Single cell analysis of tumor-microenvironment based on TIL functions

• Project/Area Number: 20H03460
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 49020:Human pathology-related
• Research Institution: Sapporo Medical University
• Principal Investigator: HIROHASHI Yoshihiko 札幌医科大学, 医学部, 准教授 (30516901)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2022: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2021: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2020: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: 腫瘍微小環境 / 細胞傷害性T細胞 / 腫瘍抗原 / 1細胞解析 / 腫瘍浸潤リンパ球 / T細胞受容体 / 単細胞解析 / 悪性黒色腫 / TCR / バイオマーカー

Outline of Research at the Start

単細胞解析は、腫瘍微小環境を解析する上で強力なツールとなる。しかしながら、単細胞解析は遺伝子発現プロファイルに基づく解析を行う為、単細胞レベルでの機能解析を不得手とする。腫瘍微小環境を作る細胞でもCTLなどのリンパ球は、遺伝子情報から機能を推定する事が出来る。T細胞受容体遺伝子配列がCTL機能を規定するからである。本研究では単細胞解析でも機能解析が可能なCTLなどの腫瘍浸潤リンパ球に主眼を置き、腫瘍微小環境を単細胞解像度で行う。具体的に、遺伝子発現情報に機能的TCR遺伝子情報をアノテーションし、CTL機能評価も合わせた上で腫瘍微小環境解析を行い、抗腫瘍効果に繋がる免疫状態微小環境を読み解く。

Outline of Final Research Achievements

In this study, we analyzed melanoma cases that were treated by PD-1 blockade and resulted in reflactory for PD-1 blockade. Tumor infiltrating lymphocytes (TILs) were analyzed by single cell RNA-sequence. T cell receptor (TCR) transduced T (TCR-T) cells were generated according to the TCR sequences and tested the specificity for autologous melanoma cells. Surprisingly, most TCR-T cells recognized melanoma cells indicating TILs were functional, but PD-1 blockade did not work. These result indicate that there are other immune escape mechanisms beside PD-L1/PD-1.

Academic Significance and Societal Importance of the Research Achievements

本研究により、悪性黒色腫症例の腫瘍浸潤リンパ球を1細胞遺伝子発現解析行った。その結果、免疫チェックポイント阻害剤(PD-1阻害剤）不応性症例の腫瘍浸潤リンパ球であっても、腫瘍細胞を認識しうることを見出した。この結果は、PD-L1/PD-1阻害治療抵抗性症例であっても、免疫療法が効く可能性を示すと同時に、PD-L1/PD-1以外の免疫チェックポイント等の免疫抑制機構が存在する事を示唆する。この解析を進める事により、PD-1阻害療法の適応を定義出来ると同時に、新たな治療法開発への rationale となる。

Research Products

• [Journal Article] Cisplatin-induced HSF1-HSP90 axis enhances the expression of functional PD-L1 in oral squamous cell carcinoma (2023)
  • Author(s): Sasaya T, Kubo T, Murata K, Mizue Y, Sasaki K, Yanagawa J, Imagawa M, Kato H, Tsukahara T, Kanaseki T, Tamura Y, Miyazaki A, Hirohashi Y, Torigoe T.
  • Journal Title: Cancer Med Volume: 12 Issue: 4 Pages: 4605-4615
  • DOI: 10.1002/cam4.5310
  • Peer Reviewed / Open Access
• [Journal Article] Cisplatin resistance driver claspin is a target for immunotherapy in urothelial carcinoma (2023)
  • Author(s): Yamada S, Miyata H, Isono M, Hori K, Yanagawa J, Murai A, Minowa T, Mizue Y, Sasaki K, Murata K, Tokita S, Nakatsugawa M, Iwabuchi S, Hashimoto S, Kubo T, Kanaseki T, Tsukahara T, Abe T, Shinohara N, Hirohashi Y, Torigoe T.
  • Journal Title: Cancer Immunology Immunotherapy Volume: Online ahead of print. Issue: 7 Pages: 2057-2065
  • DOI: 10.1007/s00262-023-03388-5
  • Peer Reviewed
• [Journal Article] SOX10 Inhibits T Cell Recognition by Inducing Expression of the Immune Checkpoint Molecule PD-L1 in A375 Melanoma Cells (2023)
  • Author(s): Sasaki K, Hirohashi Y, Murata K, Minowa T, Nakatsugawa M, Murai A, Mizue Y, Kubo T, Kanaseki T, Tsukahara T, Iwabuchi S, Hashimoto S, Uhara H, Ishida-Yamamoto A, Torigoe T.
  • Journal Title: Anticancer Res Volume: 43 Issue: 4 Pages: 1477-1484
  • DOI: 10.21873/anticanres.16296
  • Peer Reviewed
• [Journal Article] Fusion with type 2 macrophages induces melanoma cell heterogeneity that potentiates immunological escape from cytotoxic T lymphocytes (2023)
  • Author(s): Minowa T, Hirohashi Y, Murata K, Sasaki K, Handa T, Nakatsugawa M, Mizue Y, Murai A, Kubo T, Kanaseki T, Tsukahara T, Iwabuchi S, Hashimoto S, Ishida-Yamamoto A, Uhara H, Torigoe T.
  • Journal Title: J Pathol Volume: Online ahead of print. Issue: 3 Pages: 304-316
  • DOI: 10.1002/path.6083
  • Peer Reviewed
• [Journal Article] GRIK2 is a target for bladder cancer stem-like cell-targeting immunotherapy (2022)
  • Author(s): Miyata H, Hirohashi Y, Yamada S, Yanagawa J, Murai A, Hashimoto S, Tokita S, Hori K, Abe T, Kubo T, Tsukahara T, Kanaseki T, Shinohara N, Torigoe T.
  • Journal Title: Cancer Immunology Immunotherapy Volume: 71 Issue: 4 Pages: 795-806
  • DOI: 10.1007/s00262-021-03025-z
  • Peer Reviewed
• [Journal Article] Tumor-infiltrating CD8 + T cells recognize a heterogeneously expressed functional neoantigen in clear cell renal cell carcinoma (2022)
  • Author(s): Matsuki M, Hirohashi Y, Nakatsugawa M, Murai A, Kubo T, Hashimoto S, Tokita S, Murata K, Kanaseki T, Tsukahara T, Nishida S, Tanaka T, Kitamura H, Masumori N, Torigoe T.
  • Journal Title: Cancer Immunology Immunotherapy Volume: 71 Issue: 4 Pages: 905-918
  • DOI: 10.1007/s00262-021-03048-6
  • Peer Reviewed
• [Journal Article] Identification of characteristic subepithelial surface granulomatosis in immune‐related adverse event‐associated enterocolitis (2021)
  • Author(s): Kubo Terufumi、Hirohashi Yoshihiko、Keira Yoshiko、Akimoto Mayuko、Ikeda Tatsuru、Kikuchi Noriaki、Iwaki Hiroyuki、Kikuchi Tomoki、Obata Masahiko、Morita Rena、Kasai Kiyoshi、Segawa Keiko、Tsukahara Tomohide、Kanaseki Takayuki、Murata Kenji、Kikuchi Yasuhiro、Shinkawa Tomoyo、Hasegawa Tadashi、Torigoe Toshihiko
  • Journal Title: Cancer Science Volume: 112 Issue: 3 Pages: 1320-1325
  • DOI: 10.1111/cas.14773
  • Peer Reviewed / Open Access
• [Journal Article] Epithelioid granulomatous lesions express abundant programmed death ligand-1 (PD-L1): a discussion of adverse events in anti-PD-1 antibody-based cancer immunotherapy. (2021)
  • Author(s): Kubo T, Hirohashi Y, Tsukahara T, Kanaseki T, Murata K, Hasegawa T, Torigoe T.
  • Journal Title: Hum Vaccin Immunother. Volume: 18 Issue: 7 Pages: 1-3
  • DOI: 10.1080/21645515.2020.1870364
  • Peer Reviewed
• [Journal Article] Neuregulin-1-β1 and γ-secretase play a critical role in sphere-formation and cell survival of urothelial carcinoma cancer stem-like cells (2021)
  • Author(s): Matsuki M, Inoue R, Murai A, Kubo T, Hashimoto S, Murata K, Kanaseki T, Tsukahara T, Nishida S, Tanaka T, Kitamura H, Masumori N, Hirohashi Y, Torigoe T.
  • Journal Title: Biochem Biophys Res Commun Volume: 552 Pages: 128-135
  • DOI: 10.1016/j.bbrc.2021.03.038
  • Peer Reviewed
• [Presentation] 癌免疫療法と腫瘍をとりまく免疫微小環境 (2022)
  • Author(s): 廣橋 良彦
  • Organizer: 第26回日本がん免疫学会
• [Presentation] 免疫応答をコントロールするがんの多様性 (2021)
  • Author(s): 廣橋 良彦
  • Organizer: 第25回日本がん免疫学会総会
• [Presentation] 腫瘍浸潤リンパ球の機能とその認識する抗原 (2021)
  • Author(s): 廣橋 良彦
  • Organizer: 和歌山悪性腫瘍研究会