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

• 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
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: クローン病 / オルガノイド / 粘膜治癒 / 体外疾患モデル / 小腸 / ヒト体外IBDモデル

Outline of Research at the Start

独自に構築したヒト細胞での体外IBDモデルを小腸に応用し、種々のクローン病モデルを構築することで特異的障害機構が解明できると着想した。申請者独自のクローン病患者層別化とクローン病体外モデルを融合させることにより、小腸特異的障害に関する鍵分子の同定を目的とする。将来的には鍵分子を標的とした治療法の開発により、小腸粘膜を直接の標的とした治療の確立が期待でき、クローン病以外の小腸難病にまで展開させることが可能である。小腸内視鏡を多く施行することで小腸粘膜評価法を構築し、且つ豊富な生検検体を活用して基礎研究まで発展させてきた申請者のみに遂行可能であり、独創的で世界的な評価に耐えうる研究と考える。

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.

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。

Research Products

• [Journal Article] Systematic review of artificial <scp>intelligence‐based</scp> image diagnosis for inflammatory bowel disease (2022)
  • Author(s): Kawamoto Ami、Takenaka Kento、Okamoto Ryuichi、Watanabe Mamoru、Ohtsuka Kazuo
  • Journal Title: Digestive Endoscopy Volume: 34 Issue: 7 Pages: 1311-1319
  • DOI: 10.1111/den.14334
• [Journal Article] Transmural remission characterized by high biologic concentrations demonstrates better prognosis in Crohn's disease (2022)
  • Author(s): Kento Takenaka, Ami Kawamoto, Yoshio Kitazume, Toshimitsu Fujii, Yumi Udagawa, Hiromichi Shimizu, Shuji Hibiya, Masakazu Nagahori, Kazuo Ohtsuka, Mamoru Watanabe, Ryuichi Okamoto
  • Journal Title: J Crohns Colitis. Volume: - Issue: 6 Pages: 855-862
  • DOI: 10.1093/ecco-jcc/jjac185
  • Peer Reviewed
• [Journal Article] Serum Leucine-Rich α2 Glycoprotein: A Novel Biomarker for Transmural Inflammation in Crohn's Disease (2022)
  • Author(s): Kento Takenaka, Yoshio Kitazume, Ami Kawamoto, Toshimitsu Fujii, Yumi Udagawa, Ryosuke Wanatabe, Hiromichi Shimizu, Shuji Hibiya, Masakazu Nagahori, Kazuo Ohtsuka, Hiroyuki Sato, Akihiro Hirakawa, Mamoru Watanabe, Ryuichi Okamoto
  • Journal Title: Am J Gastroenterol. Volume: - Issue: 6 Pages: 1028-1035
  • DOI: 10.14309/ajg.0000000000002127
  • Peer Reviewed
• [Journal Article] Deep neural network for video colonoscopy of ulcerative colitis: a cross-sectional study (2022)
  • Author(s): Kento Takenaka, Toshimitsu Fujii, Ami Kawamoto, Kohei Suzuki, Hiromichi Shimizu, Chiaki Maeyashiki, Osamu Yamaji, Maiko Motobayashi, Akira Igarashi, Ryoichi Hanazawa, Shuji Hibiya, Masakazu Nagahori, Eiko Saito, Ryuichi Okamoto, Kazuo Ohtsuka, Mamoru Watanabe
  • Journal Title: Lancet Gastroenterol Hepatol Volume: 7 Issue: 3 Pages: 230-237
  • DOI: 10.1016/s2468-1253(21)00372-1
  • Peer Reviewed / Open Access
• [Journal Article] Higher concentrations of cytokine blockers are needed to obtain small bowel mucosal healing during maintenance therapy in Crohn's disease (2021)
  • Author(s): Kento Takenaka, Ami Kawamoto, Shuji Hibiya, Kohei Suzuki, Toshimitsu Fujii, Maiko Motobayashi, Hiromichi Shimizu, Masakazu Nagahori, Eiko Saito, Ryuichi Okamoto, Mamoru Watanabe, Kazuo Ohtsuka
  • Journal Title: Aliment Pharmacol Ther Volume: 54 Issue: 8 Pages: 1052-1060
  • DOI: 10.1111/apt.16551
  • Peer Reviewed
• [Journal Article] Deep Neural Network Accurately Predicts Prognosis of Ulcerative Colitis Using Endoscopic Images (2021)
  • Author(s): Kento Takenaka, Kazuo Ohtsuka, Toshimitsu Fujii, Shiori Oshima, Ryuichi Okamoto, Mamoru Watanabe
  • Journal Title: Gastroenterology Volume: 160 Issue: 6 Pages: 2175-2177
  • DOI: 10.1053/j.gastro.2021.01.210
  • Peer Reviewed / Open Access
• [Journal Article] Serum leucine-rich α-2 glycoprotein; a novel biomarker for small bowel mucosal activity in Crohn’s Disease. (2021)
  • Author(s): Kawamoto T, Takenaka K, Hibiya S, Ohtsuka K, Okamoto R, Watanabe M
  • Journal Title: Clin Gastroenterol Hepatol. Volume: 20 Issue: 5 Pages: e1196-e1200
  • DOI: 10.1016/j.cgh.2021.06.036
  • Peer Reviewed
• [Journal Article] Intravenous tacrolimus is a superior induction therapy for acute severe ulcerative colitis compared to oral tacrolimus (2021)
  • Author(s): Hiromichi Shimizu, Toshimitsu Fujii, Kenji Kinoshita, Ami Kawamoto, Shuji Hibiya, Kento Takenaka, Eiko Saito, Masakazu Nagahori, Kazuo Ohtsuka, Mamoru Watanabe, Ryuichi Okamoto
  • Journal Title: BMC Gastroenterol Volume: 21 Issue: 1 Pages: 494-494
  • DOI: 10.1186/s12876-021-02043-6
  • Peer Reviewed / Open Access
• [Journal Article] Combined endocytoscopy with pit pattern diagnosis in ulcerative colitis‐associated neoplasia: Pilot study (2021)
  • Author(s): Kudo Shin‐ei、Maeda Yasuharu、Okumura Taishi、Matsudaira Shingo、Ishigaki Tomoyuki、Sasanuma Seiko、Mori Yuichi、Kudo Toyoki、Hayashi Takemasa、Wakamura Kunihiko、Miyachi Hideyuki、Baba Toshiyuki、Ohtsuka Kazuo
  • Journal Title: Digestive Endoscopy Volume: - Issue: 1 Pages: 133-143
  • DOI: 10.1111/den.13964
  • Peer Reviewed
• [Journal Article] Crohn disease: magnetic resonance enterocolonography features of endoscopic ulcer stages reclassified with the healing process and the relationships to prognoses (2021)
  • Author(s): Yoshio Kitazume, Kento Takenaka, Kazuo Ohtsuka, Koichiro Kimura, Yusuke Ogihara, Tomoyuki Fujioka, Junichi Tsuchiya, Toshimitsu Fujii, Mamoru Watanabe, Ukihide Tateisi
  • Journal Title: Jpn J Radiol Volume: 39 Issue: 5 Pages: 459-476
  • DOI: 10.1007/s11604-020-01082-5
  • NAID: 210000160272
  • Peer Reviewed
• [Journal Article] Colonic strictures mimicking Crohn’s disease (2021)
  • Author(s): Takenaka Kento、Kimura Maiko、Ohtsuka Kazuo
  • Journal Title: Gut Volume: 70 Issue: 2 Pages: 233-387
  • DOI: 10.1136/gutjnl-2019-320172
  • Peer Reviewed
• [Journal Article] High b-value computed diffusion-weighted imaging for differentiating bowel inflammation in Crohn's disease (2020)
  • Author(s): Yoshio Kitazume, Junichi Tsuchiya, Kento Takenaka, Kazuo Ohtsuka, Koichiro Kimura, Ryosuke Watanabe, Tomoyuki Fujioka, Toshimitsu Fujii, Mamoru Watanabe, Ukihide Tateisi
  • Journal Title: Eur J Radiol Volume: 133 Pages: 109362-109362
  • DOI: 10.1016/j.ejrad.2020.109362
  • Peer Reviewed
• [Journal Article] Objective evaluation for treat to target in Crohn’s disease (2020)
  • Author(s): Takenaka Kento、Kitazume Yoshio、Fujii Toshimitsu、Tsuchiya Kiichiro、Watanabe Mamoru、Ohtsuka Kazuo
  • Journal Title: Journal of Gastroenterology Volume: 55 Issue: 6 Pages: 579-587
  • DOI: 10.1007/s00535-020-01678-8
  • Peer Reviewed / Open Access