Establishment of mouse models and organoids corresponding to colorectal cancer molecular subtypes and their clinical application

• Project/Area Number: 18K08694
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 55020:Digestive surgery-related
• Research Institution: Hiroshima University (2019-2020); Department of Clinical Research, National Hospital Organization Kure Medical Center (2018)
• Principal Investigator: Hinoi Takao 広島大学, 病院(医), 特任教授 (10444689)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 大腸癌のマウスモデル / 大腸癌のサブタイプ分類 / 分子腫瘍学 / FAP / RASシグナル伝達 / 大腸癌マウスモデル / CMS分類 / オルガノイド / サブクラス分類 / 大腸癌 / マウスモデル

Outline of Final Research Achievements

Consensus molecular subtypes(CMS) is the new colorectal cancer classification system based on gene expression profiles. Previously, we generated CMS1 mouse model characterized by MSI-high and CMS2 model characterized by CIN phenotype. In this study we generated GEMMs which recapitulate CMS3 (the metabolic phenotype) model characterized by the dysregulation of metabolic pathways with the activation of KRAS pathway and CMS4(mesenchymal subtype) model. In CMS3 model, we studied colonic neoplasia that has KrasG122V/BrafV600E mutation with Apc inactivation and compared gene expression profiles among Kras/Braf WT, Kras-mutated, and Braf-mutated mouse colon tumors to seek new molecular targets corresponding to the KRAS-BRAF-MAPK axis. We found Greb1 was the most upregulated gene in Braf-mutated tumors, suggesting the potential target of Wnt and RAS signaling pathway. We also established GEMM mouse with CMS4 subtypes and organoids, indicating the phenotype of CMS4 with promoted carcinogenesis.

Academic Significance and Societal Importance of the Research Achievements

大腸癌は罹患数が最も多い癌であり女性の癌死亡の第１位であり対策が急務である。大腸癌の臨床像や治療法を４つ亜型（サブタイプ）に分類したMolecular Consensus Subtype（MCS）の1-4型が欧米でも採用されつつある。私共は各CMS分類を再現する新規大腸癌マウスモデルを作製しており、これまでCMS1とCMS2を確立した。本研究では、治療抵抗性のCMS3とCMS4モデルを確立した。CMS3モデルではGreb1が新規治療標的として同定された。CMS4モデルでも、腫瘍の特性や治療法の検索が可能となった。これらのマウスモデルは大腸癌の基礎研究や新規治療法の探索に有用と思われる。

Research Products

• [Journal Article] Oncogenic mutation in RAS‐RAF axis leads to increased expression of GREB1, resulting in tumor proliferation in colorectal cancer (2020)
  • Author(s): Kochi Masatoshi、Hinoi Takao、Niitsu Hiroaki、Miguchi Masashi、Saito Yasufumi、Sada Haruki、Sentani Kazuhiro、Sakamoto Naoya、Oue Naohide、Tashiro Hirotaka、Sotomaru Yusuke、Yasui Wataru、Ohdan Hideki
  • Journal Title: Cancer Science Volume: 111 Issue: 10 Pages: 3540-3549
  • DOI: 10.1111/cas.14558
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A case of Turcot’s syndrome type 1 with loss of immunoexpression of MSH6 in colon cancer and liver metastasis due to secondary somatic mutation in coding mononucleotide (C)8 tract: a case report (2020)
  • Author(s): Akabane Shintaro、Hinoi Takao、Akagi Kiwamu、Yamamoto Hideki、Sada Haruki、Shimizu Yosuke、Shimizu Wataru、Sudo Takeshi、Onoe Takashi、Ishiyama Kohei、Suzuki Takahisa、Tazawa Hirofumi、Hadano Naoto、Misumi Toshihiro、Kojima Masato、Kubota Haruna、Taniyama Daiki、Kuraoka Kazuya、Tashiro Hirotaka
  • Journal Title: BMC Medical Genetics Volume: 21 Issue: 1 Pages: 141-141
  • DOI: 10.1186/s12881-020-01079-x
  • Peer Reviewed / Open Access
• [Journal Article] Synbiotics Suppress Colitis-Induced Tumorigenesis in a Colon-Specific Cancer Mouse Model (2019)
  • Author(s): Saito Y, Hinoi T, Adachi T, Miguchi M, Niitsu H, Kochi M, Sada H, Sotomaru Y, Sakamoto N, Sentani K, Oue N, Yasui W, Tashiro H, Ohdan H.
  • Journal Title: PLoS One Volume: 14 Issue: 6 Pages: 0216393-0216393
  • DOI: 10.1371/journal.pone.0216393
  • Peer Reviewed / Open Access
• [Journal Article] Clinicopathological significance of RCAN2 production in gastric carcinoma. (2019)
  • Author(s): Hattori Y, Sentani K, Shinmei S, Oo HZ, Hattori T, Imai T, Sekino Y, Sakamoto N, Oue N, Niitsu H, Hinoi T, Ohdan H, Yasui W.
  • Journal Title: Histopathology Volume: 74 Issue: 3 Pages: 430-442
  • DOI: 10.1111/his.13764
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Signal regulatory protein alpha blockade potentiates tumoricidal effects of macrophages on gastroenterological neoplastic cells in syngeneic immunocompetent mice (2018)
  • Author(s): Abe Tomoyuki、Tanaka Yuka、Piao Jinlian、Tanimine Naoki、Oue Naohide、Hinoi Takao、Garcia Noel Verjan、Miyasaka Masayuki、Matozaki Takashi、Yasui Wataru、Ohdan Hideki
  • Journal Title: Annals of Gastroenterological Surgery Volume: 2 Issue: 6 Pages: 451-462
  • DOI: 10.1002/ags3.12205
  • Peer Reviewed / Open Access
• [Presentation] BRAF mutation leads to increased expression of GeneX (GREB1) resulting in tumor proliferation in colorectal cancer (2020)
  • Author(s): Masatoshi Kochi, Takao Hinoi, Yasufumi Saito, Hiroaki Niitsu, Haruki Sada, Naoya Sakamoto, Kazuhiro Sentani, Naohide Oue, Wataru Yasui, Hideki Ohdan
  • Organizer: 第79回日本癌学会学術総会
• [Presentation] 日本人の遺伝性びまん性胃がんにおけるCDH1遺伝子生殖細胞系列変異 (2020)
  • Author(s): 山田英孝、胡徳、石野佳子、太田力、岩泉守哉、檜井孝夫、櫻井晃洋、田中屋宏爾、織田信弥、古庄知己、菅野康吉、吉田輝彦、椙村春彦
  • Organizer: 第79回日本癌学会学術総会
• [Presentation] Identification of novel colon cancer biomarkers by a new complex genetically modified mouse models (2019)
  • Author(s): 檜井孝夫
  • Organizer: 第74回 日本消化器外科学会総会
• [Presentation] 大腸上皮特異的転写活性配列を応用した新規大腸癌発癌マウスモデルの確立と発癌関連遺伝子の機能解析 (2019)
  • Author(s): 檜井孝夫
  • Organizer: 第28回 日本癌病態治療治療研究会
  • Invited
• [Presentation] 新規遺伝子改変大腸癌マウスモデルから確立したPtenハプロ不全型大腸癌オルガノイ ドの作成とその機能解析 (2019)
  • Author(s): 佐田春樹、檜井孝夫、坂本直也、仙谷和弘、大上直秀、安井弥、田代尊裕、大段秀樹
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] The functional analysis of Pten haploinsufficient colon cancer derived from mouse model using organoids (2019)
  • Author(s): Sada H, Hinoi T, Kochi M, Niitsu H, Ishikawa A, Sakamoto N, Sentani K, Oue N, Yasui W, Tashiro H, Ohdan H
  • Organizer: 第30回 日本消化器癌発生学会総会
• [Book] 遺伝性大腸癌診療ガイドライン 2020年版 (2020)
  • Author(s): 大腸癌研究会遺伝性大腸癌診療ガイドライン作成委員会、石田 秀行, 冨田 尚裕, 山口 達郎, 田中屋宏爾, 赤木 究, 石川 秀樹, 川崎 優子, 隈元 謙介, 下平 秀樹, 関根 茂樹, 高山 哲治, 田中 敏明, 田村 和朗, 田村智英子, 千野 晶子, 土井 悟, 中島 健, 中山 佳子, 長谷川博俊, 檜井 孝夫, 平沢 晃, 宮倉 安幸
  • Total Pages: 152
  • Publisher: 金原出版
  • ISBN: 4307204158
• [Book] 遺伝性大腸癌診療ガイドライン (2020)
  • Author(s): 大腸癌研究会 家族性腫瘍委員会
  • Total Pages: 124
  • Publisher: 金原出版株式会社