Establishment of transplantation therapy using intestinal stem cells for large animals.

• Project/Area Number: 25670372
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Keio University
• Principal Investigator: SATO Toshiro 慶應義塾大学, 医学部, 特任准教授 (70365245)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 腸管上皮細胞 / 粘膜再生治療 / 下部消化管学 / オルガノイド / 炎症性腸疾患

Outline of Final Research Achievements

Inflammatory bowel disease is characterized by chronic mucosal inflammation and mainly treated with immunomodulatory reagents. Recently it was found that mucosal healing is a therapeutic goal to predict long-term remission. Nonetheless, there is little evidence of efficacy for mucosal regenerative therapy. We developed a novel intestinal stem cell culture system, in which isolated stem cells form stereotypic organoid structgures for long-term. Using this technology, we attempted to establish mucosal regenerative therapy for large animals. Taking advantage of our large animal facility, we established pig organoids from endoscopic biopsy specimens. The pig intestinal organoids were endoscopically transplanted on the epithelium-removed mucosa and engraft for 7-10 days. Further optimization may lead realization of regenerative medicine for inflammatory bowel disease.

Research Products

• [Journal Article] Modelling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids. (2015)
  • Author(s): Matano M,Date S, Shimokawa M, Takano A, Fujii M, Ohta Y, Watanabe T, Kanai T, Sato T.
  • Journal Title: Nature Medicine Volume: 21(3) Issue: 3 Pages: 256-262
  • DOI: 10.1038/nm.3802
  • Peer Reviewed / Open Access
• [Journal Article] Classical Th1 cells obtain colitogenicity by co-existence of RORγt-expressing T cells in experimental colitis (2014)
  • Author(s): Saigusa K, Hisamatsu T, Handa T, Sujino T, Mikami Y, Hayashi A, Mizuno S, Takeshita K, Sato T, Matsuoka K, Kanai T
  • Journal Title: Inflamm Bowel Dis Volume: 20 Issue: 10 Pages: 1820-1827
  • DOI: 10.1097/mib.0000000000000149
  • Peer Reviewed
• [Journal Article] Cross-talk between RORgammat+ innate lymphoid cells and intestinal macrophages induces mucosal IL-22 production in Crohn's disease. (2014)
  • Author(s): 1.Mizuno S, Mikami Y, Kamada N, Handa T, Hayashi A, Sato T, Matsuoka K, Matano M, Ohta Y, Sugita A, Koganei K, Sahara R, Takazoe M, Hisamatsu T, Kanai T.
  • Journal Title: Inflamm Bowel Dis Volume: 20(8) Issue: 8 Pages: 1426-1434
  • DOI: 10.1097/mib.0000000000000105
  • Peer Reviewed
• [Journal Article] KLF5 regulates the integrity and oncogenicity of intestinal stem cells. (2014)
  • Author(s): Nakaya T, Ogawa S, Manabe I, Tanaka M, Sanada M, Sato T, Taketo MM, Nakao K, Clevers H, Fukayama M, Kuroda M, Nagai R.
  • Journal Title: Cancer Res Volume: in press
  • Peer Reviewed
• [Journal Article] Macrophages and dendritic cells emerge in the liver during intestinal inflammation and predjspose the liver to inflammation. (2014)
  • Author(s): Mikami Y, Mizuno S, Nakamoto N, Hayashi A, Sujino T, Sato T, Kamada N, Matsuoka K, Hisamatsu T, Ebinuma H, Hibi T, Yoshimura A, Kanai T.
  • Journal Title: PLoS One Volume: 9 Issue: 1 Pages: 1-1
  • DOI: 10.1371/journal.pone.0084619
  • Peer Reviewed
• [Journal Article] Establishment of gastro-intestinal epithelia-organoids. (2014)
  • Author(s): Mahe MM, Aihara E, Sato T, Shroyer NF.
  • Journal Title: Current Protocols in Molecular Biology Volume: in press
  • Peer Reviewed
• [Journal Article] Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis. (2013)
  • Author(s): Huch M, Bonfanti P, Boj SF, Sato T, Loomans CJ, van de Wetering M, Sojoodi M, Li VS, Schuijers J, Gracanin A, Ringnalda F, Begthel H, Hamer K, Mulder J, van Es JH, de Koning E, Vries RG, Heimberg H, Clevers H.
  • Journal Title: EMBO J. Volume: 32(20) Issue: 20 Pages: 2708-21
  • DOI: 10.1038/emboj.2013.204
  • Peer Reviewed
• [Journal Article] Continuous low-dose irradiation by I-125 seeds induces apoptosis of gastric cancer cells regardless of histological origin. (2013)
  • Author(s): Takabayashi K, Kashiwagi K, Kawata T, Sato T, Matsuoka K, Hisamatsu T, Takaishi H, Hibi T, Ogata H, Yahagi N, Kitagawa Y, Shigematsu N, Kanai T.
  • Journal Title: Cancer Biol Ther. Volume: 15(1) Issue: 1 Pages: 81-8
  • DOI: 10.4161/cbt.26610
  • Peer Reviewed
• [Journal Article] A Single Strain of Clostridium butyricum Induces Intestinal I L-10-Producing Macrophages to Suppress Acute Exlperimental Colitis in Mice. (2013)
  • Author(s): Hayashi A, Sato T, Kamada N, Mikami Y, Matsuoka K, Hisamatsu T, Hlbi T, Roers A, Yagita H, Ohteki T, Yoshimura A, Kanai T.
  • Journal Title: Cell Host Microbe Volume: 13 Issue: 6 Pages: 711-22
  • DOI: 10.1016/j.chom.2013.05.013
  • Peer Reviewed
• [Journal Article] Mucosal healing with oral tacrolimus is associated with favorable medium- and long-term prognosis in steroid-refractory/dependent ulcerative colitis patients. (2013)
  • Author(s): Miyoshi J, Matsuoka K, Inoue N, Hisamatsu T, Ichikawa R, Yajima T, Okamoto S, Naganuma M, Sato T, Kanai T, Ogata H, Iwao Y, Hibi
  • Journal Title: J Crohns Colitis Volume: 7 Issue: 12 Pages: e609-e614
  • DOI: 10.1016/j.crohns.2013.04.018
  • Peer Reviewed
• [Journal Article] Establishment of novel prediction system of intestinal absorption in humans using human intestinal tissues. (2013)
  • Author(s): Miyake M, Toguchi H, Nishibayashi T, Higaki K, Sugita A, Koganei K, Kamada N, Kitazume MT, Hisamatsu T, Sato T, Okamoto S, Kanai T, Hibi T.
  • Journal Title: J Pharm Sci. Volume: 102(8) Issue: 8 Pages: 2564-71
  • DOI: 10.1002/jps.23609
  • Peer Reviewed
• [Journal Article] TGR5 signalling inhibits the production of pro-inflammatory cytokines by in vitro differentiated inflammatory and intestinal macrophages in Crohn's disease. (2013)
  • Author(s): Yoneno K, Hisamatsu T, Shimamura K, Kamada N, Ichikawa R, Kitazume MT, Mori M, Uo M, Namikawa Y, Matsuoka K, Sato T, Koganei K, Sugita A, Kanai T, Hibi T.
  • Journal Title: Immunology Volume: 139(1) Issue: 1 Pages: 19-29
  • DOI: 10.1111/imm.12045
  • Peer Reviewed
• [Journal Article] IL-22-Producing RORyt-Dependent Innate Lymphoid Cells Play a Novel Protective Role in Murine Acute Hepatitis. (2013)
  • Author(s): Matsumoto A, Kanai T, Mikami Y, Chu PS, Nakamoto N, Ebinuma H, Saito H, Sato T, Yagita H, Hibi T.
  • Journal Title: PLoS One Volume: 8 Issue: 4 Pages: e62853-e62853
  • DOI: 10.1371/journal.pone.0062853
  • Peer Reviewed
• [Journal Article] Dendritic cells administered intrarectally penetrate the intestinal barrier to break intestinal tolerance via Th2-medeiated colitis in mice. (2013)
  • Author(s): Handa T, Kanai T, Sato T, Mikami Y, Sujino T, Hayashi. A, Mizuno S, Matsumoto A, Hisamatsu T, Hibi T.
  • Journal Title: Immunology Letters Volume: 150 Issue: 1-2 Pages: 123-9
  • DOI: 10.1016/j.imlet.2013.01.010
  • Peer Reviewed
• [Journal Article] In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. (2013)
  • Author(s): Huch M, Dorrell C, Boj SF, van Es JH, Li VSW, van de Wetering M, Sato T, Hamer K, Sasaki N, Finegold MJ, Haft A, Vries R, Grompe M, Clevers H.
  • Journal Title: Nature Volume: 494(7436) Issue: 7436 Pages: 247-50
  • DOI: 10.1038/nature11826
  • Peer Reviewed
• [Journal Article] Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. (2013)
  • Author(s): Sato T, Clevers H.
  • Journal Title: Science Volume: 340(6137) Issue: 6137 Pages: 1190-4
  • DOI: 10.1126/science.1234852
  • Peer Reviewed
• [Journal Article] Primary mouse small intestinal epithelial cell cultures. (2013)
  • Author(s): Sato T, Clevers H.
  • Journal Title: Methods in Molecular Biology. Volume: 945 Pages: 319-28
  • DOI: 10.1007/978-1-62703-125-7_19
  • ISBN: 9781627031240, 9781627031257
  • Peer Reviewed
• [Presentation] ミニ組織培養技術の大腸がん研究への応用 (2015)
  • Author(s): 佐藤俊朗
  • Organizer: 第1回TRC がん幹細胞シリーズセミナー
  • Place of Presentation: 愛媛大学
  • Year and Date: 2015-02-17
• [Presentation] ヒト腸管上皮感細胞培養の応用：ニッチシグナルと大腸がん (2014)
  • Author(s): 佐藤俊朗
  • Organizer: 第1回六甲医学研究会
  • Place of Presentation: 六甲山ホテル
  • Year and Date: 2014-11-07 – 2014-11-08
• [Presentation] The impact of intestinal Organoid culture system on colorectal cancer research. (2014)
  • Author(s): Toshiro Sato
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-26
• [Presentation] Establishment and Applications of Intestinal Stem Cell Culture system; Stem Cell Niche and Cancer. (2014)
  • Author(s): Toshiro Sato
  • Organizer: The 33rd Sapporo International Cancer Symposium.
  • Place of Presentation: 札幌
  • Year and Date: 2014-06-27
• [Presentation] 細胞外環境による腸管上皮幹細胞制御とがん化機構 (2014)
  • Author(s): 佐藤俊朗, 太田悠木
  • Organizer: 第14回日本抗加齢医学会総会
  • Place of Presentation: 大阪国際会議場
  • Year and Date: 2014-06-08
• [Presentation] Establishment and Applications of Intestinal Stem Cell Culture system; Stem Cell Niche and Cancer. (2014)
  • Author(s): Toshiro Sato
  • Organizer: GI Researh Academy
  • Place of Presentation: 東京（ロイヤルパークホテル）
  • Year and Date: 2014-06-06
• [Presentation] The Self-renewing Mechanism of Intestinal Stem Cell: Niche and Cancer (2014)
  • Author(s): 7.Toshiro Sato, Yuki Ohta, Ai Takano, Shoichi Date, Mamiko Matano, Mariko Shimokawa.
  • Organizer: 47th Annual Meeting of the Japanese Society of Developmental Biologists and Asia Pacific Developmental Biology Network
  • Place of Presentation: 名古屋（WINC愛知）
  • Year and Date: 2014-05-28
• [Presentation] Establishment and Applications of Intestinal Stem Cell Culture System (2014)
  • Author(s): Toshiro Sato
  • Organizer: Annual International Workshop on Mucosal Immunology and Vaccine for Young Investigators.
  • Place of Presentation: 東京大学
  • Year and Date: 2014-05-23
• [Presentation] 腸管幹細胞とそのニッチ (2014)
  • Author(s): 佐藤俊朗
  • Organizer: 京都大学再生医科学研究所公開シンポジウム
  • Place of Presentation: 京都大学
  • Year and Date: 2014-05-13
• [Presentation] 消化管上皮幹細胞培養の基盤と応用 (2014)
  • Author(s): 佐藤俊朗，Hans Clevers, 金井隆典
  • Organizer: 第100回日本消化器病学会総会
  • Place of Presentation: 東京国際フォーラム
  • Year and Date: 2014-04-25
• [Presentation] 消化器ミニ組織培養の開発とその再生医療応用 (2014)
  • Author(s): 佐藤俊朗
  • Organizer: 第51回日本臨床分子医学会学術集会
  • Place of Presentation: 東京国際フォーラム
  • Year and Date: 2014-04-11
• [Presentation] 幹細胞ニッチシグナルによる腸管上皮幹細胞制御機構
  • Author(s): 佐藤俊朗
  • Organizer: 細胞が集団になって初めて発現する機能．
  • Place of Presentation: 大阪大学蛋白質研究所セミナー
• [Presentation] Molecular mechanism of intestinal stem cell self-renewal: Stem cells and their niche signals.
  • Author(s): Toshiro Sato.
  • Organizer: The graduate course in Molecular and Developmental Biology (Prof. Noah Shroyer & Prof. James Wells)
  • Place of Presentation: Cincinnati Children's USA
• [Presentation] Molecular mechanism of intestinal stem cell self-renewal: Stem cells and their niche signals.
  • Author(s): Toshiro Sato.
  • Organizer: University of Michigan Center for Organogenesis Seminar. (Prof. Linda Samuelson & Prof. Jason Spence)
  • Place of Presentation: University of Michigan Center USA
• [Presentation] Molecular mechanism of intestinal carcinogenesis; Stem cells and their niche signals.
  • Author(s): Toshiro Sato, Ai Takano, Shoichi Date, Mami, Matano, Mariko Shimokawa, Takanori Kanai
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: 日本（横浜）
• [Presentation] Molecular mechanism of intestinal stem cell self-renewal:Stem cells and their niche signals.
  • Author(s): Toshiro Sato, Mami Matano, Shoichi Date, Ai Takano, Mariko Shimokawa.
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 日本（横浜）
• [Presentation] Establishment of intestinal stem cell culture: basic and application.
  • Author(s): Toshiro Sato
  • Organizer: European Cancer Stem Cells Research meeting.
  • Place of Presentation: Cardiff University. UK
  • Invited
• [Presentation] Establishment of patient-derived colorectal cancer/adenoma organoid culture system: an application to tubulology.
  • Author(s): Toshiro Sato
  • Organizer: 1st International Meeting for Epithelial Tubulology
  • Place of Presentation: 日本（札幌）
• [Presentation] Long-term culture system for human intestinal stem cells and cancer.
  • Author(s): Toshiro Sato
  • Organizer: American Association for Cancer Research (AACR) Annual Meeting 2013.
  • Place of Presentation: Washington DC.USA
  • Invited
• [Book] Wntシグナルによる消化器上皮幹細胞制御 (2014)
  • Author(s): 佐藤俊朗
  • Publisher: 中外医学社
• [Book] ヒト消化器上皮培養によるオーダーメイド治療への可能性 (2014)
  • Author(s): 太田悠木，佐藤俊朗
  • Publisher: 先端医学社
• [Book] 糖尿病・代謝・内分泌 (2014)
  • Author(s): 佐藤俊朗
  • Publisher: Annual Review
• [Book] 腸の再生機構．幹細胞研究と再生医療．中内啓光編 (2013)
  • Author(s): 南木康作,藤井正幸,佐藤俊朗
  • Publisher: 南江堂
• [Book] 大腸がん幹細胞の機能解析法．がん基盤生物学-革新的シーズ育成に向けて- 清木元治 編 (2013)
  • Author(s): 股野麻未,佐藤俊朗
  • Publisher: 南江堂
• [Book] 腸管上皮幹細胞の加齢・老化とそれに伴う関連疾患．内分泌・糖尿病・代謝内科 (2013)
  • Author(s): 半田単己，佐藤俊朗
  • Publisher: 科学評論社
• [Book] Wntシグナルを利用した消化器組織再生技術“オルガノイド培養”の開発 細胞工学 (2013)
  • Author(s): 佐藤俊朗
  • Publisher: 学研メディカル秀潤社
• [Book] 腸管上皮細胞の老化 Anti-Aging Medicine. (2013)
  • Author(s): 佐藤俊朗
  • Publisher: メディカルレビュー社
• [Book] 腸管上皮幹細胞ニッチによる幹細胞制御機構．分子消化器病 (2013)
  • Author(s): 佐藤俊朗
  • Publisher: 先端医学社