大腸上皮の癌化に伴う管腔形成異常メカニズムの解明

• Project Area: Regulation of polarity signaling during morphogenesis, remodeling, and breakdown of epithelial tubular structure
• Project/Area Number: 24112523
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Keio University
• Principal Investigator: 佐藤 俊朗 慶應義塾大学, 医学部, 特任准教授 (70365245)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥14,690,000 (Direct Cost: ¥11,300,000、Indirect Cost: ¥3,390,000); Fiscal Year 2013: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000); Fiscal Year 2012: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: 三次元培養 / 大腸がん / 幹細胞 / 大腸癌

Research Abstract

我々は，ヒト臨床検体を用い，腸管上皮幹細胞ならびに大腸がん上皮細胞の3次元培養法（オルガノイド培養）を開発した. 本研究計画では樹立した大腸がんオルガノイドを用いて，上皮細胞の管腔構造形成の異常メカニズムの解明を目的としている．平成24年度は大腸がん症例数を増やし，様々な臨床ステージの大腸がんのオルガノイド培養を樹立した．また，マウスへの異種移植により，in vivoでの管腔形成の評価が可能になった．こうした技術開発より，ヒト腸管上皮の管腔形成異常をin vitro，in vivoの双方で解析するシステムが確立された．さらに，オルガノイドの遺伝子変異の網羅的解析により，管腔構造形成異常を誘導する遺伝子について解明していきたい．
ウィルスベクタ－などの遺伝子操作技術が向上し，より自由度の高い，オルガノイドの遺伝子操作が可能になった．正常上皮細胞からの人工大腸がん誘導，細胞周期・幹細胞の蛍光可視化が実現しつつあり，今後，こうした技術を基に，腸管上皮細胞の時空間的な動態を数理学的モデルも含めて解明していきたい．

Current Status of Research Progress

Reason

25年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

25年度が最終年度であるため、記入しない。

Research Products

• [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 M, 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
• [Journal Article] Dll1(+) secretory progenitor cells revert to stem cells upon crypt damage. (2012)
  • Author(s): van Es JH, Sato T, van de Wetering M, Lyubimova A, Yee Nee AN, Gregorieff A, Sasaki N, Zeinstra L, van den Born M, Korving J, Martens AC, Barker N, van Oudenaarden A, Clevers H.
  • Journal Title: Nat Cell Biol. Volume: 14 Issue: 10 Pages: 1099-1104
  • DOI: 10.1038/ncb2581
  • 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] On the biomechanics of stem cell niche formation in the gut - modelling growing organoids. (2012)
  • Author(s): Buske P, Przybilla J, Loeffler M, Sachs N, Sato T, Clevers H, Galle J.
  • Journal Title: FEBS J Volume: 279 Pages: 3475-87
  • Peer Reviewed
• [Journal Article] Cdx2 determines the fate of postnatal intestinal endoderm (2011)
  • Author(s): Stringer EJ, Duluc I, Saandi T, Davidson I, Bialecka M, Sato T, Barker N, Clevers H, Pritchard CA, Winton DJ, Wright NA, Freund JN, Deschamps J, Beck F
  • Journal Title: Development Volume: 139 Issue: 3 Pages: 465-474
  • DOI: 10.1242/dev.070722
  • Peer Reviewed
• [Presentation] Long-term culture system for human intestinal stem cells and cancer. Methods Workshop: Stem Cells, Enteroids and Organoids: A New Era for In Vitro Models of the Intestine. (2013)
  • Author(s): Toshiro Sato
  • Organizer: American Association for Cancer Research (AACR) Annual Meeting 2013.
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2013-04-06
  • Invited
• [Presentation] Establish of human intestinal organoid culture system: Application to inflammation and cancer. (2013)
  • Author(s): Toshiro Sato
  • Organizer: The 2nd JSGE International Topic Conference.
  • Place of Presentation: Kagoshima Japan
  • Year and Date: 2013-03-22
  • Invited
• [Presentation] Paneth細胞による腸管上皮幹細胞制御機構． (2013)
  • Author(s): 佐藤俊朗
  • Organizer: 第99回日本消化器病学会総会
  • Place of Presentation: 鹿児島
  • Year and Date: 2013-03-22
  • Invited
• [Presentation] Dymamic intestinal stem cell regulation by Paneth cells. (2012)
  • Author(s): 佐藤俊朗
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-11
  • Invited
• [Presentation] Are we ready for using stem cell therapy for IBD (2012)
  • Author(s): Toshiro Sato
  • Organizer: Asian IBD Symposium Seoul 2012.
  • Place of Presentation: Seoul
  • Year and Date: 2012-11-02
  • Invited
• [Presentation] Development of human intestinal stem cell culture system. Symposium 4 Gastrointestinal Stem Cell. (2012)
  • Author(s): Toshiro Sato
  • Organizer: International Ulcer Week 2012
  • Place of Presentation: Tokyo
  • Year and Date: 2012-07-12
  • Invited
• [Presentation] ヒト腸管上皮幹細胞の同定と培養法の確立 (2012)
  • Author(s): 佐藤俊朗
  • Organizer: 第98回日本消化器病学会総会
  • Place of Presentation: 東京
  • Year and Date: 2012-04-19
• [Presentation] Development of Gut Stem Cell Culture. (2012)
  • Author(s): Toshiro Sato
  • Organizer: 第45回日本発生生物学会・第64回日本細胞生物学会.
  • Place of Presentation: Kobe
  • Invited
• [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: 大阪大学蛋白質研究所セミナー．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
• [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 stem cell self-renewal: Stem cells and their niche signals.
  • Author(s): Toshiro Sato, Ai Takano, Shoichi Date, Mami, Matano, Mariko Shimokawa
  • Organizer: 第72回 日本癌学会学術総会
  • Place of Presentation: 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: 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: 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．Annual Review 糖尿病・代謝・内分泌 (2014)
  • Author(s): 佐藤俊朗
  • Publisher: 中外医学社
• [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): 半田一己，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] 腸管上皮幹細胞の老化．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
• [Book] Methods in Molecular Biology. (2013)
  • Author(s): Sato T
  • Publisher: A product of Humana Press
• [Book] Intestinal Stem Cell Culture. (2012)
  • Author(s): Sato T
  • Publisher: 日本炎症・再生医学会
• [Book] Annual Review 消化器 (2012)
  • Author(s): Sato T
  • Publisher: 中外医学社
• [Patent(Industrial Property Rights)] CULTURE MEDIA FOR STEM CELLS． (2012)
  • Inventor(s): Sato T, Clevers H et al
  • Industrial Property Rights Holder: KNAW
  • Industrial Property Rights Type: 特許
  • Filing Date: 2012-12-13
  • Overseas