Regulation of membrane-anchored serine protease activities and its significance in epithelial pathophysiology

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K06964
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49010:Pathological biochemistry-related
• Research Institution: University of Miyazaki
• Principal Investigator: Kawaguchi Makiko 宮崎大学, 医学部, 助教 (90405598)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: HAI-2 / matriptase / prostasin / EpCAM

Outline of Final Research Achievements

In this study, we generated conditional Spint2 knockout mouse model based on the Cre recombinase and LoxP system. We found that spint2 knockout mouse showed severe epithelial damage in the whole intestinal tracts. The intestinal epithelium showed enhanced exfoliation, villous atrophy, enterocyte tufts and elongated crypts. Organoid crypt culture indicated that Spint2 ablation induced Epcam cleavage with decreased claudin-7 levels and resulted in organoid rupture. These organoid changes could be rescued by addition of serine protease inhibitors and matriptase selective inhibitor as well as by co-deletion of prostasin. These results indicate that HAI-2 is an essential cellular inhibitor for maintaining intestinal epithelium architecture.

Free Research Field

実験病理学

Academic Significance and Societal Importance of the Research Achievements

本研究では、HAI-2コンディショナルKOマウスを作製し、HAI-2をコードするSPINT2遺伝子の変異が原因であるヒトの先天性ナトリウム下痢症と類似の表現型を呈することを明らかにした。また、HAI-2 KOマウスやマウスから樹立したHAI-2欠損オルガノイドは、この疾患の病態の解明や新たな治療法開発やスクリーニングのツールとしても有用であることを示した。