難治性がんサブタイプに対するCAR-T治療開発への多重ゲノム編集技術の応用

2021 Fiscal Year Annual Research Report

• Project/Area Number: 20K21627
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 田中 真二 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
• Keywords: ゲノム編集 / CAR-T / 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 Annual 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。申請者は挑戦的研究(萌芽)H30-31によりゲノム編集システムを搭載した高効率レンチウイルスベクターを開発し、多重ゲノム編集により同時に複数の遺伝子改変を導入する革新的手法に成功した。本研究では、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。
当研究室にてmesothelin scFv, CD19 scFvを各々搭載した第三世代CARベクターを構築し、C57BL/6マウスから分離したT細胞へ導入してmesothelin-CAR-T細胞、CD19-CAR-T細胞を作成した。C57BL/6由来のmesothelin導入マウス膵癌細胞株、CD19導入マウスCdh1:Trp53変異スキルス胃癌細胞株を多重ゲノム編集にて作成し、各CAR-T細胞が特異的に反応して殺細胞作用を示すことを確認した。NSGマウスを用いた移植腫瘍モデルにより、CAR-T 細胞のin vivo腫瘍縮小効果を検出したが、C57BL/6マウスを用いた同系統移植腫瘍モデルでは、CAR-T 細胞の腫瘍縮小効果が減弱することを見出した。そこで、マウスCAR-T細胞にsgRNA GeCKOライブラリーを導入してin vivo CRISPRスクリーニングを施行し、3種類の治療抵抗性遺伝子候補を同定した。多重ゲノム編集により３種類全てノックアウトしたCAR-T細胞を作成した結果、同系統移植腫瘍モデルにおける治療効果が著しく増強されることを明らかにした。さらに多重RNA編集を用いたノックダウンにて同治療効果が得られることを見出し、より安全性が高いCAR-T治療開発を推進することが可能となった。

Remarks

分子腫瘍医学研究室ホームページ
http://www.tmd.ac.jp/grad/monc/

Research Products

• [Int'l Joint Research] Brown University/MD Anderson Cancer Center(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Brown University/MD Anderson Cancer Center
• [Int'l Joint Research] Institut Cochin INSERM(フランス)
  • Country Name: FRANCE
  • Counterpart Institution: Institut Cochin INSERM
• [Int'l Joint Research] Seoul ASAN Medical Center/Seoul National University(韓国)
  • Country Name: KOREA (REP. OF KOREA)
  • Counterpart Institution: Seoul ASAN Medical Center/Seoul National University
• [Int'l Joint Research] Peking University(中国)
  • Country Name: CHINA
  • Counterpart Institution: Peking University
• [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 Pages: 83-95
  • DOI: 10.1007/s10120-021-01226-0
  • 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 Pages: 13
  • DOI: 10.3892/ijo.2022.5303
  • Peer Reviewed
• [Journal Article] Hormonal tumor mapping for liver metastases of gastroenteropancreatic neuroendocrine neoplasms: a novel therapeutic strategy (2022)
  • 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 Volume: 148 Pages: 697-706
  • DOI: 10.1007/s00432-021-03650-2
  • 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 (2022)
  • 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 Volume: 148 Pages: 931-941
  • DOI: 10.1007/s00432-021-03660-0
  • Peer Reviewed
• [Journal Article] Curative Surgery and Ki-67 Value Rather than Tumor Differentiation Predict the Survival of Patients with High-grade Neuroendocrine Neoplasms (2022)
  • 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: - Pages: in press
  • DOI: 10.1097/SLA.0000000000004495
  • 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: - Pages: in press
  • 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 Pages: 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] Surgery after sunitinib administration to improve survival of patients with advanced pancreatic neuroendocrine neoplasms (2021)
  • Author(s): Murase Y, Kudo A, Akahoshi K, Maekawa A, Ishikawa Y, Ueda H, Ogawa K, Ono H, Tanaka S, Tanabe M.
  • Journal Title: Annals of Gastroenterological Surgery Volume: 5 Pages: 692-700
  • DOI: 10.1002/ags3.12458
  • 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 Pages: 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 Pages: e0249885-
  • DOI: 10.1371/journal.pone.0249885
  • Peer Reviewed / Open Access
• [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 T, Ono H, Fujii M, Y, Akahoshi K, Ogura T, Ogawa K, Ban D, Kudo A, Tanaka S, Tanabe M.
  • Journal Title: PLOS ONE Volume: 16 Pages: e0252917
  • DOI: 10.1371/journal.pone.0252917
  • Peer Reviewed / Open Access
• [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, Ban D, Kudo A, Akiyama Y, Tanaka S, Ito H, Tanabe M.
  • Journal Title: Scientific Reports Volume: 11 Pages: 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 T, Shirai Y, Shimada Y, Hamura R, Yanagaki M, Takada N, Horiuchi T, Haruki K, Furukawa K, Tsuboi K, Okamoto Y, Shimada S, Tanaka S, Ohashi T, Ikegami T.
  • Journal Title: Cancer Science Volume: 112 Pages: 4570-4579
  • DOI: 10.1111/cas.15123
  • Peer Reviewed
• [Journal Article] Inhibitor library screening identifies ispinesib as a new potential chemotherapeutic agent for pancreatic cancers (2021)
  • Author(s): Murase Y, Ono H, Ogawa K, Yoshioka R, Ishikawa Y, Ueda H, Akahoshi K, Ban D, Kudo A, Tanaka S, Tanabe M.
  • Journal Title: Cancer Science Volume: 112 Pages: 4641-4654
  • DOI: 10.1111/cas.15134
  • Peer Reviewed
• [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
• [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/