炎症性腸疾患における腸上皮自然炎症調節機構の破綻

• Project Area: Homeostatic inflammation: Molecular basis and dysregulation
• Project/Area Number: 22117508
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 渡辺 守 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (10175127)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000); Fiscal Year 2011: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2010: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
• Keywords: 炎症性腸疾患 / 新規治療法 / 自然炎症 / 粘膜免疫 / 腸上皮細胞

Research Abstract

本研究は申請者らが独自に研究を展開してきた腸管上皮細胞による粘膜免疫の機能調節に注目し、自然炎症の調節異常がもたらす炎症性腸疾患の発症メカニズム解明と新規治療法の開発基盤を目指すものである。その解析には細胞株を用いた解析系よりも腸管上皮細胞の初代培養細胞が理想的である。我々は当該研究期間において、大腸組織から単離した上皮細胞の初代培養系を確立することに世界で初めて成功した(Nat Med 2012)。その培養技術の特徴は、マウス大腸組織から正常上皮を単一細胞として単離し、3次元的にsphereを形成させながら長期に亘って継代培養が可能なものである。
本研究期間で我々はこの技術をさらに応用し、腸管上皮細胞によるエンドサイトーシスおよびトランスサイトーシスの解析系を構築することに成功した。この解析系の樹立によって、これまで困難であった腸管上皮細胞に特徴的な膜輸送の生理的アルゴリズム解析が可能になったといえ、今後さまざまな免疫学的解析に応用できると期待する。
さらに現在我々は、Lgr5陽性上皮細胞から吸収上皮細胞、神経内分泌細胞、杯細胞など各々の系統に分化した細胞における自然炎症調節機構を詳細に解析している段階である。こうした研究結果は、これまで明確に把握し得なかった腸管上皮細胞における自然免疫応答を、より生理的で正確に解析できる重要な手がかりになると期待できる。
またこうして得られる解析結果をもとに、既に我々が確立した腸管上皮の移植実験系に応用し、in vivoにおける評価に直ちに結びつけることが可能であると我々は確信している。

Research Products

• [Journal Article] Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5+stem cell (2012)
  • Author(s): Yui S, Nakamura T, Sato T, Nemoto Y, Mizutani T, Zheng X, Ichinose S, Nagaishi T, Okamoto R, Tsuchiya K, Clevers H, and Watanabe M
  • Journal Title: Nature Medicine Volume: 18 Issue: 4 Pages: 618-623
  • DOI: 10.1038/nm.2695
  • Peer Reviewed
• [Journal Article] Real-time analysis of P-glycoprotein-mediated drug transport across primary intestinal epithelium three-dimensionally cultured in vitro (2012)
  • Author(s): Mizutani T, Nakamura T, et al
  • Journal Title: Biochem Biophys Res Commun Volume: 419 Issue: 2 Pages: 238-243
  • DOI: 10.1016/j.bbrc.2012.01.155
  • Peer Reviewed
• [Journal Article] The development of colitogenic CD4+ T cells is regulated by IL-7 in collaboration with natural killer cell function in a murine model of colitis (2012)
  • Author(s): Yamaji O, Totsuka T
  • Journal Title: J Immunol Volume: 188 Issue: 6 Pages: 2524-2536
  • DOI: 10.4049/jimmunol.1100371
  • Peer Reviewed
• [Journal Article] Luminal CD4+ T cells penetrate gut epithelial monolayers and egress from lamina propria to blood circulation (2011)
  • Author(s): Nemoto Y, Watanabe M
  • Journal Title: Gastroenterology Volume: 141 Issue: 6 Pages: 2130-2139
  • DOI: 10.1053/j.gastro.2011.08.035
  • Peer Reviewed
• [Journal Article] Upregulated IL-7Rα expression on colitogenic memory CD4+ T cells may participate in the development and persistence of chronic colitis (2011)
  • Author(s): Shinohara, T., et al
  • Journal Title: J.Immunol. Volume: 186 Issue: 4 Pages: 2623-2632
  • DOI: 10.4049/jimmunol.1000057
  • Peer Reviewed
• [Journal Article] Suppression of Hath1 gene expression directly regulated by Heslvia Notch signaling is associated with goblet cell depletion in ulcerative colitis (2011)
  • Author(s): Zheng X, Nakamura T, et al
  • Journal Title: Inflamm Bowel Dis Volume: 17 Issue: 11 Pages: 2251-2260
  • DOI: 10.1002/ibd.21611
  • Peer Reviewed
• [Journal Article] Suppression of hathl gene expression directly regulated byhesl via notch signaling is associated with goblet cell depletion in ulcerative colitis. (2011)
  • Author(s): Zheng X, Watanabe M, et al.
  • Journal Title: Inflamm Bowel Dis. Volume: (in press)
  • Peer Reviewed
• [Journal Article] Longitudinal cell formation in the entire human small intestine is correlated with the localization of Hath1 and Klf4. (2011)
  • Author(s): Iwasaki M, Watanabe M, et al.
  • Journal Title: J Gastroenterol Volume: 46 Pages: 191-202
  • NAID: 10029027237
  • Peer Reviewed
• [Journal Article] Upretulated IL-7 receptor (alpha) expression on colitogenic memory CD4+ T cells may participate in the development and persistence of chronic colitis. (2011)
  • Author(s): Shinohara T, Watanabe M, et al.
  • Journal Title: Journal of Immunology Volume: 186 Pages: 2623-2632
  • Peer Reviewed
• [Journal Article] Delta-like 1 expression promotes goblet cell differentiation in Notch-inactivated human colonic epithelial cells. (2010)
  • Author(s): Akiyama J, Watanabe M, et al.
  • Journal Title: Biochem Biophys Res Commun. Volume: 393 Pages: 662-667
  • Peer Reviewed
• [Journal Article] IL-2 is positively involved in the development of colitogenic CD4+ IL-7R alpha high memory T cells in chronic colitis. (2010)
  • Author(s): Kameyama K, Watanabe M, et al.
  • Journal Title: Eur J Immunol. Volume: 40 Pages: 2423-2436
  • Peer Reviewed
• [Journal Article] Cell culture and in-vivo analyses of plaque-derived cytopathic hepatitis C virus mutants. (2010)
  • Author(s): Mishima K, Watanabe M, et al.
  • Journal Title: Virology. Volume: 405 Pages: 361-369
  • Peer Reviewed
• [Presentation] Epithelial regeneration by cultured colonic cells expanded from a single adult Lgr5+ stem cell (2012)
  • Author(s): Nakamura T, Watanabe M
  • Organizer: The 6th Korea-Japan IBD Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2012-01-28
• [Presentation] Natural killer cells suppress a murine model of colitis by targeting the early stage of T eell development (2011)
  • Author(s): Nagaishi T, Yamaji O, Watanabe M, et al.
  • Organizer: International Congress of Mucosal Immunology 2011
  • Place of Presentation: Paris
  • Year and Date: 2011-07-07
• [Presentation] Regeneration of damaged colonic tissue by transplanted colonic epithelial stem cells maintained and expanded in vitro (2011)
  • Author(s): Yui S, Nakamura T, Nagaishi T, Watanabe M, et al.
  • Organizer: GI Research Academy 2011
  • Place of Presentation: Kyoto
  • Year and Date: 2011-06-17
• [Presentation] Regeneration of damaged colonic tissue by transplantation of colonic epithelial stem cells maintained and expanded in vitro (2011)
  • Author(s): Yui S, Nakamura T, Watanabe M, et al.
  • Organizer: DDW 2011
  • Place of Presentation: Chicago
  • Year and Date: 2011-05-07
• [Presentation] NK cells regulate CD62L^-CD44^- T cell subset in the development of pathogenic T cells in a murine model of colitis (2011)
  • Author(s): Yamaji O, Nagaishi T, Watanabe M, et al.
  • Organizer: DDW 2011
  • Place of Presentation: Chicago
  • Year and Date: 2011-05-07
• [Presentation] TNF-α/NF-κB pathways in intestinal epithelial cells may be directly involved in colitis-associated carcinogenesis. (2010)
  • Author(s): Nagaishi T, Watanabe M, et al.
  • Organizer: The 4^<th> Japan and US Collaboration Conference in Gastroenterology
  • Place of Presentation: Tokyo
  • Year and Date: 2010-11-18
• [Presentation] TNF pathway in intestinal epithelial cells may be directly involved in colitis-associated carcinogenesis. (2010)
  • Author(s): Nagaishi T, Watanabe M, et al.
  • Organizer: The 14^<th> International Congress of Immunology
  • Place of Presentation: Kobe
  • Year and Date: 2010-08-25
• [Presentation] Myosin light chain kinase is associated with disruption of epithelial tight junction in an animal model of colitis-associated tumor. (2010)
  • Author(s): Nagaishi T, Watanabe M, et al.
  • Organizer: Federation of Clinical Immunology Society 2010
  • Place of Presentation: Boston
  • Year and Date: 2010-06-25
• [Presentation] Myosin light chain kinase is associated with disruption of epithelial tight junction in colitis-associated tumor. (2010)
  • Author(s): Nagaishi T, Watanabe M, et al.
  • Organizer: Digestive Disease Week 2010
  • Place of Presentation: New Orleans
  • Year and Date: 2010-05-02
• [Presentation] A long-term, mesenchyme-free in vitro culture system of colonic epithelial cells retaining stem cell compartment with proliferation/differentiation properties. (2010)
  • Author(s): Nakamura T, Watanabe M, et al.
  • Organizer: Digestive Disease Week 2010
  • Place of Presentation: New Orleans
• [Presentation] A Long-term, fully-defined culture system for colonic epithelial cells that allows efficient expansion of stem cell. (2010)
  • Author(s): Nakamura T, Nagaishi T, Watanabe M, et al.
  • Organizer: The 1st JSGE International Topic Conference
  • Place of Presentation: Kamakura
• [Book] 炎症性腸疾患:腸管粘膜免疫の特殊性 (2010)
  • Author(s): 永石宇司、渡辺守
  • Publisher: 医学書院
• [Book] 臨床粘膜免疫学:潰瘍性大腸炎 (2010)
  • Author(s): 永石宇司、渡辺守
  • Publisher: シナジー
• [Patent(Industrial Property Rights)] 大腸上皮幹細胞の単離・培養技術とこれを用いた大腸上皮移植技術 (2011)
  • Inventor(s): 渡辺守・中村哲也
  • Industrial Property Rights Holder: 国立大学法人東京医科歯科大学
  • Industrial Property Number: 2011-236469
  • Filing Date: 2011-10-27