Development of organoid-based carcinogenesis model of diffuse-type gastric cancer using murine primary gastric cells

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K19380
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Chiba Cancer Center (Research Institute)
• Principal Investigator: Maru Yoshiaki 千葉県がんセンター(研究所), 発がん研究グループ 発がん制御研究部, 研究員 (30742754)
• Co-Investigator(Renkei-kenkyūsha): MATSUURA Tetsuya 横浜市立大学付属病院, 消化器内科 (10784845)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: 胃がん / オルガノイド / マウス

Outline of Final Research Achievements

While long-term culture of primary gastric organoids with serum-free media was difficult at first, we were finally able to conduct long-term culture by optimization of medium composition. We then conducted lentiviral gene transduction into these gastric organoids and inoculated transduced organoids into dorsal skin of nude mice to examine tumorigenicity of frequently mutated genes in human gastric cancer (TP53, CDH1 and KRAS). Specifically, we cultured gastric cells isolated from conditional knockout and knock-in mice for Trp53 and mutant Kras, respectively, and transduced gastric organoids with Cre-recombinase to induce Trp53 deletion and/or Kras activation. Furthermore, we induced Cdh1 suppression by shRNA. Reconstitution of certain combinations of genetic alterations induced histologically different tumors. In conclusion, we here demonstrated that gastric organoids can be transformed by in vitro gene transduction.

Free Research Field

分子腫瘍学