Basic research on the immunohistological categorization of tumor microenvironment

2017 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 金沢大学, 医薬保健学総合研究科, 特任教授 (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) ; 中津川 宗秀 札幌医科大学, 医学部, 助教 (70448596)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: Tumor microenvironment / Tumor immune escape / Immune response / Tumor antigen / Cancer stem cell / Immunohistochemistry / Single cell analysis

Outline of Annual Research Achievements

（１）がん組織微小環境の単細胞遺伝子発現解析：消化器がん、腎細胞がん、乳がん、婦人科がん、悪性黒色腫の各新鮮がん組織を単細胞に分離し、NGSによって網羅的単細胞遺伝子発現解析を実施した。データをクラスタリング解析し、がん細胞・免疫細胞・間質細胞の3グループに分類。発現遺伝子の質的・量的相違と特徴によってがん組織微小環境のクラスタリング解析を実施し、がん組織の部位による組織微小環境の相違を分子レベルで明らかにすることに成功した(Scientific Rep. 2017)。さらに、本来がん細胞だけが発現しているはずの遺伝子が、間質に浸潤している免疫細胞に取り込まれて発現していることを発見。がん細胞の違いが、組織内に浸潤している免疫細胞の多様性を生み、がん細胞が宿主の免疫応答を制御するメカニズムと関係していることが推察された。
（２）免疫細胞の単細胞遺伝子発現解析：がん組織に浸潤するT細胞および単球に発現する免疫関連遺伝子の発現を単細胞遺伝子発現解析によって分析した。同じ種類の免疫細胞であっても、がん組織の部位によって発現しているサイトカインの種類や量に違いがあることを見出した。これらの遺伝子発現パターンによって、T-cell Inflamed typeとT-cell excluded typeとに分類することができ、T-cell Inflamed typeに特徴的な分子を見出した。
（３）がん組織微小環境の個性を区別する免疫組織学マーカーの確立：遺伝子発現解析結果をもとに、ホルマリン固定標本の免疫組織染色によってがん組織微小環境を類型化する研究を実施中である。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

各種がん組織の単細胞遺伝子発現解析はほぼ計画通り行われ、予想外の興味深い解析結果が得られている。当初の計画には含まれていなかった悪性黒色腫と乳がんの解析も実施することができた。その反面、婦人科がんのサンプルと消化器がん、特に大腸がんのサンプルは不足しており、今後さらに増やす必要がある。免疫組織染色もほぼ予定通り実施され、がん組織微小環境の類型化に向けた研究は進展しつつある。CD8陽性T細胞の浸潤数を定量化したImmunoscoreに関する国際共同研究成果は、国際医学雑誌Lancetに掲載されることとなった。

Strategy for Future Research Activity

単細胞遺伝子発現解析技術は開発途上の技術であるため、さらなる改良による解析スピードと解析精度の向上が必要である。今後は解析サンプル数を増やして仮説を検証することが必要である。また、単細胞T細胞抗原受容体遺伝子解析については、現在の技術ではシークエンスが難しく、技術的な問題をクリアしなければならない。今後1年以内の解決を目指す。また、Digital PathologyとA.I.の新技術を活用し、免疫組織染色結果の定量化と標準化を目指す。

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/Oregon Health and Science University(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Mayo Clinic/Providence Portland Medical Center/Oregon Health and Science University
• [Int'l Joint Research] University of Toronto/Princess Margaret Cancer Centre/Ontario Cancer Institute(Canada)
  • Country Name: Canada
  • Counterpart Institution: University of Toronto/Princess Margaret Cancer Centre/Ontario Cancer Institute
• [Int'l Joint Research] Istituto Nazionale per lo Studio/University Hospital of Siena/Humanitas Clinical and Research Center(Italy)
  • Country Name: Italy
  • Counterpart Institution: Istituto Nazionale per lo Studio/University Hospital of Siena/Humanitas Clinical and Research Center
• [Int'l Joint Research] University of Erlangen(Germany)
  • Country Name: Germany
  • Counterpart Institution: University of Erlangen
• [Int'l Joint Research]
  • # of Other Countries: 10
• [Journal Article] The Antigen ASB4 on Cancer Stem Cells Serves as a Target for CTL Immunotherapy of Colorectal Cancer (2018)
  • Author(s): Miyamoto Sho、Kochin Vitaly、Kanaseki Takayuki、Hongo Ayumi、Tokita Serina、Kikuchi Yasuhiro、Takaya Akari、Hirohashi Yoshihiko、Tsukahara Tomohide、Terui Takeshi、Ishitani Kunihiko、Hata Fumitake、Takemasa Ichiro、Miyazaki Akihiro、Hiratsuka Hiroyoshi、Sato Noriyuki、Torigoe Toshihiko
  • Journal Title: Cancer Immunology Research Volume: 6 Pages: 358～369
  • DOI: 10.1158/2326-6066.Cir-17-0518
  • Peer Reviewed
• [Journal Article] Alveolar macrophages drive hepatocellular carcinoma lung metastasis by generating leukotriene B4. (2018)
  • Author(s): Nosaka, T., Baba, T., Tanabe, Y., Sasaki S., Nishimura, T., Imamura, Y, Yurino, H., Hashimoto, S., Arita M., Nakamoto, Y., Mukaida, N.
  • Journal Title: J Immunol. Volume: 200(5) Pages: 1839-1852
  • DOI: 10.4049/jimmunol.1700544
  • Peer Reviewed
• [Journal Article] Lack of IL-6 in the tumor microenvironment augments type-1 immunity and increases the efficacy of cancer immunotherapy. (2017)
  • Author(s): Ohno, Y., Toyoshima, Y., Yurino, H., Monma, N., Xiang, H., Sumida, K., Kaneumi, S., Terada, S., Hashimoto, S., Ikeo, K., Homma, S., Kawamura, H., Takahashi, N., Taketomi, A., Kitamura, H.
  • Journal Title: Cancer Sci. Volume: 108(10) Pages: 1959-1966
  • DOI: 10.1111/cas.13330
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive single-cell transcriptome analysis reveals heterogeneity in endometrioid adenocarcinoma tissues. (2017)
  • Author(s): Hashimoto, S., Tabuchi, Y., Yurino, H., Hirohashi, Y., Deshimaru, S., Asano, T., Mariya, T., Oshima, K., Takamura, Y., Ukita, Y., Ametani, .A, Kondo N, Monma, N., Takeda, T., Misu, S., Okayama, T., Ikeo, K., Saito, T., Kaneko, S., Suzuki, Y., Hattori, M., Matsushima, K., Torigoe, T.
  • Journal Title: Scientific Rep. Volume: 7(1) Pages: 14225
  • DOI: 10.1038/s41598-017-14676-3
  • Peer Reviewed / Open Access
• [Journal Article] Brother of the regulator of the imprinted site (BORIS) variant subfamily 6 is a novel target of lung cancer stem-like cell immunotherapy (2017)
  • Author(s): Horibe, R. Hirohashi, Y. Asano, T. Mariya, T. Suzuki, T. Takaya, A. Saijo, H. Shionoya, Y. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Watanabe, K. Atsuyama, E. Toji, S. Hirano, H. Hasegawa, T. Takahashi, H. Sato, N. Torigoe, T.
  • Journal Title: PLoS One Volume: 12 Pages: 0171460
  • DOI: 10.1371/journal.pone.0171460
  • Peer Reviewed / Open Access
• [Journal Article] GRIK2 has a role in the maintenance of urothelial carcinoma stem-like cells, and its expression is associated with poorer prognosis (2017)
  • Author(s): Inoue, R. Hirohashi, Y. Kitamura, H. Nishida, S. Murai, A. Takaya, A. Yamamoto, E. Matsuki, M. Tanaka, T. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Sato, N. Masumori, N. Torigoe, T.
  • Journal Title: Oncotarget Volume: 8 Pages: 28826-28839
  • DOI: 10.18632/oncotarget.16259
  • Peer Reviewed
• [Journal Article] HLA-A24 ligandome analysis of colon and lung cancer cells identifies a novel cancer-testis antigen and a neoantigen that elicits specific and strong CTL responses (2017)
  • Author(s): Kochin, V. Kanaseki, T. Tokita, S. Miyamoto, S. Shionoya, Y. Kikuchi, Y. Morooka, D. Hirohashi, Y. Tsukahara, T. Watanabe, K. Toji, S. Kokai, Y. Sato, N. Torigoe, T.
  • Journal Title: Oncoimmunology Volume: 6 Pages: 1293214
  • DOI: 10.1080/2162402x.2017.1293214
  • Peer Reviewed
• [Journal Article] Mismatch Repair Protein Deficiency Is a Risk Factor for Aberrant Expression of HLA Class I Molecules: A Putative "Adaptive Immune Escape" Phenomenon (2017)
  • Author(s): Kubo, T. Hirohashi, Y. Matsuo, K. Sonoda, T. Sakamoto, H. Furumura, K. Tsukahara, T. Kanaseki, T. Nakatsugawa, M. Hirano, H. Furuhata, T. Takemasa, I. Hasegawa, T. Torigoe, T.
  • Journal Title: Anticancer Res Volume: 37 Pages: 1289-1295
  • DOI: 10.21873/anticanres.11446
  • Peer Reviewed
• [Journal Article] Identification of cancer-stem cell antigens and development of CTL-mediated cancer immunotherapy (2017)
  • Author(s): Miyamoto, S. Kanaseki, T. Hirohashi, Y. Tsukahara, T. Kikuchi, Y. Sato, N. Torigoe, T.
  • Journal Title: Japanese journal of clinical immunology Volume: 40 Pages: 40-47
  • DOI: 10.2177/jsci.40.40
• [Journal Article] Identification of antigenic peptides from novel renal cancer stem-like cell antigen, DNAJB8 (2017)
  • Author(s): Nishizawa, S. Hirohashi, Y. Kusumoto, H. Wakamiya, T. Iguchi, T. Yamashita, S. Iba, A. Kikkawa, K. Kohjimoto, Y. Torigoe, T. Hara, I.
  • Journal Title: Biochem Biophys Res Commun Volume: 494 Pages: 693-699
  • DOI: 10.1016/j.bbrc.2017.10.146
  • Peer Reviewed
• [Journal Article] Induction and Analysis of Cytotoxic T-Lymphocytes that Recognize Autologous Oral Squamous Cell Carcinoma (2017)
  • Author(s): Okamoto, J. Y. Miyazaki, A. Kobayashi, J. I. Miyamoto, S. Sasaki, T. Kanaseki, T. Hirohashi, Y. Torigoe, T. Hiratsuka, H.
  • Journal Title: Anticancer Res Volume: 37 Pages: 4889-4897
  • DOI: 10.21873/anticanres.11898
  • Peer Reviewed
• [Journal Article] Implication of chemo-resistant memory T cells for immune surveillance in patients with sarcoma receiving chemotherapy (2017)
  • Author(s): Shibayama, Y. Tsukahara, T. Emori, M. Murata, K. Mizushima, E. Hirohashi, Y. Kanaseki, T. Nakatsugawa, M. Kubo, T. Yamashita, T. Sato, N. Torigoe, T.
  • Journal Title: Cancer Sci Volume: 108 Pages: 1739-1745
  • DOI: 10.1111/cas.13319
  • Peer Reviewed / Open Access
• [Journal Article] Loss of tapasin in human lung and colon cancer cells and escape from tumor-associated antigen-specific CTL recognition (2017)
  • Author(s): Shionoya, Y. Kanaseki, T. Miyamoto, S. Tokita, S. Hongo, A. Kikuchi, Y. Kochin, V. Watanabe, K. Horibe, R. Saijo, H. Tsukahara, T. Hirohashi, Y. Takahashi, H. Sato, N. Torigoe, T.
  • Journal Title: Oncoimmunology Volume: 6 Pages: 1274476
  • DOI: 10.1080/2162402x.2016.1274476
  • Peer Reviewed
• [Journal Article] Immune-related adverse events of immune checkpoint inhibitors (2017)
  • Author(s): Tadano, H. Torigoe, T.
  • Journal Title: Japanese journal of clinical immunology Volume: 40 Pages: 102-108
  • DOI: 10.2177/jsci.40.102
• [Journal Article] Identification and functional analysis of variants of a cancer/testis antigen LEMD1 in colorectal cancer stem-like cells (2017)
  • Author(s): Takeda, R. Hirohashi, Y. Shen, M. Wang, L. Ogawa, T. Murai, A. Yamamoto, E. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Nishidate, T. Okita, K. Kutomi, G. Sato, N. Takemasa, I. Torigoe, T.
  • Journal Title: Biochem Biophys Res Commun Volume: 485 Pages: 651-657
  • DOI: 10.1016/j.bbrc.2017.02.081
  • Peer Reviewed
• [Journal Article] Cancer-associated oxidoreductase ERO1-alpha promotes immune escape through up-regulation of PD-L1 in human breast cancer (2017)
  • Author(s): Tanaka, T. Kutomi, G. Kajiwara, T. Kukita, K. Kochin, V. Kanaseki, T. Tsukahara, T. Hirohashi, Y. Torigoe, T. Okamoto, Y. Hirata, K. Sato, N. Tamura, Y.
  • Journal Title: Oncotarget Volume: 8 Pages: 24706-24718
  • DOI: 10.18632/oncotarget.14960
  • Peer Reviewed
• [Journal Article] LY6/PLAUR domain containing 3 has a role in the maintenance of colorectal cancer stem-like cells (2017)
  • Author(s): Wang, L. Hirohashi, Y. Ogawa, T. Shen, M. Takeda, R. Murai, A. Yamamoto, E. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Nishidate, T. Okita, K. Kutomi, G. Sato, N. Takemasa, I. Torigoe, T.
  • Journal Title: Biochem Biophys Res Commun Volume: 486 Pages: 232-238
  • DOI: 10.1016/j.bbrc.2017.02.112
  • Peer Reviewed
• [Journal Article] Phosphorylation of HSF1 at serine 326 residue is related to the maintenance of gynecologic cancer stem cells through expression of HSP27 (2017)
  • Author(s): Yasuda, K. Hirohashi, Y. Mariya, T. Murai, A. Tabuchi, Y. Kuroda, T. Kusumoto, H. Takaya, A. Yamamoto, E. Kubo, T. Nakatsugawa, M. Kanaseki, T. Tsukahara, T. Tamura, Y. Hirano, H. Hasegawa, T. Saito, T. Sato, N. Torigoe, T.
  • Journal Title: Oncotarget Volume: 8 Pages: 31540-31553
  • DOI: 10.18632/oncotarget.16361
  • Peer Reviewed
• [Presentation] CTL responses against a cancer-stem cell specific antigen, ASB4, effectively prevent colorectal cancer formation (2017)
  • Author(s): Kanaseki T, Miyamoto S, Tokita S, Torigoe T.
  • Organizer: THIRD CRI-CIM-EATI-AACR International Cancer Immunotherapy Conference
  • Int'l Joint Research
• [Presentation] Molecular Mechanism of Cancer Cell Hormesis (2017)
  • Author(s): 鳥越俊彦、廣橋良彦
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] Phosphorylation of HSF1 at Ser326 residue is associated with the maintenance of gynecologic cancer stem cells. (2017)
  • Author(s): Toshihiko Torigoe, Yoshihiko Hirohashi, Kazuyo Yasuda, Aiko Murai, Tsuyoshi Saito, Noriyuki Sato
  • Organizer: The Eighth International Congress on Stress Proteins in Biology and Medicine
  • Int'l Joint Research
• [Presentation] 病理学を基盤としたがん免疫療法研究 (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 日本癌学会2017ランチョンセミナー
  • Invited
• [Presentation] Basic methodology for immuno-oncology research (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 日本癌学会2017 モーニングレクチャー
  • Invited
• [Presentation] 免疫チェックポイント阻害薬の光と影：自己免疫炎症性副作用について (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 日本臨床免疫学会2017 アニュアルエビデンスレビュー
  • Invited
• [Presentation] A harmonized evaluation system for expression of HLA class I antigens in formalin-fixed paraffin embedded cancer tissues (2017)
  • Author(s): Toshihiko Torigoe, Naoya Nakamura, Eri Atsuyama, Shingo Toji, Yuji Heike, Nobuyoshi Hiraoka
  • Organizer: The Society for Immunotherapy of Cancer's (SITC) 32nd Annual Meeting
  • Int'l Joint Research
• [Presentation] 腫瘍免疫病理学の基礎と臨床 up-to-date (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 日本免疫学会2017 ランチョンセミナー
  • Invited
• [Presentation] Landscape of HLA class I ligand peptides (2017)
  • Author(s): Takayuki Kanaseki
  • Organizer: The 36th Sapporo International Cancer Symposium
  • Int'l Joint Research / Invited
• [Presentation] Immunothrapeutic strategy targeting cancer-stem cells (2017)
  • Author(s): Yoshihiko Hirohashi
  • Organizer: The 36th Sapporo International Cancer Symposium
  • Int'l Joint Research / Invited
• [Presentation] A novel population of stem-like memory T-cells (2017)
  • Author(s): Tomohide Tsukahara
  • Organizer: The 36th Sapporo International Cancer Symposium
  • Int'l Joint Research / Invited
• [Presentation] Landscape of natural HLA ligand peptides (2017)
  • Author(s): Toshihiko Torigoe, Takayuki Kanaseki, Kochin Vitaly
  • Organizer: 第21回日本がん免疫学会総会シンポジウム
  • Invited
• [Presentation] Single cell transcriptome analysis for dissecting tumor microenvironment (2017)
  • Author(s): Toshihiko Torigoe
  • Organizer: The 32nd Nagoya International Cancer Treatment Symposium
  • Int'l Joint Research / Invited
• [Presentation] がん免疫療法の基礎と臨床 (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 第29回北海道癌治療研究会特別講演
  • Invited
• [Presentation] がん免疫の分子病理学 (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 第106回日本病理学会総会分子病理診断講習会
  • Invited
• [Presentation] 免疫チェックポイント阻害剤最新知見 (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 第11回日本緩和医療薬学会年会メディカルセミナー
  • Invited
• [Presentation] 新たな時代を迎えたがん免疫療法 (2017)
  • Author(s): 鳥越俊彦
  • Organizer: 第71回日本交通医学会総会ランテョンセミナー
  • Invited
• [Book] がん免疫療法 Cancer Immunotherapy (2017)
  • Author(s): 上田龍三、鵜殿平一郎、河上裕、鳥越俊彦、西川博嘉
  • Total Pages: 72
  • Publisher: メディカルレビュー社
  • ISBN: 2432-8138
• [Book] 次世代がん治療 (2017)
  • Author(s): 塚原智英、鳥越俊彦
  • Total Pages: 386
  • Publisher: (株)エヌ・ティー・エス
  • ISBN: 978-4-86043-503-5 C3045
• [Remarks] 札幌医科大学医学部病理学第一講座＜研究Research＞
  • URL: http://web.sapmed.ac.jp/patho1/research/index.html