The analysis of the regulation in intestinal differentiation by thecomprehensive search of Hath1 target genes

2008 Fiscal Year Final Research Report

• Project/Area Number: 19590717
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: TSUCHIYA Kiichirou Tokyo Medical and Dental University, 医学部附属病院, 講師 (40376786)
• Co-Investigator(Kenkyū-buntansha): WATANABE Mamoru 東京医科歯科大学, 大学院・医歯学総合研究科・消化器病態学, 教授 (10175127) ; OKADA Eriko 東京医科歯科大学, 医学部附属病院, 医員 (20376784) ; FUKAO Taro 慶應義塾大学, 医学部, 助教 (20401127)
• Co-Investigator(Renkei-kenkyūsha): WATANABE Mamoru 東京医科歯科大学, 大学院・医歯学総合研究科・消化器病態学, 教授 (10175127)
• Project Period (FY): 2007 – 2008
• Keywords: Atoh1 / Hath1 / 小腸 / 分化制御 / 標的遺伝子

Research Abstract

本研究は申請者が独自に見いだした腸管上皮細胞分化マスター遺伝子であるbHLH型転写因子Hath1の機能に着目し、Hath1標的遺伝子の網羅的検索から特定の細胞種への分化機構の解明を目的とし、粘膜再生誘導を主眼とした難治性慢性腸炎の新規治療法開発への基盤とするものである。本研究では当初の研究計画に示した項目につき、下記に示すごとく大きな研究成果が得られた。1) Notch, Wntシグナル下でのHath1標的遺伝子の網羅的検索においては、腸管上皮由来細胞株にテトラサイクリン誘導Hath1発現系を確立し、Hath1発現による遺伝子発現変化をマイクロアレイを用いて網羅的に検索したところ、複数の候補遺伝子の抽出できた。中でも杯細胞形質、パネート細胞形質に関与する遺伝子の検出を確認した。2) Hath1標的遺伝子による特定細胞種への分化誘導解析においては、レンチウイルスベクターを用い腸管上皮細胞由来株に遺伝子導入する系を構築した。3) ヒト炎症性腸疾患患者の腸管上皮細胞異常分布によるHath1標的遺伝子の発現解析においては、ダブルバルーン内視鏡により全小腸を観察出来た患者の生検検体を用い、小腸の部位別に遺伝子発現解析を行った。Hath1遺伝子は口側から肛門側へより強い発現を認め、それに伴い腸管上皮細胞の中で杯細胞だけ増加を認めた。Hath1遺伝子増加は杯細胞形質発現を標的とすることが示唆された。以上よりHath1発現系を構築しHath1標的遺伝子の候補遺伝子を抽出するとともに、実際の小腸粘膜における発現検討により腸管粘膜構築との関連性を明らかにするなど多大な成果を挙げた。

Research Products

• [Journal Article] Requirement of Notch activation duringregeneration of the intestinal epithelia (2009)
  • Author(s): Okamoto R, Tsuchiya K, Nemoto Y, AkiyamaJ, Nakamura T, Kanai T, Watanabe M
  • Journal Title: Am JPhysiol GI & Liver 296 Pages: 23-35
  • Peer Reviewed
• [Journal Article] Proteasomal degradation of Atoh1 by aberrant Wnt signaling maintains the undifferentiated state ofcolon cancer (2008)
  • Author(s): Aragaki M, Tsuchiya K, Okamoto R, Yoshioka S, Nakamura T, Sakamoto N, KanaiT, Watanabe M
  • Journal Title: Biochem Biophys Res Commun 368 Pages: 923-929
  • Peer Reviewed
• [Journal Article] Inducible expression of microRNA-194 is regulated by HNF-1 during intestinal epithelial cell differentiation (2008)
  • Author(s): Hino K, Tsuchiya K, Taro Fukao, KotaroKiga, Ryuichi Okamoto, Tkanori Kanai, Watanabe M
  • Journal Title: RNA 14 Pages: 1433-1442
  • Peer Reviewed
• [Journal Article] Negative feedback regulation of colitogenic CD4+ T cells by increased granulopoiesis (2008)
  • Author(s): Nemoto Y, Kanai T, Tohda S, Totsuka T, Okamoto R, Tsuchiya K, Nakamura T, Sakamoto N, Fukuda T, Miura O, Yagita H, Watanabe M
  • Journal Title: Inflamm Bowel Dis 14 Pages: 1491-1503
  • Peer Reviewed
• [Journal Article] Systemic, but not intestinal, IL-7 is essential for the persistence ofchronic colitis (2008)
  • Author(s): Tomita T, Kanai T, Nemoto Y, Totsuka T, Okamoto R, Tsuchiya K, Sakamoto N, Watanabe M
  • Journal Title: J Immunol 180 Pages: 383-390
  • Peer Reviewed
• [Journal Article] MyD88-dependent pathway in T cells directly modulates the expansion and survival of colitogenic CD4+ T cells in chronic colitis (2008)
  • Author(s): Tomita T, Kanai T, Fujii T, Nemoto Y, Okamoto R, Tsuchiya K, Totsuka T, SakamotoN, Akira S, Watanabe M
  • Journal Title: J Immunol 180 Pages: 5291-5299
  • Peer Reviewed
• [Journal Article] Colitogenic CD4^+ effector-memory T cells actively recirculate in chronic coliticmice (2008)
  • Author(s): Tomita T, Kanai T, Nemoto Y, Fujii T, Okamoto R, Tsuchiya K, Nakamura T, Sakamoto N, Totsuka T, Watanabe M
  • Journal Title: Inflamm Bowel Dis 14 Pages: 1630-1640
  • Peer Reviewed
• [Journal Article] Intestinal lamina propria retaining CD4+CD25+ regulatory T cells is a suppressive site of intestinal inflammation (2007)
  • Author(s): Makita S, Kanai T, Nemoto Y, Totsuka T, Okamoto R, Tsuchiya K, Yamamoto M, Kiyono H, Watanabe M
  • Journal Title: J Immunol 178 Pages: 4937-4946
  • Peer Reviewed
• [Journal Article] Bone marrow retaining colitogenic CD4+ T cells may be a pathogenic reservoir for chronic colitis (2007)
  • Author(s): Nemoto Y, Kanai T, Makita S, Okamoto R, Totsuka T, Takeda K, Watanabe M
  • Journal Title: Gastroenterology 132 Pages: 176-189
  • Peer Reviewed
• [Journal Article] IL-7 is essential for the development and the persistence of chronic colitis (2007)
  • Author(s): Totsuka T, Kanai T, Nemoto Y, Makita S, Okamoto R, Tsuchiya K, Watanabe M
  • Journal Title: J Immunol 178 Pages: 4737-4748
  • Peer Reviewed
• [Journal Article] Reciprocal targeting of Hath1 and β-catenin by Wnt-glycogen synthase kinase 3β in human colon cancer (2007)
  • Author(s): Tsuchiya K, Nakamura T, Okamoto R, Kanai T, Watanabe M
  • Journal Title: Gastroenterology 132 Pages: 208-220
  • Peer Reviewed
• [Presentation] Dysregulated differentiation of intestinal epithelia in UC (2008)
  • Author(s): Okamoto R, Tsuchiya K, Nakamura T, Kanai T, Watanabe M
  • Organizer: 3rd Japan-Korea IBD Symposium
  • Place of Presentation: Korea(Seoul)
  • Year and Date: 2008-09-20
• [Presentation] Notch signaling suppresses the transcriptional activity of Hath1 Gene, resulting in the undifferentiated form of human intestinal epithelial cells (2008)
  • Author(s): Tsuchiya K, Inoue K, Aragaki M, Okamoto R, Nakamura T, Kanai T, Watanabe M
  • Organizer: DDW 2008
  • Place of Presentation: San Diego
  • Year and Date: 2008-05-20
• [Presentation] Stabilization of Atoh1 protein induces Mucin2 gene expression in human colon cancer cells (2007)
  • Author(s): Tsuchiya K, Okamoto R, Kanai T, Watanabe M
  • Organizer: DDW 2007
  • Place of Presentation: Washington, D.C.
  • Year and Date: 2007-05-23
• [Presentation] IL-7 is essential for the development and persistence of chronic colitis (2007)
  • Author(s): Totsuka T, Kanai T, Nemoto Y, Tomita T, Makita S, Totsuka T, Watanabe M
  • Organizer: DDW 2007
  • Place of Presentation: Washington, D.C.
  • Year and Date: 2007-05-22
• [Presentation] Activated notch signaling suppresses generation of goblet cells in the human intestinal mucosa (2007)
  • Author(s): Okamoto R, Tsuchiya K, Kanai T, Watanabe M
  • Organizer: DDW 2007
  • Place of Presentation: Washington, D.C.
  • Year and Date: 2007-05-20
• [Presentation] Blockade of TNF-α inhibits tumor progression in colitis-associated cancer (2007)
  • Author(s): Onizawa M, Kanai T, Nemoto Y, Oshima S, Makita S, Okamoto R, Totsuka T, Yagita H, Watanabe M
  • Organizer: DDW 2007
  • Place of Presentation: Washington, D.C.
  • Year and Date: 2007-05-20