Basic research on the immunohistological categorization of tumor microenvironment

2019 Fiscal Year Annual Research Report

• Project/Area Number: 17H01540
• Research Institution: Sapporo Medical University
• Principal Investigator: 鳥越 俊彦 札幌医科大学, 医学部, 教授 (20301400)
• Co-Investigator(Kenkyū-buntansha): 橋本 真一 和歌山県立医科大学, 先端医学研究所, 教授 (00313099) ; 廣橋 良彦 札幌医科大学, 医学部, 准教授 (30516901) ; 塚原 智英 札幌医科大学, 医学部, 准教授 (20404634) ; 金関 貴幸 札幌医科大学, 医学部, 講師 (50531266) ; 久保 輝文 札幌医科大学, 医学部, 助教 (90580019)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: シングルセル遺伝子解析 / T細胞抗原受容体 / がん抗原 / 免疫組織染色 / バイオマーカー / 免疫チェックポイント阻害剤 / がん免疫療法 / 腫瘍組織微小環境

Outline of Annual Research Achievements

１：がん組織の単細胞遺伝子発現解析：消化器がん、腎細胞がん、乳がんの各新鮮がん組織を用いて、組織シングルセル遺伝子発現解析を実施。クラスター解析によって、がん細胞・免疫細胞・間質細胞それぞれの多様性を分析し、がん組織をT-cell Inflamed type、T-cell excluded type, Immune desert typeの３カテゴリーに分類した。
２．腫瘍浸潤T細胞のTCR遺伝子解析：がん組織に浸潤するT細胞の抗原受容体遺伝子のシークエンスを行い、T細胞クロナリティー解析を行った。T-cell Inflamed typeではT細胞クロナリティーの多様性が低く、末梢血とは異なるT細胞クローンの存在が確認された。また、腎細胞がんから得られた腫瘍浸潤T細胞クローンが認識するがん特異抗原の同定に成功した。
３．組織微小環境(TME)の個性診断に有用な免疫組織マーカーの確立：組織シングルセル遺伝子発現解析結果を基にして、がん組織TMEの個性診断に有用なバイオマーカーを探索し、ホルマリン固定標本の免疫組織染色法を最適化した。HLA-class I、HLA-DR, CD8, TIA-1, PD-L1, PMS2, MSH6の各マーカーによって、TMEを分類することができた。
４．TMEの個性診断と免疫治療への応用 ：尿路上皮がんに焦点を絞り、免疫チェックポイント阻害剤(ICB)感受性のがんと抵抗性のがんについて、上記TME分類との相関性を後向きコホート試験によって検証した。その結果、上記の免疫組織染色法によるTME分類によって、ICBの治療効果を予想できることが判明した。現在、頭頚部がんと消化器がんを対象に、同様の検証を行っている。
５．以上の研究の一部は、約16カ国の国際共同研究として実施した。

Research Progress Status

令和元年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和元年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] INSERM/University of Paris Descartes/University of Pierre et Marie Curie(フランス)
  • Country Name: FRANCE
  • Counterpart Institution: INSERM/University of Paris Descartes/University of Pierre et Marie Curie
  • # of Other Institutions: 1
• [Int'l Joint Research] Mayo Clinic/Providence Portland Medical Center(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Mayo Clinic/Providence Portland Medical Center
• [Int'l Joint Research] University of Toronto/University Health Network/Princess Margaret Hospital(カナダ)
  • Country Name: CANADA
  • Counterpart Institution: University of Toronto/University Health Network/Princess Margaret Hospital
• [Int'l Joint Research] University Nijmegen Medical Center(オランダ)
  • Country Name: NETHERLANDS
  • Counterpart Institution: University Nijmegen Medical Center
• [Int'l Joint Research] University of Erlangen(ドイツ)
  • Country Name: GERMANY
  • Counterpart Institution: University of Erlangen
• [Int'l Joint Research]
  • # of Other Countries: 12
• [Journal Article] Osteosarcoma-initiating cells show high aerobic glycolysis and attenuation of oxidative phosphorylation mediated by LIN28B (2020)
  • Author(s): Mizushima, E. Tsukahara, T. Emori, M. Murata, K. Akamatsu, A. Shibayama, Y. Hamada, S. Watanabe, Y. Kaya, M. Hirohashi, Y. Kanaseki, T. Nakatsugawa, M. Kubo, T. Yamashita, T. Sato, N. Torigoe, T.
  • Journal Title: Cancer Science Volume: 111 Pages: 36-46
  • DOI: 10.1111/cas.14229
  • Peer Reviewed
• [Journal Article] Peptide vaccinations elicited strong immune responses that were reboosted by anti-PD1 therapy in a patient with myxofibrosarcoma (2020)
  • Author(s): Tsukahara, T. Watanabe, K. Murata, K. Takahashi, A. Mizushima, E. Shibayama, Y. Kameshima, H. Hatae, R. Ohno, Y. Kawahara, R. Murai, A. Nakatsugawa, M. Kubo, T. Kanaseki, T. Hirohashi, Y. Terui, T. Asanuma, H. Hasegawa, T. Sato, N. Torigoe, T.
  • Journal Title: Cancer immunology immunotherapy Volume: 69 Pages: 189-197
  • DOI: 10.1007/s00262-019-02455-0
  • Peer Reviewed
• [Journal Article] Gli signaling pathway modulates fibroblast activation and facilitates scar formation in pulmonary fibrosis. (2019)
  • Author(s): Tsukui, T., Ueha, S., Shichino, S., Hashimoto, S., Nakajima, T.,Shiraishi, K., Kihara, M., Kiyonari, H., Inagak, Y., Matsushima K.
  • Journal Title: Biochem Biophys Res Commun. Volume: S0006-291X(19) Pages: 30879-4
  • DOI: 10.1016/j.bbrc.2019.05.011.
  • Peer Reviewed
• [Journal Article] Engraftment and proliferation potential of embryonic lung tissue cells in irradiated mice with emphysema. (2019)
  • Author(s): Shiraishi, K., Shichino, S., Tsukui, T., Hashimoto, S., Ueha, S., Matsushima, K.
  • Journal Title: Sci Rep. Volume: 9 Pages: 3657
  • DOI: 10.1038/s41598-019-40237-x.
  • Peer Reviewed
• [Journal Article] TCR repertoire analysis reveals mobilization of novel CD8+ T cell clones into the Cancer-Immunity Cycle following anti-CD4 antibody administration. (2019)
  • Author(s): Aoki, H., Ueha, S., Shichino, S., Ogiwara, S., Hashimoto, S., Kakimi, K., Ito, S., Matsushima, K.
  • Journal Title: Front Immunol. Volume: 9 Pages: 3185
  • DOI: 10.3389/fimmu.2018.03185.
  • Peer Reviewed
• [Journal Article] Mesenchymal-epithelial interactome analysis reveals essential factors required for fibroblast-free alveolosphere formation. (2019)
  • Author(s): Shiraishi, K., Shichino, S., Ueha, S., Nakajima, T., Hashimoto, S., Yamazaki, S., Matsushima, K.
  • Journal Title: iScience Volume: 11 Pages: 318-333
  • Peer Reviewed
• [Journal Article] Notch signaling facilitates hepatitis B virus covalently closed circular DNA transcription via cAMP response element-binding protein with E3 ubiquitin ligase-modulation. (2019)
  • Author(s): Kawaguchi, K., Wang, Z., Honda, M., Hashimoto, S., hirasaki, T., Okada, H., Orita, N., Shimakami, T., Yamashita, T., Sakai, Y., Mizukoshi, E., Murakami, S., Kaneko, S.
  • Journal Title: Sci Rep. Volume: 9 Pages: 1621
  • DOI: 10.1038/s41598-018-38139-5.
  • Peer Reviewed
• [Journal Article] “Nx1-Seq (Well Based Single-Cell Analysis System)” (2019)
  • Author(s): Shinichi Hashimoto
  • Journal Title: Adv Exp Med Biol. Volume: 1129 Pages: 51-61
  • DOI: 10.1007/978-981-13-6037-4_4
  • Peer Reviewed
• [Journal Article] “Single-Cell Transcriptome Analysis in Tumor Tissues” (2019)
  • Author(s): Sadahiro Iwabuchi and Shinichi Hashimoto.
  • Journal Title: Transcriptome Analysis, Intechopen Volume: - Pages: -
  • DOI: 10.5772/intechopen.84558.
  • Peer Reviewed
• [Journal Article] Upstream Position of Proline Defines Peptide-HLA Class I Repertoire Formation and CD8(+) T Cell Responses (2019)
  • Author(s): Hongo, A. Kanaseki, T. Tokita, S. Kochin, V. Miyamoto, S. Hashino, Y. Codd, A. Kawai, N. Nakatsugawa, M. Hirohashi, Y. Sato, N. Torigoe, T.
  • Journal Title: J Immunol Volume: 202 Pages: 2849-2855
  • DOI: 10.4049/jimmunol.1900029
  • Peer Reviewed
• [Journal Article] Severe cytokine release syndrome resulting in purpura fulminans despite successful response to nivolumab therapy in a patient with pleomorphic carcinoma of the lung: a case report (2019)
  • Author(s): Honjo, O. Kubo, T. Sugaya, F. Nishizaka, T. Kato, K. Hirohashi, Y. Takahashi, H. Torigoe, T.
  • Journal Title: J Immunother Cancer Volume: 7 Pages: 97
  • DOI: 10.1186/s40425-019-0582-4
  • Peer Reviewed
• [Journal Article] Proteogenomic discovery of cancer antigens: Neoantigens and beyond (2019)
  • Author(s): Kanaseki, T. Tokita, S. Torigoe, T.
  • Journal Title: Pathology international Volume: 69 Pages: 511-518
  • DOI: 10.1111/pin.12841
  • Peer Reviewed
• [Journal Article] Proteogenomics: advances in cancer antigen research (2019)
  • Author(s): Kanaseki, T. Torigoe, T.
  • Journal Title: Immunological Medicine Volume: 42 Pages: 65-70
  • DOI: 10.1080/25785826.2019.1640500
  • Peer Reviewed
• [Journal Article] Efficacy of combined radiotherapy and anti-programmed death 1 therapy in acral and mucosal melanoma (2019)
  • Author(s): Kato, J. Hida, T. Someya, M. Sato, S. Sawada, M. Horimoto, K. Fujioka, M. Minowa, T. Matsui, Y. Tsuchiya, T. Kitagawa, M. Nakata, K. Sakata, K. I. Torigoe, T. Uhara, H.
  • Journal Title: The Journal of dermatology Volume: 46 Pages: 328-333
  • DOI: 10.1111/1346-8138.14805
  • Peer Reviewed
• [Journal Article] ABCG2 expression is related to low 5-ALA photodynamic diagnosis (PDD) efficacy and cancer stem cell phenotype, and suppression of ABCG2 improves the efficacy of PDD (2019)
  • Author(s): Kawai, N. Hirohashi, Y. Ebihara, Y. Saito, T. Murai, A. Saito, T. Shirosaki, T. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Shichinohe, T. Li, L. Hirano, S. Torigoe, T.
  • Journal Title: PloS one Volume: 14 Pages: e0216503
  • DOI: 10.1371/journal.pone.0216503
  • Peer Reviewed
• [Journal Article] Differential bronchial epithelial response regulated by ΔNp63: a functional understanding of the epithelial shedding found in asthma (2019)
  • Author(s): Kubo, T. Tsujiwaki, M. Hirohashi, Y. Tsukahara, T. Kanaseki, T. Nakatsugawa, M. Hasegawa, T. Torigoe, T.
  • Journal Title: Lab Invest Volume: 99 Pages: 158-168
  • DOI: 10.1038/s41374-018-0132-6
  • Peer Reviewed
• [Journal Article] Immunohistological analysis of pancreatic carcinoma after vaccination with survivin 2B peptide: Analysis of an autopsy series (2019)
  • Author(s): Kubo, T. Tsurita, G. Hirohashi, Y. Yasui, H. Ota, Y. Watanabe, K. Murai, A. Matsuo, K. Asanuma, H. Shima, H. Wada, S. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Mizuguchi, T. Hirata, K. Takemasa, I. Imai, K. Sato, N. Torigoe, T.
  • Journal Title: Cancer Science Volume: 110 Pages: 2386-2395
  • DOI: 10.1111/cas.14099
  • Peer Reviewed
• [Journal Article] Identification of novel human leukocyte antigen-A*11:01-restricted cytotoxic T-lymphocyte epitopes derived from osteosarcoma antigen papillomavirus binding factor (2019)
  • Author(s): Li, D. Toji, S. Watanabe, K. Torigoe, T. Tsukahara, T.
  • Journal Title: Cancer Science Volume: 110 Pages: 1156-1168
  • DOI: 10.1111/cas.13973
  • Peer Reviewed
• [Journal Article] Randomized phase II trial of survivin 2B peptide vaccination for patients with HLA-A24-positive pancreatic adenocarcinoma (2019)
  • Author(s): Shima, H. Tsurita, G. Wada, S. Hirohashi, Y. Yasui, H. Hayashi, H. Miyakoshi, T. Watanabe, K. Murai, A. Asanuma, H. Tokita, S. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Nakae, Y. Sugita, O. Ito, Y. M. Ota, Y. Kimura, Y. Kutomi, G. Hirata, K. Mizuguchi, T. Imai, K. Takemasa, I. Sato, N. Torigoe, T.
  • Journal Title: Cancer Science Volume: 110 Pages: 2378-2385
  • DOI: 10.1111/cas.14106
  • Peer Reviewed
• [Journal Article] Clonal analysis revealed functional heterogeneity in cancer stem-like cell phenotypes in uterine endometrioid adenocarcinoma (2019)
  • Author(s): Tabuchi, Y. Hirohashi, Y. Hashimoto, S. Mariya, T. Asano, T. Ikeo, K. Kuroda, T. Mizuuchi, M. Murai, A. Uno, S. Kawai, N. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Saito, T. Torigoe, T.
  • Journal Title: Exp Mol Pathol Volume: 106 Pages: 78-88
  • DOI: 10.1016/j.yexmp.2018.11.013
  • Peer Reviewed
• [Journal Article] Development of a T-cell receptor multimer with high avidity for detecting a naturally presented tumor-associated antigen on osteosarcoma cells (2019)
  • Author(s): Watanabe, K. Tsukahara, T. Toji, S. Saitoh, S. Hirohashi, Y. Nakatsugawa, M. Kubo, T. Kanaseki, T. Kameshima, H. Terui, T. Sato, N. Torigoe, T.
  • Journal Title: Cancer Science Volume: 110 Pages: 40-51
  • DOI: 10.1111/cas.13854
  • Peer Reviewed
• [Presentation] １細胞トランスクリプトーム手法と炎症細胞解析への適用 (2020)
  • Author(s): 橋本 真一
  • Organizer: 情報計算化学生物学会（CBI学会）関西部会
  • Invited
• [Presentation] がん幹細胞と免疫微小環境 (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 第108回日本病理学会総会
  • Invited
• [Presentation] Immunotherapeutic Strategy Targeting Cancer Stem Cells:Characterization of Specific Antigens and Fibroblastic Niche (2019)
  • Author(s): Toshihiko Torigoe
  • Organizer: The New York Academy of Sciences: Frontiers in Cancer Immunotherapy
  • Int'l Joint Research
• [Presentation] A novel immunohistological profiling method for cancer tissue, Immunohistogram, that would contribute to personalized precision medicine (2019)
  • Author(s): Toshihiko Torigoe
  • Organizer: 17th International Congress of Immunology
  • Int'l Joint Research
• [Presentation] Single-cell Gene Expression Analysis in Tumor Microenvironment (2019)
  • Author(s): Shinichi Hashimoto
  • Organizer: EMBO single cell work shop
  • Int'l Joint Research / Invited
• [Presentation] １細胞解析による生物学と疾患の新たな理解 (2019)
  • Author(s): 橋本 真一
  • Organizer: 第36回和漢医薬学会学術大会
  • Invited
• [Presentation] Cell type and marker gene prediction for single-cell transcriptome analysis (2019)
  • Author(s): Shinichi Hashimoto
  • Organizer: 1st International Symposium on Inflammation Cellular Sociology
  • Int'l Joint Research / Invited
• [Presentation] Comparison of the gene expression profile in the different cultured condition by single cell transcriptome analysis. (2019)
  • Author(s): Sadahiro Iwabuchi, Shinichi Hashimoto
  • Organizer: Cold Spring Harbor Laboratory meeting,The Biology of Genomes
  • Int'l Joint Research
• [Presentation] Single-cell RNA sequencing identifies the common and unique cell population between in an in vitro and in vivo cultured condition. (2019)
  • Author(s): Iwabuchi Sadahiro, Shinichi Hashimoto
  • Organizer: Cold Spring Harbor Laboratory meeting,Single Cell Analysis
  • Int'l Joint Research
• [Presentation] プロテオゲノミクスによるがん抗原の解明と免疫療法の開発 (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 日本医学会総会2019中部
  • Invited
• [Presentation] Proteogenomics によるネオアンチゲンの探索 (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 千里ライフサイエンスセミナー：がんシリーズ第７回がん微小環境
  • Invited
• [Presentation] 尿路上皮癌におけるがん免疫療法が切り開く未来：biomarkerがもたらすprecision medicine (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 第32回日本老年泌尿器科学会
  • Invited
• [Presentation] 免疫チェックポイント阻害剤の免疫病理学 (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 第33回膠原病臨床病理研究会
  • Invited
• [Presentation] Novel immunodiagnostic and immunotherapeutic strategies targeting cancer stem cells (2019)
  • Author(s): Toshihiko Torigoe
  • Organizer: The 38th Sapporo International Cancer Symposium
  • Int'l Joint Research / Invited
• [Presentation] がん幹細胞と免疫の相互作用 (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 第5回Immuno-Oncology Forum
  • Invited
• [Presentation] HLA ligandome analysis reveals an antigen processing signature required for HLA class I presentation and CD8+ T cell responses. (2019)
  • Author(s): Kanaseki T, Hongo A, Tokita S, Torigoe T.
  • Organizer: CRI-EATI-CIMT-AACR 5th International Cancer Immunotherapy Conference
  • Int'l Joint Research
• [Presentation] Implication of chemo-resistant memory T cells for immune surveillance in patients with sarcoma receiving chemotherapy. (2019)
  • Author(s): Tsukahara T, Shibayama Y, Murata K, Mizushima E, Hamada S, Torigoe T.
  • Organizer: CRI-EATI-CIMT-AACR 5th International Cancer Immunotherapy Conference
  • Int'l Joint Research
• [Presentation] A determinant leading to CD8+ T-cell induction in neoantigens with a single amino acid substitution (2019)
  • Author(s): Shinkawa T, Kanaseki T, Tokita S, Torigoe T.
  • Organizer: CRI-EATI-CIMT-AACR 5th International Cancer Immunotherapy Conference
  • Int'l Joint Research
• [Presentation] Identification of a new type of neoantigen derived from splice peptides (2019)
  • Author(s): Kato K, Nakatsugawa M, Kanaseki T, Tokita S, Torigoe T.
  • Organizer: CRI-EATI-CIMT-AACR 5th International Cancer Immunotherapy Conference
  • Int'l Joint Research
• [Presentation] がん免疫療法の免疫病理学Up-to-Date (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 第11回埼玉南西部地区がん化学療法研究会
  • Invited
• [Presentation] 免疫チェックポイント阻害剤副作用の特徴 (2019)
  • Author(s): 鳥越 俊彦
  • Organizer: 第47回日本臨床免疫学会総会
  • Invited
• [Presentation] Proteogenomic approach for natural HLA class I ligand peptides of cancer cells. (2019)
  • Author(s): Torigoe T, Kanaseki T, Hirohashi Y, Tsukahara T, Nakatsugawa M, Kubo T, Shinkawa T.
  • Organizer: EMBL Conference: Cancer Genomics
  • Int'l Joint Research
• [Presentation] Immunopathological properties of fibroblastic niche for cancer stem cells. (2019)
  • Author(s): Torigoe T, Hirohashi Y
  • Organizer: SITC 34th Annual Meeting
  • Int'l Joint Research
• [Presentation] Analysis of the molecular mechanism for cancer cell hormesis. (2019)
  • Author(s): Torigoe T, Hirohashi Y
  • Organizer: 9th International Congress on Stress Responses in Biology and Medicine
  • Int'l Joint Research
• [Patent(Industrial Property Rights)] がん幹細胞特異的抗体 (2019)
  • Inventor(s): 塚原智英，鳥越俊彦
  • Industrial Property Rights Holder: 塚原智英，鳥越俊彦
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: PCT/JP2015/039400
  • Overseas