third- or fourth-line therapy for wild-type EGFR NSCLC showed numerically better clinical outcomes than erlotinib. UMIN000005308.
  • K. Shiba, Y. Fujita, H. Miyazawa, K. Muramatsu, M. Watanabe, M. Nishimura, S. Shinkuma, T. Nomura, W. Nishie, J. Taguchi, I. Kinoshita, H. Shimizu
    JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 31 (7) E324 - E326 0926-9959 2017/07 [Refereed][Not invited]
  • Masahisa Jinushi, Tomoko Morita, Zhihang Xu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Hideo Yagita, Yutaka Kawakami
    PLOS ONE 12 (7) e0179357  1932-6203 2017/07 [Refereed][Not invited]
     
    Autophagy is a vital process controlling the lysosomal degradation of cellular organelles and thereby regulating tissue homeostasis in an environment-dependent fashion. Recent studies have unveiled the critical role of tumor cell-derived autophagy in regulating pro-tumor and anti-tumor processes depending on different stages and tumor microenvironments. However, the precise mechanism whereby autophagy regulates tumor progression remains largely unclear. Since myeloid cells contribute to tumor progression and metastasis, we evaluated the role of myeloid cell-specific autophagy in the regulation of tumor progression. We found that the number and size of metastatic lesions were smaller in myeloid cell-specific autophagy-deficient mice. Furthermore, autophagy-mediated regulation of TGF-beta in myeloid cells was associated with the induction of epithelial-mesenchymal transition (EMT), which increases the invasive and metastatic potentials of tumor cells. Myeloid-derived autophagy also plays a critical role in impairing antitumor immune responses and promoting the survival and accumulation of M2 macrophages in tumor tissues in a CSF-1 and TGF-beta-dependent manner. Taken together, our findings elucidate previously unrecognized mechanisms by which myeloid cells promote tumor progression through autophagy-mediated regulation of malignancy and immune tolerance.
  • Jun Sakakibara-Konishi, Yasuyuki Ikezawa, Satoshi Oizumi, Junko Kikuchi, Eiki Kikuchi, Hidenori Mizugaki, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    INTERNATIONAL JOURNAL OF CLINICAL ONCOLOGY 22 (2) 257 - 268 1341-9625 2017/04 [Refereed][Not invited]
     
    Inhibition of Notch by gamma-secretase inhibitor (GSI) has been shown to have an antitumor effect in Notch-expressing non-small cell lung cancer (NSCLC) and to induce apoptosis through modulation of Bcl-2 family proteins. In particular, Bim, a BH3-only member of the Bcl-2 family of proteins, has an important role in the induction of apoptosis in NSCLC when cells are treated with GSI. ABT-737, a BH3-only mimetic, targets the pro-survival Bcl-2 family and also induces apoptosis. The Notch-expressing NSCLC cell lines H460, A549, H1793, and HCC2429 were used. The combined antitumor effect of GSI and ABT-737 was evaluated using the MTT proliferation assay in vitro and in xenograft mouse models. The expression of the Notch pathway and Bcl-2 family was analyzed using Western blotting analysis when cells were treated with a single drug treatment or a combination treatment. GSI XX or ABT-737 alone inhibited cell proliferation in a dose-dependent manner, and combination drug treatment showed a synergistic antitumor effect in vitro. In vivo, this drug combination significantly suppressed tumor proliferation compared to the single drug treatment. Phospho-Bcl-2 was downregulated and Bax was upregulated by both the single and combination drug treatments. Bim was induced by a single drug treatment and was enhanced by combination treatment. Combination treatment-induced apoptosis was decreased by Bim inhibition, suggesting that the antitumor effect of the drug combination was dependent on Bim. Based on our data, we propose that the combination treatment is a promising strategy for NSCLC therapy.
  • Yoshitaka Saito, Masaki Kobayashi, Takehiro Yamada, Kumiko Kasashi, Rio Honma, Satoshi Takeuchi, Yasushi Shimizu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Ken Iseki
    SUPPORTIVE CARE IN CANCER 25 (2) 481 - 487 0941-4355 2017/02 [Refereed][Not invited]
     
    Magnesium supplementation is an effective protective method against cisplatin-induced nephrotoxicity (CIN); however, there are few reports regarding the mechanism of its nephroprotective effect. The aim of this study was to determine whether premedication with intravenous magnesium prevents CIN and to determine the relationship between its nephroprotective effect and serum magnesium level.Fifty-eight patients with head and neck cancer who received cisplatin, docetaxel, and 5-fluorouracil (DCF) were retrospectively investigated. Grade 2 or more serum creatinine elevation was defined as CIN. The incidence of CIN was compared between a magnesium sulfate (20 mEq, 2.46 g) premedication group and a non-magnesium group during the first cycle and in all cycles.CIN did not occur in any patients receiving magnesium premedication but did occur in 5 of 29 patients during the first cycle and in 6 patients during all subsequent cycles in patients who did not receive magnesium premedication. Furthermore, the variation of creatinine clearance was significantly worse in the non-magnesium group than in the magnesium premedication group from baseline. There was no difference in adverse effects or response rate between the two groups. Univariate analysis suggested that magnesium premedication significantly reduced the risk of CIN. On the other hand, serum magnesium depletion was seen in both groups to equal degrees despite supplementation.Intravenous magnesium premedication has a protective effect on cisplatin-induced nephrotoxicity without the influence on the serum magnesium level. Magnesium premedication is a simple nephroprotective method that does not influence other adverse effects or rate of response to chemotherapy.
  • Intravenous magnesium premedication has prophylactic efficacy against cisplatin-induced nephrotoxicity without an influence on serum magnesium level.
    Saito Y, Kobayashi M, Yamada T, Kasashi K, Honma R, Takeuchi S, Shimizu Y, Kinoshita I, Dosaka-Akita H, Iseki K
    Support Care Cancer. 25 481 - 487 2017 [Refereed][Not invited]
  • Kazuhiro Usui, Shunichi Sugawara, Masaru Nishitsuji, Yuka Fujita, Akira Inoue, Atsuto Mouri, Hiroshi Watanabe, Hiroshi Sakai, Ichiro Kinoshita, Yoshihito Ohhara, Makoto Maemondo, Hiroshi Kagamu, Koichi Hagiwara, Kunihiko Kobayashi
    LUNG CANCER 99 131 - 136 0169-5002 2016/09 [Refereed][Not invited]
     
    Background: Vascular endothelial growth factor (VEGF) plays a pivotal role in the pathogenesis of malignant pleural effusion (MPE). Here, a multicenter phase II trial to evaluate bevacizumab in non-squamous non-small cell lung carcinoma patients with MPE was conducted. Methods: Patients having MPE with no prior treatment and performance status of 0-2 received carboplatin (area under the curve: AUC 6; up to 6 cycles) and pemetrexed (500 mg/m(2)) with bevacizumab (15 mg/kg) every 3 weeks. The primary endpoint was the control rate of MPE without pleurodesis at 8 weeks after treatment. VEGF levels in plasma and MPE were measured by enzyme immunoassay. Results: Of 30 patients entered (median 66 years; 24 males; adenocarcinoma; 4 epidermal growth factor receptor: EGFR mutations), 28 patients (2 withdrawn patients) were given a median of 4 cycles of carboplatin, and 68% of the patients received maintenance pemetrexed with bevacizumab (median 8 cycles). At eight weeks, MPE was controlled without pleurodesis in 93% of treated patients (95% confidence interval: 77-99%). At the median follow-up time of 12.8 months, 78.6% of the cases required no pleurodesis. Response rate was 46%, and median progression-free survival (PFS) and overall survival (OS) were 8.2 months and 18.6 months, respectively. Toxicities of grade >= 3 included neutropenia (28.6%), thrombocytopenia (28.6%), proteinuria (3.6%), and hypertension (3.6%). Assessment of VEGF levels before treatment indicated that patients with low VEGF (<1000 pg/ml) in MPE frequently needed pleurodesis (p = 0.011), and that high VEGF (>= 100 pg/ml) in plasma was indicative of poor prognosis in the context of PFS (p = 0.012). Conclusion: The combination of bevacizumab with carboplatin and pemetrexed demonstrated efficacy with acceptable toxicities in patients with MPE. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
  • Yoshihito Ohhara, Naoki Fukuda, Satoshi Takeuchi, Rio Honma, Yasushi Shimizu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita
    WORLD JOURNAL OF GASTROINTESTINAL ONCOLOGY 8 (9) 642 - 655 1948-5204 2016/09 [Refereed][Not invited]
     
    Colorectal cancer (CRC) is a significant cause of cancer-related morbidity and mortality all over the world. Improvements of cytotoxic and biologic agents have prolonged the survival in metastatic CRC (mCRC), with a median overall survival of approximately 2 years and more in the past two decades. The biologic agents that have proven clinical benefits in mCRC mainly target vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR). In particular, bevacizumab targeting VEGF and cetuximab and panitumumab targeting EGFR have demonstrated significant survival benefits in combination with cytotoxic chemotherapy in the first-line, second-line, or salvage setting. Aflibercept, ramucirumab, and regorafenib are also used in second-line or salvage therapy. Recent retrospective analyses have shown that KRAS or NRAS mutations were negative predictive markers for anti-EGFR therapy. Based on the evidence from large randomized clinical trials, personalized therapy is necessary for patients with mCRC according to their tumor biology and characteristics. The aim of this paper was to summarize the results of the major randomized clinical trials and highlight the benefits of the molecular targeted agents in patients with mCRC.
  • Miyauchi Eisaku, Usui Kazuhiro, Sugawara Shunichi, Nishitsuji Masaru, Fujita Yuka, Mouri Atsuto, Watanabe Hiroshi, Sakai Hiroshi, Kinoshita Ichiro, Hagiwara Koichi
    ANNALS OF ONCOLOGY 27 0923-7534 2016/07/01 [Refereed][Not invited]
  • Taichi Takashina, Ichiro Kinoshita, Junko Kikuchi, Yasushi Shimizu, Jun Sakakibara-Konishi, Satoshi Oizumi, Masaharu Nishimura, Hirotoshi Dosaka-Akita
    CANCER SCIENCE 107 (7) 955 - 962 1347-9032 2016/07 [Refereed][Not invited]
     
    Recent discoveries have revealed that human cancer involves aberrant epigenetic alterations. We and others have previously shown that the histone methyltransferase EZH2, the catalytic subunit of polycomb repressive complex 2 (PRC2), is frequently overexpressed in non-small-cell lung cancer (NSCLC) and that an EZH2 inhibitor, 3-deazaneplanocin A, inhibits the proliferation of NSCLC cells. Transcriptional silencing by EZH2 was recently shown to be required for the activity of histone deacetylases (HDACs) that interact with another PRC2 protein, EED. To develop a more effective epigenetic therapy for NSCLC, we determined the effects of co-treatment with 3-deazaneplanocin A and the HDAC inhibitor vorinostat (SAHA) in NSCLC cells. The co-treatment synergistically suppressed the proliferation of all tested NSCLC cell lines, regardless of their epidermal growth factor receptor (EGFR) status. The synergistic effect was associated with slightly decreased histone H3 lysine 27 trimethylation, modestly increased histone acetylation, and the depletion of EZH2 and other PRC2 proteins. The co-treatment resulted in an accumulation of p27Kip1, decrease in cyclin A, and increased apoptotic fraction in an additive/synergistic manner. Interestingly, the co-treatment strongly suppressed EGFR signaling, not only in EGFR-wild-type NSCLC cells, but also in EGFR-mutant cells, mainly through dephosphorylation of EGFR. Furthermore, the co-treatment suppressed the in vivo tumor growth of EGFR-mutant, EGFR-tyrosine kinase-resistant H1975 cells more effectively than did each agent alone, without visible toxicity. These results suggest that the combined pharmacological targeting of EZH2 and HDACs may provide more effective epigenetic therapeutics for NSCLC.
  • 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.
  • Yoshihiro Komohara, Tomoko Morita, Dorcas A. Annan, Hasita Horlad, Koji Ohnishi, Sohsuke Yamada, Toshiyuki Nakayama, Shohei Kitada, Shinya Suzu, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Koichi Akashi, Motohiro Takeya, Masahisa Jinushi
    CANCER IMMUNOLOGY RESEARCH 3 (9) 999 - 1007 2326-6066 2015/09 [Refereed][Not invited]
     
    Clear cell renal cell carcinoma (ccRCC) is one of most common cancers in urogenital organs. Although recent experimental and clinical studies have shown the immunogenic properties of ccRCC as illustrated by the clinical sensitivities to various immunotherapies, the detailed immunoregulatory machineries governing the tumorigenicity of human ccRCC remain largely obscure. In this study, we demonstrated the clinical significance and functional relevance of T-cell immunoglobulin and mucin domain-containing molecule-3 (TIM-3) expressed on tumor cells and myeloid cells in patients with ccRCC. TIM-3 expression was detected on cancer cells and CD204(+) tumor-associated macrophages (TAM), and higher expression level of TIM-3 was positively correlated with shorter progression-free survival (PFS) in patients with ccRCC. We found that TIM-3 expression was detected on a large number of tumors, and there was significant correlation between an increased number of TAMs and high expression level of TIM-3 in patients with ccRCC. Furthermore, TIM-3 rendered RCC cells with the ability to induce resistance to sunitinib and mTOR inhibitors, the standard regimen for patients with ccRCC, as well as stem cell activities. TIM-3 expression was induced on CD14(+) monocytes upon long-term stimulation with RCC cells, and TIM-3-expressing myeloid cells play a critical role in augmenting tumorigenic activities of TIM-3-negative RCC cells. More importantly, treatment with anti-TIM-3 mAb suppressed its tumorigenic effects in in vitro and in vivo settings. These findings indicate the coordinated action of TIM-3 in cancer cells and in myeloid cells regulates the tumorigenicity of human RCC. (C) 2015 AACR.
  • Akira Inoue, Shunichi Sugawara, Makoto Maemondo, Yoshiaki Mori, Satoshi Oizumi, Masao Harada, Kageaki Taima, Naoto Morikawa, Takashi Ishida, Ichiro Kinoshita, Hiroshi Watanabe, Toshiro Suzuki, Taku Nakagawa, Ryota Saito, Toshihiro Nukiwa
    LUNG CANCER 89 (1) 61 - 65 0169-5002 2015/07 [Refereed][Not invited]
     
    Purpose: Amrubicin and re-challenge of platinum doublet are both effective treatments for sensitive-relapsed small-cell lung cancer (SCLC). However, no comparative study of these treatments has been reported. This randomized study was conducted to select the most suitable regimen for future evaluation. Patients and methods: SCLC patients who had relapsed more than 90 days after their first-line platinum-doublet regimen were randomized to receive amrubicin (40 mg/m(2), days 1-3) or re-challenge with platinum doublet. Primary endpoint was objective response rate (ORR), with secondary endpoints of progression-free survival (PFS), overall survival and toxicity profiles. We assumed that an ORR of 50% indicates potential usefulness, while that of 30% would constitute the lower limit of interest (alpha 0.1; beta 0.1). Initial estimated accrual was 28 patients to each arm. Results: From February 2008 to June 2013,60 patients were enrolled and 57 patients (27 amrubicin and 30 re-challenge) were found to be evaluable for efficacy and safety. The ORR and PFS were 67% (90% confidence interval, 52-82) and 5.4 months in the amrubicin group, and 43% (90% confidence interval, 28-58) and 5.1 months in the re-challenge group, respectively. Although grade 3 febrile neutropenia was observed in 19% of patients in the amrubicin group, these episodes were transient and manageable. Non-hematological toxicities were generally moderate and no treatment-related death was observed in either group. Conclusion: Only amrubicin met the primary endpoint. Moreover, amrubicin demonstrated superior efficacy over re-challenge of platinum with acceptable levels of toxicity. Further evaluation of amrubicin for sensitive-relapsed SCLC is warranted. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
  • Takatsugu Mizumachi, Akihiro Homma, Tomohiko Kakizaki, Tomohiro Sakashita, Satoshi Kano, Hiromitsu Hatakeyama, Kazuhiko Tsuchiya, Koichi Yasuda, Rikiya Onimaru, Hiroki Shirato, Jun Taguchi, Yasushi Shimizu, Ichiro Kinoshita, Hirotoshi Akita, Satoshi Fukuda
    INTERNATIONAL JOURNAL OF CLINICAL ONCOLOGY 20 (3) 431 - 437 1341-9625 2015/06 [Refereed][Not invited]
     
    The aim of this study was to evaluate the feasibility of induction docetaxel, cisplatin, and 5-fluorouracil chemotherapy followed by concurrent weekly cisplatin chemoradiotherapy for patients with locally advanced head and neck squamous cell carcinoma (HNSCC). Between 2010 and 2013, 30 patients with Stage IV HNSCC were treated in Hokkaido University Hospital with three cycles of induction chemotherapy (docetaxel 75 mg/m(2) and cisplatin 75 mg/m(2), day 1; and 5-fluorouracil 750 mg/m(2)/day 120 h continuous infusion, every 3 weeks) followed by concurrent weekly cisplatin (40 mg/m(2), on weeks 1, 2, 3, 5, 6 and 7) chemoradiotherapy. Three courses of induction chemotherapy were performed in 25 patients (83 %) with grade 3-4 toxicities during induction chemotherapy observed in 22 patients (73 %). The major toxicities were hematologic, with 22 cases (73 %) showing grade 3-4 neutropenia. Radiotherapy was completed (70 Gy) in 29 patients (97 %), while a total of 19 patients (63 %) completed five (13 patients) or six (6 patients) courses of chemotherapy. During concurrent chemoradiotherapy, no grade 4 hematological toxicities were observed. Grade 4 dermatitis was observed in one patient, and grade 3 mucositis was observed in 12 patients. There were no treatment-related deaths during the induction chemotherapy or concurrent chemoradiotherapy. The 1- and 2-year progression-free survival rates and the 1- and 2-year overall survival rates were 86 %, 72 %, and 89 %, 81 %, respectively. Sequential therapy composed of induction chemotherapy followed by concurrent weekly cisplatin chemoradiotherapy is feasible, showing encouraging results in patients with locally advanced HNSCC. Concurrent weekly cisplatin chemoradiotherapy following induction chemotherapy appears to be a suitable alternative to three-weekly high-dose cisplatin therapy.
  • Tatsuro Fukuhara, Makoto Maemondo, Akira Inoue, Kunihiko Kobayashi, Shunichi Sugawara, Satoshi Oizumi, Hiroshi Isobe, Akihiko Gemma, Masao Harada, Hirohisa Yoshizawa, Ichiro Kinoshita, Yuka Fujita, Yasuo Saijo, Koichi Hagiwara, Satoshi Morita, Toshihiro Nukiwa
    LUNG CANCER 88 (2) 181 - 186 0169-5002 2015/05 [Refereed][Not invited]
     
    Introduction: Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI) treatment is the standard therapy for non-small cell lung cancer (NSCLC) harbouring EGFR-activating mutations. The NEJ002 phase 3 clinical trial demonstrated the efficacy of EGFR-TKI; gefitinib was significantly superior in both progression-free survival (PFS) and objective response rate (ORR) than carboplatin plus paclitaxel. However, several cases showed no response. In this study, we performed further analysis of the characteristics of these non-responders. Methods: Available data from NEJ002 on maximum changes in tumour size were obtained from 103 cases (90.4%) and 110 cases (96.5%) in the carboplatin paclitaxel and gefitinib groups, respectively. Waterfall plots of maximum tumour size changes were created for non-responders. Results: Five (4.9%) and 9 (8.2%) cases in the carboplatin paclitaxel and gefitinib groups were non-responders, respectively. The mean pack years of the non-responders in the carboplatin paclitaxel and gefitinib groups were 0.33 and 31.7, respectively. The ORR of total smokers (61.5%) and heavy smokers (over 40 pack years, 52.6%) in the gefitinib group were significantly lower compared to people who have never smoked (80.0%) (P = 0.044 and P=0.020, respectively). Smoker cases also showed a tendency towards lower PFS and overall survival (05). In addition, the EGFR common mutation types did not affect PFS and OS in gefitinib-treated cases in NEJ002. However, in this study, the ORR and waterfall plots showed that gefitinib-treated non-responders who had a deletion in exon 19 in the EGFR gene exhibited a tendency towards a higher response compared to those with a L858R mutation. Conclusions: NSCLC patients with a smoking history or the EGFR L858R mutation may demonstrate a poorer response to gefitinib treatment. 0 2015 The Authors. Published by Elsevier Ireland Ltd.
  • Rio Honma, Ichiro Kinoshita, Eiji Miyoshi, Utano Tomaru, Yoshihiro Matsuno, Yasushi Shimizu, Satoshi Takeuchi, Yuka Kobayashi, Kichizo Kaga, Naoyuki Taniguchi, Hirotoshi Dosaka-Akita
    ONCOLOGY 88 (5) 298 - 308 0030-2414 2015 [Refereed][Not invited]
     
    Objecitive: Fucosyltransferase 8 (FUT8), the only enzyme responsible for the core alpha 1,6-fucosylation of asparagine-linked oligosaccharides of glycoproteins, is a vital enzyme in cancer development and progression. We examined FUT8 expression in non-small cell lung cancers (NSCLCs) to analyze its clinical significance. We also examined the expression of guanosine diphosphate-mannose-4,6-dehydratase (GMD), which is imperative for the synthesis of fucosylated oligosaccharides. Methods: Using immunohistochemistry, we evaluated the expression of FUT8 and GMD in relation to patient survival and prognosis in potentially curatively resected NSCLCs. Results: High expression of FUT8 was found in 67 of 129 NSCLCs (51.9%) and was significantly found in non-squamous cell carcinomas (p = 0.008). High expression of FUT8 was associated with poor survival (p = 0.03) and was also a significant and independent unfavorable prognostic factor in patients with potentially curatively resected NSCLCs (p = 0.047). High expression of GMD was significantly associated with high FUT8 expression (p = 0.04). Conclusions: High expression of FUT8 is associated with an unfavorable clinical outcome in patients with potentially curatively resected NSCLCs, suggesting that FUT8 can be a prognostic factor. The analysis of FUT8 expression and its core fucosylated products may provide new insights for the therapeutic targets of NSCLCs. (C) 2015 S. Karger AG, Basel
  • Takafumi Jinushi, Yoshihiko Shibayama, Ichiro Kinoshita, Satoshi Oizumi, Masahisa Jinushi, Tadahiro Aota, Toshiyuki Takahashi, Shoichi Horita, Hirotoshi Dosaka-Akita, Ken Iseki
    CANCER MEDICINE 3 (6) 1544 - 1552 2045-7634 2014/12 [Refereed][Not invited]
     
    A component of polycomb repressor complex 2, enhancer of zeste homolog 2 (EZH2), plays an important role in tumor malignancy and metastasis, while milk fat globule-epidermal growth factor-factor 8 (MFGE8) plays a key role in tumor progression and prognosis. MicroRNAs (miRs) are also critically involved in various physiological and pathological processes. We here evaluated the relationship between overall survival (OS) in colorectal cancer patients and the expression of onco-miRs and miRs, which may target EZH2 and MFGE8. Plasma and formalin-fixed paraffin-embedded (FFPE) samples were obtained from 71 colorectal cancer patients. The expression levels of miRs complementary to EZH2 and MFGE8 mRNA and cancer malignancies were evaluated. The miRs analyzed were as follows: miR-16, miR-21, miR-26a, miR-34a, miR-98, miR-101-3p, miR-101-5p, miR-124-5p (also known as miR-124*), miR-126-3p, miR-126-5p, miR-210, miR-217, and miR-630. The plasma expression levels of MFGE8 in completely resected patients were significantly lower than those in unresectable patients. Lower miR-26a expression levels were correlated with a higher probability of OS. Higher miR-124-5p expression levels in plasma and FFPE samples were correlated with a higher probability of OS. The transfection of mimic miR-124-5p into WiDr and COLO201 cells inhibited the expression of structural maintenance of chromosomes 4 (SMC4) mRNA. Our results indicate that miR-124-5p may target the tumorigenesis gene, SMC4, which suggests that expression levels of miR-124-5p in plasma and FFPE samples; therefore, the expression of MFGE8, miR-26a, and miR-124-5p in plasma may be used as biomarkers to determine the prognosis of colorectal cancer patients.
  • Kenya Kanazawa, Hiroshi Yokouchi, Xintao Wang, Takashi Ishida, Yuka Fujita, Satoru Fujiuchi, Toshiyuki Harada, Masao Harada, Kei Takamura, Satoshi Oizumi, Ichiro Kinoshita, Yutaka Katsuura, Osamu Honjo, Tetsuya Kojima, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Mitsuru Munakata, Masaharu Nishimura
    CANCER CHEMOTHERAPY AND PHARMACOLOGY 74 (6) 1149 - 1157 0344-5704 2014/12 [Refereed][Not invited]
     
    This phase II study evaluated the response rate (RR) and safety of combination therapy with carboplatin (CBDCA) and pemetrexed (PEM) in Japanese patients with non-squamous non-small cell lung cancer (non-sq NSCLC). Further, the relationship between therapy efficacy/toxicity and genetic polymorphisms associated with PEM metabolism was analyzed. Forty-one patients received CBDCA at a dose targeting an area under the concentration-time curve of 5 mg/mL x min and PEM of 500 mg/m(2) on day 1 every 3 weeks. Single-nucleotide polymorphisms of the thymidylate synthase (TYMS) coding gene, the variable number of tandem repeat (VNTR) in the TYMS, and the methylenetetrahydrofolate reductase (MTHFR) coding gene were analyzed. The overall RR was 36.6 %. Median progression-free survival and median survival time were 4.7 months [95 % confidence interval (CI) 3.9-5.6 months] and 16.2 months (95 % CI 6.1-26.2 months), respectively. Epidermal growth factor receptor gene mutations were detected in 6 patients (14.6 %). The VNTR in the TYMS significantly correlated with anemia (p = 0.047) and thrombocytopenia (p = 0.038). This combination therapy was effective and tolerable in patients with advanced non-sq NSCLC. The VNTR in the TYMS appears to be a predictive factor for anemia and thrombocytopenia in patients treated with this regimen.
  • Expression of α1,6-fucosyltransferase is associated with prognosis and histology in non-small cell lung cancers
    Honma R, Kinoshita I, Miyoshi E, Tomaru U, Matsuno Y, Shimizu Y, Takeuchi S, Kobayashi Y, Kaga K, Taniguchi N, Akita DH
    2014/10 [Refereed][Not invited]
  • Camille Ragin, Monisola Obikoya-Malomo, Sungjin Kim, Zhengjia Chen, Rafael Flores-Obando, Denise Gibbs, Chihaya Koriyama, Francisco Aguayo, Jill Koshiol, Neil E. Caporaso, Giovanna E. Carpagnano, Marco Ciotti, Hirotoshi Dosaka-Akita, Masashi Fukayama, Akiteru Goto, Demetrios A. Spandidos, Vassilis Gorgoulis, Danielle A. M. Heideman, Robert A. A. van Boerdonk, Kenzo Hiroshima, Reika Iwakawa, Nikolaos G. Kastrinakis, Ichiro Kinoshita, Suminori Akiba, Maria T. Landi, H. Eugene Liu, Jinn-Li Wang, Ranee Mehra, Fadlo R. Khuri, Wan-Teck Lim, Taofeek K. Owonikoko, Suresh Ramalingam, Emmanuela Sarchianaki, Kari Syrjanen, Ming-Sound Tsao, Jenna Sykes, Siew Wan Hee, Jun Yokota, Apostolos Zaravinos, Emanuela Taioli
    CARCINOGENESIS 35 (6) 1267 - 1275 0143-3334 2014/06 [Refereed][Not invited]
     
    Human papillomavirus (HPV) is the etiologic risk factor for cervical cancer. Some studies have suggested an association with a subset of lung tumors, but the etiologic link has not been firmly established. We performed an international pooled analysis of cross-sectional studies (27 datasets, n = 3249 patients) to evaluate HPV DNA prevalence in lung cancer and to investigate viral presence according to clinical and demographic characteristics. HPV16/18 were the most commonly detected, but with substantial variation in viral prevalence between geographic regions. The highest prevalence of HPV16/18 was observed in South and Central America, followed by Asia, North America and Europe (adjusted prevalence rates = 22, 5, 4 and 3%, respectively). Higher HPV16 prevalence was noted in each geographic region compared with HPV18, except in North America. HPV16/18-positive lung cancer was less likely observed among White race (adjusted odds ratio [OR] = 0.33, 95% confidence interval [CI] = 0.12-0.90), whereas no associations were observed with gender, smoking history, age, histology or stage. Comparisons between tumor and normal lung tissue show that HPV was more likely to be present in lung cancer rather than normal lung tissues (OR = 3.86, 95% CI = 2.87-5.19). Among a subset of patients with HPV16-positive tumors, integration was primarily among female patients (93%, 13/14), while the physical status in male cases (N = 14) was inconsistent. Our findings confirm that HPV DNA is present in a small fraction of lung tumors, with large geographic variations. Further comprehensive analysis is needed to assess whether this association reflects a causal relationship.
  • Tsunaki Yamashina, Muhammad Baghdadi, Akihiro Yoneda, Ichiro Kinoshita, Shinya Suzu, Hirotoshi Dosaka-Akita, Masahisa Jinushi
    CANCER RESEARCH 74 (10) 2698 - 2709 0008-5472 2014/05 [Refereed][Not invited]
     
    Resistance to anticancer therapeutics greatly affects the phenotypic and functional properties of tumor cells, but how chemoresistance contributes to the tumorigenic activities of cancer stem-like cells remains unclear. In this study, we found that a characteristic of cancer stem-like cells from chemoresistant tumors (CSC-R) is the ability to produce a variety of proinflammatory cytokines and to generate M2-like immunoregulatory myeloid cells from CD14(+) monocytes. Furthermore, we identified the IFN-regulated transcription factor IRF5 as a CSC-R-specific factor critical for promoting M-CSF production and generating tumorigenic myeloid cells. Importantly, myeloid cells primed with IRF5(+) CSC-R facilitate the tumorigenic and stem cell activities of bulk tumors. Importantly, the activation of IRF5/M-CSF pathways in tumor cells were correlated with the number of tumor-associated CSF1 receptor(+) M2 macrophages in patients with non-small lung cancer. Collectively, our findings show how chemoresistance affects the properties of CSCs in their niche microenvironments. (C)2014 AACR.
  • Satoshi Oizumi, Shunichi Sugawara, Koichi Minato, Toshiyuki Harada, Akira Inoue, Yuka Fujita, Makoto Maemondo, Hirohisa Yoshizawa, Kazuhiko Ito, Akihiko Gemma, Masaru Nishitsuji, Masao Harada, Hiroshi Isobe, Ichiro Kinoshita, Satoshi Morita, Kunihiko Kobayashi, Koichi Hagiwara, Minoru Kurihara, Toshihiro Nukiwa
    JOURNAL OF CLINICAL ONCOLOGY 32 (15) 0732-183X 2014/05 [Refereed][Not invited]
  • Hidenori Mizugaki, Jun Sakakibara-Konishi, Junko Kikuchi, Jun Moriya, Kanako C. Hatanaka, Eiki Kikuchi, Ichiro Kinoshita, Satoshi Oizumi, Hirotoshi Dosaka-Akita, Yoshihiro Matsuno, Masaharu Nishimura
    INTERNATIONAL JOURNAL OF CLINICAL ONCOLOGY 19 (2) 254 - 259 1341-9625 2014/04 [Refereed][Not invited]
     
    CD133 is a membrane glycoprotein containing five transmembrane loops. Previous reports suggest that a CD133-positive subpopulation of multipotent cells with extensive proliferative and self-renewal characteristics has biological features of a cancer stem cell. In addition, the presence of CD133-positive cells was associated with a significantly poorer prognosis for some solid tumors, compared to those with CD133-negative cells. However, the clinicopathological significance of CD133 in non-small cell lung cancer (NSCLC) remains controversial. We conducted immunohistochemical assessment of 161 NSCLCs surgically resected at Hokkaido University Hospital between 1982 and 1994 to evaluate correlations between CD133 expression and various clinicopathological features. CD133 expression was significantly correlated with pathological stages (pStages) II, III, and IV for the various NSCLC types analyzed and was an independent factor for unfavorable prognosis in this population (hazard ratio = 3.157, P = 0.015). CD133 expression was correlated with pStage and was predictive of unfavorable prognosis in patients with pStages II, III, and IV NSCLC. These results suggest the possibility of using CD133 as a novel prognostic marker in these patients.
  • Muhammad Baghdadi, Akihiro Yoneda, Tsunaki Yamashina, Hiroko Nagao, Yoshihiro Komohara, Shigenori Nagai, Hisaya Akiba, Marc Foretz, Hironori Yoshiyama, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Motohiro Takeya, Benoit Viollet, Hideo Yagita, Masahisa Jinushi
    IMMUNITY 39 (6) 1070 - 1081 1074-7613 2013/12 [Refereed][Not invited]
     
    Phagocytosis of apoptotic cells by myeloid cells has been implicated in the maintenance of immune homeostasis. In this study, we found that T cell immunoglobulin-and mucin domain-containing molecule-4 (TIM-4) repressed tumor-specific immunity triggered by chemotherapy-induced tumor cell death. TIM-4 was found to be highly expressed on tumor-associated myeloid cells such as macrophages (TAMs) and dendritic cells (TADCs) and danger-associated molecular patterns (DAMPs) released from chemotherapy-damaged tumor cells induced TIM-4 on tumor-associated myeloid cells recruited from bone marrow-derived precursors. TIM-4 directly interacted with AMPK alpha 1 and activated autophagy-mediated degradation of ingested tumors, leading to reduced antigen presentation and impaired CTL responses. Consistently, blockade of the TIM-4-AMPK alpha 1-autophagy pathway augmented the antitumor effect of chemotherapeutics by enhancing tumor-specific CTL responses. Our finding provides insight into the immune tolerance mediated by phagocytosis of dying cells, and targeting of the TIM-4-AMPK alpha 1 interaction constitutes a unique strategy for augmenting antitumor immunity and improving cancer chemotherapy.
  • A. Inoue, K. Kobayashi, M. Maemondo, S. Sugawara, S. Oizumi, H. Isobe, A. Gemma, M. Harada, H. Yoshizawa, I. Kinoshita, Y. Fujita, S. Okinaga, H. Hirano, K. Yoshimori, T. Harada, Y. Saijo, K. Hagiwara, S. Morita, T. Nukiwa
    ANNALS OF ONCOLOGY 1 24 (1) 54 - 59 0923-7534 2013/01 [Refereed][Not invited]
     
    NEJ002 study, comparing gefitinib with carboplatin (CBDCA) and paclitaxel (PTX; Taxol) as the first-line treatment for advanced non-small cell lung cancer (NSCLC) harboring an epidermal growth factor receptor (EGFR) mutation, previously reported superiority of gefitinib over CBDCA/PTX on progression-free survival (PFS). Subsequent analysis was carried out mainly regarding overall survival (OS). For all 228 patients in NEJ002, survival data were updated in December, 2010. Detailed information regarding subsequent chemotherapy after the protocol treatment was also assessed retrospectively and the impact of some key drugs on OS was evaluated. The median survival time (MST) was 27.7 months for the gefitinib group, and was 26.6 months for the CBDCA/PTX group (HR, 0.887; P = 0.483). The OS of patients who received platinum throughout their treatment (n = 186) was not statistically different from that of patients who never received platinum (n = 40). The MST of patients treated with gefitinib, platinum, and pemetrexed (PEM) or docetaxel (DOC, Taxotere; n = 76) was around 3 years. No significant difference in OS was observed between gefitinib and CBDCA/PTX in the NEJ002 study, probably due to a high crossover use of gefitinib in the CBDCA/PTX group. Considering the many benefits and the risk of missing an opportunity to use the most effective agent for EGFR-mutated NSCLC, the first-line gefitinib is strongly recommended.
  • Toshiyuki Harada, Satoshi Oizumi, Kenichiro Ito, Kei Takamura, Eiki Kikuchi, Tomoya Kuda, Shunichi Sugawara, Aya Suzuki, Makoto Maemondo, Yuka Fujita, Ichiro Kinoshita, Akira Inoue, Fumihiro Hommura, Yutaka Katsuura, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Masaharu Nishimura
    The oncologist 18 (4) 439 - 45 1083-7159 2013 [Refereed][Not invited]
     
    Amrubicin, a third-generation synthetic anthracycline agent, has favorable clinical activity and acceptable toxicity for the treatment of patients with non-small cell lung cancer (NSCLC) and small cell lung cancer. We conducted this study to evaluate the efficacy and safety of amrubicin for advanced NSCLC patients as a third- or fourth-line therapy. Eligible patients had recurrent or refractory advanced NSCLC after second- or third-line therapy. Patients received amrubicin, 35 mg/m(2) i.v. on days 1-3 every 3 weeks. The primary endpoint was the disease control rate (DCR). Secondary endpoints were the overall survival (OS) time, progression-free survival (PFS) time, response rate, and toxicity profile. Of the 41 patients enrolled, 26 received amrubicin as a third-line and 15 received it as a fourth-line therapy. The median number of treatment cycles was two (range, 1-9). Objective responses were complete response (n = 0), partial response (n = 4), stable disease (n = 21), progressive disease (n = 15), and not evaluable (n = 1), resulting in a DCR of 61.0% (95% confidence interval, 46.0%-75.9%). The overall response rate was 9.8% (95% confidence interval, 0.6%-18.8%). The median PFS interval was 3.0 months, median OS time was 12.6 months, and 1-year survival rate was 53.7%. Grade 3 or 4 hematological toxicities were neutropenia (68%), anemia (12%), thrombocytopenia (12%), and febrile neutropenia (17%). Nonhematological toxicities were mild and reversible. No treatment-related deaths were observed. Amrubicin showed significant clinical activity with manageable toxicities as a third- or fourth-line therapy for patients with advanced NSCLC. This study provides relevant data for routine practice and future prospective trials evaluating third- or fourth-line treatment strategies for patients with advanced NSCLC.
  • Junko Kikuchi, Taichi Takashina, Ichiro Kinoshita, Eiki Kikuchi, Yasushi Shimizu, Jun Sakakibara-Konishi, Satoshi Oizumi, Victor E. Marquez, Masaharu Nishimura, Hirotoshi Dosaka-Akita
    LUNG CANCER 78 (2) 138 - 143 0169-5002 2012/11 [Refereed][Not invited]
     
    EZH2 (enhancer of zeste homolog 2) is the catalytic subunit of PRC2 (polycomb repressive complex 2), which mediates histone methyltransferase activity and functions as transcriptional repressor involved in gene silencing. EZH2 is involved in malignant transformation and biological aggressiveness of several human malignancies. We previously demonstrated that non-small cell lung cancers (NSCLCs) also overexpress EZH2 and that high expression of EZH2 correlates with poor prognosis. Growing evidence indicates that EZH2 may be an appropriate therapeutic target in malignancies, including NSCLCs. Recently, an S-adenosyl-L-homocysteine hydrolase inhibitor, 3-Deazaneplanocin A (DZNep), has been shown to deplete and inhibit EZH2. The aim of this study was to determine the effect of DZNep in NSCLC cells. Knockdown of EZH2 by small-interfering RNA (siRNA) resulted in decreased growth of four NSCLC cell lines. MTT assays demonstrated that DZNep treatment resulted in dose-dependent inhibition of proliferation in the NSCLC cell lines with a half maximal inhibitory concentration (IC50) ranging from 0.08 to 0.24 mu M. Immortalized but non-cancerous bronchial epithelial and fibroblast cell lines were less sensitive to DZNep than the NSCLC cell lines. Soft agarose assays demonstrated that anchorage-independent growth was also reduced in all three NSCLC cell lines that were evaluated using this assay. Flow cytometry analysis demonstrated that DZNep induced apoptosis and G1 cell cycle arrest in NSCLC cells, which was partially associated with cyclin A decrease and p27(Kip1) accumulation. DZNep depleted cellular levels of EZH2 and inhibited the associated histone H3 lysine 27 trimethylation. These results indicated that an epigenetic therapy that pharmacologically targets EZH2 via DZNep may constitute a novel approach to treatment of NSCLCs. (C) 2012 Elsevier Ireland Ltd. All rights reserved.
  • Shigeki Chiba, Muhammad Baghdadi, Hisaya Akiba, Hironori Yoshiyama, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Yoichiro Fujioka, Yusuke Ohba, Jacob V. Gorman, John D. Colgan, Mitsuomi Hirashima, Toshimitsu Uede, Akinori Takaoka, Hideo Yagita, Masahisa Jinushi
    NATURE IMMUNOLOGY 13 (9) 832 - 842 1529-2908 2012/09 [Refereed][Not invited]
     
    The mechanisms by which tumor microenvironments modulate nucleic acid mediated innate immunity remain unknown. Here we identify the receptor TIM-3 as key in circumventing the stimulatory effects of nucleic acids in tumor immunity. Tumor-associated dendritic cells (DCs) in mouse tumors and patients with cancer had high expression of TIM-3. DC-derived TIM-3 suppressed innate immune responses through the recognition of nucleic acids by Toll-like receptors and cytosolic sensors via a galectin-9-independent mechanism. In contrast, TIM-3 interacted with the alarmin HMGB1 to interfere with the recruitment of nucleic acids into DC endosomes and attenuated the therapeutic efficacy of DNA vaccination and chemotherapy by diminishing the immunogenicity of nucleic acids released from dying tumor cells. Our findings define a mechanism whereby tumor microenvironments suppress antitumor immunity mediated by nucleic acids.
  • Jun Sakakibara-Konishi, Satoshi Oizumi, Junko Kikuchi, Eiki Kikuchi, Hidenori Mizugaki, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    BMC CANCER 12 286  1471-2407 2012/07 [Refereed][Not invited]
     
    Background: The BH3-only members of the Bcl-2 protein family have been proposed to play a key role in the control of apoptosis and in the initiation of the apoptotic pathways. In this study, we evaluated the expression of Bim, Noxa, and Puma in non-small cell lung cancer (NSCLC). Methods: A total of 135 surgically resected NSCLCs were immunohistochemically assessed for Bim, Noxa, and Puma expression. The immunoscores were determined, and then its correlation with either the clinicopathological variables or the survival outcomes were analyzed. Results: Immunohistochemical reactivity for Bim, Noxa, and Puma was detected in the cytoplasm of the tumor cells. Bim expression was associated with several clinicopathological factors, including sex (p < 0.001), smoking habit (p = 0.03), pathological histology (p = 0.001), pathological T stage (p = 0.03), pathological disease stage (p = 0.02), and differentiation of tumor (p < 0.001). Multivariate logistic regression analysis showed a significant correlation between low Bim expression and squamous cell carcinoma (p = 0.04), in addition to a correlation between high Bim expression and well differentiated tumors (p = 0.02). Analysis of cellular biological expression demonstrated a link between low Bim expression and high Ki67. While Noxa expression was also shown to be correlated with both smoking habit (p = 0.02) and the pathological histology (p = 0.03), there was no strong association observed between the expression and the clinical features when they were examined by a multivariate logistic regression analysis. No correlations were noted between Puma expression and any of the variables. Our analyses also indicated that the expression levels of the BH3-only proteins were not pertinent to the survival outcome. Conclusions: The current analyses demonstrated that Bim expression in the NSCLCs was associated with both squamous cell carcinoma histology and tumor proliferation.
  • Satoshi Oizumi, Kunihiko Kobayashi, Akira Inoue, Makoto Maemondo, Shunichi Sugawara, Hirohisa Yoshizawa, Hiroshi Isobe, Masao Harada, Ichiro Kinoshita, Shoji Okinaga, Terufumi Kato, Toshiyuki Harada, Akihiko Gemma, Yasuo Saijo, Yuki Yokomizo, Satoshi Morita, Koichi Hagiwara, Toshihiro Nukiwa
    ONCOLOGIST 17 (6) 863 - 870 1083-7159 2012/06 [Refereed][Not invited]
     
    Background. For non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations, first-line gefitinib produced a longer progression-free survival interval than first-line carboplatin plus paclitaxel but did not show any survival advantage in the North East Japan 002 study. This report describes the quality of life (QoL) analysis of that study. Methods. Chemotherapy-naive patients with sensitive EGFR-mutated, advanced NSCLC were randomized to receive gefitinib or chemotherapy (carboplatin and paclitaxel). Patient QoL was assessed weekly using the Care Notebook, and the primary endpoint of the QoL analysis was time to deterioration from baseline on each of the physical, mental, and life well-being QoL scales. Kaplan-Meier probability curves and log-rank tests were employed to clarify differences. Results. QoL data from 148 patients (72 in the gefitinib arm and 76 in the carboplatin plus paclitaxel arm) were analyzed. Time to defined deterioration in physical and life well-being significantly favored gefitinib over chemotherapy (hazard ratio [HR] of time to deterioration, 0.34; 95% confidence interval [CI], 0.23-0.50; p < .0001 and HR, 0.43; 95% CI, 0.28-0.65; p < .0001, respectively). Conclusion. QoL was maintained much longer in patients treated with gefitinib than in patients treated with standard chemotherapy, indicating that gefitinib should be considered as the standard first-line therapy for advanced EGFR-mutated NSCLC in spite of no survival advantage. The Oncologist 2012;17:863- 870
  • Masahisa Jinushi, Shigeki Chiba, Muhammad Baghdadi, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Koyu Ito, Hironori Yoshiyama, Hideo Yagita, Toshimitsu Uede, Akinori Takaoka
    Cancer research 1 72 (1) 56 - 65 0008-5472 2012/01/01 [Refereed][Not invited]
     
    Although the tumor microenvironment plays a critical role in tumor progression and metastasis, the relationship between chemotherapy resistance and modulation of the tumor microenvironment remains unclear. Here, we report a novel mechanism showing how constitutive DNA damage signals in therapy-resistant tumor cells suppress antitumor immunity in an integrin-αvβ3-dependent manner. Integrin-αvβ3 was upregulated on various therapy-resistant tumor cells through chronic activation of ATM/Chk2-and NFκB-mediated pathways. Inhibiting tumor-specific expression of integrin-αvβ3 improved therapeutic responses to anticancer drugs by stimulating endogenous host immune systems. Mechanistic investigations revealed that tumor-specific integrin-αvβ3 expression targeted dendritic cells, facilitating their ability to phagocytose viable therapy-resistant tumor cells and thereby impaired their ability to cross-prime antigen-specific T lymphocytes. Together, our results clarify the detrimental effects of constitutive DNA damage signals to chemosensitivity and antitumor immunity. Furthermore, these findings suggest that integrin-αvβ3 targeting may benefit patients' refractory to current anticancer regimens by defeating DNA damage signaling-induced immune escape.
  • Chiba S, Baghdadi M, Akiba H, Kinoshita I, Yoshiyama H, Hirashima M, Dosaka-Akita H, Uede T, Takaoka A, Yagita H, Jinushi M
    Nat Immunol 13 (9) 832 - 842 2012 [Not refereed][Not invited]
  • Jun Sakakibara-Konishi, Satoshi Oizumi, Ichiro Kinoshita, Naofumi Shinagawa, Junko Kikuchi, Mototsugu Kato, Tetsuya Inoue, Norio Katoh, Rikiya Onimaru, Hiroki Shirato, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    LUNG CANCER 74 (2) 248 - 252 0169-5002 2011/11 [Refereed][Not invited]
     
    Introduction: Although paclitaxel with carboplatin and thoracic radiotherapy has improved survival for patients with locally advanced unresectable non-small cell lung cancer (NSCLC), the optimal dose of paclitaxel has not been well defined in Japan. This study was conducted to determine the maximum tolerated dose (MTD) and recommended dose (RD) of paclitaxel in combination with carboplatin and concurrent real-time tumor-tracking thoracic radiation therapy (thoracic RTRT). Patients and methods: Previously untreated patients with histologically confirmed, locally advanced unresectable NSCLC were eligible. Before treatment, gold markers were inserted into the lung and the mediastinum of all patients. RTRT comprised a total of 66 Gy at 2 Gy/fraction, 5 days/week, for 7 weeks. Patients received paclitaxel at a starting dose of 40 mg/m(2) followed by carboplatin at a fixed area under the curve (AUC) of 2, as a weekly regimen with RTRT. The dose of paclitaxel was escalated by 5 mg/m(2) per level. Results: Eight patients with locally advanced unresectable NSCLC were enrolled and treated with two dose levels of paclitaxel (40 mg/m(2) and 45 mg/m(2)), carboplatin (AUC = 2) and RTRT. No dose limiting toxicities (DLTs) were observed at Level 1 (paclitaxel, 40 mg/m(2) and carboplatin, AUC = 2). At Level 2 (paclitaxel, 45 mg/m(2) and carboplatin, AUC = 2), two of five patients experienced DLTs, in the form of esophagitis and discontinuation of chemotherapy more than twice. The MTD and RD of paclitaxel were thus defined as 45 mg/m(2) and 40 mg/m(2), respectively. Conclusions: This phase I study was well tolerated and the RD of paclitaxel and carboplatin with RTRT is 40 mg/m(2) at AUC = 2, respectively. Further studies are warranted to evaluate the efficacy of this regimen. (C) 2011 Elsevier Ireland Ltd. All rights reserved.
  • Satoshi Takeuchi, Songji Zhao, Yuji Kuge, Yan Zhao, Ken-Ichi Nishijima, Toshiyuki Hatano, Yasushi Shimizu, Ichiro Kinoshita, Nagara Tamaki, Hirotoshi Dosaka-Akita
    ONCOLOGY REPORTS 26 (3) 725 - 730 1021-335X 2011/09 [Refereed][Not invited]
     
    We investigated whether F-18-fluorothymidine-positron-emission tomography/computed tomography (F-18-FLT-PET/CT) is useful for the evaluation of the very early response to anti-epidermal growth factor receptor (EGFR) antibody cetuximab therapy in human lung cancer xenografts. A human tumor xenograft model was established with a human non-small cell lung cancer cell line. The mice were randomly assigned to four groups: tumor growth follow-up, ex vivo study, PET/CT imaging and non-treated control. Mice were administered saline as control or cetuximab on day 1. An immunohistochemical study with Ki-67 was performed. Tumor volume treated with cetuximab was kept significantly smaller than control after day 8, although there was no difference on day 3. On day 3, F-18-FLT distribution was higher in the tumor than in other tissues, and was significantly decreased by treatment with cetuximab. On PET/CT imaging, F-18-FLT distribution in the tumor was clearly visualized, and the maximum standardized uptake value (SUV) was significantly decreased after treatment with cetuximab (p<0.01). Ki-67 expression was also significantly decreased on day 3 (p=0.01). These results suggest that F-18-FLT-PET/CT can be a useful predictor to determine the response to molecular targeted drugs such as cetuxima.b at an earlier time point than the change of tumor size.
  • Masahisa Jinushi, Shigeki Chiba, Hironori Yoshiyama, Kenkichi Masutomi, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Hideo Yagita, Akinori Takaoka, Hideaki Tahara
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 108 (30) 12425 - 12430 0027-8424 2011/07 [Refereed][Not invited]
     
    Recent evidence has unveiled the critical role of tumor cells with stem cell activities in tumorigenicity and drug resistance, but how tumor microenvironments regulate cancer stem/initiating cells (CSCs) remains unknown. We clarified the role of tumor-associated macrophages (TAMs) and their downstream factor milk-fat globule-epidermal growth factor-VIII (MFG-E8) in the regulation of CSC activities. Bone marrow chimeric systems and adoptive cell transfers elucidated the importance of MFG-E8 from TAMs in conferring to CSCs with the ability to promote tumorigenicity and anticancer drug resistance. MFG-E8 mainly activates signal transducer and activator of transcription-3 (Stat3) and Sonic Hedgehog pathways in CSCs and further amplifies their anticancer drug resistance in cooperation with IL-6. Thus, the pharmacological targeting of key factors derived from tumor-associated inflammation provides a unique strategy to eradicate therapy-resistant tumors by manipulating CSC activities.
  • Junko Kikuchi, Ichiro Kinoshita, Yasushi Shimizu, Eiki Kikuchi, Kayoko Takeda, Hiroyuki Aburatani, Satoshi Oizumi, Jun Konishi, Kichizo Kaga, Yoshihiro Matsuno, Michael J. Birrer, Masaharu Nishimura, Hirotoshi Dosaka-Akita
    LUNG CANCER 72 (2) 229 - 237 0169-5002 2011/05 [Refereed][Not invited]
     
    Background: Minichromosome maintenance (MCM) proteins 2-7 form a complex essential for the initiation of DNA replication. In the process to screen expression changes related to growth suppression of non-small cell lung cancer (NSCLC) cells by a cJun dominant-negative mutant, we found that reduced expression of MCM4 was correlated with this growth suppression. Method: We determined the relevance of MCM4 in proliferation of NSCLC by downregulating its expression with small-interfering RNA in three NSCLC cell lines. We then immunohistochemically analyzed MCM4 expression in 156 surgically resected NSCLCs to correlate clinicopathologic characteristics. Results: MCM4 downregulation reduced proliferation in two cell lines. MCM4 expression was higher in cancer cells than in adjacent normal bronchial epithelial cells (p < 0.001). High MCM4 expression was correlated with male gender, heavy smoking, poorer differentiation and non-adenocarcinoma histology (p < 0.001, respectively). High MCM4 expression was also correlated with proliferation markers, Ki-67 and cyclin E expression (p < 0.001, respectively). MCM4 expression was not associated with survival. Conclusion: MCM4 may play an essential role in the proliferation of some NSCLC cells. Taken together with higher expression in NSCLCs and its correlation with clinicopathologic characteristics such as non-adenocarcinoma histology. MCM4 may have potential as a therapeutic target in certain population with NSCLCs. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
  • Satoshi Takeuchi, Rio Honma, Jun Taguchi, Toraji Amano, Yasushi Shimizu, Ichiro Kinoshita, Kanako Kubota, Yoshihiro Matsuno, Hirotoshi Dosaka-Akita
    Case Reports in Oncology 4 (2) 260 - 266 1662-6575 2011/05 [Refereed][Not invited]
     
    High-grade neuroendocrine carcinoma differs from usual neuroendocrine carcinoma, and its prognosis is dismal. In this case report, a case of high-grade neuroendocrine carcinoma that improved with bevacizumab plus modified FOLFOX6 as the fourth-line chemotherapy is presented. A 29-year-old male with a huge liver tumor was diagnosed with high-grade neuroendocrine carcinoma originating from the liver. Multiple liver and bone metastases were found one month after surgery. He was treated with three chemotherapy regimens used for the management of small-cell lung cancer with extensive disease. However, none of them could be maintained because of tumor progression. He was then treated with bevacizumab plus modified FOLFOX6 as the fourth-line regimen. Dramatic tumor shrinkage was obtained, and a partial response was achieved. This case suggests that high-grade neuroendocrine carcinoma can be treated with bevacizumab in combination with cytotoxic chemotherapy. Copyright © 2011 S. Karger AG, Basel.
  • Kenji Akie, Satoshi Oizumi, Shigeaki Ogura, Naofumi Shinagawa, Eiki Kikuchi, Shinichi Fukumoto, Masao Harada, Ichiro Kinoshita, Tetsuya Kojima, Toshiyuki Harada, Yuka Fujita, Yoshinobu Ohsaki, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Masaharu Nishimura
    ONCOLOGY 81 (2) 84 - 90 0030-2414 2011 [Not refereed][Not invited]
     
    Objective: Platinum-free regimens can represent an alternative for advanced non-small cell lung cancer (NSCLC) if similar efficacy is provided with better tolerability. This study evaluated the efficacy and safety of combined irinotecan and S-1 for chemotherapy-naive advanced NSCLC. Methods: Chemotherapy consisted of 4-week cycles of intravenous irinotecan (100 mg/m(2), days 1 and 15) and oral S-1 (80 mg/m(2), days 1-14). The primary endpoint was response rate, while secondary endpoints were overall survival, progression-free survival (PFS), and safety. Results: A total of 112 cycles was administered to 40 patients (median 3 cycles; range 1-6 cycles). Twelve patients showed partial response and 17 patients had stable disease, representing a response rate of 30% and a disease control rate of 72.5%. Median survival time and median PFS were 16.1 and 4.8 months, respectively. Hematological toxicities of grade 3 or 4 were neutropenia (32.5%) and anemia (5.0%). The most common nonhematological toxicities of grade 3 or 4 included diarrhea (15.0%) and anorexia (17.5%). Patients homo-or heterozygous for UGTA1A*6 tended to show a higher incidence of grade 3 diarrhea (p = 0.055). Conclusion: The combination of irinotecan and S-1 offers good efficacy and tolerability for previously untreated advanced NSCLC. Copyright (C) 2011 S. Karger AG, Basel
  • Kayoko Takeda, Ichiro Kinoshita, Yasushi Shimizu, Yoshihiro Matsuno, Toshiaki Shichinohe, Hirotoshi Dosaka-Akita
    ANTICANCER RESEARCH 31 (1) 263 - 270 0250-7005 2011/01 [Not refereed][Not invited]
     
    Background: The Wnt target LGR5 has been recently identified as a murine intestinal stem cell marker. Its role during each stage of human colorectal tumorigenesis remains to be determined. Patients and Methods: LGR5 expression was investigated by immunohistochemical analysis in 17 low-grade and 13 high-grade intraepithelial neoplasias and 30 adenocarcinomas. Results: The number of LGR5-positive cells increased tumor, with clustering of the cells to form patches. An apparent difference in their distribution among the tumorigenesis stages was observed. LGR5 expression in luminal surface showed a negative association with the progressive grade of tumors, while that in lower crypt exhibited a positive association with grade. In adenocarcinomas, LGR5 expression in luminal surface was negatively associated with pStage, while it was almost invariably high in lower crypt during advanced pStage disease. Conclusion: These results suggest that the shifts in the distribution of LGR5-positive cells towards the lower crypt play a role in the development and progression of colorectal cancer.
  • Jun Taguchi, Ichiro Kinoshita, Hirotoshi Akita
    Japanese Journal of Cancer and Chemotherapy 38 (4) 518 - 523 0385-0684 2011 [Refereed][Not invited]
     
    Superior vena cava (SVC) syndrome is one of the complication of malignant neoplasia, which often occurs in clinical practice. It is caused by obstruction of the SVC by invasion, extrinsic compression by adjacent pathologic processes, or by internal thrombus. They sometimes coexist. The increased venous pressure in the upper body caused by this syndrome results in edema of the head, neck, and arms, which is visually striking but generally of little clinical consequence. But edema may cause a functional compromise of the larynx or pharynx, causing dyspnea, stridor, cough, hoarseness, and dysphagia. Cerebral edema may lead to cerebral ischemia, confusion, coma, and possibly death. We therefore recognize the SVC syndrome as an oncologic emergency. The most common malignant cause of this disease is lung cancer. It is necessary to plan a management strategy after consideration of staging and the histopathologic diagnosis. The most useful imaging study for this disease is CT scanning of the chest with administration of contrast material. Management of the SVC syndrome associated with malignant disease includes both treatment of the cancer and relief of the symptoms of obstruction. Major therapeutic modalities are supportive care and medical management, including chemotherapy, radiotherapy, placement of intravascular stent, and surgery. The presence of the SVC syndrome does not reduce the likelihood of a cure for the underlying malignant condition, and should not compromise the choice of appropriate therapy.
  • Sakakibara-Konishi J, Oizumi S, Kinoshita I, Shinagawa N, Kikuchi J, Kato M, Inoue T, Katoh N, Onimaru R, Shirato H, Dosaka-Akita H, Nishimura M
    Lung Cancer 74 (2) 248 - 52 1872-8332 2011 [Refereed][Not invited]
  • 18F‐Fluorothymidine(FLT)による抗EGFR抗体薬の早期治療効果予測
    竹内啓, 趙松吉, 趙松吉, 久下裕司, 趙莞, 西嶋剣一, 波多野利行, 清水康, 木下一郎, 玉木長良, 秋田弘俊
    日本臨床腫瘍学会学術集会プログラム・抄録集 9th 367  2011 [Not refereed][Not invited]
  • 大腸発癌における腸管幹細胞マーカーLgr5発現の腫瘍内分布の変化(Distinctive distribution of expression of Lgr5, an intestinal stem cell marker in each stage of colorectal tumorigenesis)
    武田 香陽子, 木下 一郎, 清水 康, 秋田 弘俊
    日本癌学会総会記事 69回 487 - 487 0546-0476 2010/08
  • Makoto Maemondo, Akira Inoue, Kunihiko Kobayashi, Shunichi Sugawara, Satoshi Oizumi, Hiroshi Isobe, Akihiko Gemma, Masao Harada, Hirohisa Yoshizawa, Ichiro Kinoshita, Yuka Fujita, Shoji Okinaga, Haruto Hirano, Kozo Yoshimori, Toshiyuki Harada, Takashi Ogura, Masahiro Ando, Hitoshi Miyazawa, Tomoaki Tanaka, Yasuo Saijo, Koichi Hagiwara, Satoshi Morita, Toshihiro Nukiwa
    NEW ENGLAND JOURNAL OF MEDICINE 362 (25) 2380 - 2388 0028-4793 2010/06 [Not refereed][Not invited]
     
    Background: Non-small-cell lung cancer with sensitive mutations of the epidermal growth factor receptor (EGFR) is highly responsive to EGFR tyrosine kinase inhibitors such as gefitinib, but little is known about how its efficacy and safety profile compares with that of standard chemotherapy. Methods: We randomly assigned 230 patients with metastatic, non-small-cell lung cancer and EGFR mutations who had not previously received chemotherapy to receive gefitinib or carboplatin-paclitaxel. The primary end point was progression-free survival; secondary end points included overall survival, response rate, and toxic effects. Results: In the planned interim analysis of data for the first 200 patients, progression-free survival was significantly longer in the gefitinib group than in the standard-chemotherapy group (hazard ratio for death or disease progression with gefitinib, 0.36; P<0.001), resulting in early termination of the study. The gefitinib group had a significantly longer median progression-free survival (10.8 months, vs. 5.4 months in the chemotherapy group; hazard ratio, 0.30; 95% confidence interval, 0.22 to 0.41; P<0.001), as well as a higher response rate (73.7% vs. 30.7%, P<0.001). The median overall survival was 30.5 months in the gefitinib group and 23.6 months in the chemotherapy group (P=0.31). The most common adverse events in the gefitinib group were rash (71.1%) and elevated aminotransferase levels (55.3%), and in the chemotherapy group, neutropenia (77.0%), anemia (64.6%), appetite loss (56.6%), and sensory neuropathy (54.9%). One patient receiving gefitinib died from interstitial lung disease. Conclusions: First-line gefitinib for patients with advanced non-small-cell lung cancer who were selected on the basis of EGFR mutations improved progression-free survival, with acceptable toxicity, as compared with standard chemotherapy. (UMIN-CTR number, C000000376.) N Engl J Med 2010;362:2380-8.
  • Junko Kikuchi, Ichiro Kinoshita, Yasushi Shimizu, Eiki Kikuchi, Jun Konishi, Satoshi Oizumi, Kichizo Kaga, Yoshihiro Matsuno, Masaharu Nishimura, Hirotoshi Dosaka-Akita
    CANCER 116 (12) 3015 - 3024 0008-543X 2010/06 [Not refereed][Not invited]
     
    BACKGROUND: The polycomb group genes Bmi1 polycomb ring finger oncogene (Bmi1) and enhancer of zeste homolog 2 (EZH2) function as transcriptional repressors involved in gene silencing and in the malignant transformation and biologic aggressiveness of several human carcinomas. In the current study, the authors evaluated Bmi1 and EZH2 protein expression in specimens of human nonsmall cell lung cancer (NSCLC). METHODS: The authors conducted an immunohistochemical assessment of 157 surgically resected NSCLCs to evaluate the correlation between Bmi1 and EZH2 expression and various features, including clinical, clinicopathologic, and biologic characteristics. RESULTS: Normal bronchial epithelia revealed abundant expression of Bmi1 and sporadic expression of EZH2. Patients who had high EZH2 expression in tumor cells had a poorer prognosis than patients who had low EZH2 expression in tumor cells all pathologic stages of NSCLC (P=.001) and in pathologic stage I NSCLC (P=.006). Multivariate analysis revealed that high EZH2 expression was a independent, unfavorable prognostic factor in patients with pathologic stage I disease (P=.048). High EZH2 expression was correlated significantly with nonadenocarcinoma histology (P=.001), moderate and poor differentiation (P=.001), advanced pathologic tumor classification (P=.02), and high Ki-67 and cyclin E labeling indices (P<.001). Bmi1 expression, in contrast, was not a significant prognostic factor and was not correlated with any clinicopathologic factors other than early pathologic tumor classification. CONCLUSIONS: Bmi1 and EZH2 had characteristic and distinctive expression in NSCLCs. High EZH2 expression was correlated with tumor aggressiveness and may provide a novel prognostic marker for NSCLCs. Cancer 2010;116:3015-24. (C) 2010 American Cancer Society.
  • Hajime Asahina, Satoshi Oizumi, Akira Inoue, Ichiro Kinoshita, Takashi Ishida, Yuka Fujita, Noriaki Sukoh, Masao Harada, Makoto Maemondo, Yasuo Saijo, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Toshihiro Nukiwa, Masaharu Nishimura
    ONCOLOGY 79 (5-6) 423 - 429 0030-2414 2010 [Not refereed][Not invited]
     
    Objective: Salvage treatment for acquired resistance to gefitinib has yet to be developed. We conducted the first prospective phase II study of gefitinib readministration in previous gefitinib responders. Methods: Gefitinib (250 mg/day) was readministered to patients with advanced/metastatic non-small cell lung cancer who had achieved objective response to initial gefitinib and subsequently received cytotoxic chemotherapy after disease progression with initial gefitinib. The primary endpoint was the objective response rate with gefitinib readministration. Secondary endpoints were disease control rate, progression-free survival (PFS), overall survival (OS), quality of life, and toxicity. Changes in lung cancer-related symptoms were evaluated using the seven-item lung cancer subscale of the questionnaire. Results: Sixteen patients were enrolled between February 2005 and January 2008. Most had received >= 3 regimens of chemotherapy. Response and disease-control rates for all patients were 0 and 44%. Median PFS and OS were 2.5 and 14.7 months, respectively. Four of 7 patients with stable disease experienced a long duration (>= 6 months) of disease control without severe toxicity. Symptom improvement was observed in 2 of 12 patients (17%) for whom quality of life was evaluable. Conclusion: Gefitinib represents a useful therapeutic option for selected previous gefitinib responders. Copyright (C) 2011 S. Karger AG, Basel
  • ヒト癌細胞移植マウスにおける抗EGFR抗体(cetuximab)による分子標的療法:FLTを用いた早期治療効果評価
    竹内啓, 趙松吉, 久下祐司, 趙莞, 西嶋剣一, 波多野利行, 清水康, 木下一郎, 玉木長良, 秋田弘俊
    日本臨床腫瘍学会学術集会プログラム・抄録集 8th 188  2010 [Not refereed][Not invited]
  • ヒト癌細胞移植マウスにおける抗VEGF抗体(Bevacizumab)による分子標的療法:FLTを用いた早期治療効果評価
    趙松吉, 久下裕司, 趙芫, 竹内啓, 波多野利行, 西嶋剣一, 木下一郎, 秋田弘毅, 玉木長良
    核医学 46 (3) S260-S261  2009 [Not refereed][Not invited]
  • J. Kikuchi, I. Kinoshita, Y. Shimizu, S. Oizumi, M. Nishimura, M. J. Birrer, H. Dosaka-Akita
    BRITISH JOURNAL OF CANCER 99 (12) 2013 - 2019 0007-0920 2008/12 [Not refereed][Not invited]
     
    c-Jun is a major constituent of AP-1 transcription factor that transduces multiple mitogen growth signals, and it is frequently overexpressed in non-small cell lung cancers (NSCLCs). Earlier, we showed that blocking AP-1 by the overexpression of a c-Jun dominant-negative mutant, TAM67, inhibited NSCLC cell growth. The phosphatidylinositol 3-kinase (PI3K)/Akt signal transduction pathway is important in transformation, proliferation, survival and metastasis of NSCLC cells. In this study, we used NCI-H1299 Tet-on clone cells that express TAM67 under the control of inducible promoter to determine the effects of inhibition of AP-1 and PI3K on cell growth. The PI3K inhibitor, LY294002, produced a dose-dependent inhibition of growth in H1299 cells and that inhibition was enhanced by TAM67. TAM67 increased dephosphorylation of Akt induced by LY294002 and reduced the TPA response element DNA-binding of phosphorylated c-Jun. TAM67 increased G1 cell cycle blockade induced by LY294002, which was partially associated with cyclin A decrease and p27(Kip1) accumulation. Furthermore, TAM67 and LY294002 act, at least additively, to inhibit anchorage-independent growth of the H1299 cells. These results suggest that AP-1 and PI3K/Akt pathways play an essential role in the growth of some NSCLC cells.
  • Kayoko Takeda, Ichiro Kinoshita, Yasushi Shimizu, Yusuke Ohba, Tomoo Itoh, Yoshihiro Matsuno, Toshiaki Shichinohe, Hirotoshi Dosaka-Akita
    BMC CANCER 8 328  1471-2407 2008/11 [Not refereed][Not invited]
     
    Background: A recent study has shown that phosphorylated c-Jun (p-c-Jun) interacts with TCF4 to form a complex that cooperatively enhances their transcriptional activity in the presence of beta-Catenin, and that their interaction is critical for mouse intestinal tumorigenesis. To determine the significance of these three proteins in human colorectal tumors, we analyzed their nuclear expression by immunohistochemistry. Methods: we analyzed their nuclear expression by immunohistochemistry using paraffin-embedded specimens of 68 resected colorectal tumors, which consisted of 19 adenomas, 14 high-grade intraepithelial neoplasia (HGINs) and 35 adenocarcinomas. We also analyzed the expression of MMP7, which has functional AP-I and TCF binding sites in its promoter. Results: Expression of p-c-Jun, TCF4 and beta-Catenin were significantly higher in adenomas than in the adjacent normal epithelia. Expression of p-c-Jun and beta-Catenin in HGINs and adenocarcinomas were also significantly higher than in the adjacent normal epithelia. p-c-Jun expression, but not TCF4 and beta-Catenin, was higher in adenomas and HGINs than in adenocarcinomas, in which p-c-Jun expression was negatively correlated with pT stage progression. Furthermore, significant correlations of expression were observed between p-c-Jun and TCF4 ( r = 0.25, p = 0.04), TCF4 and beta-Catenin ( r = 0.30, p = 0.01), p-c-Jun and MMP7 ( r = 0.26, p = 0.03), and TCF4 and MMP7 ( r = 0.39, p = 0.0008), respectively. Conclusion: These results suggest that nuclear expression of p-c-Jun, TCF4 and beta-Catenin have important roles in human colorectal tumor development and that p-c-Jun may play a pivotal role in the earlier stages of tumor development.
  • 菊地 順子, 木下 一郎, 清水 康, 武田 香陽子, 油谷 浩幸, 大泉 聡, 西村 正治, 秋田 弘俊
    肺癌 (NPO)日本肺癌学会 48 (5) 449 - 449 0386-9628 2008/10
  • 非小細胞肺癌においてMinichromosome maintenance(MCM)4タンパク質は細胞増殖と分化のマーカーである(Minichromosome maintenance protein 4 as a marker for proliferation and differentiation in non-small cell lung cancer)
    菊地 順子, 木下 一郎, 清水 康, 武田 香陽子, 油谷 浩幸, 大泉 聡史, 西村 正治, 秋田 弘俊
    日本癌学会総会記事 67回 102 - 102 0546-0476 2008/09
  • Akita H, Kinoshita I
    Gan to kagaku ryoho. Cancer & chemotherapy 5 35 720 - 724 0385-0684 2008/05 [Refereed][Not invited]
  • Y. Shimizu, I. Kinoshita, J. Kikuchi, K. Yamazaki, M. Nishimura, M. J. Birrer, H. Dosaka-Akita
    BRITISH JOURNAL OF CANCER 98 (5) 915 - 922 0007-0920 2008/03 [Not refereed][Not invited]
     
    cJun, a major constituent of AP-1 transcription factor transducing multiple mitogen growth signals, is frequently overexpressed in non-small cell lung cancers (NSCLCs). The purpose of this study is to determine the effects of AP-1 blockade on the growth of NSCLC cells using a cJun dominant-negative mutant, TAM67. Transiently transfected TAM67 inhibited AP-1 transcriptional activity in NSCLC cell lines, NCI-H1299 (H1299), A549 and NCI-H520 (H520). The colony-forming efficiency of H1299 and A549 was reduced by TAM67, while that of H520 was not. To elucidate the effects of TAM67 on the growth of H1299, we established H1299 clone cells that expressed TAM67 under the control of a doxycycline-inducible promoter. In the H1299 clone cells, the induced TAM67 inhibited anchorage-dependent growth by promoting G1 cell-cycle block, but not by apoptosis. The induced TAM67 decreased the expression of a cell-cycle regulatory protein, cyclin A. TAM67 also inhibited anchorage-independent growth of these cells. Furthermore, TAM67 reduced growth of established xenograft tumours from these cells in nude mice. These results suggest that AP-1 plays an essential role in the growth of at least some of NSCLC cells.
  • ヒト大腸癌切除標本におけるpcJun、TCF4及びβカテニンの核発現の臨床病理学的意義(Clinicopathologic significance of nuclear expression of pcJun, TCF4 and beta-Catenin in resected human colorectal tumors)
    武田 香陽子, 木下 一郎, 清水 康, 秋田 弘俊
    日本癌学会総会記事 66回 265 - 265 0546-0476 2007/08
  • Hiroshi Yokouchi, Koichi Yamazaki, Ichiro Kinoshita, Jun Konishi, Hajime Asahina, Noriaki Sukoh, Masao Harada, Kenji Akie, Shigeaki Ogura, Takashi Ishida, Mitsuru Munakata, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Masaharu Nishimura
    BMC CANCER 7 51  1471-2407 2007/03 [Not refereed][Not invited]
     
    Background: Gefitinib, an oral agent of epidermal growth factor receptor tyrosine kinase inhibitor, has a certain efficacy against non-small cell lung cancer (NSCLC). Several predictive factors of gefitinib sensitivity have been well described. However, few studies have investigated the clinical features of gefitinib-responders. In the present study, we analyzed the response and disease progression of primary and metastatic lesions to gefitinib in responders and the results of gefitinib readministration following temporary cessation of gefitinib upon progression of initial gefitinib treatment and other treatments. Method: We retrospectively evaluated the clinical courses of 27 NSCLC patients who received gefitinib and achieved either a complete or partial response. Results: The best-response rate and disease-control rate against the initial chemotherapy for the gefitinib-responders were 27.3% and 77.3%, respectively. Favorable efficacy was observed in the primary lesion and metastases to the lung, liver and brain, while there was no obvious effect on bone metastasis. The primary lesion and intrapulmonary metastasis were the sites of major recurrence. Median progression-free survival was 13.8 months, median duration of gefitinib treatment was 17.0 months and median overall survival was 29.2 months. Some of the patients who experienced disease progression after responding to gefitinib were again sensitive to readministration of gefitinib following temporary cessation of gefitinib and other treatments. Conclusion: Patients may still be expected to have prolonged survival if they once responded to gefitinib and then underwent various subsequent treatments followed by readministration of gefitinib. These findings might provide valuable information for the management of gefitinib-responders.
  • Nobuyuki Hakuma, Tomoko Betsuyaku, Ichiro Kinoshita, Tomoo Itoh, Kichizo Kaga, Satoshi Kondo, Masaharu Nishimura, Hirotoshi Dosaka-Akita
    ONCOLOGY 72 (3-4) 197 - 204 0030-2414 2007 [Not refereed][Not invited]
     
    Objective: Extracellular matrix metalloproteinase inducer (EMMPRIN) is a highly glycosylated transmembrane protein that is widely present on the surface of various tumor cells, assisting in tumor progression by stimulating the production of several matrix metalloproteinases in adjacent stromal cells. However, its clinical relevance remains to be evaluated in lung cancers. Therefore, we aimed to investigate the relationship between EMMPRIN expression in non-small cell lung cancer (NSCLC) and clinicopathological characteristics and prognosis. Methods: EMMPRIN expression was semi-quantified by immunohistochemistry with anti-human EMMPRIN monoclonal antibody in 208 surgically resected NSCLCs and was analyzed statistically in relation to various characteristics. Results: EMMPRIN expression was seen in most NSCLC samples (92%). High levels of EMMPRIN expression were significantly associated with differentiation and pT(1) stage in adenocarcinomas. There were no significant differences in overall survival between patients with tumors having high and low levels of EMMPRIN expression in pathological stage I NSCLCs (5-year survival rates, 69 vs. 60%). Conclusions: EMMPRIN was preferentially expressed in most NSCLCs. High levels of expression were associated with early T stage and well-differentiated adenocarcinoma, and were not a prognostic factor in NSCLC. Copyright (C) 2007 S. Karger AG, Basel.
  • H. Asahina, K. Yamazaki, I. Kinoshita
    BRITISH JOURNAL OF CANCER 96 (2) 400 - 400 0007-0920 2007/01 [Not refereed][Not invited]
  • Masaru Hasegawa, Tomoko Betsuyaku, Nobuya Yoshida, Yasuyuki Nasuhara, Ichiro Kinoshita, Satoshi Ohta, Tomoo Itoh, Pyong Woo Park, Masaharu Nishimura
    RESPIROLOGY 12 (1) 140 - 143 1323-7799 2007/01 [Refereed][Not invited]
     
    Multicentric Castleman's disease (MCD) is a rare and often incurable lymphoproliferative disorder. It is typically a systemic illness, but occasionally manifests primarily as a pulmonary parenchymal disease with massive infiltration of CD138 (syndecan-1)-positive plasma cells. This is the first report to demonstrate a marked elevation of soluble CD138, despite the absence of plasma cells, in BAL fluid in an MCD patient with pulmonary involvement. This finding suggests that the quantitative measurement of soluble CD138 in BAL fluid may reflect plasma cell infiltration and disease activity in the lungs of patients with MCD.
  • Hajime Asahina, Koichi Yamazaki, Ichiro Kinoshita, Hiroshi Yokouchi, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    LUNG CANCER 54 (3) 419 - 422 0169-5002 2006/12 [Not refereed][Not invited]
     
    Mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR) are associated with clinical responsiveness to EGFR-tyrosine kinase inhibitors (EGFR-TKIs) in patients with non-small cell, lung cancers (NSCLCs). However, certain rare EGFR mutations including S7681 are reported to confer less in vitro sensitivity to gefitinib, an EGFR-TKI, than major mutations such as exon 19 deletions and L858R and even the wild-type counterpart. Here, we report the first case of adenocarcinoma of the lung in which the patient had rare mutations S7681 and V769L and was treated with gefitinib. Disease progressed during 6 weeks of treatment. This case suggests that in vitro sensitivity to gefitinib correlates with distinct clinical responsiveness to gefitinib in various types of EGFR mutations. (C) 2006 Elsevier Ireland Ltd. All rights reserved.
  • Hajime Asahina, Koichi Yamazaki, Ichiro Kinoshita, Hiroshi Yokouchi, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    LUNG CANCER 54 (3) 419 - 422 0169-5002 2006/12 [Refereed][Not invited]
     
    Mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR) are associated with clinical responsiveness to EGFR-tyrosine kinase inhibitors (EGFR-TKIs) in patients with non-small cell, lung cancers (NSCLCs). However, certain rare EGFR mutations including S7681 are reported to confer less in vitro sensitivity to gefitinib, an EGFR-TKI, than major mutations such as exon 19 deletions and L858R and even the wild-type counterpart. Here, we report the first case of adenocarcinoma of the lung in which the patient had rare mutations S7681 and V769L and was treated with gefitinib. Disease progressed during 6 weeks of treatment. This case suggests that in vitro sensitivity to gefitinib correlates with distinct clinical responsiveness to gefitinib in various types of EGFR mutations. (C) 2006 Elsevier Ireland Ltd. All rights reserved.
  • Eiki Kikuchi, Ichiro Kinoshita, Koichi Yamazaki, Tomoo Itoh, Tadamichi Shimizu, Hiroshi Shimizu, Masaharu Nishimura
    RESPIROLOGY 11 (6) 826 - 829 1323-7799 2006/11 [Refereed][Not invited]
     
    A 39-year-old Japanese woman presented with a swollen right hand and a right-sided pneumothorax. Chest CT revealed bilateral multiple pulmonary thin-walled cysts measuring <= 1 cm in diameter and small nodules. An initial skin biopsy led to a misdiagnosis of metastatic adenocarcinoma, as tumour cells were positive for cytokeratin, epithelial membrane antigen, carcinoembryonic antigen and cancer antigen 125. However, chemotherapy proved ineffective, and the skin biopsy was repeated. A final diagnosis of epithelioid sarcoma (ES) was made. Open lung biopsy showed that the pulmonary nodules represented metastases of ES. Although the pulmonary cyst walls did not contain tumour cells, bronchiolar wall adjacent to the cysts had been infiltrated by tumour cells. These findings suggested that pulmonary cysts, a rare form of pulmonary metastases from soft tissue sarcomas, had developed through a ball-valve effect of metastatic tumour in small airways. However, presence of cancer antigen 125 hindered obtaining a correct diagnosis of ES.
  • H. Asahina, K. Yamazaki, I. Kinoshita, N. Sukoh, M. Harada, H. Yokouchi, T. Ishida, S. Ogura, T. Kojima, Y. Okamoto, Y. Fujita, H. Dosaka-Akita, H. Isobe, M. Nishimura
    BRITISH JOURNAL OF CANCER 95 (8) 998 - 1004 0007-0920 2006/10 [Not refereed][Not invited]
     
    Retrospective analysis has shown that activating mutations in exons 18-21 of the epidermal growth factor receptor ( EGFR) gene are a predictor of response to gefitinib. We conducted a phase II trial to evaluate the efficacy and safety of gefitinib as first-line therapy for advanced non-small cell lung cancer (NSCLC) with EGFR mutations. Patients with stage IIIB or IV chemotherapy- naive NSCLC with EGFR mutation were treated with 250 mg gefitinib daily. For mutational analysis, DNA was extracted from paraffin-embedded tissues and EGFR mutations were analysed by direct sequence of PCR products. Twenty (24%) of the 82 patients analysed had EGFR mutations ( deletions in or near E746-A750, n = 16; L858R, n 4). Sixteen patients were enrolled and treated with gefitinib. Twelve patients had objective response and response rate was 75% (95% CI, 48-93%). After a median follow-up of 12.7 months ( range, 3.1-16.8 months), 10 patients demonstrated disease progression, with median progression-free survival of 8.9 months ( 95% CI, 6.7-11.1 months). The median overall survival time has not yet been reached. Most of the toxicities were mild. This study showed that gefitinib is very active and well tolerated as first-line therapy for advanced NSCLC with EGFR mutations.
  • Megumi Nakadate, Koichi Yamazaki, Jun Konishi, Ichiro Kinoshita, Noriaki Sukoh, Masao Harada, Kenji Akie, Shigeaki Ogura, Takashi Ishida, Mitsuru Munakata, Hirotoshi Dosaka-Akita, Hiroshi Isobe, Masaharu Nishimura
    ANTICANCER RESEARCH 26 (5B) 3767 - 3772 0250-7005 2006/09 [Not refereed][Not invited]
     
    Background: The optimal schedule of taxane administration has been an area of active interest in several clinical trials. Patients and Methods: To evaluate the efficacy and toxicity of carboplatin and weekly paclitaxel combination chemotherapy, a phase H study was conducted for chemo-naive, advanced non-small cell lung cancer (NSCLC) patients. Patients received paclitaxel 100 mg/m(2) on days 1, 8 and 15, and carboplatin with the target dose of area under the curve of 6 on day I every 28 days. Results: Forty patients were enrolled. Overall response rate and survival at one year by intent-to-treat analyses was 35% and 57.5%, respectively. The median survival time was 12.2 months. Twenty-two patients (56%) had grade 3 or greater neutropenia. Grade 3 sensory and motor neuropathy were seen in one patient (3%). Conclusion: Carboplatin and weekly paclitaxel combination chemotherapy is an active and feasible regimen for patients with advanced NSCLC.
  • Hiroshi Yokouchi, Koichi Yamazaki, Hajime Asahina, Masahiko Shigemura, Takanori Moriyama, Kazuo Takaoka, Jun Moriya, Tomoo Itoh, Ichiro Kinoshita, Hirotoshi Dosaka-Akita, Yutaka Tsutsumi, Masaharu Nishimura
    ANTICANCER RESEARCH 26 (4B) 2821 - 2827 0250-7005 2006/07 [Not refereed][Not invited]
     
    Background: Few studies have successfully established an amylase-producing lung cancer cell line or have examined its cytological, biochemical and biological features. Patients and Methods: Cancer cells, isolated from pleural effusion using a gradient method, were cultivated. Results: Amylase production from the newly established cell line was confirmed by positive staining for a-amylase and increased amylase levels in the culture supernatant. Electron microscopy revealed zymogen granule-like structures. Sialylation of salivary-type amylase was confirmed directly from the cell line by examining the neuraminidase sensitivity and amylase elution profile under high-performance liquid chromatography. Neither EGFR or KRAS mutation were found. Conclusion: This cell line offers a useful tool for analyzing the pathogenesis and pathophysiology of amylase-producing lung cancers. Moreover, it might be useful for probing the metastasis and invasiveness of lung cancer cells and for developing an early diagnostic method based on sialylated salivary-amylase production.
  • N Shinagawa, K Yamazaki, Kinoshita, I, S Ogura, M Nishimura
    RESPIRATION 73 (1) 90 - 94 0025-7931 2006 [Refereed][Not invited]
     
    Background: Fiber-optic flexible bronchoscopy (FFB) is a frequently performed procedure for the diagnosis and treatment of pulmonary disorders. However, hypoxemia occasionally occurs during FFB. Objectives: We attempted to examine the causes of arterial oxygen desaturation during FFB. Methods: We studied 336 patients who underwent FFB without intervention between June 1, 2001 and September 30, 2002. Arterial oxygen saturation (SpO(2)) was continuously monitored using oximetry with a recording system. We analyzed the relationship between a reduction in SpO(2) during FFB and various clinical parameters or background lung diseases. Results: Of the 336 patients, 73 (22%) had an episode of oxygen desaturation (SpO(2) < 90% over 10 s). Of patients over 80 years old, 55% had an episode of oxygen desaturation, which was significantly higher than 27% observed in the patients of 80 and less than 80 years old (p < 0.05). Patients with pulmonary fibrosis had a higher risk of desaturation (55%) compared to patients with other complications or patients without any complication ( p < 0.05). Multivariable analysis revealed that both age and pulmonary fibrosis were independent predictors of oxygen desaturation. However, the majority of the patients (94%) did not require routine oxygen supplementation. Conclusion: Although FFB is safe and does not require oxygen supplementation in most cases, age over 80 years and pulmonary fibrosis are high risk factors for significant oxygen desaturation during FFB. Copyright (C) 2006 S. Karger AG, Basel.
  • N Hakuma, Kinoshita, I, Y Shimizu, K Yamazaki, K Yoshida, M Nishimura, H Dosaka-Akita
    CANCER RESEARCH 65 (23) 10776 - 10782 0008-5472 2005/12 [Not refereed][Not invited]
     
    E1AF/PEA3, an Ets family transcription factor, is frequently overexpressed in non-small-cell lung cancers (NSCLCs). Overexpression of E1AF increases motility and invasion of VMRC-LCD and NCI-H226 NSCLC cells, which lack endogenous E1AF expression, and the effect is synergistically increased by hepatocyte growth factor (HGF). The.small GTPase Rho/Rho-associated kinase (ROCK) pathway is also involved in motility and invasion. To determine the role of the Rho/ROCK pathway in malignant phenotypes induced by E1AF, we analyzed VMRC-LCD cells transfected with an EIAF expression vector (LCD-E1AF cells) or with empty vector (LCD-vector cells). LCD-E1AF cells had more GTP-bound (active) Rho than LCD-vector cells and Rho activation was synergistically increased by HGF. The Rho activation by E1AF and HGF was also shown in NCI-H226 cells. Phosphorylation of myosin light chain (MLC), a downstream effector of ROCK signaling, was higher in LCD-E1AF cells than in LCD-vector cells, especially under HGF treatment. A specific ROCK inhibitor, Y27632, strongly suppressed MLC phosphorylation, cell motility, and invasion. In nude mice implanted s.c. and intrapulmonarily, LCD-E1AF cells made more local tumors than LCD-vector cells (six of six versus one of seven mice and four of seven versus one of seven mice, respectively). Three of the four mice with lung tumors from LCD-E1AF cells had lymph node metastases whereas the mouse with LCD-vector tumors did not. LCD-E1AF tumors showed higher MLC phosphorylation than LCD-vector tumors. These results suggest that EIAF activates the Rho/ROCK pathway in an HGF-enhanced manner and its activation is important in E1AF-induced motility and invasion as well as tumorigenesis and metastasis in NSCLC cells. (Cancer Res 2005; 65(23): 10776-82).
  • N Hakuma, Kinoshita, I, Y Shimizu, K Yamazaki, K Yoshida, M Nishimura, H Dosaka-Akita
    CANCER RESEARCH 65 (23) 10776 - 10782 0008-5472 2005/12 [Refereed][Not invited]
     
    E1AF/PEA3, an Ets family transcription factor, is frequently overexpressed in non-small-cell lung cancers (NSCLCs). Overexpression of E1AF increases motility and invasion of VMRC-LCD and NCI-H226 NSCLC cells, which lack endogenous E1AF expression, and the effect is synergistically increased by hepatocyte growth factor (HGF). The.small GTPase Rho/Rho-associated kinase (ROCK) pathway is also involved in motility and invasion. To determine the role of the Rho/ROCK pathway in malignant phenotypes induced by E1AF, we analyzed VMRC-LCD cells transfected with an EIAF expression vector (LCD-E1AF cells) or with empty vector (LCD-vector cells). LCD-E1AF cells had more GTP-bound (active) Rho than LCD-vector cells and Rho activation was synergistically increased by HGF. The Rho activation by E1AF and HGF was also shown in NCI-H226 cells. Phosphorylation of myosin light chain (MLC), a downstream effector of ROCK signaling, was higher in LCD-E1AF cells than in LCD-vector cells, especially under HGF treatment. A specific ROCK inhibitor, Y27632, strongly suppressed MLC phosphorylation, cell motility, and invasion. In nude mice implanted s.c. and intrapulmonarily, LCD-E1AF cells made more local tumors than LCD-vector cells (six of six versus one of seven mice and four of seven versus one of seven mice, respectively). Three of the four mice with lung tumors from LCD-E1AF cells had lymph node metastases whereas the mouse with LCD-vector tumors did not. LCD-E1AF tumors showed higher MLC phosphorylation than LCD-vector tumors. These results suggest that EIAF activates the Rho/ROCK pathway in an HGF-enhanced manner and its activation is important in E1AF-induced motility and invasion as well as tumorigenesis and metastasis in NSCLC cells. (Cancer Res 2005; 65(23): 10776-82).
  • M Katabami, H Donninger, F Hommura, VD Leaner, Kinoshita, I, JFB Chick, MJ Birrer
    JOURNAL OF BIOLOGICAL CHEMISTRY 280 (17) 16728 - 16738 0021-9258 2005/04 [Not refereed][Not invited]
     
    Overexpression of c-Jun enables Rat1a cells to grow in an anchorage-independent manner. We used an inducible c-Jun system under the regulation of doxycycline in Rat1a cells to identify potential c-Jun target genes necessary for c-Jun-induced anchorage-independent growth. Induction of c-Jun results in sustained expression of cyclin A in the nonadherent state with only minimal expression in the absence of c-Jun. The promoter activity of cyclin A2 was 4-fold higher in Rat1a cells in which c-Jun expression was induced compared with the control cells. Chromatin immunoprecipitation demonstrated that c-Jun bound directly to the cyclin A2 promoter. Mutation analysis of the cyclin A2 promoter mapped the c-Jun regulatory site to an ATF site at position - 80. c-Jun was able to bind to this site both in vitro and in vivo, and mutation of this site completely abolished promoter activity. Cyclin A1 was also elevated in c-Jun-overexpressing Rat1a cells; however, c-Jun did not regulate this gene directly, since it did not bind directly to the cyclin A1 promoter. Suppression of cyclin A expression via the introduction of a cyclin A antisense sequences significantly reduced the ability of c-Jun-overexpressing Rat1a cells to grow in an anchorage-independent fashion. Taken together, these results suggest that cyclin A is a target of c-Jun and is necessary but not sufficient for c-Jun-induced anchorage-independent growth. In addition, we demonstrated that the cytoplasmic oncogenes Ras and Src transcriptionally activated the cyclin A2 promoter via the ATF site at position - 80. Using a dominant negative c-Jun mutant, TAM67, we showed that this transcriptional activation of cyclin A2 requires c-Jun. Thus, our results suggest that c-Jun is a mediator of the aberrant cyclin A2 expression associated with Ras/Src-induced transformation.
  • M Katabami, H Donninger, F Hommura, VD Leaner, Kinoshita, I, JFB Chick, MJ Birrer
    JOURNAL OF BIOLOGICAL CHEMISTRY 280 (17) 16728 - 16738 0021-9258 2005/04 [Refereed][Not invited]
     
    Overexpression of c-Jun enables Rat1a cells to grow in an anchorage-independent manner. We used an inducible c-Jun system under the regulation of doxycycline in Rat1a cells to identify potential c-Jun target genes necessary for c-Jun-induced anchorage-independent growth. Induction of c-Jun results in sustained expression of cyclin A in the nonadherent state with only minimal expression in the absence of c-Jun. The promoter activity of cyclin A2 was 4-fold higher in Rat1a cells in which c-Jun expression was induced compared with the control cells. Chromatin immunoprecipitation demonstrated that c-Jun bound directly to the cyclin A2 promoter. Mutation analysis of the cyclin A2 promoter mapped the c-Jun regulatory site to an ATF site at position - 80. c-Jun was able to bind to this site both in vitro and in vivo, and mutation of this site completely abolished promoter activity. Cyclin A1 was also elevated in c-Jun-overexpressing Rat1a cells; however, c-Jun did not regulate this gene directly, since it did not bind directly to the cyclin A1 promoter. Suppression of cyclin A expression via the introduction of a cyclin A antisense sequences significantly reduced the ability of c-Jun-overexpressing Rat1a cells to grow in an anchorage-independent fashion. Taken together, these results suggest that cyclin A is a target of c-Jun and is necessary but not sufficient for c-Jun-induced anchorage-independent growth. In addition, we demonstrated that the cytoplasmic oncogenes Ras and Src transcriptionally activated the cyclin A2 promoter via the ATF site at position - 80. Using a dominant negative c-Jun mutant, TAM67, we showed that this transcriptional activation of cyclin A2 requires c-Jun. Thus, our results suggest that c-Jun is a mediator of the aberrant cyclin A2 expression associated with Ras/Src-induced transformation.
  • Y Ishibashi, H Dosaka-Akita, E Miyoshi, M Shindoh, M Miyamoto, Kinoshita, I, H Miyazaki, T Itoh, S Kondo, M Nishimura, N Taniguchi
    ONCOLOGY 69 (4) 301 - 310 0030-2414 2005 [Not refereed][Not invited]
     
    Objective: N-Acetylglucosaminyltransferase V (GnT-V) is a key enzyme in the formation of branching asparagine-linked oligosaccharides and is linked to tumor invasion and metastasis in colon and breast cancers. In normal esophageal epithelium, beta 1,6-branched asparagine-linked oligosaccharides synthesized by GnT-V are seen in the basal cell layers but not in the superficial cell layers, and its presence has been shown in invasive esophageal cancers. However, neither GnT-V expression nor its clinical significance has been previously examined in human normal, premalignant and malignant esophageal tissues. Methods: GnT-V expression was studied by immunohistochemistry using a specific monoclonal antibody in 121 surgically resected specimens of esophageal squamous cell carcinomas (SCCs) and adjacent tissues, and was analyzed statistically in relation to various characteristics. Results: GnT-V expression was observed in none (0%) of the 19 normal epithelial tissues, 1 (2%) of the 43 hyperplastic tissues, 30 (54%) of the 56 mildly dysplastic tissues, 27 (63%) of the 43 moderately dysplastic tissues, 21 (44%) of the 48 in situ SCCs and 29 (26%) of the 110 invasive SCCs ( p < 0.005). GnT-V expression was observed significantly more frequently in mildly and moderately dysplastic tissues when compared with normal epithelial and hyperplastic tissues ( p < 0.005), and its frequency was decreased in in situ and invasive SCCs ( p < 0.005). GnT-V expression was frequently observed in SCCs of small size and without distant metastasis or lymph node metastasis. Conclusions: Increased expression of GnT-V is associated with the early event of esophageal tumorigenesis. Copyright (C) 2005 S. Karger AG, Basel.
  • 「気管支肺胞洗浄所見の変化を追跡したgefitinibによる間質性肺炎の1例」
    『日本呼吸器学会雑誌』 43巻 466 - 70 2005 [Not refereed][Not invited]
  • J Konishi, K Yamazaki, Kinoshita, I, H Isobe, S Ogura, S Sekine, T Ishida, R Takashima, M Nakadate, S Nishikawa, T Hattori, H Asahina, M Imura, E Kikuchi, J Kikuchi, N Shinagawa, H Yokouchi, M Munakata, H Dosaka-Akita, M Nishimura
    ANTICANCER RESEARCH 25 (1B) 435 - 441 0250-7005 2005/01 [Refereed][Not invited]
     
    Background: Gefitinib is an oral agent that inhibits the tyrosine kinase of epidermal growth factor receptor (EGFR), which had antitumor activity in patients with previously treated non-small cell lung cancer (NSCLC). We analyzed the efficacy, toxicity and overall survival time of gefitinib treatment in patients with NSCLC. Patients and Methods: One hundred and twenty-two patients with NSCLC, who received gefitinib between 2002 and 2004 in our institutes, were evaluated retrospectively. Results: The objective response rate was 24.6%. The variables identified as significant in univariate analysis included gender and smoking habit. The median overall survival time was 14.4 months. Significant variables associated with improved survival included good performance status (PS), female, adenocarcinoma and never smoked status, while never smoked status and good PS were independent prognostic factors in multivariate analysis. Four patients (3.3%) developed interstitial pneumonitis associated with gefitinib. Conclusion: Gefitinib showed favorable antitumor activity in females, never smokers and adenocarcinoma.
  • Y Ishibashi, H Dosaka-Akita, E Miyoshi, M Shindoh, M Miyamoto, Kinoshita, I, H Miyazaki, T Itoh, S Kondo, M Nishimura, N Taniguchi
    ONCOLOGY 69 (4) 301 - 310 0030-2414 2005 [Refereed][Not invited]
     
    Objective: N-Acetylglucosaminyltransferase V (GnT-V) is a key enzyme in the formation of branching asparagine-linked oligosaccharides and is linked to tumor invasion and metastasis in colon and breast cancers. In normal esophageal epithelium, beta 1,6-branched asparagine-linked oligosaccharides synthesized by GnT-V are seen in the basal cell layers but not in the superficial cell layers, and its presence has been shown in invasive esophageal cancers. However, neither GnT-V expression nor its clinical significance has been previously examined in human normal, premalignant and malignant esophageal tissues. Methods: GnT-V expression was studied by immunohistochemistry using a specific monoclonal antibody in 121 surgically resected specimens of esophageal squamous cell carcinomas (SCCs) and adjacent tissues, and was analyzed statistically in relation to various characteristics. Results: GnT-V expression was observed in none (0%) of the 19 normal epithelial tissues, 1 (2%) of the 43 hyperplastic tissues, 30 (54%) of the 56 mildly dysplastic tissues, 27 (63%) of the 43 moderately dysplastic tissues, 21 (44%) of the 48 in situ SCCs and 29 (26%) of the 110 invasive SCCs ( p < 0.005). GnT-V expression was observed significantly more frequently in mildly and moderately dysplastic tissues when compared with normal epithelial and hyperplastic tissues ( p < 0.005), and its frequency was decreased in in situ and invasive SCCs ( p < 0.005). GnT-V expression was frequently observed in SCCs of small size and without distant metastasis or lymph node metastasis. Conclusions: Increased expression of GnT-V is associated with the early event of esophageal tumorigenesis. Copyright (C) 2005 S. Karger AG, Basel.
  • E Kikuchi, K Yamazaki, N Sukoh, J Kikuchi, H Asahina, M Imura, Y Onodera, N Kurimoto, Kinoshita, I, M Nishimura
    EUROPEAN RESPIRATORY JOURNAL 24 (4) 533 - 537 0903-1936 2004/10 [Not refereed][Not invited]
     
    The usefulness of endobronchial ultrasonography (EBUS) with guide-sheath (GS) as a guide for transbronchial biopsy (TBB) for diagnosing peripheral pulmonary lesions (PPL)s and for improving diagnostic accuracy was evaluated in this study. EBUS-GS-guided TBB was performed in 24 patients with 24 PPLs of less than or equal to30 mm in diameter (average diameter=18.4 mm). A 20-MHz radial-type ultrasound probe, covered with GS was inserted via a working bronchoscope channel and advanced to the PPL in order to produce an EBUS image. The probe with the GS was confirmed to reach the lesion by EBUS imaging and X-ray fluoroscopy. When the lesion was not identified on the EBUS image, the probe was removed and a curette was used to lead the GS to the lesion. After localising the lesion, the probe was removed, and TBB and bronchial brushing were performed via the GS. Nineteen peripheral lesions (79.2%) were visualised by EBUS. All patients whose PPLs were visible on EBUS images subsequently underwent an EBUS-GS-guided diagnostic procedure. A total of 14 lesions (58.3%) were diagnosed. Even when restricted to PPLs <20 mm in diameter, the diagnostic sensitivity was 53%. In conclusion, endobronchial ultrasonography with guide sheath-guided transbronchial biopsy was feasible and effective for diagnosing peripheral pulmonary lesions.
  • Junko Kikuchi, Koichi Yamazaki, Ichiro Kinoshita, Hajime Asahina, Mikado Imura, Eiki Kikuchi, Jun Konishi, Naofumi Shinagawa, Hiromitsu Oki, Hirotoshi Dosaka-Akita, Masaharu Nishimura
    Japanese Journal of Clinical Oncology 34 (9) 505 - 509 0368-2811 2004/09 [Not refereed][Not invited]
     
    Objectives: This study was designed to determine the maximum tolerated dose of paclitaxel administered weekly in combination with carboplatin and to assess its dose limiting toxicity and preliminary activity in patients with previously untreated, advanced non-small-cell lung cancer. Methods: Carboplatin was administered at a fixed dose that maintained a n area under the curve of 6. Paclitaxel was given over 1 h once a week for 3 weeks starting at 60 mg/m2 and escalated in 10 mg/m2 increments. Results: Twenty-one patients were treated with six dose levels (60, 70, 80, 90, 100, 110 mg/m2) of paclitaxel. The dose limiting toxicity was infection and the maximum tolerated dose was 110 mg/m2. Nine of 21 (42.9%) patients demonstrated a therapeutic response. Conclusion: Weekly paclitaxel and carboplatin were well toler ated. Based on our results, 100 mg/m2 of paclitaxel for 3 weeks of a 4-week cycle, in combination with carboplatin, was recommended for a phase II study. © 2004 Foundation for Promotion of Cancer Research.
  • J Kikuchi, K Yamazaki, Kinoshita, I, H Asahina, M Imura, E Kikuchi, J Konishi, N Shinagawa, H Oki, H Dosaka-Akita, M Nishimura
    JAPANESE JOURNAL OF CLINICAL ONCOLOGY 34 (9) 505 - 509 0368-2811 2004/09 [Refereed][Not invited]
     
    Objective: This study was designed to determine the maximum tolerated dose of paclitaxel administered weekly in combination with carboplatin and to assess its dose limiting toxicity and preliminary activity in patients with previously untreated, advanced non-small-cell lung cancer. Methods: Carboplatin was administered at a fixed dose that maintained an area under the curve of 6. Paclitaxel was given over 1 h once a week for 3 weeks starting at 60 mg/m(2) and escalated in 10 mg/m(2) increments. Results: Twenty-one patients were treated with six dose levels (60, 70, 80, 90, 100, 110 mg/m(2)) of paclitaxel. The dose limiting toxicity was infection and the maximum tolerated dose was 110 mg/m(2). Nine of 21 (42.9%) patients demonstrated a therapeutic response. Conclusion: Weekly paclitaxel and carboplatin were well tolerated. Based on our results, 100 mg/m(2) of paclitaxel for 3 weeks of a 4-week cycle, in combination with carboplatin, was recommended for a phase II study.
  • J Konishi, K Yamazaki, M Azuma, Kinoshita, I, H Dosaka-Akita, M Nishimura
    CLINICAL CANCER RESEARCH 10 (15) 5094 - 5100 1078-0432 2004/08 [Refereed][Not invited]
     
    Purpose: B7-H1/PD-L1 (B7-H1) and B7-DC/PD-L2 (B7-DC) are ligands for the receptor PD-1, which is known to negatively regulate T-cell activation. In the present study, we investigated the expression of B7-H1 and B7-DC in tumor specimens of non-small cell lung cancer and their relationships with clinicopathological variables and postoperative survival. Furthermore, we examined the correlation between B7-H1 expression on tumor cells and the number of tumor-infiltrating lymphocytes (TILs) or PD-1 expression on TILs. Experimental Design: The expression of B7-H1 and B7-DC in 52 surgically resected specimens of non-small cell lung cancer was evaluated immunohistochemically. Results: Expression of B7-H1 and B7-DC was focally observed in all non-small cell lung cancer tumor specimens. No relationship was found between the expression of B7-H1 or B7-DC and clinicopathological variables or postoperative survival. However, in the same sections evaluated, significantly fewer TILs were identified in B7-H1-positive tumor regions than in B7-H1-negative tumor regions in a subset of five patients (P = 0.01). Moreover, the percentage of TILs expressing PD-1 was significantly lower in B7-H1-positive tumor regions than in B7-H1-negative tumor regions (P = 0.02). Conclusions: The expression of B7-H1 on tumor cells in local areas reciprocally correlated with the number of TILs, and this may contribute to negative regulation in antitumor immune responses in non-small cell lung cancer.
  • J Konishi, K Yamazaki, K Chikai, K Nagashima, K Sakai, Kinoshita, I, H Dosaka-Akita, M Nishimura
    INTERNAL MEDICINE 43 (7) 602 - 606 0918-2918 2004/07 [Not refereed][Not invited]
     
    We report a case of a 71-year-old man who presented with cerebellar dysfunction. He was diagnosed as having squamous cell carcinoma of the lung (T2N3M0, Stage HI,). No anti-onconeural antibodies were found in his serum. Cerebral spinal fluid (CSF) examination showed mild mononuclear pleocytosis alone. Magnetic resonance imaging (MRI) of the brain and spinal cord revealed no abnormalities. At autopsy, there was complete disappearance of Purkinje cells with reactive astrocytosis. These findings are compatible with paraneoplastic cerebellar degeneration (PCD). To our knowledge, no case of PCD has been reported previously in patients with squamous cell carcinoma of the lung.
  • J Konishi, K Yamazaki, K Chikai, K Nagashima, K Sakai, Kinoshita, I, H Dosaka-Akita, M Nishimura
    INTERNAL MEDICINE 43 (7) 602 - 606 0918-2918 2004/07 [Refereed][Not invited]
     
    We report a case of a 71-year-old man who presented with cerebellar dysfunction. He was diagnosed as having squamous cell carcinoma of the lung (T2N3M0, Stage HI,). No anti-onconeural antibodies were found in his serum. Cerebral spinal fluid (CSF) examination showed mild mononuclear pleocytosis alone. Magnetic resonance imaging (MRI) of the brain and spinal cord revealed no abnormalities. At autopsy, there was complete disappearance of Purkinje cells with reactive astrocytosis. These findings are compatible with paraneoplastic cerebellar degeneration (PCD). To our knowledge, no case of PCD has been reported previously in patients with squamous cell carcinoma of the lung.
  • Akita H D, Miyoshi E, Suzuki O, Itoh T, Kinoshita I, Yamazaki K, Nishimura M, Katoh H, Taniguchi N
    Clninical Cancer Res. 10, 1773-1779 2004/05 [Not refereed][Not invited]
  • K Takamura, Y Nasuhara, M Kobayashi, T Betsuyaku, Y Tanino, Kinoshita, I, E Yamaguchi, S Matsukura, RP Schleimer, M Nishimura
    AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY 286 (4) L777 - L785 1040-0605 2004/04 [Not refereed][Not invited]
     
    Retinoic acid (RA) is known to accelerate wound healing and induce cell differentiation. All-trans RA (ATRA) exerts its effect by binding retinoic acid receptors, which are members of the nuclear receptor family. We investigated whether RA can alter expression of eotaxin, a potent eosinophil chemoattractant that is regulated by the transcription factors signal transducer and activator of transcription 6 (STAT6) and NF-kappaB. We examined the effects of RA on eotaxin expression in a human bronchial epithelial cell line BEAS-2B. ATRA and its stereodimer 9-cis retinoic acid (9-cis RA) inhibited IL-4-induced release of eotaxin at 10(-6) M by 78.0 and 52.0%, respectively (P < 0.05). ATRA and 9-cis RA also significantly inhibited IL-4-induced eotaxin mRNA expression at 10(-6) M by 52.3 and 53.5%, respectively (P < 0.05). In contrast, neither ATRA nor 9-cis RA had any effects on TNF-alpha-induced eotaxin production. In transfection studies using eotaxin promoter luciferase plasmids, the inhibitory effect of ATRA on IL-4-induced eotaxin production was confirmed at the transcriptional level. Interestingly, ATRA had no effects on IL-4-induced tyrosine phosphorylation, nuclear translocation, or DNA binding activity of STAT6. Activating protein-1 was not involved in ATRA-mediated transrepression of eotaxin with IL-4 stimulation. The mechanism of the inhibitory effect of ATRA on IL-4-induced eotaxin production in human bronchial epithelial cells has not been elucidated but does not appear to be due to an effect on STAT6 activation. These findings raise the possibility that RA may reduce eosinophilic airway inflammation, one of the prominent pathological features of allergic diseases such as bronchial asthma.
  • K Takamura, Y Nasuhara, M Kobayashi, T Betsuyaku, Y Tanino, Kinoshita, I, E Yamaguchi, S Matsukura, RP Schleimer, M Nishimura
    AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY 286 (4) L777 - L785 1040-0605 2004/04 [Refereed][Not invited]
     
    Retinoic acid (RA) is known to accelerate wound healing and induce cell differentiation. All-trans RA (ATRA) exerts its effect by binding retinoic acid receptors, which are members of the nuclear receptor family. We investigated whether RA can alter expression of eotaxin, a potent eosinophil chemoattractant that is regulated by the transcription factors signal transducer and activator of transcription 6 (STAT6) and NF-kappaB. We examined the effects of RA on eotaxin expression in a human bronchial epithelial cell line BEAS-2B. ATRA and its stereodimer 9-cis retinoic acid (9-cis RA) inhibited IL-4-induced release of eotaxin at 10(-6) M by 78.0 and 52.0%, respectively (P < 0.05). ATRA and 9-cis RA also significantly inhibited IL-4-induced eotaxin mRNA expression at 10(-6) M by 52.3 and 53.5%, respectively (P < 0.05). In contrast, neither ATRA nor 9-cis RA had any effects on TNF-alpha-induced eotaxin production. In transfection studies using eotaxin promoter luciferase plasmids, the inhibitory effect of ATRA on IL-4-induced eotaxin production was confirmed at the transcriptional level. Interestingly, ATRA had no effects on IL-4-induced tyrosine phosphorylation, nuclear translocation, or DNA binding activity of STAT6. Activating protein-1 was not involved in ATRA-mediated transrepression of eotaxin with IL-4 stimulation. The mechanism of the inhibitory effect of ATRA on IL-4-induced eotaxin production in human bronchial epithelial cells has not been elucidated but does not appear to be due to an effect on STAT6 activation. These findings raise the possibility that RA may reduce eosinophilic airway inflammation, one of the prominent pathological features of allergic diseases such as bronchial asthma.
  • Taniguchi N, Shinagawa N, Kinoshita I, Nasuhara Y, Yamazaki K, Yamaguchi E, Akita H, Nishimura M
    Nihon Kokyuki Gakkai zasshi = the journal of the Japanese Respiratory Society 2 42 158 - 163 1345-9538 2004/02 [Refereed][Not invited]
  • 「パクリタキセル投与との関連が示唆された薬剤性肺炎の1例」
    『日本呼吸器学会雑誌』 42 158 - 163 2004 [Not refereed][Not invited]
  • Masayuki Suga, Koichi Yamazaki, Kunio Hamada, Ichiro Kinoshita, Hirotoshi Akita, Masaharu Nishimura
    Japanese Journal of Lung Cancer 44 (1) 61 - 66 0386-9628 2004 [Not refereed][Not invited]
     
    Background. Remitting seronegative symmetrical synovitis with pitting edema (RS3PE) syndrome is characterized as a sudden onset symmetrical polyarthritis together with pitting edema of the hands, feet and legs which mainly affects the elderly. This syndrome responds to low dose steroids, and does not relapse. Recently, some cases of malignancy, although rarely lung cancer, have been reported in association with RS3PE syndrome. Case. An 83-year-old man had been followed up with RS3PE syndrome since April 2002. He was referred to our hospital because of an abnormal shadow in the left upper lung field in January 2003. Chest computed tomography (CT) revealed a 2.5 cm nodule in the S1+2 segment of the left lobe. He was diagnosed as having squamous cell carcinoma of the lung, T4N0M1, stage IV. He was treated with radiation therapy for brain metastasis and he is now being followed up as an outpatient. Conclusion. We report a rare case of squamous cell carcinoma of the lung in association with RS3PE syndrome. Considering previous report in the past literature, RS3PE syndrome may have been a prodrome of lung cancer in this case.
  • O Wakabayashi, K Yamazaki, S Oizumi, F Hommura, Kinoshita, I, S Ogura, H Dosaka-Akita, M Nishimura
    CANCER SCIENCE 94 (11) 1003 - 1009 1347-9032 2003/11 [Not refereed][Not invited]
     
    We investigated intratumoral tumor-infiltrating lymphocytes (TILs), including CD4(+) and CD8(+) T cells, in non-small cell lung cancers (NSCLCs) and their relationships with clinicopathological variables and post-operative survival. Tumor specimens from 178 NSCLCs were consecutively obtained by surgery at the Hokkaido University Medical Hospital between 1976 and 1994. CD8(+) T cells, CD4(+) T cells and Ki-67/CD8(+) T cells were visualized immunohistochemically, and counted within cancer cell nests and in cancer stroma. CD8(+) T cells and CD4(+) T cells were observed at higher frequencies within cancer cell nests in moderately and poorly differentiated tumors compared with well differentiated tumors (P<0.01), and in tumors with high Ki-67 expression compared with low Ki-67 expression (P<0.01), that showed severe cellular atypia and a higher growth rate. Patients with higher numbers of CD8(+) T cells within cancer cell nests showed significantly shorter survival times compared to those with lower numbers of CD8(+) T cells within cancer cell nests (5-year survival rates, 47% and 60%, respectively; P=0.03). Moreover, patients with higher labeling index of Ki-67/CD+ T cells showed significantly shorter survival than those with lower labeling index of Ki-67/CD8(+) T cells within cancer cell nests (5-year survival rates, 41% and 69%, respectively; P=0.02), and the labeling index of Ki-67/CD8(+) T cells within cancer cell nests was found to be a significant and independent unfavorable prognostic factor by multivariate analysis (P=0.01). On the other hand, higher numbers of CD4(+) T cells in cancer stroma, but not within cancer cell nests, were correlated with longer survival times in patients with NSCLC (5-year survival rates, 64% and 43%, respectively; P=0.04). CD4(+) T cells in cancer stroma might reflect immune responses against cancer cells, while CD8(+) T cells do not appear to work as effectors in tumor tissues of NSCLC. Moreover, the higher labeling index of Ki-67/ CD8(+) T cells within cancer cell nests is a strong indicator of unfavorable clinical outcome.
  • O Wakabayashi, K Yamazaki, S Oizumi, F Hommura, Kinoshita, I, S Ogura, H Dosaka-Akita, M Nishimura
    CANCER SCIENCE 94 (11) 1003 - 1009 1347-9032 2003/11 [Refereed][Not invited]
     
    We investigated intratumoral tumor-infiltrating lymphocytes (TILs), including CD4(+) and CD8(+) T cells, in non-small cell lung cancers (NSCLCs) and their relationships with clinicopathological variables and post-operative survival. Tumor specimens from 178 NSCLCs were consecutively obtained by surgery at the Hokkaido University Medical Hospital between 1976 and 1994. CD8(+) T cells, CD4(+) T cells and Ki-67/CD8(+) T cells were visualized immunohistochemically, and counted within cancer cell nests and in cancer stroma. CD8(+) T cells and CD4(+) T cells were observed at higher frequencies within cancer cell nests in moderately and poorly differentiated tumors compared with well differentiated tumors (P<0.01), and in tumors with high Ki-67 expression compared with low Ki-67 expression (P<0.01), that showed severe cellular atypia and a higher growth rate. Patients with higher numbers of CD8(+) T cells within cancer cell nests showed significantly shorter survival times compared to those with lower numbers of CD8(+) T cells within cancer cell nests (5-year survival rates, 47% and 60%, respectively; P=0.03). Moreover, patients with higher labeling index of Ki-67/CD+ T cells showed significantly shorter survival than those with lower labeling index of Ki-67/CD8(+) T cells within cancer cell nests (5-year survival rates, 41% and 69%, respectively; P=0.02), and the labeling index of Ki-67/CD8(+) T cells within cancer cell nests was found to be a significant and independent unfavorable prognostic factor by multivariate analysis (P=0.01). On the other hand, higher numbers of CD4(+) T cells in cancer stroma, but not within cancer cell nests, were correlated with longer survival times in patients with NSCLC (5-year survival rates, 64% and 43%, respectively; P=0.04). CD4(+) T cells in cancer stroma might reflect immune responses against cancer cells, while CD8(+) T cells do not appear to work as effectors in tumor tissues of NSCLC. Moreover, the higher labeling index of Ki-67/ CD8(+) T cells within cancer cell nests is a strong indicator of unfavorable clinical outcome.
  • VD Leaner, Kinoshita, I, MJ Birrer
    ONCOGENE 22 (36) 5619 - 5629 0950-9232 2003/08 [Not refereed][Not invited]
     
    To investigate the role of individual Jun proteins in cell growth and transformation, we have used a doxycycline-inducible retroviral vector to regulate their expression in rat fibroblasts. AP-1 complexes enriched with cJun and JunB result in morphological alterations and anchorage-independent cell growth consistent with a transformation-like phenotype, whereas complexes enriched with JunD had an antiproliferative effect. These results suggest that genes regulated by both cJun and JunB are potentially involved in transformation and that they can be distinguished from those regulated by AP-1 complexes containing JunD. To identify genes regulated by cJun and JunB that may have a role in anchorage-independent growth, we investigated differential gene expression by each of the Jun family members using the Affymetrix Rat oligonucleotide microarray, RG_U34A containing approximately 8000 genes. Differentially regulated genes were identified and grouped for correlation with regulation by the different Jun proteins. A total of 33 candidate genes were found to be differentially regulated by both cJun and JunB and not by JunD. These genes have roles in cell metabolism, growth, signal transduction, migration and adhesion. We validated the differential regulation by cJun and JunB of 10 candidate genes by Northern blot analysis. Of these, eight were further characterized as potential direct targets of AP-1 regulation based on Northern blot results showing differential regulation that correlate with cJun expression. Our results show that inducible cJun and JunB expression result in anchorage-independent growth of Rat1a cells, distinct from JunD-expressing cells. This model system and a functional genomic approach enabled us to differentiate AP-1-regulated genes involved in transformation from AP-1-regulated genes known as bystander genes. This approach significantly reduces the number of bystanders and allows for the targeting of genes specifically involved in transformation.
  • T Kojima, K Yamazaki, Y Tamura, S Ogura, K Tani, J Konishi, N Shinagawa, Kinoshita, I, N Hizawa, E Yamaguchi, H Dosaka-Akita, M Nishimura
    HUMAN GENE THERAPY 14 (8) 715 - 728 1043-0342 2003/05 [Not refereed][Not invited]
     
    Granulocyte-macrophage colony-stimulating factor (GM-CSF)-based cancer cell vaccines have been shown to be potent inducers of antitumor immunity in several murine models, but the antitumor effects on established tumors have been minimal. Conversely, the major role of the heat shock protein gp96, localized in the endoplasmic reticulum ( ER), is to act as a molecular chaperone to assist the folding of nascent polypeptide chains in the ER. gp96 derived from tumor cells elicits specific protective immunity against parental tumors, presumably through the transport of tumor-specific peptides to antigen-presenting cells and the maturation of dendritic cells ( DCs). However, the therapeutic effects of tumor-derived gp96 on established tumors have not been promising. The present study analyzes the therapeutic effects of GM-CSF gene-transduced Lewis lung cancer (LLC/GM) cells combined with LLC-derived gp96 on established wild-type LLC tumors in immunocompetent C57BL/6 mice. Therapy with either irradiated LLC/GM cells or LLC-derived gp96 barely affected established LLC tumor growth. The antitumor effect was significantly enhanced when 1 mug of LLC-derived gp96 was administered together with 1 3 106 irradiated LLC/GM cells (p < 0.05). The antitumor effects of irradiated LLC/GM cells and LLC-derived gp96 required mainly CD8(+) T cells. Spleen cells obtained from mice vaccinated with irradiated LLC/GM cells and LLC-derived gp96 showed specific CD8 cytotoxic activities against LLC cells (specific lysis rate of approximately 28%). This antibody response was not associated with a synergic effect of the combination therapy. Moreover, draining lymph nodes from mice immunized with irradiated LLC/GM cells and LLC-derived gp96 contained more migrating mature CD11c(+) cells ( higher levels of CD86 and major histocompatibility complex [MHC] class II molecules) compared with those from any other immunization protocols. These results suggest that the combination of irradiated LLC/GM cells and tumor-derived gp96 has potential as a new immunogene therapeutic strategy against lung cancer.
  • Kinoshita, I, Leaner, V, M Katabami, RG Manzano, P Dent, A Sabichi, MJ Birrer
    ONCOGENE 22 (18) 2710 - 2722 0950-9232 2003/05 [Not refereed][Not invited]
     
    cJun is a major component of the transcription factor AP-1 and mediates a diverse set of biologic properties including proliferation, differentiation, and apoptosis. To identify cJun-responsive genes, we inducibly expressed cJun in Rat-1a cells and observed two distinct phenotypes: changes in cellular morphology with adherent growth and anchorage-independent growth. The biologic effects of cJun were entirely reversible demonstrating that they require the continued presence of cJun. To determine the genes, which mediate the biologic effects of cJun, we employed multiple methods including differential gene analysis, suppression subtractive hybridization, and cDNA microarrays. We identified 38 cJun-responsive genes including three uncharacterized genes under adherent and/or nonadherent conditions. Half of the known 36 genes were cytoskeleton- and adhesion-related genes, suggesting a major role of cJun in the regulation of the genes related to cell morphology. As proof of the principle that this approach could identify genes whose upregulation was necessary for nonadherent growth, we investigated one gene, stathmin whose upregulation by cJun was observed only under these conditions. Although overexpression of stathmin did not result in nonadherent growth, inhibition of stathmin protein expression by antisense oligonucleotides in cJun-induced Rat-1a cells prevented nonadherent growth. These results suggest that stathmin plays an essential role in anchorage-independent growth by cJun and may be a potential target for specific inhibitors for AP-1-dependent processes involved in carcinogenesis.
  • T Harada, S Ogura, K Yamazaki, Kinoshita, I, T Itoh, H Isobe, K Yamashiro, H Dosaka-Akita, M Nishimura
    CANCER SCIENCE 94 (4) 394 - 399 1347-9032 2003/04 [Not refereed][Not invited]
     
    Chemoresistance is a major problem in the chemotherapy of non-small cell lung cancers (NSCLCs). Several mechanisms are thought to be involved in drug resistance, including those associated with apoptosis, drug transport and detoxification. Here, we investigated the predictive value of P53, Bcl-2 and lung resistance-related protein (LRP) expression for response to platinum-based chemotherapy, using transbronchial biopsy (TBB) specimens from patients with NSCLC. We evaluated TBB specimens from 57 patients with NSCLC who had not previously been treated with either chemotherapy or radiotherapy before TBB, and who were treated with systemic platinum-based chemotherapy. The specimens included 33 adenocarcinomas, 22 squamous cell carcinomas and two large cell carcinomas. One to 6 courses of chemotherapy were administered. Expression of P53, Bcl-2 and LRP was analyzed by immunohistochemistry using TBB specimens. Positive expression of P53, Bcl-2 and LRP was observed in 28 (49%), 41 (71%) and 42 (73%) of the 57 NSCLCs, respectively. P53 expression correlated significantly with response to chemotherapy in nonsquamous cell carcinomas, including adenocarcinomas and large cell carcinomas (response rates, 38% and 6% for patients with P53-positive and P53-negative tumors, respectively, P=0.03). LRP expression significantly correlated inversely with response to chemotherapy in squamous cell carcinomas (response rates, 33% and 100% for patients with LRP-positive and LRP-negative tumors, respectively, P=0.02). Bcl-2 expression did not correlate with response to chemotherapy. These findings indicate that immunostaining for P53 and LRP using TBB specimens may be useful for dividing patients with NSCLC into chemoresponsive and chemoresistant groups.
  • Watanabe K, Shimizu Y, Oizumi S, Shinagawa N, Kinoshita I, Yamazaki K, Onodera Y, Saito H, Yamaguchi E, Dosaka-Akita H, Nishimura M
    Nihon Kokyuki Gakkai zasshi = the journal of the Japanese Respiratory Society 2 41 107 - 111 1345-9538 2003/02 [Refereed][Not invited]
  • WATANABE Kaori, SHIMIZU Yasushi, OIZUMI Satoshi, SHINAGAWA Naofumi, KINOSHITA Ichiro, YAMAZAKI Koichi, ONODERA Yuya, SAITO Hiroshi, YAMAGUCHI Etsuro, DOSAKA-AKITA Hirotoshi, NISHIMURA Masaharu
    日本呼吸器学会雑誌 41 (2) 107 - 111 1343-3490 2003 [Not refereed][Not invited]
  • Hakuma Nobuyuki, Yamazaki Koichi, Fukumoto Shinichi, Yokouchi Hiroshi, Harada Toshiyuki, Wakabayashi Osamu, Kinoshita Ichiro, Ogura Shigeaki, Dosaka-Akita Hirotoshi, Nishimura Masaharu
    The Journal of the Japan Society for Respiratory Endoscopy 特定非営利活動法人 日本呼吸器内視鏡学会 25 (2) 114 - 117 0287-2137 2003 [Not refereed][Not invited]
     
    Background, Argon plasma coagulation (APC) has been mainly used to treat gastrointestinal hemorrhage, but has recently also been applied to the treatment of airway stenosis. Case. A 48-year-old woman who had undergone right nephrectomy because of renal cell carcinoma at age 40 complained of dyspnea and cough in April 2000. Bronchoscopic examination revealed severe stenosis of the left main bronchus due to endobronchial metastasis of renal cell carcinoma and stenosis of the right truncus intermedius due to extrinsic compression caused by right hilar lymph nodal metastasis. The tumor in the left main bronchus was removed by forceps under bronchoscopy with the use of APC. Then, a Z stent could be successfully inserted into the right truncus intermedius, which led to the dramatic improvement of her symptoms. Conclusion. The use of APC was effective for the treatment of airway stenosis in this case. (JJSB. 2003 ; 25 : 114-117)
  • J Konishi, K Yamazaki, E Tsukamoto, N Tamaki, Y Onodera, T Otake, T Morikawa, Kinoshita, I, H Dosaka-Akita, M Nishimura
    RESPIRATION 70 (5) 500 - 506 0025-7931 2003 [Not refereed][Not invited]
     
    Background: Accurate staging of mediastinal and hilar lymph nodes is a critical factor determining operability in patients with non-small cell lung cancer (NSCLC). Positron emission tomography with 2-[F-18]fluoro-2-deoxy-D-glucose as a tracer (FDG-PET) has recently been reported to be more effective in detecting tumor involvement in mediastinal and hilar lymph nodes than computed tomography (CT). Objective: In this study, we analyzed the accuracy of FDG-PET in mediastinal and hilar lymph node staging in patients with NSCLC and the factors associated with false-positive or false-negative FDG-PET findings in mediastinal and hilar lymph node staging. Methods: Fifty-four patients with NSCLC who underwent preoperative analysis including chest CT and whole-body FDG-PET were evaluated retrospectively. Using FDG-PET, lesions were considered to be positive if a definite, localized area of higher uptake, excluding physiologic uptake, than in surrounding normal tissue was present. On CT findings, lymph nodes were considered to be positive if they were 110 mm in short-axis diameter, except subcarinal lymph nodes (#7), which were considered to be positive if they were >15 mm in short-axis diameter. All patients underwent surgical resection of primary tumors and mediastinal and hilar lymph nodes between 1999 and 2001 in our institute. Resected lymph nodes were histologically examined for the existence of tumor cells. Results: A total of 306 lymph nodes were resected and used for analysis. The sensitivity, specificity, positive predictive value and negative predictive value of FDG-PET were 73, 98, 70 and 98%, while those of CT were 55, 96, 55 and 96%, respectively. When preoperative nodal staging was compared with post-operative histopathological staging, 44 patients (81%) were correctly staged, 7 (13%) were overstaged and 3 (6%) were understaged by FDG-PET, while 39 patients (72%) were correctly staged, 8 (15%) were overstaged and 7 (13%) were understaged by CT. All 7 overstaged patients by FDG-PET had other pulmonary complications, including interstitial pneumonitis (n = 2), previous pulmonary tuberculosis (n = 3), silicosis (n = 1) and emphysema ( n = 1), although they were not in the active stage. In 3 understaged patients by FDG-PET, lymph nodes were also undetectable by CT. Conclusion: FDG-PET is superior to CT in mediastinal and hilar lymph node staging of patients with NSCLC. However, care should be taken in lymph node staging for patients who have other pulmonary complications, including interstitial pneumonitis, previous pulmonary tuberculosis and silicosis. Copyright (C) 2003 S. Karger AG, Basel.
  • J Konishi, K Yamazaki, E Tsukamoto, N Tamaki, Y Onodera, T Otake, T Morikawa, Kinoshita, I, H Dosaka-Akita, M Nishimura
    RESPIRATION 70 (5) 500 - 506 0025-7931 2003 [Refereed][Not invited]
     
    Background: Accurate staging of mediastinal and hilar lymph nodes is a critical factor determining operability in patients with non-small cell lung cancer (NSCLC). Positron emission tomography with 2-[F-18]fluoro-2-deoxy-D-glucose as a tracer (FDG-PET) has recently been reported to be more effective in detecting tumor involvement in mediastinal and hilar lymph nodes than computed tomography (CT). Objective: In this study, we analyzed the accuracy of FDG-PET in mediastinal and hilar lymph node staging in patients with NSCLC and the factors associated with false-positive or false-negative FDG-PET findings in mediastinal and hilar lymph node staging. Methods: Fifty-four patients with NSCLC who underwent preoperative analysis including chest CT and whole-body FDG-PET were evaluated retrospectively. Using FDG-PET, lesions were considered to be positive if a definite, localized area of higher uptake, excluding physiologic uptake, than in surrounding normal tissue was present. On CT findings, lymph nodes were considered to be positive if they were 110 mm in short-axis diameter, except subcarinal lymph nodes (#7), which were considered to be positive if they were >15 mm in short-axis diameter. All patients underwent surgical resection of primary tumors and mediastinal and hilar lymph nodes between 1999 and 2001 in our institute. Resected lymph nodes were histologically examined for the existence of tumor cells. Results: A total of 306 lymph nodes were resected and used for analysis. The sensitivity, specificity, positive predictive value and negative predictive value of FDG-PET were 73, 98, 70 and 98%, while those of CT were 55, 96, 55 and 96%, respectively. When preoperative nodal staging was compared with post-operative histopathological staging, 44 patients (81%) were correctly staged, 7 (13%) were overstaged and 3 (6%) were understaged by FDG-PET, while 39 patients (72%) were correctly staged, 8 (15%) were overstaged and 7 (13%) were understaged by CT. All 7 overstaged patients by FDG-PET had other pulmonary complications, including interstitial pneumonitis (n = 2), previous pulmonary tuberculosis (n = 3), silicosis (n = 1) and emphysema ( n = 1), although they were not in the active stage. In 3 understaged patients by FDG-PET, lymph nodes were also undetectable by CT. Conclusion: FDG-PET is superior to CT in mediastinal and hilar lymph node staging of patients with NSCLC. However, care should be taken in lymph node staging for patients who have other pulmonary complications, including interstitial pneumonitis, previous pulmonary tuberculosis and silicosis. Copyright (C) 2003 S. Karger AG, Basel.
  • H Dosaka-Akita, Kinoshita, I, K Yamazaki, H Izumi, T Itoh, H Katoh, M Nishimura, K Matsuo, Y Yamada, K Kohno
    BRITISH JOURNAL OF CANCER 87 (7) 751 - 755 0007-0920 2002/09 [Not refereed][Not invited]
     
    N-acetylgalactosaminyl transferase-3 (GalNAc-T3) is an enzyme involved in the initial glycosylation of mucin-type O-linked proteins. In the present study, we used immunohistochemistry to examine GalNAc-T3 expression in 2 15 surgically resected non-small cell lung cancers. We analysed the biological and clinical importance of GalNAc-T3 expression, especially with regard to its potential as a prognostic factor. We found that normal bronchial epithelial cells, bronchial gland cells, and alveolar pneumocytes showed cytoplasmic immunostaining for GalNAc-T3. Low expression of GalNA&T3, observed in 93 of 215 tumours (43.4%), was found more frequently in tumours from smokers than those from nonsmokers (P = 0.001), in squamous cell carcinomas than nonsquamous cell carcinomas (P < 0.0001), and in moderately and poorly differentiated tumours than well differentiated tumours (P = 0.0002). Multivariate logistic regression analysis showed that an association of low GalNAc-T3 expression with squamous cell carcinomas was the only one significant relationship of GalNA&T3 expression with various factors (P < 0.0001). Moreover, tumours losing GalNAc-T3 expression had a significantly higher Ki-67 labelling index than tumours retaining GalNA&T3 expression (P = 0.0003). Patients with low GalNAc-T3 expression survived a significantly shorter time than patients with high GalNAc-T3 expression in 103 pStage I non-small cell lung cancers (5-year survival rates, 58% and 78%, respectively; P = 0.02 by log-rank test) as well as in 6 1 pStage I nonsquamous cell carcinomas (5-year survival rates, 63% and 85%, respectively; P = 0.03). Low GalNAc-T3 expression was an unfavourable prognostic factor in pStage I non-small cell lung cancers (hazards ratio, 2.04; P = 0.03), and in pStage I nonsquamous cell carcinomas (hazards ratio, 2,70; P = 0.03). These results suggest that GalNAc-T3 is a new marker of non-small cell lung cancers with specificity for histology and prognosis. (C) 2002 Cancer Research UK.
  • RG Manzano, LM Montuenga, M Dayton, P Dent, Kinoshita, I, S Vicent, GJ Gardner, PM Nguyen, YH Choi, J Trepel, N Auersperg, MJ Birrer
    ONCOGENE 21 (28) 4435 - 4447 0950-9232 2002/06 [Not refereed][Not invited]
     
    Although early stage ovarian cancer can be effectively treated with surgery and chemotherapy, the majority of cases present with advanced disease, which remains essentially incurable. Unfortunately, little is known about the genes important for the development and progression of this disease. In this study, the expression of 68 phosphatases was determined in immortalized ovarian epithelial cells (IOSE) and compared to ovarian cancer cell lines. CL100, a dual specificity phosphatase, displayed 10-25-fold higher expression in normal compared to malignant ovarian cell lines. Immunohistochemical staining of normal ovaries and 68 ovarian cancer specimens confirmed this differential expression. Re-expression of CL100 in ovarian cancer cells decreased adherent and nonadherent cell growth and induced phenotypic changes including loss of filopodia and lamellipodia with an associated decrease in cell motility. Induced expression of CL100 in ovarian cancer cells suppressed intraperitoneal tumor growth in nude mice. These results show for the first time that CL100 expression is altered in human ovarian cancer, that CL100 expression changes cell morphology and motility, and that it suppresses intraperitoneal growth of human ovarian epithelial cancer. These data suggest that down-regulation of CL100 may play a role in the progression of human ovarian cancer.
  • RG Manzano, LM Montuenga, M Dayton, P Dent, Kinoshita, I, S Vicent, GJ Gardner, PM Nguyen, YH Choi, J Trepel, N Auersperg, MJ Birrer
    ONCOGENE 21 (28) 4435 - 4447 0950-9232 2002/06 [Refereed][Not invited]
     
    Although early stage ovarian cancer can be effectively treated with surgery and chemotherapy, the majority of cases present with advanced disease, which remains essentially incurable. Unfortunately, little is known about the genes important for the development and progression of this disease. In this study, the expression of 68 phosphatases was determined in immortalized ovarian epithelial cells (IOSE) and compared to ovarian cancer cell lines. CL100, a dual specificity phosphatase, displayed 10-25-fold higher expression in normal compared to malignant ovarian cell lines. Immunohistochemical staining of normal ovaries and 68 ovarian cancer specimens confirmed this differential expression. Re-expression of CL100 in ovarian cancer cells decreased adherent and nonadherent cell growth and induced phenotypic changes including loss of filopodia and lamellipodia with an associated decrease in cell motility. Induced expression of CL100 in ovarian cancer cells suppressed intraperitoneal tumor growth in nude mice. These results show for the first time that CL100 expression is altered in human ovarian cancer, that CL100 expression changes cell morphology and motility, and that it suppresses intraperitoneal growth of human ovarian epithelial cancer. These data suggest that down-regulation of CL100 may play a role in the progression of human ovarian cancer.
  • F Hommura, K Furuuchi, K Yamazaki, S Ogura, Kinoshita, I, M Shimizu, T Moriuchi, H Katoh, M Nishimura, H Dosaka-Akita
    CANCER 94 (3) 752 - 758 0008-543X 2002/02 [Not refereed][Not invited]
     
    BACKGROUND. beta-Catenin has been shown to function as a Wnt signaling molecule to stimulate cyclin D1 expression and cell growth in several kinds of tumors. METHODS. The authors immunohistochemically examined specimens of 217 surgically resected primary nonsmall cell lung carcinomas (NSCLCs) for beta-catenin expression and classified them semiquantitatively into three categories, including those with high, moderate, and low scores of expression. RESULTS. High, moderate, and low scores of expression were found in 37 (17.1%), 145 (66.8%), and 35 (16.1%) tumors, respectively. beta-Catenin expression was not correlated with cyclin D1 expression, but was positively correlated with the Ki-67 cell growth fraction (P = 0.04). The direct sequencing analysis for the beta-catenin gene mutation of 13 specimens of 217 tumors for the current study revealed no mutations. The relation between survival and beta-catenin expression was evaluated in 148 potentially curatively resected tumors with pathologic Stages I-IIIA. A trend toward better survival was found in patients with tumors having higher scores. In multivariate analysis, high beta-catenin expression was a significant and independent favorable prognostic factor (hazards ratio, 0.31; P = 0.007) as was pathologic stage. Analyzed by cell type, in nonsquamous cell carcinomas, patients with tumors having high scores survived a significantly longer time than those with tumors having moderate or low scores (5-year survival rates, 84%, 55%, and 32%, respectively; P = 0.02), and high beta-catenin expression tended to be a favorable prognostic factor (hazards ratio, 0.32; P = 0.052). CONCLUSIONS. These results indicate that, in NSCLCs, increased expression of beta-catenin can predict favorable Prognosis of patients with resected tumors, suggesting that accumulation of beta-catenin has no or little oncogenic effect via activation of the Wnt pathway, unlike in colon carcinomas or hepatomas. Cancer 2002;94:752-8. (C) 2002 American Cancer Society.
  • F Hommura, H Dosaka-Akita, Kinoshita, I, T Mishina, H Hiroumi, S Ogura, H Katoh, Y Kawakami
    BRITISH JOURNAL OF CANCER 81 (4) 696 - 701 0007-0920 1999/10 [Not refereed][Not invited]
     
    The predictive value of expression of p16(INK4A), retinoblastoma (Rb) and p53 proteins for prognosis was evaluated in 76 patients with non-small-cell lung cancers (NSCLCs) that were potentially curatively resected between 1990 and 1995, using the results of immunostaining analyses of these proteins as reported in our previous study (Kinoshita et al, 1996). Of these NSCLCs, 22 (29%) lacked p16 protein expression and eight (11%) Rb protein, while 30 (39%) showed positive (altered) p53 protein expression. Survival of patients with p16-negative tumours was not significantly different from that of patients with p16-positive tumours (5-year survival rates 67% and 72% respectively, P = 0.8), nor was survival of patients with Rb-negative tumours significantly different from that of patients with Rb-positive tumours (5-year survival rates 42% and 69% respectively, P = 0.9). Moreover, survival of patients with p16/Rb-negative (either p16- or Rb-negative) tumours was not significantly different from that of patients with p16/Rb-positive (both p16- and Rb-positive) tumours (5-year survival rates 67% and 68% respectively, P = 0.7). In contrast, survival of patients with p53-positive (altered) tumours tended to be shorter than that of patients with p53-negative (unaltered) tumours (5-year survival rates 56% and 78% respectively, P = 0.06). In univariate analysis of potential prognostic factors, p16, Rb and p16/Rb proteins were not significant prognostic factors in the present cohort of potentially curatively resected NSCLCs. Altered p53 protein status tended to be a negative prognostic factor (P = 0.06 by the univariate analysis). These results indicate that loss of p16 protein alone, or in combination with loss of Rb protein, does not predict the clinical outcome of patients with resected NSCLCs. (C) 1999 Cancer Research Campaign.
  • T Mishina, H Dosaka-Akita, Kinoshita, I, F Hommura, T Morikawa, H Katoh, Y Kawakami
    BRITISH JOURNAL OF CANCER 80 (8) 1289 - 1295 0007-0920 1999/06 [Not refereed][Not invited]
     
    Cyclin D1, like p16(INK4) (p16) and retinoblastoma (RB) proteins, participates in the cell cycle control at the GI-S transition. We have previously demonstrated altered p16 and RE protein status in non-small-cell lung cancers (NSCLCs) and their potential synergistic effect with altered p53 protein on proliferative activity (Kinoshita et al (1996) Cancer Res 56. 5557-5562). In the present study, cyclin D1 expression was studied by immunohistochemistry in the same cohort of 111 resected NSCLCs as in our previous study, and the amount of the cyclin D1 gene was analysed by Southern blot analysis in 29 NSCLCs. Cyclin D1 expression was analysed in relation to the status of p53, p16 and RE proteins, and proliferative activity determined by the Ki-67 index. It was also analysed in relation to survival of 77 patients with NSCLCs which were potentially curatively resected between 1990 and 1995. We found that: (1) cyclin D1 was expressed in 13 (11.7%) of 111 NSCLCs; (2) the cyclin D1 gene was neither significantly amplified nor rearranged; (3) cyclin D1 expression significantly correlated with altered p53 protein expression (P = 0.04), whereas it did not correlate with p16 and RE protein status, (4) proliferative activity tended to be higher in cyclin D1-positive (+) tumours than in cyclin D1-negative (-) tumours, although this difference was not statistically significant (P = 0.08); and (5) patients with cyclin D1+ tumours survived longer than patients with cyclin D1- tumours (5-year survival rates, 89% and 64% respectively, by the Kaplan-Meier method; P = 0.045 by the log-rank test), and cyclin D1 expression tended to be a favourable prognostic factor (P = 0.08 in univariate analysis). These findings suggest the involvement of cyclin D1 in the development and progression of NSCLCs, their proliferative activity and clinical outcome of NSCLC patients.
  • H Dosaka-Akita, F Hommura, H Fujita, Kinoshita, I, M Nishi, T Morikawa, H Katoh, Y Kawakami, N Kuzumaki
    CANCER RESEARCH 58 (2) 322 - 327 0008-5472 1998/01 [Not refereed][Not invited]
     
    Most lung and bladder cancers have been shown to be associated with smoking. We have previously demonstrated the frequent loss of gelsolin expression and its tumor suppressor activity in bladder cancer (M. Tanaka et al., Cancer Res., 55: 3228-3232, 1995). Here, we examined gelsolin expression in 12 cultured non-small cell lung cancer (NSCLC) cell lines. Furthermore, we analyzed gelsolin expression in relation to patients' smoking habits in 88 surgically resected NSCLCs to investigate whether gelsolin could be a molecular target for tobacco-induced carcinogenesis of lung cancer. All 12 NSCLC cell lines showed low-to-undetectable expression of the gelsolin gene, compared to that in normal lung tissue, by Northern blot analysis. On the other hand, Southern blot analysis of genomic DNA did not show any gross rearrangements or deletions of the gene in the NSCLC cell lines. Western blot analysis of gelsolin expression showed low-to-undetectable gelsolin expression in all 12 NSCLC cell lines, compared to normal lung tissue. Immunocytochemical analysis of gelsolin expression in NSCLC cell lines showed results that were consistent with those obtained by Western blot analysis, using normal bronchial epithelial cells as a positive control: two cell lines with lower gelsolin expression by Western blot analysis had reduced but positive cytoplasmic immunostaining of gelsolin, compared with primary normal bronchial epithelial cells, whereas no such immunostaining was observed in two cell lines with much lower or undetectable gelsolin expression by Western blot analysis. Therefore, gelsolin expression was analyzed in surgically resected NSCLCs by immunohistochemistry. Reduced or undetectable gelsolin expression was observed in 48 of 88 (55%) resected NSCLCs. Such altered gelsolin expression significantly correlated with heavy smoking of patients (greater than or equal to 20 pack-years; P = 0.008 by the chi(2) test and P = 0.03 by multivariate logistic regression analysis), whereas there was no significant correlation between gelsolin expression and histological type, pathological tumor-node-metastasis (pTNM) stage, or survival. These findings suggest that the frequent loss of gelsolin expression may be involved in the development of NSCLCs as a potential molecular target of tobacco-induced carcinogenesis.
  • H DosakaAkita, SX Hu, M Fujino, M Harada, Kinoshita, I, HJ Xu, N Kuzumaki, Y Kawakami, WF Benedict
    CANCER 79 (7) 1329 - 1337 0008-543X 1997/04 [Not refereed][Not invited]
     
    BACKGROUND. Inactivation of the retinoblastoma (Rb) gene has been documented in various types of cancer, including lung cancer. Alterations of the p53 and ms genes are also common features in the molecular biology of lung carcinoma, and tile authors of this article have reported previously on the prognostic significance of both of them. In the present study, the authors evaluated the prognostic signifi cance of the loss of Rb protein expression alone, then performed a combined analysis of Rb protein and ms p21 status (Rb/ras) as well as an analysis of Rb and p53 protein status (Rb/p53) in patients with nonsmall cell lung cancer (NSCLC). METHODS. Ninety-one patients with NSCLC underwent potentially curative resection between 1977 and 1988, 65 of whom received postoperative combination chemotherapy. Tumor specimens were analyzed for Rb protein expression by immunohistochemistry. Univariate and multivariate analyses were performed to assess the association between Rb protein expression and survival. RESULTS. Nineteen (21%) of the 91 NSCLCs showed negative Rb protein expression. Positive or negative Rb protein expression (Rb+ or Rb-) as an individual factor was not statistically correlated with survival or prognosis in this cohort of NSCLC patients, although a tendency among Rb- patients to do worse was observed. The authors then combined the Rb protein status with previously studied results of ms p21 and p53 protein expression in the same tumor specimens, and compared the prognosis between the individuals with theoretically the best pattern of gene expression in their tumors and those with theoretically the worst pattern of expression, i.e., Rb+/ras- versus Rb-/ras+ and Rb+/p53-versus Rb-/p53+. In patients with adenocarcinoma, those with Rb-/ras+ tumors survived for a significantly shorter period after surgery (13% 5-year survival) than those with Rb+/ras- tumors (82% 5-year survival) (P = 0.01). Similarly, patients with Rb-/p53+ tumors survived for a significantly shorter period (20% 5-year survival) compared with those who had Rb+/p53- tumors (73% 5-year survival) (P = 0.008). Rb/ras status was a significant prognostic factor (P = 0.02 by univariate analysis, P = 0.048 by multivariate analysis), and Rb/p53 status tended to be significant as a prognostic factor (P = 0.04 by univariate analysis, P = 0.08 by multivariate analysis). In patients with squamous cell carcinoma, neither Rb/ras nor Rb/p53 status was a significant prognostic factor in this cohort. CONCLUSIONS. These results suggest that combined immunohistochemical analyses of Rb and ms p21 proteins and of Rb and p53 proteins may indicate their potentially synergistic effects on survival and prognosis. These analyses may also be useful for stratifying patients with adenocarcinoma of the lung into different prognostic groups and identifying populations with different risks of recurrence. Larger prospective studies with Stage I NSCLC patients are necessary to confirm the current findings. (C) 1997 American Cancer Society.
  • Kinoshita, I, H DosakaAkita, T Mishina, K Akie, M Nishi, H Hiroumi, F Hommura, Y Kawakami
    CANCER RESEARCH 56 (24) 5557 - 5562 0008-5472 1996/12 [Not refereed][Not invited]
     
    p16(INK4) protein (p16) and retinoblastoma protein (pRB), like p53 protein, are important tumor suppressors that regulate the cell cycle. We immunohistochemically examined fresh-frozen specimens of 114 resected non-small cell lung cancers (NSCLCs) for loss of p16 and pRB expression, together with aberrant accumulation of p53 protein and the proliferative activity determined by the Ki-67 index. Three pRB-positive tumors were uninterpretable for p16 status. Of the remaining 111 tumors, 30 (27%) lacked p16 expression, and 10 (9%) lost pRB expression, No tumors showed coincident loss of both proteins, supporting the hypothesis that they function in a single pathway. Of 25 tumors, including 4 p16-negative tumors, examined by Southern blot analysis, only 2 p16-negative tumors were considered to have reduced gene dosage consistent with possible homozygous deletion of the CDKN2 gene encoding p16, suggesting that immunohistochemistry is a sensitive and suitable method to screen for p16 alteration. Loss of p16 expression did not correlate with any clinical factors or p53 status, whereas loss of pRB expression correlated with heavy smoking (P=0.03 by Fisher's exact test and P=0.01 by the multivariate logistic regression analysis). Proliferative activity was considerably higher in p53-positive tumors than in p53-negative tumors (P <0.001). Loss of p16 or pRB expression was associated with a further increase in proliferative activity in the p53-positive tumors (P=0.009) but not with proliferative activity in the p53-negative tumors. These results suggest that alteration of the p16/pRB pathway is relatively frequently involved in the development and progression of NSCLCs and that its effect on the proliferative activity is potentially synergistic with altered p53 protein.
  • M FUJINO, H DOSAKAAKITA, M HARADA, H HIROUMI, KINOSHITA, I, K AKIE, Y KAWAKAMI
    CANCER 76 (12) 2457 - 2463 0008-543X 1995/12 [Not refereed][Not invited]
     
    Background. Alterations of the p53 gene are one of the most common genetic changes in various types of cancer, including lung cancer. Abnormalities in the ras genes, including point mutations and overexpression, are another common feature in the molecular biology of lung cancer and are associated with a poorer prognosis. The authors' purpose was to determine expression of the mutated p53 gene in nonsmall cell lung cancer (NSCLC) specimens that were studied for expression of ras p21 and to document whether altered p53 expression was also an important factor for survival. Methods. Ninety-six patients with NSCLC underwent surgical resection between 1977 and 1985, 63 of whom received postoperative combination chemotherapy. None received radiation therapy. Tumor specimens were analyzed for altered p53 expression by immunohistochemistry. Univariate and multivariate analyses were performed to assess the association between p53 expression and survival. Results. Fifty-six (58%) of 96 tumor specimens showed altered p53 expression, and 91 patients were analyzed for survival, Altered p53 expression did not correlate with clinicopathologic characteristics except for postsurgical pathologic tumor (pT) classification. The patients with altered p53 expression survived for a significantly shorter period after surgery than those without p53 expression, including all patients who underwent resection and potentially curative resection (P = 0.02 and P = 0.048, respectively, generalized Wilcoxon test). Multivariate analysis showed independent prognostic significance for altered p53 expression (hazard ratio [HR] = 1.72, P = 0.04) and surgical cure (HR = 4.69, P < 0.001). The combined analysis of mutated p53 and ras p21 expression in the same tumor specimens revealed that patients with p53- and ras p21-negative tumors survived the longest among those with different p53 and ras p21 features (P = 0.005, generalized Wilcoxon test). Conclusion. Altered p53 expression is a significant and independent negative prognostic factor for patients with surgically resected NSCLC. Combined immunohistochemical analysis of mutated p53 and ras p21 expression can divide patients with NSCLC into more accurate prognostic groups. If the current findings can be confirmed in larger prospective studies, combined immunohistochemical analysis of mutated p53 and ras p21 expression can be a useful clinical tool for stratifying patients with NSCLC into accurate prognostic groups and for identifying the population with a different risk of recurrence.
  • M FUJINO, H DOSAKAAKITA, M KATO, KINOSHITA, I, K AKIE, Y KAWAKAMI
    AMERICAN JOURNAL OF CLINICAL PATHOLOGY 104 (3) 319 - 324 0002-9173 1995/09 [Not refereed][Not invited]
     
    Malignant neoplasms possess multiple genetic abnormalities. Among those, p53 gene abnormalities are the most frequent in human neoplasms. To screen for p53 abnormalities, both immunohistochemistry (IHC) and the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) methods are commonly used, but neither can detect all kinds of p53 abnormalities. In this study, by examining 35 lung cancer specimens simultaneously by the two methods, 12 abnormal cases (34%) were detected by IHC and 9 abnormal cases (26%) were detected by PCR-SSCP. Six abnormal (17%) and 20 normal (57%) cases showed concordant results between the methods, although 9 cases (26%) showed discordance, including 6 IHC-positive cases (17%) and 3 SSCP-positive cases (9%). Because it is known that some p53 abnormalities are detected only by IHC or only by PCR-SSCP, discordant cases should be assessed as possessing abnormalities. Fifteen cases (43%) were finally assessed as abnormal, These results suggest that the two methods together can increase the sensitivity of screening for p53 abnormalities.
  • H DOSAKAAKITA, K AKIE, H HIROUMI, KINOSHITA, I, Y KAWAKAMI, A MURAKAMI
    CANCER RESEARCH 55 (7) 1559 - 1564 0008-5472 1995/04 [Not refereed][Not invited]
     
    We evaluated the antiproliferative effect of L-myc antisense DNA in NCI-H209, a human small cell lung cancer (SCLC) cell line overexpressing the L-myc gene. The synthetic DNA used in the present study was oligodeoxynucleoside phosphorothioate, which showed rapid incorporation into NCI-H209 cells and localized mainly in the cell nucleus and weakly in the cytoplasm. The exposure of this cell line to L-myc antisense DNA covering the translational initiation site of L-myc proteins inhibited the cell proliferation in a dose-dependent sequence-specific manner. Furthermore, the growth inhibition by this antisense DNA was correlated with the level of L-myc expression in three SCLC cell lines, NCI-H209, NCI-H510, and NCI-H82. In Western blot analysis, expression of the L-myc proteins was down-regulated in the antisense-treated cells compared with control-treated cells in NCI-H209. Together with unique characteristics of the L-myc gene, including: (a) a frequently amplified and overexpressed state in SCLC; and (b) very restricted and low-level expression in human adult tissues, the present data indicate that L-myc is a good candidate for the target gene for antisense DNA therapy based on molecular biological diagnosis in SCLC.
  • KINOSHITA, I, H DOSAKAAKITA, M SHINDOH, M FUJINO, K AKIE, M KATO, K FUJINAGA, Y KAWAKAMI
    BRITISH JOURNAL OF CANCER 71 (2) 344 - 349 0007-0920 1995/02 [Not refereed][Not invited]
     
    To provide an accurate evaluation of the association of human papillomavirus (HPV) with lung cancer, 36 cases of lung cancer were analysed for HPV DNAs by polymerase chain reaction (PCR) with dot-blot and Southern blot analyses, and for the transcripts from the E6-E7 transforming region by in situ hybridisation (ISH). HPV-18 DNA was detected in three (8%) of 36 specimens; histologically, in one (10%) of 10 squamous cell carcinomas and two (9%) of 22 adenocarcinomas. Neither HPV-16 nor -33 DNA was detected in any cases examined. Expression of E6-E7 mRNA was confirmed in the cases which contained HPV-18 DNA. HPV-18 may play an important role in the development and progression of cancer in some cases of both squamous cell carcinoma and adenocarcinoma of the lung.
  • H DOSAKAAKITA, M SHINDOH, M FUJINO, KINOSHITA, I, K AKIE, M KATOH, Y KAWAKAMI
    AMERICAN JOURNAL OF CLINICAL PATHOLOGY 102 (5) 660 - 664 0002-9173 1994/11 [Not refereed][Not invited]
     
    Among the most common mutations in human lung cancer are those affecting the p53 gene. The expression of p53 in the nucleus is considered an immunohistochemical reflection of the nuclear accumulation of mutant p53 protein, which is coded by the p53 gene with missense mutation and has a prolonged half-life. In the present study, p53 expression detected by means of immunohistochemistry occurred frequently in human lung cancer and was associated with histologic subtypes. The alteration in the p53 gene was found to be a relatively early genetic event in the development and progression of lung cancer and to be maintained in the process of metastasis: abnormal p53 expression was found in both the early and late clinical stages, and identical p53 expression was detected consistently among primary and metastatic lesions from the same patients. Furthermore, an observed association between abnormal p53 expression and the patients' smoking history suggests that the p53 gene could be a common target of tobacco-associated carcinogenesis in lung cancer.

MISC

Books etc

  • 入門腫瘍内科学
    篠原出版新社 2009

Association Memberships

  • American Society of Clinical Oncology   世界肺癌学会   日本乳癌学会   日本分子標的治療学会   日本臨床腫瘍学会   日本臨床細胞学会   日本呼吸器学会   日本気管支学会   日本癌治療学会   日本癌学会   アメリカ癌学会   北海道癌談話会   北海道癌治療研究会   日本肺癌学会   アジア太平洋呼吸器学会   日本内科学会   

Works

  • スライドセミナー内科系：日本臨床細胞学会北海道支部会
    2009

Research Projects

  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2022/04 -2025/03 
    Author : 菊地 順子, 畑中 佳奈子, 天野 虎次, 畑中 豊, 木下 一郎
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 2021/04 -2024/03 
    Author : 榊原 純, 木下 一郎
     
    Prox1の小細胞肺癌（SCLC）における腫瘍原性に与える影響についての研究；SCLC細胞株のProx1の発現を最初に確認した。4つの(SCLC細胞株（MS-1、HCC827、SBC-3,SBC-5）を使用しProx1の発現をウエスタンブロットを用いてタンパク発現を確認したところMS-1、HCC827でProx1の発現を認め残りの2つのSBC-3とSBC-5のSCLC細胞株の発現は低かった。さらにPCRでもProx-1の発現を確認しタンパク発現とmRNAの発現が同様の結果であることを確認した。このため発現抑制のための実験にはMS-1とHCC827を使用することにした。Prox1の機能解析のために2つの細胞株でsiRNAを用いてProx1の発現が抑制されていることを確認した。増殖能についてMTT assayを行ったところProx1の抑制によりコントロールと比較して細胞増殖は増加した。clnogenic assayも行いMTT assayの結果と同様にProx1の抑制によりcolony数が増加した。さらにinvasion assayとmigration assayをtranswell chamberを用いて確認したところProx1の抑制によりコントロールと比較して細胞遊走能、浸潤能ともに増加した。 Prox1とNotch pathwayの関連についての検討：Prox1の抑制時にNotch 関連タンパク(Notch1-4、HES-1、HEY-1）の発現をウエスタンブロットにて確認したがコントロールと比較してProx1抑制時にNotch関連タンパク発現に変化を認めなかった。さらにPCRでmRNAについても検討したがProx1の発現の抑制時に変化を認めなかった。 CDDP耐性株におけるProx1の影響：当科でCDDP耐性株を当科で樹立しておりCDDP耐性株（MS-1）においてProx1の発現を確認したところProx1の発現が低下しており薬剤耐性との関与が考えられた。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2020/04 -2023/03 
    Author : 木下 一郎
     
    エピジェネティクスを標的にした新たながん治療が期待される中、クロマチン再構成を担うSWI/SNF複合体構成因子の異常が、ヒストンH3K27メチル化酵素EZH2に対する阻害薬のバイオマーカーとなる可能性が示された。初年度の研究で、SWI/SNFの状態の異なる種々の細胞株におけるEZH2阻害薬と、ヒストンH3K4脱メチル化酵素KDM5の抗腫瘍効果を検討したが、検討した細胞株におけるEZH2阻害薬Tazemetostat単剤での細胞障害活性が低く、KDM4阻害薬との併用効果も認められなかった。一方、種々のエピジェネティクス治療薬と、関連する因子の分子標的治療薬の併用効果を検討し、複数の細胞株で、Clonogenic法におけるHDAC阻害薬vorinostatとCDK4/6阻害薬palbociclibの相乗効果を見出した。 令和3年度は、両薬剤の相乗効果をソフトアガロース法によるクローン形成能によって解析した。Clonogenic法と同様、SWI/SNFの異常のある細胞株を主体とした5種類の細胞株に相乗効果を認め、4種類の細胞株には認めなかった。遺伝子変異やRNA発現データをCancer Cell Line Encyclopedia (CCLE)から入手し、相乗効果を認めた細胞グループと認めなかった細胞グループでの違いを比較した。相乗効果のある5細胞株の中4細胞株に共通する遺伝子変異を4つ認めたが、全てに共通する遺伝子異常は認めなかった。一方、RNA発現についてVolcano plotで検討したところ、相乗効果の有無により発現比の顕著な遺伝子を複数認めた。このうち、相乗効果のある細胞株全てに高発現している遺伝子も見出された。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2018/04 -2021/03 
    Author : sakakibara jun
     
    To investigate the role of Numb in tumorigenesis of lung adenocarcinoma and squamous cell carcinoma, we firstly performed loss-of-function and gain-of-function assays. Moreover, Numb expression was investigated in surgically resected lung adenocarcinoma and squamous cell carcinoma tissues by immunohistochemistry and correlations with prognosis were analyzed. Numb suppressed the proliferation, migration, and invasion of adenocarcinoma cells. In contrast, Numb promoted the proliferation, migration, and invasion of squamous cell carcinoma cells. High Numb expression was associated with favorable prognosis in patients with lung adenocarcinoma, but not in those with squamous cell carcinoma. Collectively, our data demonstrate that Numb plays distinct roles in lung adenocarcinoma and squamous cell carcinoma. In lung adenocarcinoma, Numb impairs tumor growth, whereas in lung squamous cell carcinoma it may promote proliferation.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2017/04 -2020/03 
    Author : Kinoshita Ichiro
     
    PBIT, a small molecule inhibitor of the JARID1a/b histone demethylases, restored the drug sensitivity of multiple drug-resistant persister cells that were established from NSCLC lines. In addition, PBIT prevented the expansion of EGFR-TKI-resistant persisters through modifying secretomes from EGFR-TKI sensitive cells by inhibiting downregulation of FRA1 expression via prevention of decreased H3K4me3 levels at FRA1 promoter regions. These findings suggest the potential efficacy of PBIT as a novel therapeutic strategy for lung cancer and have particularly important implications in restoring drug resistance in NSCLC, given that patients with activating EGFR mutations who are given EGFR-targeted therapies commonly develop resistance.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2014/04 -2017/03 
    Author : Kinoshita Ichiro, JINUSHI Masahisa, AKITA Hirotoshi
     
    EGFR-tyrosine kinase inhibitor (TKI) resistant non-small lung cancer (NSCLC) cell H1975 had 1% side population fraction, which showed increased tumorigenicity, elevated expression of stem cell markers and histone H3K27 trimethyl transferase EZH2, suggesting that EZH2 may play a role in maintenance of cancer stem cells. The combined treatment with inhibitors of the EZH2 and the histone deacetylases HDACs showed synergistic growth suppressive effects, suppression of EGFR signaling, and decrease in the in vivo tumor growth of H1975 cells, suggesting that the combined pharmacological targeting of the histone modification enzymes may provide more effective epigenetic therapeutics for NSCLCs including those with EGFR-TKI-resistant mutations.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    Date (from‐to) : 2014/04 -2016/03 
    Author : Akita Hirotoshi, Ohba Yusuke, Kinoshita Ichiro
     
    EGFR-TKI resistant cells were isolated in three non-small cell lung cancer (NSCLC) cell lines, using FRET biosensor and flow cytometer. cDNA microarray analysis showed common gene expression changes among the resistant cells. Annotation analysis revealed three up-regulated pathways and five down-regulated pathways. The ABC transporter system was included in the genes and pathways, which were found to be changed in relation to the acquired resistance, suggesting that this system could be involved in the resistance to EGFR-TKI in NSCLCs.
  • 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 : SAKAKIBARA Jun, KINOSHITA Ichiro
     
    Inhibition of Notch by gamma-secretase inhibitor (GSI) has been shown to have an antitumor effect in Notch expressing non-small cell lung cancer (NSCLC) and induce apoptosis through modulation of Bcl-2 family proteins. ABT-737, a BH3-only mimetic, targets the prosurvival Bcl-2 family and also induces apoptosis. GSI XX or ABT-737 alone inhibited cell proliferation in a dose dependent manner and combination drug treatment showed a synergistic antitumor effect in Notch expressing NSCLC in vitro. In vivo, this drug combination significantly suppressed tumor proliferation compared to single drug treatment. Phospho-Bcl-2 was down-regulated and Bax was up-regulated by both the single and combination drug treatments. Bim was induced by single drug treatment and was enhanced by combination treatment. Combination treatment-induced apoptosis was decreased by Bim inhibition, suggesting that the antitumor effect of the drug combination was dependent on Bim.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2011 -2013 
    Author : KINOSHITA Ichiro, SHIMIZU Yasushi
     
    Sphere forming capacity under serum-free culture, indicative of cancer stem cells, were decreased by cJun inhibition in non-small lung cancer (NSCLC) cells, while sphere forming cells did not show enhanced tumorigenicity in SCID mice. Meanwhile, an inhibitor of histone methyltransferase EZH2 involved in stem cell maintenance via epigenetic regulation suppressed growth of NSCLC cells. An inhibitor of histone deacetylase (HDAC), another epigenetic regulator, showed a synergistic growth suppressive effect with the EZH2 inhibitor. Combined epigenetic therapy with an EZH2 inhibitor and an HDAC inhibitor may represent an effective approach for NSCLCs.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2010 -2012 
    Author : AKITA Hirotoshi, KINOSHITA Ichiro
     
    High 〓1-6 fucosyltransferase (〓1-6 FucT) expression is more frequently found in adenocarcinomas than squamous cell carcinomas, and is correlated with advanced pN status in adenocarcinomas. High 〓1,6-FT expression is associated with an unfavorable clinical outcome in potentially curatively resected and pStage I adenocarcinomas. These findings suggest that 〓1,6-FT and its core-fucosylated products could be promisingbiomarkers and therapeutic targets for NSCLCs, especially for adenocarcinomas.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2008 -2010 
    Author : KINOSHITA Ichiro, AKITA Hirotoshi
     
    We identified 27 genes suppressed by blockade of AP-1, an oncogenic transcription factor, in lung cancer cells sensitive to the AP-1 blockade using microarray analysis. Among them, knockdown of MCM 4, a component of DNA replication licensing complex, by siRNA reduced cell growth in multiple lung cancer cells. Immunohistochemical analysis in surgically resected lung cancers demonstrated that MCM4 expression was correlated with proliferation markers including Ki-67. MCM4 may play an important role for growth of lung cancers and be a novel molecular target for lung cancer.
  • 日本学術振興会：科学研究費助成事業 萌芽研究
    Date (from‐to) : 2006 -2007 
    Author : 秋田 弘俊, 三善 英知, 木下 一郎
     
    本研究では、糖転移酵素α1,6-フコース転移酵素に着目して、非小細胞肺癌における発現異常(発現の低下・喪失)を免疫組織化学法で解析し、臨床病理学的因子、患者予後、細胞増殖能、各種分子マーカー発現、他の糖転移酵素発現との関係を解析して、非小細胞肺癌におけるα1,6-フコース転移酵素の発現異常の臨床病理学的意義を明らかにすることを目的とした。 非小細胞肺癌手術摘出腫瘍220腫瘍を材料として、α1,6‐フコース転移酵素の発現について特異抗体を用いて免疫組織化学法(Streptavidin biotin法)で解析した。肺癌組織における解析と同時に、肺葉切除された同一手術摘出材料内に存在する正常肺・気管支組織おける発現や随伴する非腫瘍性疾患肺組織における発現を解析した。α1,6‐フコース転移酵素に対する特異抗体は、大阪大学大学院医学研究科生化学・分子生物学教室が開発したものを用い、抗体の希釈濃度は1600倍とした。抗原賦活法としてオートクレーブ法を用いたところ、メチルグリーンで十分な核染色が得られず、ヘマトキシリンでは核染色が得られた。 α1,6-フコース転移酵素の発現は、正常気管支上皮細胞、正常気管支腺細胞では認められた。非小細胞肺癌においては、組織型では腺癌に比べて扁平上皮癌で発現の低下が認められた。また腺癌のなかでは、分化度では高分化なものに比べて低分化なもので発現の低下が認められ、臨床病理学的な意義が示唆された。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2004 -2006 
    Author : AKITA Hirotoshi, KINOSHITA Ichiro, YAMAZAKI Koichi
     
    The aim of this research is to identify molecular therapeutic targets and Biomarkers for the diagnosis and stratification in lung cancer. We searched for genes specifically overexpressed in lung cancer through microarray analysis, and identified seven genes, including AKRB1B10, in squamous cell carcinoma (SCC) of the lung. AKR1B10 was overexpressed in most cases with SCC, which is closely associated with smoking, and in many adenocarcinomas from smokers. These results suggest that AKR1B10 is a potential diagnostic marker specific to smokers' non-small cell lung cancers (NSCLCs) and might be involved in tobacco-related carcinogenesis. We next studied on the biomarker in molecular targeting therapy using EGF receptor (EGFR)-tyrosine kinase inhibitors (TKIs). Retrospective analyses have shown that activating mutations in exons 18-21 of the EGFR gene are a predictor of response to EGFR-TKIs. We conducted a phase II study to evaluate the efficacy and safety of gefitinib, one of EGFR-TKIs, as first-line therapy for advanced NSCLCs with EGFR mutations. For mutation analysis, DNA was extracted from paraffin-embedded tissues and EGFR mutations were analysed by direct sequence of PCR products. Twenty (24%) of the 82 patients analysed had EGFR mutations. Sixteen patients were enrolled and treated with gefitinib. Twelve patients had objective response and response rate was 75%. This study has shown that mutations (deletions in exon 19 and L858R point mutation in exon 21) of the EGFR gene are a predictor of response to EGFR-TKIs.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2004 -2005 
    Author : KINOSHITA Ichiro
     
    AP-1, a transcription factor transducing multiple mitogen growth signals, may be a useful target for gene and molecular target therapy. A previous study has demonstrated that cJun, a major constituent of AP-1, is frequently overexpressed in non-small cell lung cancers (NSCLCs). Therefore, in this study, we investigated the effect of AP-1 blockade on the growth of NSCLC cells using a cJun dominant negative mutant, TAM67. Transiently transfected TAM67 inhibited AP-1 transcriptional activity in NSCLC cell lines, H520 and H1299 cells. Colony forming efficiency of H1299 was much reduced by TAM67, while that of H520 was not. To elucidate the effect of TAM67 on the growth of H1299, we established H1299 clones that expressed TAM67 under the control of doxycycline-inducible promoter. Luciferase assay confirmed that the induction of TAM67 decreased AP-1 activity. MTT assay demonstrated that TAM67 inhibited cell growth. Flow cytometry analysis showed that TAM67 induced G1 cell cycle arrest. TAM67 also inhibited anchorage-independent growth determined by soft agarose assay, and Furthermore, we demonstrated that tumor growth was inhibited under the condition of expressing TAM67 in nude mice. These results suggest that AP-1 transcriptional activity plays an essential role in the growth of at least a part of NSCLC cells and that AP-1 can be a potential target for the treatment of NSCLCs.
  • 肺癌のAP-1を標的にした遺伝子治療に関する基礎的研究 転写因子E1AF/PEA3に関する研究 肺癌のEGFRの異常とGefitinib感受性

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