Identification of human colorectal cancer stem cells.

• Project/Area Number: 24689036
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial 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)
• Research Collaborator: MATANO Mami 慶應義塾大学, 医学部, 研究員 (20439889) ; TAKANO Ai 慶應義塾大学, 医学部, 研究員 (50647584)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥25,740,000 (Direct Cost: ¥19,800,000、Indirect Cost: ¥5,940,000); Fiscal Year 2013: ¥12,090,000 (Direct Cost: ¥9,300,000、Indirect Cost: ¥2,790,000); Fiscal Year 2012: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
• Keywords: 大腸がん / 幹細胞 / 腸管上皮幹細胞 / WNT / LGR5

Research Abstract

We have established intestinal organoids from human intestinal epithelium and applied genome engineering system to organoids. With this approach, we introduced multiple driver mutations into human intestinal organoids to elucidate the mechanism of colorectal carcinogenesis. Furthermore, we knocked-in reporter to LGR5 locus of intestinal organoids using genome editing. Owing to the genetic silencing of Lgr5 promoter, we are attempting to employ other stem cell genetic markers. We also established xenotransplantation system with engineered organoids or colorectal cancer organoids. The organoids were readily engrafted and form tumor recapitulating clinical colorectal cancer. Our results suggest that engineered organoid system provide new insights into the understanding of human colorectal cancer. The platform will be useful not only to elucidate cell biological mechanism of colorectal cancer stem cells but also to apply genetic mutation based drug discovery and personalized medicine.

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 MM, Nakao K, Clevers H, Fukayama M, Kuroda M, Nagai R
  • Journal Title: Cancer Res Volume: (in press)PMID Pages: 24626089-24626089
• [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)
• [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] 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] 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] 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: Immunol Lett Volume: 150 Pages: 123-9
• [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] 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] ミニ組織技術を用いた大腸がん幹細胞制御機構の解明 (2014)
  • Author(s): 佐藤俊朗
  • Organizer: がん研究分野の特性等を踏まえた支援活動公開シンポジウム
  • Place of Presentation: 一橋講堂,竹橋
  • Year and Date: 2014-01-30
• [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 (2013)
  • 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
  • Year and Date: 2013-11-28
• [Presentation] Molecular mechanism of intestinal stem cell self-renewal : Stem cells and their niche signals (2013)
  • 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
  • Year and Date: 2013-11-13
• [Presentation] Molecular mechanism of intestinal stem cell self-renewal : Stem cells and their niche signals (2013)
  • 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
  • Year and Date: 2013-11-12
• [Presentation] Molecular mechanism of intestinal carcinogenesis; Stem cells and their niche signals (2013)
  • Author(s): Toshiro Sato, Ai Takano, Shoichi Date, Mami Matano, Mariko Shimokawa, Takanori Kanai
  • Organizer: Symposia 12, Aberrant signal transduction and therapeutic strategy for molecular target, 第72回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2013-10-03
• [Presentation] 腸管上皮幹細胞培養技術の確立～幹細胞ニッチと自己複製機構の解明～ (2013)
  • Author(s): 佐藤俊朗
  • Organizer: 第6回Symphony
  • Place of Presentation: 飯田橋
  • Year and Date: 2013-09-22
• [Presentation] Molecular mechanism of intestinal stem cell self-renewal : Stem cells and their niche signals (2013)
  • Author(s): Toshiro Sato, Mami Matano, Shoichi Date, Ai Takano, Mariko Shimokawa
  • Organizer: International Symposium 3. Molecular mechanisms for the growth and differentiation of tissue-specific stem cells in mammals, 第86回日本生化学会大会
  • Place of Presentation: 横浜
  • Year and Date: 2013-09-11
• [Presentation] Establishment of intestinal stem cell culture : basic and application (2013)
  • Author(s): Toshiro Sato
  • Organizer: European Cancer Stem Cells Research meeting
  • Place of Presentation: Cardiff Univ. UK
  • Year and Date: 2013-07-25
  • Invited
• [Presentation] Establishment of patient-derived colorectal cancer/adenoma organoid culture system : an application to tubulology (10.1016/j.imlet.2013.01.010) (2013)
  • Author(s): Toshiro Sato
  • Organizer: 1st International Meeting for Epithelial Tubulology
  • Place of Presentation: Sapporo
  • Year and Date: 2013-06-22
• [Presentation] 腸管上皮幹細胞の自己複製機構:幹細胞ニッチと発がん (2013)
  • Author(s): 佐藤俊朗
  • Organizer: 第13回GIリサーチフォーラム
  • Place of Presentation: 東北大学
  • Year and Date: 2013-06-19
• [Presentation] 消化器幹細胞培養による組織再生技術基礎から応用 (2013)
  • Author(s): 佐藤俊朗
  • Organizer: 徳島大学疾患酵素学研究センターセミナー
  • Place of Presentation: 徳島大学
  • Year and Date: 2013-05-23
• [Presentation] Long-term culture system for human intestinal stem cells and cancer (2013)
  • Author(s): Toshiro Sato
  • Organizer: Methods Workshop : Stem Cells, Enteroids and Organoids : A New Era for In Vitro Models of the Intestine. American Association for Cancer Research (AACR) Annual Meeting 2013
  • Place of Presentation: Washington DC.USA
  • Year and Date: 2013-04-06
  • Invited
• [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(Invited Speaker)
  • Year and Date: 2013-04-06
• [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(Invited Speaker)
  • Year and Date: 2013-03-23
• [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シンポジウム14.消化器領域における幹細胞研究の進歩
  • Place of Presentation: 鹿児島
  • Year and Date: 2013-03-23
• [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 ワークショップ 幹細胞制御機構 : ニッチはどこまで解明されたか?
  • Place of Presentation: 神戸(招待講演)
  • Year and Date: 2012-12-11
• [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(Invited Speaker)
  • Year and Date: 2012-11-03
• [Presentation] Development of human intestinal stem cell culture system. Symposium 4 Gastrointestinal Stem Cell. (2012)
  • Author(s): Toshiro Sato
  • Organizer: International Ulcer Week 2012 (第40回日本潰瘍学会・第14回国際潰瘍学会)
  • Place of Presentation: Tokyo(Invited Speaker)
  • Year and Date: 2012-07-12
• [Presentation] ヒト腸管上皮幹細胞の同定と培養法の確立 (2012)
  • Author(s): Toshiro Sato
  • Organizer: 第98回日本消化器病学会総会.シンポジウム2 : 消化管stem cellの新たな展開
  • Place of Presentation: Tokyo(Invited Speaker)
  • Year and Date: 2012-04-19
• [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
• [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: 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 Univ. 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. Methods Workshop: Stem Cells, Enteroids and Organoids: A New Era for In Vitro Models of the Intestine.
  • 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] 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 (2013)
  • Author(s): 南木康作,藤井正幸,佐藤俊朗 (中内啓光編)
  • 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- (2013)
  • Author(s): 股野麻未,佐藤俊朗(清木元治編)
  • Publisher: 南江堂
• [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] 先端医学社 (2013)
  • Author(s): 佐藤俊朗
  • Publisher: 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
• [Patent(Industrial Property Rights)] Culture medium for epithelial stem cells and organoids comprising the stem cells (2014)
  • Inventor(s): Toshiro Sato, Johannes Carolus Clevers
  • Industrial Property Rights Holder: Koninklijke Nederlandse Akademie van Wetenschappen
  • Industrial Property Rights Type: 特許
  • Filing Date: 2014-02-04