Analysis of intestinal mucosal repair by Notch-activated stem cells

• Project/Area Number: 23790777
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: OKAMOTO Ryuichi 東京医科歯科大学, 医歯学総合研究科, 准教授 (50451935)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 炎症性腸疾患 / Notchシグナル / 腸管上皮 / 上皮幹細胞 / 粘膜再生 / 腸管上皮初代培養 / オルガノイド / Notch活性化 / 細胞移植

Research Abstract

Through our studies aimed to develop a primary culture system of the intestinal epithelial cells, in which Notch-activated stem cell can be easily visualized, we succeeded to establish a culture method using the LGR5-EGFP mice. Using those mice, we found that the intestinal organoids that were maintained in our culture method contains high number of stem cells. In addition, transplantation of those organiods to a colitic mice model resulted in re-construction of the damaged epithelia by the organoids, which was maintained for a sustained time period. These results establish the basis for cell transplantation-mediated regeneration therapy for intestinal diseases.

Research Products

• [Journal Article] The acquisition of malignant potential in colon cancer is regulated by the stabilization of Atonal homolog 1 protein. (2013)
  • Author(s): Kano Y
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 432 Issue: 1 Pages: 175-181
  • DOI: 10.1016/j.bbrc.2013.01.034
  • Peer Reviewed / Open Access
• [Journal Article] Wnt5a Signaling Mediates Biliary Differentiation of Fetal Hepatic Stem/Progenitor Cells (2013)
  • Author(s): Kakinuma S, Kiyohashi K, Kamiya A, Sakamoto N, Nitta S, Yamanaka H, YoshinoK, Fujiki J, Murakawa M, Kusano-Kitazume A, Shimizu H, Okamoto R, Azuma S, Nakagawa M, Asahina Y, Tanimizu N, Kikuchi A, Nakauchi H, and *WatanabeM
  • Journal Title: Hepatology Volume: (in press) Issue: 6 Pages: 2502-2513
  • DOI: 10.1002/hep.26293
  • Peer Reviewed
• [Journal Article] Bone marrow-mesenchymal stem cells are a major source of interleukin-7 and sustain colitis by forming the niche for colitogenic CD4+ memory T cells. (2013)
  • Author(s): Nemoto Y, Nakamura T, Watanabe M et al
  • Journal Title: Gut. Volume: 62 Issue: 8 Pages: 1142-1152
  • DOI: 10.1136/gutjnl-2012-302029
  • 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] 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] Endoscopic ultrasound with double-balloon endoscopy for the diagnosis of inverted Meckel'sdiverticulum: a case report (2012)
  • Author(s): Araki A, Tsuchiya K, Oshima S, Okada E,Suzuki S, Akiyama JM, Fujii T, OkamotoR, Watanabe M
  • Journal Title: J Med Case Rep Volume: 6(1) Issue: 1 Pages: 328-328
  • DOI: 10.1186/1752-1947-6-328
  • 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] 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
• [Presentation] 炎症性腸疾患における上皮再生機構の解明と粘膜再生治療への応用 (2013)
  • Author(s): 岡本隆一、村野竜朗、渡辺守
  • Organizer: 第99回日本消化器病学会総会
  • Place of Presentation: 鹿児島
  • Year and Date: 2013-03-23
• [Presentation] 腸管上皮Notch-Hes1シグナルによるIL-22誘導性抗菌応答の制御 (2013)
  • Author(s): 村野竜朗、岡本隆一、渡辺守
  • Organizer: 第31回サイトプロテクション研究会
  • Place of Presentation: 京都
  • Year and Date: 2013-03-15
• [Presentation] Notch signaling regulates expression of Bestrophin-4, a novelenterocyte-specific HCO3-/Cl- channel, in human intestinal epithelial cells (2012)
  • Author(s): Go Ito, Ryuichi Okamoto, Satoru Fujii, Hiromichi Shimizu, Tatsuro Murano, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: UEGW2012
  • Place of Presentation: Amsterdam
  • Year and Date: 2012-11-23
• [Presentation] Notch signaling and TNF-alpha synergistically up-regulate OLFM4expression in human intestinal epithelial cells (2012)
  • Author(s): Ryuichi Okamoto, Go Ito, Satoru Fujii, Hiromichi Shimizu, Tatsuro Murano, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: UEGW2012
  • Place of Presentation: Amsterdam
  • Year and Date: 2012-11-23
• [Presentation] Identification of Notch ligand expressing cells in normal and inflamed intestinal mucosa (2012)
  • Author(s): Hiromichi Shimizu, Ryuichi Okamoto, Tatsuro Murano, Go Ito, Satoru Fujii, Morio-Akiyama Junko, Kiichiro Tsuchiya, Tetsuya Nakamura, MamoruWatanabe
  • Organizer: Asian IBD Symposium Seoul 2012.
  • Place of Presentation: Seoul
  • Year and Date: 2012-11-03
• [Presentation] Notch pathway and TNF-a synergistically up-regulatesOLFM4 expression in the inflamed mucosa of the human intestine (2012)
  • Author(s): Ryuichi Okamoto, Morio-AkiyamaJunko, Hiromichi Shimizu, Tatsuro Murano, Go Ito, Satoru Fujii, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: Asian IBD Symposium Seoul 2012.
  • Place of Presentation: Seoul
  • Year and Date: 2012-11-03
• [Presentation] Establishment of novel bio-screening methods using cultured intestinal epithelial cells (2012)
  • Author(s): Okamoto R
  • Organizer: The 1st International Symposium on Chemical Biology of Natural Products: Target ID and Regulation of Bioactivity.
  • Place of Presentation: Kyoto
  • Year and Date: 2012-10-31
• [Presentation] The acquisition of cancer stemness in colon cancer by the Atoh1 protein stabilization (2012)
  • Author(s): Kano Y, Tsuchiya K, Horita N, Zheng X, Okamoto R, Nakamura T, Watanabe M
  • Organizer: ISSCR2012.
  • Place of Presentation: Yokohama
  • Year and Date: 2012-06-14
• [Presentation] Regeneration of damaged colon epithelium by transplanted colon Lgr5+ stem cells maintained and expanded in vitro (2012)
  • Author(s): Yui S, Nakamura T, Nemoto Y, Mizutani T, Fukuda M, Nozaki K, Yamauchi Y, Mochizuki W, Zheng X, Nagaishi T, Okamoto R, Tsuchiya K, Watanabe M
  • Organizer: 第10回幹細胞シンポジウム.
  • Place of Presentation: 淡路島
  • Year and Date: 2012-06-01
• [Presentation] Colitogenic effector memory CD4+ T cells develop TH1/TH17 mediated interstitial pneumonia independent to intestinal bacterial antigens (2012)
  • Author(s): Nemoto Y, Kanai T, Okamoto R, Tsuchiya K, Nakamura T, Matsumoto S, Watanabe M
  • Organizer: AGA.
  • Place of Presentation: San Diego
  • Year and Date: 2012-05-22
• [Presentation] Notch signaling regulates expression of Gelsolin superfamily genes, Gelsolin and Scinderin, and promotes re-assembly of actin cytoskelton in human intestinal epithelial cells (2012)
  • Author(s): Ryuichi Okamoto,Tatsuro Murano, Go Ito, Satoru Fujii, Hiromichi Shimizu, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: DDW 2012
  • Place of Presentation: SanDiego
  • Year and Date: 2012-05-21
• [Presentation] Real-time analysis of p-glycoprotein-mediated drugtransport across primary intestinal epithelial cellsthree-dimensionally cultured in vitro (2012)
  • Author(s): Mizutani T, Nakamura T, Morikawa R, Fukuda M, Mochizuki W, Yamauchi Y, Nozaki K, Yui S, Okamoto R, Tsuchiya K, Watanabe M
  • Organizer: DDW2012.
  • Place of Presentation: San Diego
  • Year and Date: 2012-05-21
• [Presentation] Hes1 promotes IL-22-mediated epithelial regeneration through enhancement of STAT3-dependent transcription in human intestinal epithelial cells (2012)
  • Author(s): Tatsuro Murano, Ryuichi Okamoto, Go Ito, Satoru Fujii, Hiromichi Shimizu, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: DDW 2012
  • Place of Presentation: SanDiego
  • Year and Date: 2012-05-19
• [Presentation] The acquisition of cancer stemness in colon cancer by the Atoh1 protein stabilization (2012)
  • Author(s): Kano Y, Tsuchiya K, Horita N, Zheng X, Okamoto R, Nakamura T, Watanabe M
  • Organizer: DDW2012.
  • Place of Presentation: San Diego
  • Year and Date: 2012-05-19
• [Presentation] Notch-Hes1 pathway and TNF- synergistically up-regulates OLFM4 expression in the inflamed mucosa of the human intestine (2012)
  • Author(s): Ryuichi Okamoto, Go Ito, SatoruFujii, Hiromichi Shimizu, Tatsuro Murano, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: 6thKorea-Japan IBD Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2012-01-28
• [Presentation] Hes1 promotes IL-22-mediated epithelial regeneration through enhancement of STAT3-dependent transcription in human intestinal epithelial cells (2012)
  • Author(s): Tatsuro Murano, Ryuichi Okamoto, Go Ito, Satoru Fujii, Hiromichi Shimizu, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: 6th Korea-Japan IBD Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2012-01-28
• [Presentation] Regeneration of damaged colonic tissue by transplanted colonic epithelial stem cells maintained and expanded in vitro. (2011)
  • Author(s): Yui S., Okamoto R. et al.
  • Organizer: GI Research Academy 2011.
  • Place of Presentation: 京都
• [Presentation] Notch-Hes1 pathway and TNF-α synergistically up-regulates OLFM4 expression in the inflamed mucosa of the human intestine. (2011)
  • Author(s): Okamoto R. et al.
  • Organizer: DDW 2011.
  • Place of Presentation: シカゴ
• [Presentation] Hes1 promotes IL-22-Mediated epithelial regeneration through enhancement of STAT3-Dpendent transcription in human intestinal epithelial cells.
  • Author(s): Murano T, Okamoto R, Nakamura T, Watanabe M et al
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] The acquisition of cancer stemness in colon cancer by the Atoh1 protein stabilization.
  • Author(s): Kano Y, Okamoto R, Nakamura T, Watanabe M et al
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Notch signaling regulates expression of Gelsolin superfamily genes, Gelsolin and Scinderin, and promotes re-assembly of actin cytoskelton in human intestinal epithelial cells.
  • Author(s): Okamoto R, Nakamura T, Watanabe M et al
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Real-time analysis of p-glycoprotein-mediated drug transport across primary intestinal epitherial cells three-dimensionally cultured in vitro.
  • Author(s): Mizutani T, Nakamura T, Okamoto R, Watanabe M et al
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Colitogenic effector memory CD4+ T cells develop TH1/TH17 mediated interstitial pneumonia independent to intestinal bacterial antigens.
  • Author(s): Nemoto Y, Okamoto R, Nakamura T, Watanabe M et al
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Notch pathway and TNF-a synergistically up-regulates OLFM4 expression in the inflamed mucosa of the human intestine.
  • Author(s): Okamoto R, Murano T, Watanabe M et al
  • Organizer: Asian IBD Symposium Seoul 2012
  • Place of Presentation: Seoul
• [Presentation] 炎症性腸疾患における上皮再生機構の解明と粘膜再生治療への応用
  • Author(s): 岡本隆一、村野竜朗、渡辺 守
  • Organizer: 第99回日本消化器病学会総会
  • Place of Presentation: 鹿児島
• [Presentation] Flagellin response via TLR5 on basolateral membrane of primary intestinal epithelial cells is regulated by Notch signaling.
  • Author(s): Tsuchiya K, Okamoto R, Nakamura T, Watanabe M et al
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Identification of Notch ligand expressing cells in normal and inflamed intestinal mucosa.
  • Author(s): Shimizu H, Okamoto R, Watanabe M et al
  • Organizer: Asian IBD Symposium Seoul 2012
  • Place of Presentation: Seoul
• [Book] Microscopic Colitis(顕微鏡的腸炎) (2012)
  • Author(s): 岡本隆一、渡辺守
  • Publisher: Modern Physician
• [Book] PD-3確定診断困難な多発小腸潰瘍の1例栄養-評価と治療 (2012)
  • Author(s): 森尾純子,荒木昭博,岡田英理子,鈴木伸治,土屋輝一郎,藤井俊光,長沼誠,長堀正和,江頭徹哉,齊藤詠子,福島啓太,河合富貴子,藤木純子,櫻井幸,根本泰宏,中川美奈,岡本隆一,東正新,柿沼晴,永石宇司,中村哲也,坂本直哉,渡辺守,河内洋,江石義信
  • Publisher: メディカルレビュー社
• [Book] 腫瘍をめぐるQ&A(Question45)Lgr5とは? (2012)
  • Author(s): 伊藤剛、岡本隆一、清水寛路、村野竜朗、水谷知裕、油井史郎、土屋輝一郎、中村哲也、渡辺守
  • Publisher: メディカルレビュー社
• [Book] 炎症と組織再生.FunctionalFood (2011)
  • Author(s): 岡本隆一、渡辺守
  • Publisher: フジメディカル
• [Book] 炎症性腸疾患の診断と治療をめぐる新展開 (2011)
  • Author(s): 岡本隆一、渡辺守
  • Publisher: メジカルビュー社
• [Book] 上皮細胞再生による腸炎治療戦略 (2011)
  • Author(s): 中村哲也、岡本隆一、渡辺守
  • Publisher: ニューサイエンス社