Crosstalk between Wnt and Notch signaling in colonic epithelium.

• Project/Area Number: 21590805
• 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: SUZUKI Shinji 東京医科歯科大学, 大学院・医歯学総合研究科, 助教 (10456212)
• Co-Investigator(Kenkyū-buntansha): NAKAMURA Tetsuya 東京医科歯科大学, 大学院・医歯学総合研究科, 寄附講座教員 (70265809) ; WATANABE Mamoru 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (10175127)
• Co-Investigator(Renkei-kenkyūsha): ARAKI Akihiro 東京医科歯科大学, 医学部附属病院, 講師 (80361690) ; NAGAHORI Masakazu 東京医科歯科大学, 医学部附属病院, 助教 (60420254)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2009: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: 大腸上皮幹細胞 / Wntシグナル / Notchシグナル/初代培養/細胞移植治療 / 大腸上皮細胞 / 細胞増殖 / 細胞分化 / Notchシグナル

Research Abstract

Wnt and Notch pathways are known to function together to regulate proliferation and differentiation of intestinal epithelium ; however, the molecular mechanism of their functional interplay remains unclear. In this study, we aimed at investigating the signaling crosstalk between these two pathways in colonic epithelium. First, by using several human colon cancer-derived cell lines, we have demonstrated that the expression of Hath1, a bHLH transcription factor known to be involved in Notch signaling, is regulated by the proteasome-mediated protein degradation that is under the control of Wnt signaling. We also showed that this post-translational regulation of Hath1 by Wnt/Notch signals is important for cell fate determination of colonic cells. Second, in order to investigate the crosstalk between Wnt/Notch signals, we have established a novel method to isolate and culture normal colonic epithelium in vitro. By this method, colonic cells continuously grow and the Lgr5+colonic stem cells preferentially expand as a result of Wnt/Notch activation. We also showed for the first time that the cultured colonic cells are able to regenerate damaged epithelium in recipient mice when transplanted. These data would built a basis for the future study to elucidate molecular mechanism that govern the interplay of Wnt/Notch signals to regulate the regenerative properties and/or malignant transformation of colonic epithelial cells.

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, Morikawa R, Fukuda M, Mochizuki W, Yamauchi Y, NozakiK, Yui S, Nemoto Y, Nagaishi T, Okamoto R, Tsuchiya K, and Watanabe M
  • Journal Title: Biochem Biophys Res Commun Volume: 419(2) Pages: 238-243
• [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] MR enterocolonography is useful for simultaneous evaluation of small and large intestinal lesions in Crohn's disease (2011)
  • Author(s): Hyun SB, Kitazume Y, Nagahori M, Toriihara A, Fujii T, Tsuchiya K, Suzuki S, Okada E, Araki A, Naganuma M, Watanabe M
  • Journal Title: Inflam Bowel Dis Volume: 17(5) Pages: 1063-1072
• [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, Tsuchiya K, Okamoto R, ZhengX, Kano Y, Okamoto E, Okada E, Araki A, Suzuki S, Sakamoto N, Kitagaki K, Akashi T, Eishi Y, Nakamura T, Watanabe M
  • Journal Title: J Gastroenterol Volume: 46(2) Pages: 191-202
  • NAID: 10029027237
• [Journal Article] Gene bookmarking accelerates the kinetics of post-mitotic transcriptional activation (2011)
  • Author(s): Zhao R, Nakamura T, Fu Y, Lazar Z, and Spector DL
  • Journal Title: Nature Cell Biology Volume: 13(11) Pages: 1295-1304
• [Journal Article] Suppression of Hath1 gene expression directly regulated by Hes1 via Notch signaling is associated with goblet cell depletion in ulcerative colitis (2011)
  • Author(s): Zheng X, Tsuchiya K, Okamoto R, Iwasaki M, Kano Y, Sakamoto N, Nakamura T, Watanabe M
  • Journal Title: Inflamm Bowel Dis Volume: 17(11) Pages: 2251-2260
• [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] MR enterocolonography is useful for simultaneous evaluation of small and large intestinal lesions in Crohn's disease (2011)
  • Author(s): Hyun SB, Kitazume Y, Suzuki S, Watanabe M, et al
  • Journal Title: Inflam Bowel Dis Volume: 17 Issue: 5 Pages: 1063-1072
  • DOI: 10.1002/ibd.21510
  • 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, Tsuchiya K, Suzuki S, Watanabe M, et al
  • Journal Title: J Gastroenterol Volume: 46 Issue: 2 Pages: 191-202
  • DOI: 10.1007/s00535-010-0346-x
  • NAID: 10029027237
  • 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] Magnetic resonance enterocolonography is useful for simultaneous evaluation of small and large intestinall esions in Crohn's disease. (2011)
  • Author(s): Hyun SB, Kitazume Y, Nagahori M, Toriihara A, Fujii T, Tsuchiya K, Suzuki S, Okada E, Araki A, Naganuma M, Watanabe M
  • Journal Title: Inflamm Bowel Dis. Volume: In press
  • Peer Reviewed
• [Journal Article] Longtudinal cell formation in the entire human small intestine is correlated with the localization of Hathl and KIM (2011)
  • Author(s): Iwasaki M, Tsuchiya K, Okamoto R, Zheng X, Kano Y, Okamoto E, Okada E, Araki A, Suzuki S, Sakamoto N, Kitagaki K, Akashi T, Eishi Y, Nakamura T, Watanabe__M
  • Journal Title: J Gastroenterol. Volume: 46 Pages: 191-202
  • Peer Reviewed
• [Journal Article] Nuclear organization and dynamics of 7SK RNA in regulating gene expression (2010)
  • Author(s): Prasanth KV, Camiolo M, Chan G, Tripathi V, Denis L, Nakamura T, Hubner M, and Spector DL
  • Journal Title: Mol Biol Cell Volume: 21(23) Pages: 4184-4196
• [Journal Article] A long nuclear retained non-coding RNA regulates synaptogenesis by modulating gene expression (2010)
  • Author(s): Bernard D, Prasanth KV, Tripathi V, Colasse S, Nakamura T, Xuan Z, Zhang MQ, Sedel F, Jourdren L, Coulpier F, Triller A, Spector DL, and Bessis A
  • Journal Title: EMBO J Volume: 29(18) Pages: 3082-3093
• [Journal Article] Delta-like 1 expression promotes goblet cell differentiation in Notch-inactivated human colonic epithelial cells (2010)
  • Author(s): Akiyama J, Okamoto R, Iwasaki M, Zheng X, Yui S, Tsuchiya K, Nakamura T, and Watanabe M
  • Journal Title: Biochem Biophys Res Commun Volume: 393(4) Pages: 662-667
• [Journal Article] Delta-like 1 expression promotes goblet cell differentiation in Notch-inactivated human colonic epithelial cells. (2010)
  • Author(s): Akiyama J, Okamoto R, Nakamura T, et al
  • Journal Title: Biochem Biophys Res Commun. 393 Pages: 662-667
  • Peer Reviewed
• [Journal Article] Signaling pathway via TNF-a/NF-kB in intestinal epithelial cells may be directly involved in colitis-associated carcinogenesis (2009)
  • Author(s): Onizawa M, Nagaishi T, Kanai T, NaganoKI, Oshima S, Nemoto Y, Yoshioka A, TotsukaT, Okamoto R, Nakamura T, Sakamoto N, Tsuchiya K, Aoki K, Ohya K, Yagita H, Watanabe M
  • Journal Title: Am J Physiol Gastrointest Liver Physiol Volume: 296(4)
• [Journal Article] Requirement of Notch activation during regeneration of the intestinal epithelia. (2009)
  • Author(s): Okamoto R, Tsuchiya K, Nemoto Y, Akiyama J, Nakamura T, Kanai T, Watanabe M
  • Journal Title: Am J Physiol Gastrointest Liver Physiol Volume: 296(1)
• [Journal Article] Single-operator double-balloon endoscopy(DBE)is as effective as dual-operator DBE. (2009)
  • Author(s): Araki A, Tsuchiya K, Okada E, Suzuki S, et al.
  • Journal Title: J Gastroenterol Hepatol 24 Pages: 770-775
  • Peer Reviewed
• [Journal Article] ダブルバルーン内視鏡で診断し得た、先天性胆道拡張症術後肝内結石症の1例 (2009)
  • Author(s): 藤井俊光、鈴木伸治, 他
  • Journal Title: Progress of Digestive Endoscopy 75 Pages: 112-113
  • NAID: 130004985267
  • Peer Reviewed
• [Journal Article] Requirement of Notch activation during regeneration of the intestinal epithelia. (2009)
  • Author(s): Okamoto R, Nakamura T, et al.
  • Journal Title: Am J Physiol Gastrointest Liver Physiol 296
  • Peer Reviewed
• [Journal Article] RANK-RANKL signaling pathway is critically involved in the function of CD4+CD25+ regulatory T cells in chronic colitis. (2009)
  • Author(s): Totsuka T, Kanai T, Nakamura T, et al
  • Journal Title: J Immunol. 182 Pages: 6079-6087
  • Peer Reviewed
• [Journal Article] Signaling pathway via TNF-alpha/NF-kappaB in intestinal epithelial cells may be directly involved in colitis-associated carcinogenesis. (2009)
  • Author(s): Onizawa M, Nagaishi T, Nakamura T, et al
  • Journal Title: Am J Physiol Gastrointest Liver Physiol 296 Pages: 850-859
  • Peer Reviewed
• [Journal Article] (2009)
  • Author(s): 鈴木伸治、渡守
  • Journal Title: 総合臨床【消化管疾患診断と治療の進歩】治療法の進歩クローン病、潰瘍性大腸炎に対する新しい治療法(永井書店) Pages: 1972-1977
• [Presentation] Epithelial regeneration by cultured coloniccells expanded from a single adult LGR5+stem cell (2012)
  • Author(s): Tetsuya Nakamura
  • Organizer: The 6th Korea-Japan IBD Symposium
  • Place of Presentation: Tokyo, Japan
  • Year and Date: 2012-01-28
• [Presentation] Epithelial regeneration by cultured colonic cells expanded from asingle adult Lgr5+ stem cell (2012)
  • Author(s): Tetsuya Nakamura
  • Organizer: The 6th Korea-Japan IBD Symposium
  • Place of Presentation: Tokyo, Japan
  • Year and Date: 2012-01-28
• [Presentation] Regeneration of damaged colon epithelium by transplanted colon Lgr5+stem cells maintained and expanded in vitro (2011)
  • Author(s): Shiro Yui, Tetsuya Nakamura, Toshiro Sato, Yasuhiro Nemoto, Tomohiro Mizutani, Xiu Zheng, Shizuko Ichinose, Takashi Nagaishi, Ryuichi Okamoto, Kiichiro Tsuchiya, Hans Clevers, and Mamoru Watanabe
  • Organizer: The 5th Japan & US Collaboration Conference in Gastroenterology
  • Place of Presentation: Tokyo, Japan
  • Year and Date: 2011-11-10
• [Presentation] Flagellin via TLR5 on basolateral membrane of primary intestinal epithelial cells(IEC) shows the role of IEC in the response to bacteria (2011)
  • Author(s): Tsuchiya K, Zheng X, Kano Y, Okamoto R, Nakamura T, Watanabe M
  • Organizer: United European Gastroenterology Week 2011
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2011-10-26
• [Presentation] 原因不明消化管出血(OGIB)症例におけるカプセル内視鏡に対するダブルバルーン内視鏡の有用性の検討 (2011)
  • Author(s): 鈴木伸治、荒木昭博、渡辺守
  • Organizer: JDDW2011
  • Place of Presentation: 福岡
  • Year and Date: 2011-10-22
• [Presentation] 術後に判明した空腸異所性膵炎の一例 (2011)
  • Author(s): 鈴木康平、鈴木伸治、渡辺守
  • Organizer: 第16回お茶の水消化器セミナー
  • Place of Presentation: 東京
  • Year and Date: 2011-08-27
• [Presentation] Regeneration of damaged colonic tissue by transplanted colonic epithelial stem cells maintained and expanded in vitro (2011)
  • Author(s): Yui S, Nakamura T, Nemoto Y, MizutaniT, Zheng X, Nagaishi T, Tsuchiya K, Watanabe M, Okamoto R, Ichinose S, Sato T, Clevers H
  • Organizer: GI Research Academy 2011
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2011-06-17
• [Presentation] 再生医療へ向けた腸管上皮研究～幹細胞体外培養と細胞移植～ (2011)
  • Author(s): 中村哲也
  • Organizer: 日本消化器病学会総会シンポジウム指定口演
  • Place of Presentation: 東京
  • Year and Date: 2011-05-14
• [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: DDW2011
  • Place of Presentation: Chicago
  • Year and Date: 2011-05-07
• [Presentation] Regeneration of Damaged Colonic Tissue by Transplantation of Colonic Epithelial Stem Cells Maintained and Expanded In Vitro (2011)
  • Author(s): Shiro Yui, Tetsuya Nakamura, Yasuhiro Nemoto, Tomohiro Mizutani, Xiu Zheng, Shizuko Ichinose, Takashi Nagaishi, Ryuichi Okamoto, Kiichiro Tsuchiya, Toshiro Sato, Hans Clevers, and Mamoru Watanabe
  • Organizer: Basic Science Plenary Symposium, Digestive Disease Week 2011
  • Place of Presentation: Chicago, USA
• [Presentation] Awell-defined culture system for colonic epithelial cells thet allows efficient enrichment of LGR5+stem cells (2010)
  • Author(s): Yui S, Nakamura T, Nemoto Y, MizutaniT, Zheng X, Ichinose S, Nagaishi T, Okamoto R, Tsuchiya K, Watanabe M
  • Organizer: The 1st JSGE International Topic Conference-Stem Cells in Digestive Organs
  • Place of Presentation: Kamakura, Japan
  • Year and Date: 2010-09-26
• [Presentation] Up-reguration of DELTA1 expression is required for goblet cell differentiation in human intestinal epithelial cells (2010)
  • Author(s): Akiyama J, Okamoto R, Tsuchiya K, Nakamura T, Watanabe M
  • Organizer: The 1st JSGE International Topic Conference-Stem Cells in Digestive Organs
  • Place of Presentation: Kamakura, Japan
  • Year and Date: 2010-09-26
• [Presentation] HES1 via notch signaling directly suppresses ATOH1/HATH1 gene transcription, resulting in the undifferentiated state of intestinal epithelial cells (2010)
  • Author(s): Zheng X, Tsuchiya K, Okamoto R, Okamoto E, Iwasaki M, Kano Yoshihito, Nakamura T, Watanabe M
  • Organizer: The 1st JSGE International Topic Conference-Stem Cells in Digestive Organs
  • Place of Presentation: Kamakura, Japan
  • Year and Date: 2010-09-26
• [Presentation] TNF-αand notch signaling synergistically up-regulate OLFM4 expression in human intestinal epithelial cells (2010)
  • Author(s): Murano T, Okamoto R, Shimizu H, Akiyama J, Tsuchiya K, Nakamura T, Watanabe M
  • Organizer: The 1st JSGE International Topic Conference-Stem Cells in Digestive Organs
  • Place of Presentation: Kamakura, Japan
  • Year and Date: 2010-09-26
• [Presentation] A long-term, fully-deffined culture system for colonic epithelial cells that alows efficient expansion of stem cell compartment (2010)
  • Author(s): Tetsuya Nakamura, Watanabe M
  • Organizer: The 1st JSGE International Topic Conference-Stem Cells in Digestive Organs
  • Place of Presentation: Kamakura, Japan
  • Year and Date: 2010-09-25
• [Presentation] Hes1 via Notch signaling directly suppresses Atoh1/Hath1 gene transcription, resulting in the goblet cell depletion of Ulcerative Colitis (2010)
  • Author(s): Zheng X, Tsuchiya K, Okamoto R, Iwasaki M, Kano Y, Nakamura T, Watanabe M
  • Organizer: Digestive Disease Week 2010
  • Place of Presentation: NewOrleans, USA
  • Year and Date: 2010-05-03
• [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): Tetsuya Nakamura, Shiro Yui, Tomohiro Mizutani, Yasuhiro Nemoto, Ryuichi Okamoto, Kiichiro Tsuchiya, Shizuko Ichinose, and Mamoru Watanabe
  • Organizer: Digestive Disease Week 2010
  • Place of Presentation: New Orleans, USA
• [Presentation] GSK3 inhibitor induces the intestinal differentiation by the protein stabilization of Atoh1 (2009)
  • Author(s): Tsuchiya K, Okamoto R, Nakamura T, Watanabe M
  • Organizer: Digestive Disease Week 2009
  • Place of Presentation: Chicago, USA
  • Year and Date: 2009-06-02
• [Presentation] GSK3 inhibitor induces the intestinal differentiation by the protein stabilization of Atoh1. (2009)
  • Author(s): Tsuchiya K, Okamoto R, Nakamura T, Watanabe M
  • Organizer: DDW2009
  • Place of Presentation: シカゴ
  • Year and Date: 2009-06-02
• [Presentation] クローン病小腸病変の評価とその治療-クローン病小腸病変の評価におけるダブルバルーン小腸内視鏡の有用性- (2009)
  • Author(s): 鈴木伸治
  • Organizer: 第95回 日本消化器病学会総会
  • Place of Presentation: 札幌
  • Year and Date: 2009-05-08
• [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
  • Filing Date: 2011-10-27