Establishment of enterohormone farm by using cultured human intestinal epithelial cells

• Project/Area Number: 15K15286
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• 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, 再生医療研究センター, 教授 (50451935)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 腸上皮 / 消化管ホルモン / 腸上皮オルガノイド

Outline of Final Research Achievements

In this study, we investigated the method to induce secretory-lineage differentiation of cultured human intestinal organoids, and also performed some functional analyses of those organoids. Consequently, we successfully established over 40-lines of human small intestinal- or colonic-organoids from endoscopic biopsy specimens. Short-term exposure to a gamma-secretase inhibitor, combined with depletion of Wnt3a, SB20190 and Nicotinamide, certainly directed human intestinal organoids towards secretory cell-lineage differentiation. We also established a quantitative analysis system to evaluate the response of human intestinal organoids to a secretagogue, PGE2, or an enterohormone, VIP, and showed their properties to regulate the body-fluid balance.

Research Products

• [Journal Article] PGE2 is a direct and robust mediator of anion/fluid secretion by human intestinal epithelial cells. (2016)
  • Author(s): Fujii S, Okamoto R, Suzuki K, Kawamoto A, Ishibashi F, Nakata T, Murano T, Ito G, Shimizu H, Mizutani T, Oshima S, Tsuchiya K, Nakamura T, Araki A, Ohtsuka K, Watanabe M
  • Journal Title: Sci. Rep. Volume: 6 Issue: 1 Pages: 36795-36795
  • DOI: 10.1038/srep36795
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [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 Issue: 1 Pages: 11-21
  • DOI: 10.1007/s00535-015-1098-4
  • Peer Reviewed / Open Access
• [Journal Article] Reduced human α-defensin 6 in non-inflamed jejunal tissue of Crohn’s disease patients (2016)
  • Author(s): Hayashi R , Tsuchiya K, Okamoto R,Watanabe M,et al
  • Journal Title: Inflammatory Bowel Diseases Volume: 22 Issue: 5 Pages: 1119-28
  • DOI: 10.1097/mib.0000000000000707
  • Peer Reviewed
• [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 Issue: 2 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 Issue: 7 Pages: 1052-1062
  • DOI: 10.1080/15548627.2015.1055439
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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 Issue: 8 Pages: 1000-7
  • DOI: 10.1111/cas.12703
  • Peer Reviewed / Open Access
• [Presentation] In vitro organoid model reveals PGE2 as a robust mediator of chloride & water secretion by human intestinal epithelial cells (2016)
  • Author(s): Satoru Fujii, Kohei Suzuki, Ami Kawamoto, Fumiaki Ishibashi, Toru Nakata, Go Ito, Hiromichi Shimizu, Tomohiro Mizutani, Kiichiro Tsuchiya, Tetsuya Nakamura, Kazuo Ohtsuka, Ryuichi Okamoto, Mamoru Watanabe
  • Organizer: DDW2016
  • Place of Presentation: サンディエゴ（米国）
  • Int'l Joint Research
• [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] 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] 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] 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