Identification of critical factor to generate human colon from small intestine

2018 Fiscal Year Annual Research Report

• Project/Area Number: 17K19513
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 渡辺 守 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (10175127)
• Project Period (FY): 2017-06-30 – 2019-03-31
• Keywords: 炎症性腸疾患 / 再生医療 / 幹細胞運命制御 / 大腸オルガノイド

Outline of Annual Research Achievements

本研究はヒト大腸上皮幹細胞の運命制御を明らかにすることにより、小腸上皮幹細胞を大腸上皮幹細胞に転換することで、ヒト小腸上皮から大腸上皮を作成することを目的とする。申請者は以前より炎症性腸疾患（IBD）の臨床・研究に従事している。IBDは慢性・難治性の炎症疾患であり、小腸・大腸に潰瘍を形成し、下痢・出血など重篤な症状を引き起こす。若年者に多く、本邦で患者数が急増していることからも根本的な対策が急務の状況である。これまでは、慢性炎症疾患であることから、免疫制御を中心とした治療・研究が行われてきた。しかし、免疫を標的とした治療のみでは粘膜が修復せず症状は一時的に改善しても再燃を繰り返すことから新しい治療法の開発が望まれている。そこで、粘膜修復を念頭に置いた腸上皮細胞の機能に着目し、上皮細胞における粘膜免疫制御や腸内細菌防御などの粘膜バリアー制御機構などを明らかとしてきた。さらに、上皮細胞幹細胞に着目し、大腸オルガノイドをマウス大腸潰瘍に移植すると大腸組織として生着するのみならず、潰瘍治癒促進効果を認めたことから大腸上皮幹細胞移植による再生医療はIBDの大腸粘膜治癒を目標とした次世代治療として確立できると考えた。現状では体性幹細胞である大腸上皮幹細胞を培養、増殖することが最善と思われ、さらに他家移植では拒絶反応の可能性もあるため自家移植が最も安全で有効な手段であると考えた。そこで大腸上皮幹細胞運命制御を明らかとし、小腸幹細胞から大腸幹細胞への人工的な転換技術を構築することで自家大腸上皮細胞の供給が可能となると着想した。その結果、同一人物由来の小腸、大腸オルガノイドを樹立し、マイクロアレイ解析により小腸特異的発現遺伝子、大腸特異的発現遺伝子を同定することに成功した。

Research Products

• [Journal Article] TP53 mutation by CRISPR system enhances the malignant potential of colon cancer. (2019)
  • Author(s): Watanabe S, Tsuchiya K, Nishimura R, Shirasaki T, Katsukura N, Hibiya S, Okamoto R, Nakamura T, Watanabe M
  • Journal Title: Mol Cancer Res. Volume: in press Pages: in press
  • DOI: 10.1158/1541-7786.MCR-18-1195.
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of a system to evaluate the therapeutic effect and the dynamics of an investigational drug on ulcerative colitis using human colonic organoids. (2019)
  • Author(s): Nishimura R, Shirasakai T, Tsuchiya K, Miyake Y, Watanabe Y, Hibiya S, Watanabe S, Nakamura T, Watanabe M
  • Journal Title: J Gastroenterol. Volume: in press Pages: in press
  • DOI: 10.1007/s00535-018-01540-y.
  • Peer Reviewed
• [Journal Article] Ubiquitin D is up-regulated by synergy of Notch signaling and TNF-α in the inflamed intestinal epithelia of IBD patients. (2019)
  • Author(s): Kawamoto A, Nagata S, Anzai S, Takahashi J, Kawai M, Hama M, Nogawa D, Yamamoto K, Kuno R, Suzuki K, Shimizu H, Hiraguri Y, Yui S, Ohshima S, Tsuchiya K, Nakamura T, Ohtsuka K, Kitagawa M, Okamoto R, Watanabe M
  • Journal Title: J Crohns Colitis. Volume: 13 Pages: 495-509
  • DOI: 10.1093/ecco-jcc/jjy180.
  • Peer Reviewed
• [Journal Article] Genetic and environmental factors drive personalized medicine for Crohn’s disease. (2019)
  • Author(s): Oshima S, Watanabe M
  • Journal Title: J Clin Invest. Volume: 128 Pages: 4758-4760
  • DOI: 10.1172/JCI124303.
  • Peer Reviewed
• [Journal Article] Single cell analysis of Crohn’s disease patient-derived small intestinal organoids reveals disease activity-dependent modification of stem cell properties. (2018)
  • Author(s): Suzuki K, Murano T, Shimizu M, Ito G, Nakata T, Fujii S, Ishibashi F, Kawamoto A, Anzai S, Kuno R, Kuwabara K, Takahashi J, Hama M, Nagata S, Hiraguri Y, Takenaka K, Yui S, Tsuchiya K, Nakamura T, Ohtsuka K, Watanabe M, Okamoto R
  • Journal Title: J Gastroenterol. Volume: 53 Pages: 1035-1047
  • DOI: 10.1007/s00535-018-1437-3.
  • Peer Reviewed
• [Journal Article] B cell activation in the cecal patches during the development of an experimental colitis model. (2018)
  • Author(s): Watabe T, Nagaishi T, Tsugawa M, Kojima Y, Jose N, Hosoya A, Onizawa M, Nemoto Y, Oshima S, Nakamura T, Karasuyama H, Adachi T, Watanabe M
  • Journal Title: Biochem Biophys res Commun. Volume: 496 Pages: 367-373
  • DOI: 10.1016/j.bbrc.2018.01.053.
  • Peer Reviewed
• [Journal Article] YAP/TAZ-dependent reprogramming of colonic epithelium links ECM remodeling to tissue regeneration. (2018)
  • Author(s): Yui S, Azzolin L, Maimets M, Pedersen M, Fordham R, Hansen S, Guiu J, Alves M, Rundsen C, Johansen J, Li Y, Madsen C, Nakamura T, Wanatabe M, Nielsen O, Schweiger P, Piccolo S, Jensen K
  • Journal Title: Cell Stem Cell Volume: 22 Pages: 35-49,
  • DOI: 10.1016/j.stem.2017.11.001.
  • Peer Reviewed
• [Journal Article] Contribution of ATOH1+ cells to the homeostasis, repair and tumorigenesis of the colonic epithelium. (2018)
  • Author(s): Ishibashi F, Shimizu H, Nakata T, Fujii S, Suzuki K, Kawamoto A, Anzai S, Kuno R, Nagata S, Ito G, Murano T, Mizutani T, Oshima S, Tsuchiya K, Nakamura T, Watanabe M, Okamoto R
  • Journal Title: Stem Cell Rep. Volume: 10 Pages: 27-42
  • DOI: 10.1016/j.stemcr.2017.11.006.
  • Peer Reviewed / Open Access
• [Journal Article] Novel polyubiquitin imaging system, PolyUb-FC, reveals that K33-linked polyubiquitin is recruited by p62/SQSTM1. (2018)
  • Author(s): Nibe Y, Oshima S, Kobayashi M, Maeyashiki C, Matsuzawa Y, Otsubo K, Matsuda H, Aonuma E, Nemoto Y, Nagaishi T, Okamoto R, Tsuchiya K, Nakamura T, Nakada S, Watanabe M
  • Journal Title: Autophagy Volume: 24 Pages: 347-358
  • DOI: 10.1080/15548627.2017.1407889.
  • Peer Reviewed
• [Presentation] TP53 mutation in human colonic organoids acquires resistance to in vitro long-term inflammation. (2019)
  • Author(s): Tsuchiya K, Watanabe S, Shirasaki T, Nishimura R, Katsukura N, Hibiya S, Okamoto R, Nakamura T, Watanabe M
  • Organizer: ECCO2019
  • Int'l Joint Research
• [Presentation] Synergy of Notch signalling and TNF-α in the inflamed intestinal epithelia of IBD patients leads to up-regulation of UBD, a ubiquitin-like protein. (2019)
  • Author(s): Kawamoto A, Nagata S, Anzai S, Takahashi J, Kawai M, Hama M, Nogawa D, Yamamoto K, Kuno R, Suzuki K, Shimizu H, Hiraguri Y, Yui S, Oshima S, Tsuchiya K, Nakamura T, Ohtsuka K, Kitagawa M, Okamoto R, Watanabe M
  • Organizer: ECCO2019
  • Int'l Joint Research
• [Presentation] UC-related and segment-specific properties of patientderived colonic organoids. (2019)
  • Author(s): Suzuki K, Shimizu H, Kawai M, Takahashi J, Anzai S, Kawamoto A, Nagata S, Hiraguri Y, Yui S, Tsuchiya K, Nakamura T, Ohtsuka K, Okamoto R, Watanabe M
  • Organizer: ECCO2019
  • Int'l Joint Research
• [Presentation] Introduction to IBD Centre at Tokyo Medical and Dental University. (2018)
  • Author(s): Watanabe M
  • Organizer: IBD Center of Excellence
  • Invited
• [Presentation] Intestinal Epithelial Stem Cell Organoid and IBD. (2018)
  • Author(s): Watanabe M
  • Organizer: 第55回日本消化器免疫学会総会 シンポジウム
  • Invited
• [Presentation] Regeneration of intestinal epithelial cell and IBD (2018)
  • Author(s): Watanabe M
  • Organizer: Falk symposium2018
  • Int'l Joint Research / Invited
• [Presentation] Downregulated APL expression in T cells in the setting of chronic colitis. (2018)
  • Author(s): Kojima Y, Nagaishi T, Watabe T, Yamada D, Tsugawa N, Jose N, Hosoya A, Shimizu M, Onizawa M, Watanabe M
  • Organizer: 第47回日本免疫学会学術集会
  • Int'l Joint Research
• [Presentation] Mice overexpressing MKL1 in macrophages experience fulminant colitis. (2018)
  • Author(s): An J, Nagaishi T, Watabe T, Naruse TK., Watanabe M, Kimura A
  • Organizer: UEGW 2018
  • Int'l Joint Research
• [Presentation] B cell-mediated ileocecal immune response is activated in the early phase of colitis development. (2018)
  • Author(s): Kojima Y, Nagaishi T, Watabe T, Tsugawa N, Jose N, Yamada D, Hosoya A, Adachi T, Watanabe M
  • Organizer: UEGW 2018
  • Int'l Joint Research
• [Presentation] Lesion-specific gene expression in the Epithelial cells of Crohn’s disease by comparing Small intestinal organoids from active and inactive Lesion in the same patient. (2018)
  • Author(s): Hibiya S, Tsuchiya K, Nishimura R, Shirasaki T, Watanabe S, Katsukura N, Okamoto R, Nakamura T, Watanabe M
  • Organizer: UEGW 2018
  • Int'l Joint Research
• [Presentation] Tp53 mutation enhances cell proliferation and stemness in human colon epithelial organoids and promotes a resistance against long-term inflammation. (2018)
  • Author(s): Watanabe S, Tsuchiya K, Shirasaki T, Hibiya S, Nishimura S, Katsukura N, Oshima S, Okamoto R, Tetsuya N, Watanabe M
  • Organizer: UEGW 2018
  • Int'l Joint Research
• [Presentation] Long-lived secretory cells residing in the mouse proximal colon serve as reserve stem cells under DNA damage-induced mucosal injury. (2018)
  • Author(s): Kuwabara K, Ishibashi F, Kawamoto A, Anzai S, Takahashi J, Nagata S, Shimizu H, Yui S, Oshima S, Tsuchiya K, Nakamura T, Watanabe M, Okamoto R
  • Organizer: UEGW 2018
  • Int'l Joint Research
• [Presentation] Ectopic expression of reg3a in the mice distal Colon is mediated by interactions between notch and Il-22 signaling pathways, and promotes tissue repair By the augmentation of EGFR signaling. (2018)
  • Author(s): Ishibashi F, Kuwabara K, Kawamoto A, Anzai S, Takahashi J, Nagata S, Shimizu H, Yui S, Oshima S, Tsuchiya K, Nakamura T, Watanabe M, Okamoto R
  • Organizer: UEGW 2018
  • Int'l Joint Research
• [Presentation] Regenerative medicine for the treatment of inflammatory bowel disease. (2018)
  • Author(s): Okamoto R, Watanabe M
  • Organizer: 5th TERMIS World Congress 2018
  • Int'l Joint Research
• [Presentation] Crohn’s disease patient-derived small intestinal organoids reveal diseasestatus related modification of stem cell properties. (2018)
  • Author(s): Suzuki K, Takahashi J, Hiraguri Y, Yui S, Shimizu H, Tsuchiya K, Nakamura T, Ohtsuka K, Okamoto R, Watanabe M:
  • Organizer: 5th TERMIS World Congress 2018
  • Int'l Joint Research
• [Presentation] Crohn’s disease patient-derived small intestinal organoids reveal disease-status related modification of stem cell properties. (2018)
  • Author(s): Suzuki K, Murano T, Hiraguri Y, Takahashi J, Shimizu H,Anzai S, Kuwabara K, Kawamoto A, Ishibashi F, Yui S, Tsuchiya K, Nakamura T, Ohtsuka K, Watanabe M, Okamoto R
  • Organizer: 5th TERMIS World Congress 2018
  • Int'l Joint Research
• [Presentation] Development of a new mouse model of short bowel syndrome that may allow for the assessment of therapeutic efficacy of heterotopic transplantation of small intestinal organoids. (2018)
  • Author(s): Matsumoto Y, Watanabe M, Nakamura T
  • Organizer: 5th TERMIS World Congress 2018
  • Int'l Joint Research
• [Presentation] Overexpression of MKL1 in Macrophages leads to the development of murine colitis. (2018)
  • Author(s): An J, Nagaishi T, Watabe T, Naruse TK., Watanabe M, Kimura A
  • Organizer: MICS 2018
  • Int'l Joint Research
• [Presentation] Mice overexpressing MKL1 in macrophages are susceptible to DSS-induced colitis. (2018)
  • Author(s): An J, Nagaishi T, Watabe T, Naruse TK., Watanabe M, Kimura A
  • Organizer: FOCIS 2018
  • Int'l Joint Research
• [Presentation] Analysis of cecal immune response in a murine model of colitis using intravital imaging. (2018)
  • Author(s): Nagaishi T, Watabe T, Tsugawa N, Kojima Y, Jose N, Yamada D, Adachi T, Watanabe M
  • Organizer: FOCIS 2018
  • Int'l Joint Research
• [Presentation] Mouse colonic secretory cells de-differentiate into intestinal stem cells and promote mucosal repair through activation of NF-KB signaling. (2018)
  • Author(s): Ishibashi F, Shimizu H, Kawamoto A, Suzuki K, Anzai S, Kuwabara K, Takahashi J, Nagata S, Oshima S, Tsuchiya K, Nakamura T, Watanabe M, Okamoto R
  • Organizer: ISSCR2018
  • Int'l Joint Research
• [Presentation] Mice overexpressing MKL1 in macrophages are susceptible to the development of colitis. (2018)
  • Author(s): An J, Nagaishi T, Watabe T, Naruse TK., Watanabe M, Kimura A
  • Organizer: DDW 2018
  • Int'l Joint Research
• [Presentation] Human colonic organoid treated with inflammatory factors might mimic the pathophysiology of epithelial cells in ulcerative colitis. (2018)
  • Author(s): Hibiya S, Tsuchiya K, Watanabe S, Nishimura R, Shirasaki T, Oshima S, Okamoto R, Nakamura T, Watanabe M
  • Organizer: DDW 2018
  • Int'l Joint Research
• [Presentation] Identification of lesion-specific epithelial function of ulcerative colitis by comparing colonic organoids from lesion and non-lesion parts of same patients. (2018)
  • Author(s): Shirasaki T, Tsuchiya K, Nishimura R, Watanabe S, Hibiya S, Katsukura N, Okamoto R, Nakamura T, Watanabe M
  • Organizer: DDW 2018
  • Int'l Joint Research
• [Presentation] Analysis of ileocecal immune response in an animal model of colitis using intra-vital imaging. (2018)
  • Author(s): Watabe T, Nagaishi T, Tsugawa N, Kojima Y, Jose N, Hosoya A, Adachi T, Watanabe M
  • Organizer: DDW 2018
  • Int'l Joint Research
• [Presentation] Single-cell level analysis of organoids derived from CD patients reveals disease-status related modifications of small intestinal stem cells. (2018)
  • Author(s): Suzuki K, Kuwabara K, Takahashi J, Anzai S, Kuno R, Kawamoto A, Ishibashi F, Nagata S, Hiraguri Y, Yui S, Tsuchiya K, Nakamura T, Ohtsuka K, Okamoto R, Watanabe M
  • Organizer: DDW 2018
  • Int'l Joint Research
• [Presentation] Fetalization of colonic epithelium medicated by YAP/TAZ links ECM remodeling to tissue regeneration. (2018)
  • Author(s): Yui S, Okamoto R, Tsuchiya K, Nakamura T, Watanabe M
  • Organizer: GI Research Academy 2018
  • Int'l Joint Research