Elucidation of mucosal healing mechanisms through the construction of a human Crohn 's disease model

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K08378
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Kazuo Ohtsuka 東京医科歯科大学, 医学部附属病院, 准教授 (00338443)
• Co-Investigator(Kenkyū-buntansha): 土屋 輝一郎 筑波大学, 医学医療系, 教授 (40376786)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: クローン病 / オルガノイド

Outline of Final Research Achievements

In this study, we aimed to integrate applicant-specific Crohn's disease patient stratification and in vitro models to obtain novel insights into small intestine-specific mucosal damage. During the study period, we isolated intestinal epithelial cells from the human small intestine and established organoids by culturing the small intestinal epithelial cells using a three-dimensional primary culture method. We confirmed the expression of inflammatory stimulus receptors in the organoids and determined pro-inflammatory factors to establish the culture conditions. After 12 weeks of inflammatory stimulation, we recovered the organoids, extracted RNA, and compared them with organoids treated with the stimulus for 3 hours, confirming the accumulation of NF-kB signaling. Furthermore, we examined the appropriate culture conditions, including matrices, from the perspective of obtaining organoids from endoscopic biopsy tissues and accumulated new findings.

Free Research Field

消化器病学

Academic Significance and Societal Importance of the Research Achievements

本研究で得られた知見・成果を基に、治療薬や病型等により層別化を行ったクローン病患者由来の小腸オルガノイドについて解析を進めることで、炎症環境のみならず生物学的製剤等による治療応答やLRG等の既存の血清バイオマーカー等と小腸上皮障害維持機構・粘膜治癒を規定する因子がいかなる関係を有するのかが明らかとなることが期待できる。さらに同疾患において小腸病変の成立・局在を規定する要因の解析・新規バイオマーカーの同定を進めることも可能となり、小腸粘膜の全層性の寛解を規定する因子等も明らかとなることが期待できる。