Application of multiplex genome editing technology to the development of CAR-T therapy for refractory cancer subtypes

• Project/Area Number: 20K21627
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Tanaka Shinji 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (30253420)
• Co-Investigator(Kenkyū-buntansha): 秋山 好光 東京医科歯科大学, 大学院医歯学総合研究科, 講師 (80262187) ; 島田 周 東京医科歯科大学, 大学院医歯学総合研究科, 助教 (20609705) ; 新部 彩乃 (樺嶋) 東京医科歯科大学, 大学院医歯学総合研究科, 助教 (20445448)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2021: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2020: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
• Keywords: CAR-T / ゲノム編集 / 免疫チェックポイント / 免疫疲弊 / 難治性がん / 同系統移植モデル / 免疫抑制因子 / RNA編集

Outline of Research at the Start

免疫疲弊には様々な免疫チェックポイントや疲弊誘導因子など複数が報告されており、どのような欠失の組み合わせが最も有効なのか解明されていない。高効率多重ゲノム編集技術は、この難題を打破することができる重要なツールであり、本研究により免疫疲弊の分子機序が解明されるだけでなく、CAR-T細胞の疲弊を回避し有効性を持続させるという臨床応用への展開が可能となる。前臨床試験には実臨床に適したモデルが必須であるが、当研究室では臨床病態を再現する正常免疫モデル構築のみならず、独自のスキルス胃癌マウスを開発している。難治性癌の治療開発に直結した研究課題として、高いオリジナリティーを認める。

Outline of Final Research Achievements

CAR-T cell therapy is expected to be applied as one of the next-generation cancer immunotherapies, but the efficacy is still limited, and there is an urgent need to develop therapies that respond to the diversity of tumor immunity. In this study, we developed a highly efficient multiplex genome editing technology and developed novel CAR-T cells for refractory cancer. We constructed mouse CAR-T cells in our laboratory and observed their remarkable anti-tumor effect on pancreatic cancer tumors in mice transplanted into immunodeficient mice. We found that the anti-tumor effect was attenuated on tumors transplanted in the syngeneic immunocompetent mice, and identified candidate genes for the resistance to CAR-T therapy. Multiplex genome editing for the candidates revealed that the therapeutic effect was enhanced in the syngeneic immunocompetent xenograft models.

Academic Significance and Societal Importance of the Research Achievements

ゲノム編集技術は研究分野のみならず、治療開発への応用が強く期待されている。本研究では、申請者が開発した高効率多重ゲノム編集技術をCAR-T細胞開発へ応用し、新規がん免疫治療に展開することを目的とする。本課題は、CAR-T細胞の生体内維持及び免疫応答の機序解明のみならず、難治性がん治療開発に直結した実践的研究である。その成果によって、免疫応答下ではCAR-Ｔ細胞に対する治療抵抗性メカニズムが存在することを見出し、治療抵抗性遺伝子候補の解析によって難治性がんを克服できる可能性が示唆されており、現在RNA編集技術の応用によって安全性が高い新規CARベクター開発を進めている。

Research Products

• [Int'l Joint Research] Brown University/MD Anderson Cancer Center(米国)
• [Int'l Joint Research] Institut Cochin INSERM(フランス)
• [Int'l Joint Research] Seoul ASAN Medical Center/Seoul National University(韓国)
• [Int'l Joint Research] Peking University(中国)
• [Int'l Joint Research] Brown University/MD Anderson Cancer Center(米国)
• [Int'l Joint Research] Cochin INSERM(フランス)
• [Int'l Joint Research] Seoul ASAN Medical Center/Seoul National University(韓国)
• [Int'l Joint Research] Peking University(中国)
• [Journal Article] Somatostatin Receptor 2 Expression Profiles and Their Correlation with the Efficacy of Somatostatin Analogues in Gastrointestinal Neuroendocrine Tumors (2022)
  • Author(s): Watanabe Hirofumi、Fujishima Fumiyoshi、Komoto Izumi、Imamura Masayuki、Hijioka Susumu、Hara Kazuo、Yatabe Yasushi、Kudo Atsushi、Masui Toshihiko、Tsuchikawa Takahiro、Sakamoto Kazuhiro、Shiga Hisashi、Nakamura Tomohiro、Nakaya Naoki、Motoi Fuyuhiko、Unno Michiaki、Sasano Hironobu
  • Journal Title: Cancers Volume: 14 Issue: 5 Pages: 775-775
  • DOI: 10.1002/ags3.12458
  • Peer Reviewed / Open Access
• [Journal Article] Genomic landscape of a mouse model of diffuse-type gastric adenocarcinoma (2022)
  • Author(s): Zhang M, Sugita I, Komura D, Katoh H, Shimada S, Inazawa J, Tanaka S, Ishikawa S.
  • Journal Title: Gastric Cancer Volume: 25 Issue: 1 Pages: 83-95
  • DOI: 10.1007/s10120-021-01226-0
  • NAID: 40022799738
  • Peer Reviewed
• [Journal Article] Functional assessment of miR-1291 in colon cancer cells (2022)
  • Author(s): Wang J, Hirose H, Yokoyama Y, Ikeshima R, Tsujimura N, Bonkobara S, Takeda K, Hata T, Inoue A, Hiraki M, Ohtsuka M, Nishida N, Takahashi H, Haraguchi N, Tanaka S, Wu X, Mori M, Yamamoto H.
  • Journal Title: International Journal of Oncology Volume: 60 Issue: 2 Pages: 13-13
  • DOI: 10.3892/ijo.2022.5303
  • Peer Reviewed
• [Journal Article] Molecular targeted drugs, comprehensive classification and preclinical models for the implementation of precision immune oncology in hepatocellular carcinoma. (2022)
  • Author(s): Shimada S, Tanaka S.
  • Journal Title: International Journal of Clinical Oncology Volume: - Issue: 7 Pages: 1101-1109
  • DOI: 10.1007/s10147-022-02174-0
  • Peer Reviewed
• [Journal Article] Intrinsic activation of β-catenin signaling by CRISPR/Cas9-mediated exon skipping contributes to immune evasion in hepatocellular carcinoma (2021)
  • Author(s): Akasu M, Shimada S, Kabashima A, Akiyama Y, Shimokawa M, Akahoshi K, Kudo A, Yamaoka S, Tanabe M, Tanaka S.
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 16732-16732
  • DOI: 10.1038/s41598-021-96167-0
  • Peer Reviewed / Open Access
• [Journal Article] Arterial chemoembolisation with cisplatin versus epirubicin for hepatocellular carcinoma (ACE 500 study): A multicentre, randomised controlled phase 2/3 trial (2021)
  • Author(s): Aramaki O, Takayama T, Moriguchi M, Sakamoto H, Yodono H, Kokudo N, Yamanaka N, Kawasaki S, Sasaki Y, Kubota K, Otsuji E, Tanaka S, Matsuyama Y, Fujii M, for the ACE500 study group.
  • Journal Title: European Journal of Cancer Volume: 157 Pages: 373-382
  • DOI: 10.1016/j.ejca.2021.08.027
  • Peer Reviewed
• [Journal Article] Laparoscopic distal pancreatectomy in a patient with aberrant splenic artery originating from the superior mesenteric artery (2021)
  • Author(s): Ishida H, Ishikawa Y, Akahoshi K, Ueda H, Morimoto K, Yamashita H, Ogawa K, Ono H, Kudo A, Tanaka S, Tanabe M.
  • Journal Title: Medicine Volume: 100 Issue: 18 Pages: e25704-e25704
  • DOI: 10.1097/md.0000000000025704
  • Peer Reviewed
• [Journal Article] Prediction of early recurrence of pancreatic ductal adenocarcinoma after resection (2021)
  • Author(s): Sugawara T, Ban D, Nishino J, Watanabe S, Ishikawa Y, Akahoshi K, Ogawa K, Ono H, Kudo A, Tanaka S, Tanabe M.
  • Journal Title: PLOS ONE Volume: 16 Issue: 4 Pages: e0249885-e0249885
  • DOI: 10.1371/journal.pone.0249885
  • Peer Reviewed / Open Access
• [Journal Article] Inhibitor library screening identifies ispinesib as a new potential chemotherapeutic agent for pancreatic cancers (2021)
  • Author(s): Murase Yoshiki、Ono Hiroaki、Ogawa Kosuke、Yoshioka Risa、Ishikawa Yoshiya、Ueda Hiroki、Akahoshi Keiichi、Ban Daisuke、Kudo Atsushi、Tanaka Shinji、Tanabe Minoru
  • Journal Title: Cancer Science Volume: 112 Issue: 6 Pages: 4641-4654
  • DOI: 10.1371/journal.pone.0252917
  • Peer Reviewed / Open Access
• [Journal Article] C646 inhibits G2/M cell cycle-related proteins and potentiates anti-tumoreffects in pancreatic cancer (2021)
  • Author(s): Hiroaki Ono, Tomotaka Kato, Yoshiki Murase, Yutaro Nakamura, Yoshiya Ishikawa, Shuichi Watanabe, Keiichi Akahoshi, Toshiro Ogura, Kosuke Ogawa, Daisuke Ban, Atsushi Kudo, Yoshimitsu Akiyama, Shinji Tanaka, Hiromichi Ito, Minoru Tanabe
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 10078-10078
  • DOI: 10.1038/s41598-021-89530-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Inhibition of acid ceramidase elicits mitochondrial dysfunction and oxidative stress in pancreatic cancer cells (2021)
  • Author(s): Taniai Tomohiko、Shirai Yoshihiro、Shimada Yohta、Hamura Ryoga、Yanagaki Mitsuru、Takada Naoki、Horiuchi Takashi、Haruki Koichiro、Furukawa Kenei、Uwagawa Tadashi、Tsuboi Kazuhito、Okamoto Yasuo、Shimada Shu、Tanaka Shinji、Ohashi Toya、Ikegami Toru
  • Journal Title: Cancer Science Volume: 112 Issue: 11 Pages: 4570-4579
  • DOI: 10.1111/cas.15123
  • Peer Reviewed / Open Access
• [Journal Article] Surgery after sunitinib administration to improve survival of patients with advanced pancreatic neuroendocrine neoplasms (2021)
  • Author(s): Murase Yoshiki、Kudo Atsushi、Akahoshi Keiichi、Maekawa Aya、Ishikawa Yoshiya、Ueda Hiroki、Ogawa Kosuke、Ono Hiroaki、Tanaka Shinji、Tanabe Minoru
  • Journal Title: Annals of Gastroenterological Surgery Volume: 5 Issue: 11 Pages: 692-700
  • DOI: 10.1111/cas.15134
  • Peer Reviewed
• [Journal Article] Hormonal tumor mapping for liver metastases of gastroenteropancreatic neuroendocrine neoplasms: a novel therapeutic strategy. (2021)
  • Author(s): Maekawa A, Kudo A, Kishino Mitsuhiro, Murase Y, Watanabe S, Ishikawa Y, Ueda Hiroki, Akahoshi K, Ogawa K, Ono H, Tanaka S, Kinowaki Y, Tanabe M.
  • Journal Title: Journal of Cancer Research and Clinical Oncology, in press Volume: - Issue: 3 Pages: 931-941
  • DOI: 10.1007/s00432-021-03650-2
  • Peer Reviewed
• [Journal Article] Cytoplasmic RRM1 activation as an acute response to gemcitabine treatment is involved in drug resistance of pancreatic cancer cells (2021)
  • Author(s): Kato Tomotaka、Ono Hiroaki、Fujii Mikiya、Akahoshi Keiichi、Ogura Toshiro、Ogawa Kosuke、Ban Daisuke、Kudo Atsushi、Tanaka Shinji、Tanabe Minoru
  • Journal Title: PLOS ONE Volume: 16 Issue: 4 Pages: 0252917-0252917
  • DOI: 10.1007/s00432-021-03660-0
  • Peer Reviewed
• [Journal Article] Molecular and immunological paradigms of hepatocellular carcinoma: Special reference to therapeutic approaches (2021)
  • Author(s): Kabashima A, Shimada S, Shimokawa M, Akiyama Y, Tanaka S.
  • Journal Title: Journal of Hepato-Biliary-Pancreatic Sciences Volume: 28 Issue: 1 Pages: 62-75
  • DOI: 10.1002/jhbp.874
  • Peer Reviewed / Open Access
• [Journal Article] Islet cell dedifferentiation is a pathologic mechanism of long-standing progression of type 2 diabetes. (2021)
  • Author(s): Amo-Shiinoki K, Tanabe K, Hoshii Y, Matsui H, Harano R, Fukuda T, Takeuchi T, Bouchi R, Takagi T, Hatanaka M, Takeda K, Okuya S, Nishimura W, Kudo A, Tanaka S, Tanabe M, Akashi T, Yamada T, Ogawa Y, Ikeda E, Nagano H, Tanizawa Y.
  • Journal Title: JCI Insight Volume: 6 Issue: 1 Pages: 143791-143791
  • DOI: 10.1172/jci.insight.143791
  • Peer Reviewed / Open Access
• [Journal Article] Importance of Intestinal Environment and Cellular Plasticity of Islets in the Development of Postpancreatectomy Diabetes. (2021)
  • Author(s): Fukuda T, Bouchi R, Takeuchi T, Amo-Shiinoki K, Kudo A, Tanaka S, Tanabe M, Akashi T, Hirayama K, Odamaki T, Igarashi M, Kimura I, Tanabe K, Tanizawa Y, Yamada T, Ogawa Y
  • Journal Title: Diabetes Care Volume: 44(4) Issue: 4 Pages: 1002-1011
  • DOI: 10.2337/dc20-0864
  • Peer Reviewed / Open Access
• [Journal Article] Curative surgery and Ki-67 value rather than tumor differentiation predict the survival of patients with high-grade neuroendocrine neoplasms. (2021)
  • Author(s): Asano D, Kudo A, Akahoshi K, Ogawa K, Ono H, Ban D, Tanaka S, Tanabe M.
  • Journal Title: Annals of Surgery, in press Volume: -
  • Peer Reviewed
• [Journal Article] Laparoscopic distal pancreatectomy in a patient with aberrant splenic artery originating from the superior mesenteric artery (2021)
  • Author(s): Ishida H, Ishikawa Y, Akahoshi K, Ueda H, Morimoto K, Yamashita H, Ogawa K, Ono H, Kudo A, Tanaka S, Tanabe M.
  • Journal Title: Medicine, in press Volume: -
  • Peer Reviewed
• [Journal Article] A novel classification of portal venous tumor invasion to predict residual tumor status after surgery in patients with pancreatic neuroendocrine neoplasms. (2021)
  • Author(s): Kato T, Kudo A, Ishikawa Y, Watanabe S, Akahoshi K, Ogawa K, Ono H, Ban D, Tanaka S, Tanabe M.
  • Journal Title: Journal of Cancer Research and Clinical Oncology, in press Volume: -
  • Peer Reviewed
• [Journal Article] C646 inhibits G2/M cell cycle-related proteins and potentiates anti-tumor effects in pancreatic cancer. (2021)
  • Author(s): Ono H, Kato T, Murase Y, Nakamura Y, Ishikawa Y, Watanabe S, Akahoshi K, Ogura T, Ogawa K, Daisuke Ban, Kudo A, Akiyama Y, Tanaka S, Ito H, Tanabe M.
  • Journal Title: Scientific Reports, in press Volume: -
  • Peer Reviewed / Open Access
• [Journal Article] Curative Surgery and Ki-67 Value Rather than Tumor Differentiation Predict the Survival of Patients with High-grade Neuroendocrine Neoplasms (2020)
  • Author(s): Asano Daisuke、Kudo Atsushi、Akahoshi Keiichi、Maekawa Aya、Murase Yoshiki、Ogawa Kosuke、Ono Hiroaki、Ban Daisuke、Tanaka Shinji、Tanabe Minoru
  • Journal Title: Annals of Surgery Volume: Publish Ahead of Print Issue: 2 Pages: e108-e113
  • DOI: 10.1097/sla.0000000000004495
  • Peer Reviewed
• [Journal Article] Loss of ARID1A induces a stemness gene ALDH1A1 expression with histone acetylation in the malignant subtype of cholangiocarcinoma. (2020)
  • Author(s): Yoshino J, Akiyama Y, Shimada S, Ogura T, Ogawa K, Ono H, Mitsunori Y, Ban D, Kudo A, Yamaoka S, Tanabe M, Tanaka S.
  • Journal Title: Carcinogenesis Volume: in press Issue: 6 Pages: 0000-0000
  • DOI: 10.1093/carcin/bgz179
  • Peer Reviewed / Open Access
• [Presentation] Surgical oncology to develop novel targeted and immunotherapies for hepato-biliary-pancreatic cancer (Award Session) (2021)
  • Author(s): Shinji Tanaka
  • Organizer: 80th Annual Meeting of Japanese Cancer Association
  • Int'l Joint Research / Invited
• [Presentation] Stemness and heterogeneity of liver cancer (2021)
  • Author(s): Shinji Tanaka
  • Organizer: JSH International Liver Conference 2021
  • Int'l Joint Research / Invited
• [Presentation] 多様性と科学の発展 (2021)
  • Author(s): 田中真二
  • Organizer: 第25回日本心療内科学会学術大会
  • Invited
• [Presentation] 肝臓外科からみた代謝関連肝癌の分子機序と治療戦略 (2021)
  • Author(s): 田中真二、島田周、秋山好光、新部彩乃、波多野恵、下川雅弘、田邉稔、小川佳宏
  • Organizer: 第7回肝臓と糖尿病・代謝研究会
  • Invited
• [Presentation] ゲノム医療・ゲノム編集時代における癌研究と治療展開（シンポジウム：基調講演） (2020)
  • Author(s): 田中真二
  • Organizer: 第56回日本肝臓学会総会
  • Invited
• [Presentation] ゲノム医療・ゲノム編集時代における難治性消化器がんの治療戦略（統合プログラム） (2020)
  • Author(s): 田中真二、絹笠祐介、田邉稔
  • Organizer: 第18回日本消化器外科学会大会
  • Invited
• [Presentation] ゲノム医療・ゲノム編集時代における複合免疫治療の開発と外科臨床研究の重要性（ワークショップ） (2020)
  • Author(s): 田中真二、吉野潤、奥野圭佑、赤須雅文、金本栄美、渡辺秀一、伴大輔、工藤篤、絹笠祐介、田邉稔
  • Organizer: 第75回日本消化器外科学会総会
• [Remarks] β-catenin活性化型肝がんの免疫回避機構を解明
  • URL: https://www.tmd.ac.jp/press-release/20210824-1/
• [Remarks] 第23回日本肝がん分子標的治療研究会（主催：オンライン）
  • URL: https://site2.convention.co.jp/kangan23/
• [Remarks] 分子腫瘍医学研究室ホームページ
  • URL: http://www.tmd.ac.jp/grad/monc/