Development of colitic mucosa repair therapy by the induction of regeneration-specific stem cell niche formation

• Project/Area Number: 25293170
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Okamoto Ryuichi 東京医科歯科大学, 再生医療研究センター, 教授 (50451935)
• Co-Investigator(Kenkyū-buntansha): Tsuchiya Kiichiro 東京医科歯科大学, 医学部附属病院消化器内科, 准教授 (40376786) ; Nakamura Tetsuya 東京医科歯科大学, 医歯学総合研究科, 寄附講座教授 (70265809)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000); Fiscal Year 2015: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2013: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: 腸管上皮 / 炎症性腸疾患 / Notchシグナル / オルガノイド / ニッチ / 幹細胞ニッチ

Outline of Final Research Achievements

Through the studies of the current project, results were acquired in the following areas: 1) Identification of Dll1+ve or Dll4+ve cells in inflamed and non-inflamed intestinal epithelium, and the functional relevance of those cells, 2) Comprehensive analysis of gene expression in Notch and TNF-α pathway activated cells, which constitute the regenerative epithelium inbt he colitic mucosa and 3) Identification of a distinct intestinal stem cell population that appears at the recovery phase of colitis.

Research Products

• [Journal Article] Role of epithelial cells in the pathogenesis and treatment of inflammatory bowel disease. (2016)
  • Author(s): Ryuichi Okamoto, Mamoru Watanabe
  • Journal Title: J. Gastroenterol Volume: 51 Pages: 11-21
  • DOI: 10.1007/s00535-015-1098-4
  • Peer Reviewed / Open Access
• [Journal Article] Co-culture with intestinal epithelial organoids allows efficient expansion and motility analysis of intraepithelial lymphocytes. (2016)
  • Author(s): Nozaki K, Mochizuki W, Matsumoto Y, Matsumoto T, Fukuda M, Mizutani T, Watanabe M, Nakamura T
  • Journal Title: J Gastroenterol. Volume: 51 Pages: 206-213
  • DOI: 10.1007/s00535-016-1170-8
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Matrix metalloproteinase-14 mediates formation of bile ducts and hepatic maturation of fetal hepatic progenitor cells. (2016)
  • Author(s): Otani S, Kakinuma S, Kamiya A, Goto F, Kaneko S, Miyoshi M, Tsunoda T, Asano Y, Kawai-Kitahata F, Nitta S, Nakata T, Okamoto R, Itsui Y, Nakagawa M, Azuma S, Asahina Y, Yamaguchi T, Koshikawa N, Seiki M, Nakauchi H, Watanabe M.
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 469 Pages: 1062-1068
  • DOI: 10.1016/j.bbrc.2015.12.105
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Perspectives for Regenerative Medicine in the Treatment of Inflammatory Bowel Diseases. (2015)
  • Author(s): Ryuichi Okamoto, Mamoru Watanabe
  • Journal Title: Digestion Volume: 92 Pages: 73-77
  • DOI: 10.1159/000438663
  • Peer Reviewed / Open Access
• [Journal Article] TNFAIP3 promotes survival of CD4 T cells by restricting MTOR and promoting autophagy. (2015)
  • Author(s): Matsuzawa Y, Oshima S, Takahara M, Maeyashiki C, Nemoto Y, Kobayashi M, Nibe Y, Nozaki K, Nagaishi T, Okamoto R, Tsuchiya K, Nakamura T, Averil Ma, Watanabe M
  • Journal Title: Autophagy Volume: 11 Pages: 1052-1062
  • DOI: 10.1080/15548627.2015.1055439
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Atonal homolog 1 protein stabilized by tumor necrosis factor α induces high malignant potential in colon cancer cell line. (2015)
  • Author(s): Keita Fukushima, Kiichiro Tsuchiya,Okamoto Ryuichi, Mamoru Watanabe,et al
  • Journal Title: Cancer Sci. Volume: 106 Pages: 1000-1007
  • DOI: 10.1111/cas.12703
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Reduced human α-defensin 6 in non-inflamed jejunal tissue of Crohn’s disease patients (2015)
  • Author(s): Hayashi R , Tsuchiya K, Okamoto R, Watanabe M.et al.
  • Journal Title: Inflammatory Bowel Diseases
  • Peer Reviewed
• [Journal Article] RIPK3 regulates p62- LC3 complex formation via the caspase-8-dependent cleavage of p62 (2015)
  • Author(s): Y. Matsuzawa , S. Oshima, Y. Nibe, M.Kobayashi, C. Maeyashiki, Y. Nemoto, T. Nagaishi, R. Okamoto R, K. Tsuchiya, T. Nakamura, M. Watanabe.
  • Journal Title: Biochem Biophys Res Commun. Volume: 456(1) Pages: 298-304
  • DOI: 10.1016/j.bbrc.2014.11.075
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt. (2014)
  • Author(s): N. Horita, K. Tsuchiya, R. Hayashi, K. Fukushima, S. Hibiya, M. Fukuda, Y. Kano, T. Mizutani, Y. Nemoto, S. Yui, R. Okamoto, T. Nakamura, M. Watanabe
  • Journal Title: Biochem Biophys Res Commun. Volume: 454 Pages: 493-499
  • DOI: 10.1016/j.bbrc.2014.10.091
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Myosin light chain kinase expression induced via tumor necrosis factor receptor 2 signaling in the epithelial cells regulates the development of colitis-associated carcinogenesis (2014)
  • Author(s): Suzuki M, Nagaishi T, Yamazaki M, Onizawa M, Watabe T, Sakamaki Y, Ichinose S, Totsuka M, Oshima S, Okamoto R, Shimonaka M, Yagita H, Nakamura T, Watanabe M.
  • Journal Title: PLoS One. Volume: 9
  • DOI: 10.1371/journal.pone.0088369
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Distinct expression patterns of Notch ligands, Dll1 and Dll4, in normal and inflamed mice intestine. (2014)
  • Author(s): Shimizu H, Okamoto R, Ito G, Nakata T, Suzuki K, Murano T, Mizutani T, Tsuchiya K, Nakamura T, Hozumi K, Watanabe M.
  • Journal Title: 2014 Volume: 2
  • DOI: 10.7717/peerj.370
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Hes1 promotes the IL-22-mediated antimicrobial response by enhancing STAT3-dependent transcription in human intestinal epithelial cells. (2014)
  • Author(s): Murano T, Okamoto R, Ito G, Nakata T, Hibiya S, Shimizu H, Fujii S, Kano Y, Mizutani T, Yui S, Akiyama-Morio J, Nemoto Y, Tsuchiya K, Nakamura T, Watanabe M
  • Journal Title: Biochem Biophys Res Commun. Volume: 443 Pages: 840-6
  • DOI: 10.1016/j.bbrc.2013.12.061
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Lineage-specific expression of bestrophin-2 and bestrophin-4 in human intestinal epithelial cells (2013)
  • Author(s): Ito G, Okamoto R, Nakamura T, Watanabe M et al
  • Journal Title: PLoS One. Volume: 8
  • DOI: 10.1371/journal.pone.0079693
  • Peer Reviewed
• [Journal Article] IL-7 promotes long-term in vitro survival of unique long-lived memory subset generated from mucosal effector memory CD4+ T cells in chronic colitis mice (2013)
  • Author(s): Takahara M, Nemoto Y, Oshima S, Matsuzawa Y, Kanai T, Okamoto R, Tsuchiya K, Nakamura T, Yamamoto K, Watanabe M.
  • Journal Title: Immunol Lett Volume: 156 Pages: 82-93
  • DOI: 10.1016/j.imlet.2013.09.001
  • NAID: 120005468269
  • 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 Pages: 1142-1152
  • DOI: 10.1136/gutjnl-2012-302029
  • Peer Reviewed
• [Journal Article] Th1/Th17-mediated interstitial pneumonia in chronic colitis mice independent of intestinal microbiota (2013)
  • Author(s): Nemoto Y, Watanabe M et al
  • Journal Title: J Immunol Volume: 190 Pages: 6616-25
  • DOI: 10.4049/jimmunol.1202930
  • Peer Reviewed
• [Presentation] Establishment of a 3D cell culture-based screening platform to identify natural products that can regulate transepithelial water transport of the human gastrointestinal tract. (2015)
  • Author(s): Satoru Fujii, Ryuichi Okamoto, Toru Nakata, Kohei Suzuki, Fumiaki Ishibashi, Ami Kawamoto,Sayaka Ohashi Segawa, Tomohiro Mizutani, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: Pacifichem2015
  • Place of Presentation: ハワイ（米国）
  • Year and Date: 2015-12-17
  • Int'l Joint Research
• [Presentation] TNF-α stabilizes atoh1 protein in colitis-associated colorectal cancer resulting in enhanced malignant potential. (2015)
  • Author(s): Tomoaki Shirasaki, Keita Fukushima, Kiichiro Tsuchiya, Shuuji Hibiya, Ryohei Hayashi, Nobukatsu Horita, Ryuichi Okamoto, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: UEGW2015
  • Place of Presentation: バルセロナ（スペイン）
  • Year and Date: 2015-10-28
  • Int'l Joint Research
• [Presentation] Contiuous stimulation with cytokines leads to irreversible activation of NF-γb signaling in colonic epithelial cells by organoid culture. (2015)
  • Author(s): Shuji Hibiya, Kiichiro Tsuchiya, Tomoaki Shirasaki, Keita Fukushima, Ryohei Hayashi, Nobukatsu Horita, Shigeru Oshima ,Ryuichi Okamoto, Tetsuya Nakamura,Mamoru Watanabe
  • Organizer: UEGW2015
  • Place of Presentation: バルセロナ（スペイン）
  • Year and Date: 2015-10-27
  • Int'l Joint Research
• [Presentation] Atoh1 protein stabilized by TNF-α acquires enhanced malignant potential in colitis-associated colorectal cancer. (2015)
  • Author(s): Tomoaki Shirasaki, Keita Fukushima, Kiichiro Tsuchiya, Shuuji Hibiya, Ryohei Hayashi, Nobukatsu Horita, Ryuichi Okamoto, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: AOCC2015
  • Place of Presentation: 北京（中国）
  • Year and Date: 2015-06-19
  • Int'l Joint Research
• [Presentation] A novel fluorescent labelling system into small intestinal organoid reveals independent long-lived intestinal stem cells in a crypt. (2015)
  • Author(s): Horita N,Tsuchiya K,Hayashi R,Fukushima K,Hibiya S,Fukuda M,Mizutani T,Okamoto R,Nakamura T and Watanabe M.
  • Organizer: AOCC2015
  • Place of Presentation: 北京（中国）
  • Year and Date: 2015-06-19
  • Int'l Joint Research
• [Presentation] Enrichment of Intestinal Stem Cells by the 3D-Culture of IBD Patient Derived Intestinal epithelium. (2015)
  • Author(s): Kohei Suzuki,Satoru Fujii,Toru Nakata,Ami Kawamoto,Fumiaki Ishibashi,Sayaka Ohashi Segawa, Tomohiro Mizutani, Kiichiro Tsuchiya,Kazuo Otsuka,Tetsuya Nakamura,Ryuichi Okamoto,Mamoru Watanabe.
  • Organizer: AOCC2015
  • Place of Presentation: 北京（中国）
  • Year and Date: 2015-06-19
  • Int'l Joint Research
• [Presentation] Continuous Stimulation With Cytokines Leads to Irreversible Accumulation of NF-kB Signaling in Colonic Epithelial Cells. (2015)
  • Author(s): Shuji Hibiya, Kiichiro Tsuchiya, Keita Fukushima, Ryohei Hayashi, Nobukatsu Horita, Shigeru Oshima, Ryuichi Okamoto, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: DDW 2015
  • Place of Presentation: ワシントン（米国）
  • Year and Date: 2015-05-19
  • Int'l Joint Research
• [Presentation] Distinct Role of Notch Ligands, DLL1 and Dll4, in Normal and in Tumor Intestinal Epithelium. (2015)
  • Author(s): Toru Nakata, Hiromichi Shimizu, Kohei Suzuki, Satoru Fujii, Go Ito, Kiichiro Tsuchiya, Tetsuya Nakamura, Ryuichi Okamoto, Katsuto Hozumi, Mamoru Watanabe.
  • Organizer: DDW 2015
  • Place of Presentation: ワシントン（米国）
  • Year and Date: 2015-05-19
  • Int'l Joint Research
• [Presentation] Atoh1 Protein Expression by TNF-a and the Acquisition of Malignant Potential in Colitis- Associated Colorectal Cancer. (2015)
  • Author(s): Keita Fukushima, Kiichiro Tsuchiya, Shuji Hibiya, Ryohei Hayashi, Nobukatsu Horita, Yoshihito Kano, Ryuichi Okamoto, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: DDW 2015
  • Place of Presentation: ワシントン（米国）
  • Year and Date: 2015-05-17
  • Int'l Joint Research
• [Presentation] Notch Ligands DLL1 and Dll4 Are Expressed by Distinct Population of Epithelial Cells in the Mice Intestine. (2015)
  • Author(s): Hiromichi Shimizu, Kohei Suzuki, Satoru Fujii, Toru Nakata, Go Ito, Tatsuro Murano, Kiichiro Tsuchiya, Tetsuya Nakamura, Ryuichi Okamoto, Katsuto Hozumi, Mamoru Watanabe.
  • Organizer: DDW 2015
  • Place of Presentation: ワシントン；米国
  • Year and Date: 2015-05-16
  • Int'l Joint Research
• [Presentation] Live imaging analysis of intraepithelial lymphocytes (IELs) co-cultured with intestinal epithelial organoids. (2015)
  • Author(s): Nozaki K, Mochizuki W, Matsumoto Y, Matsumoto T, Fukuda M, Mizutani T, Watanabe M, Nakamura T
  • Organizer: Cold Spring Harbor Meeting "Fundamental Immunology and Its Therapeutic Applications"
  • Place of Presentation: ニューヨーク（米国）
  • Year and Date: 2015-04-16
  • Int'l Joint Research
• [Presentation] Notch signaling and TNF-α synergistically up-regulates expression of OLFM4 in human intestinal epithelial cells. (2014)
  • Author(s): Go Ito, Ryuichi Okamoto, Tasturo Murano, Hiromichi Shimizu, Satoru Fujii, Toru Nakata, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe .
  • Organizer: 第8回Japan-US Collabaration Conference in Gastroenterology
  • Place of Presentation: Intercontinental TOKYO BAY (東京都港区海岸1-16-2)
  • Year and Date: 2014-11-21
• [Presentation] Notch signaling regulates expression of gelsolin superfamily genes, gelsolim and scinderin and promotes re-assembly of action cytoskeleton in human intestinal epithelial cells. (2014)
  • Author(s): Ryuichi Okamoto, Go Ito, Hiromichi Shimizu, Satoru Fujii , Toru Nakata, Kohei Suzuki, Mamoru Watanabe.
  • Organizer: UEGW2014
  • Place of Presentation: Vienna (Austria)
  • Year and Date: 2014-10-22
• [Presentation] Notch Signaling and TNF-alpha synergistically promotes intracellular protein accumulation of olfm4 in the inflamed mucosa of ulcerative colitis. (2014)
  • Author(s): Go Ito, Ryuichi Okamoto, Hiromichi Shimizu, Satoru Fujii, Toru Nakata, Kohei Suzuki, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: UEGW2014
  • Place of Presentation: Vienna (Austria)
  • Year and Date: 2014-10-21
• [Presentation] Notch signaling and TNF-α synergistically promotes intracellular protein expression of OLFM4 in human intestinal epithelial cells. (2014)
  • Author(s): Go Ito, Ryuichi Okamoto, Hiromichi Shimizu, Satoru Fujii, Toru Nakata, Kohei Suzuki, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe.
  • Organizer: The 2nd Annual Meeting of Asian Organization for Crohn's and Colitis
  • Place of Presentation: Seoul (Korea)
  • Year and Date: 2014-06-21
• [Presentation] Distinct Expression Patterns of Notch Ligands, DLL1 and DLL4, in Normal and Inflamed Mice Intestine. (2014)
  • Author(s): Hiromichi Shimizu, Ryuichi Okamoto, Go Ito, Satoru Fujii, Toru Nakata, Kohei Suzuki, Tatsuro Murano, Tomohiro Mizutani, Kiichiro Tsuchiya, Tetsuya Nakamura, Katsuto Hozumi, Mamoru Watanabe.
  • Organizer: The 2nd Annual Meeting of Asian Organization for Crohn's and Colitis
  • Place of Presentation: Seoul (Korea)
  • Year and Date: 2014-06-21
• [Presentation] Complete conversion of crypt progenitor cells into Atoh1-positive cells by targeted deletion of Dll1 and Dll4 in Lgr5-positive intestinal stem cells
  • Author(s): Hiromichi Shimizu, Ryuichi Okamoto, Satoru Fujii, Go Ito, Tatsuro Murano, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: DDW 2013
  • Place of Presentation: オーランド、米国
• [Presentation] Notch signaling regulates expression of Bestrophin-4, a novel enterocyte-specific HCO3-/Cl- channel, in human intestinal epithelial cells.
  • Author(s): Go Ito, Ryuichi Okamoto, Satoru Fujii, Hiromichi Shimizu, Tatsuro Murano, Kiichiro Tsuchiya, Tetsuya Nakamura, Mamoru Watanabe
  • Organizer: DDW 2013
  • Place of Presentation: オーランド、米国
• [Presentation] Hes1 promotes IL-22-mediated antimicrobial response through enhancement of STAT3-dependent transcription in human intestinal epithelial cells
  • Author(s): Ryuichi Okamoto, Tatsuro Murano, Hiromichi Shimizu, Go Ito, Satoru Fujii, Kiichiro Tsuchiya, Tetsuya Nakamura, and Mamoru Watanabe
  • Organizer: The 1st Annual Meeting of Asian Organization for Crohn's and Colitis
  • Place of Presentation: 東京、ラフォーレ東京
• [Presentation] COMPLETE CONVERSION OF CRYPT PROGENITOR CELLS INTO ATOH1-POSITIVE CELLS BY TARGETED DELETION OF DLL1 AND DLL4 IN LGR5-POSITIVE INTESTINAL STEM CELLS
  • Author(s): H. Shimizu, R. Okamoto, T. Nakata, S. Fujii, G. Ito, T. Murano, K. Tsuchiya, T. Nakamura, M. Watanabe
  • Organizer: UEGW 2013
  • Place of Presentation: ベルリン、ドイツ
• [Presentation] IL-22-MEDIATED ANTIMICROBIAL RESPONSE IS REGULATED BY HES1 VIA STAT3-DEPENDENT TRANSCRIPTION IN HUMAN INTESTINAL EPITHELIAL CELLS
  • Author(s): R. Okamoto, T. Murano, H. Shimizu, G. Ito, S. Fujii, T. Nakata, K. Tsuchiya, T. Nakamura, M. Watanabe
  • Organizer: UEGW 2013
  • Place of Presentation: ベルリン、ドイツ
• [Presentation] LINEAGE-SPECIFIC EXPRESSION OF BESTROPHIN-2 AND BESTROPHIN-4 IS REGULATED BY NOTCH SIGNALING IN HUMAN INTESTINAL EPITHELIAL CELLS
  • Author(s): G. Ito , R. Okamoto, T. Murano, H. Shimizu, K. Tsuchiya, T. Nakamura, M. Watanabe
  • Organizer: UEGW 2013
  • Place of Presentation: ベルリン、ドイツ