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ヒト大腸上皮培養による大腸癌幹細胞の分化破綻機構解析

Publicly

Project AreaDevelopment of Novel Treatment Strategies Targeting Cancer Stem Cells
Project/Area Number 25130704
Research Category

Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionTokyo Medical and Dental University

Principal Investigator

土屋 輝一郎  東京医科歯科大学, 医歯(薬)学総合研究科, 寄附講座准教授 (40376786)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000)
Fiscal Year 2014: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2013: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Keywords炎症発癌モデル / Atoh1 / 幹細胞可視化 / 癌幹細胞 / ニッチ形成
Outline of Annual Research Achievements

本計画は独自に開発した大腸上皮細胞の初代培養系を応用し癌化モデルによる正常幹細胞由来癌幹細胞の形成を空間的・経時的に解析することで癌幹細胞の特異性、機能維持機構を解明することを目的とする。本年度は、マウス由来大腸上皮の初代培養を行い、in vitroにおける炎症発がんモデルを確立するため、初代培養細胞にAOM処理を行い、さらに種々のサイトカインにて刺激するなど条件検討を行った。幹細胞可視化に関しては、レンチウイルスを用いて蛍光タンパクであるmCherryを初代培養細胞に導入することにより、1幹細胞の可視化に成功した。小腸初代培養細胞においてTLR5は基底側に限局して発現しており、フラジェリン添加によりサイトカインを産生することが明らかとなった。NFkB p65の免疫染色によりNFkBシグナルが作動する細胞を同定し、幹細胞も炎症刺激によりシグナルが作動することを明らかとした。また炎症発がんの特徴である粘液産生がんに着目し、粘液産生に寄与するAtoh1蛋白の機能解析を行い、粘液産生のみならずがん幹細胞性質、抗がん剤耐性機構まで関与することを明らかとし、TNFαがAtoh1蛋白を安定化させることにより、粘液形質を獲得することを発見した。炎症性腸疾患患者のがん部と非がん部の免疫染色を行い、炎症発がんではAtoh1が発現し粘液形質を維持することを明らかとした。
以上の成果は当初の研究計画をおおむね順調に遂行しており、ヒト大腸初代培養の確立および幹細胞可視化への条件検討は炎症発がんのモデルだけでなく、種々の病態、疾患予測に有用であると期待される。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Report

(2 results)
  • 2014 Annual Research Report
  • 2013 Annual Research Report

Research Products

(25 results)

All 2015 2014 2013

All Journal Article Presentation

  • [Journal Article] Suppressing TGFβ Signaling in Regenerating Epithelia in an Inflammatory Microenvironment Is Sufficient to Cause Invasive Intestinal Cancer2015

    • Author(s)
      Oshima H, Nakayama M, Han TS, Naoi K, Ju X, Maeda Y, Robine S, Tsuchiya K, Sato T, Sato H, Taketo MM, Oshima M.
    • Journal Title

      Cancer Research

      Volume: 75 Pages: 766-776

    • DOI

      10.1158/0008-5472.can-14-2036

    • NAID

      120005593962

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] RIPK3 regulates p62- LC3 complex formation via the caspase-8-dependent cleavage of p622015

    • Author(s)
      Y. Matsuzawa , S. Oshima, Y. Nibe, M.Kobayashi, C. Maeyashiki, Y. Nemoto, T. Nagaishi, R. Okamoto R, K. Tsuchiya, T. Nakamura, M. Watanabe.
    • Journal Title

      Biochem Biophys Res Commun.

      Volume: 456(1) Pages: 298-304

    • DOI

      10.1016/j.bbrc.2014.11.075

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Associations of HLA class I alleles in Japanese patients with Crohn's disease.2015

    • Author(s)
      102)Oryoji D, Hisamatsu T, Tsuchiya K, Umeno J, Ueda S, Yamamoto K, Matsumoto T, Watanabe M, Hibi T, Sasazuki T.
    • Journal Title

      Genes Immun.

      Volume: 16 Pages: 54-56

    • DOI

      10.1038/gene.2014.61

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt.2014

    • Author(s)
      N. Horita, K. Tsuchiya, R. Hayashi, K. Fukushima, S. Hibiya, M. Fukuda, Y. Kano, T. Mizutani, Y. Nemoto, S. Yui, R. Okamoto, T. Nakamura, M. Watanabe
    • Journal Title

      Biochem Biophys Res Commun.

      Volume: 454 Pages: 493-499

    • DOI

      10.1016/j.bbrc.2014.10.091

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Distinct expression patterns of Notch ligands, Dll1 and Dll4, in normal and inflamed mice intestine.2014

    • Author(s)
      Shimizu H, Okamoto R, Ito G, Nakata T, Suzuki K, Murano T, Mizutani T, Tsuchiya K, Nakamura T, Hozumi K, Watanabe M.
    • Journal Title

      2014

      Volume: 2

    • DOI

      10.7717/peerj.370

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Hes1 promotes the IL-22-mediated antimicrobial response by enhancing STAT3-dependent transcription in human intestinal epithelial cells.2014

    • Author(s)
      Murano T, Okamoto R, Ito G, Nakata T, Hibiya S, Shimizu H, Fujii S, Kano Y, Mizutani T, Yui S, Akiyama-Morio J, Nemoto Y, Tsuchiya K, Nakamura T, Watanabe M
    • Journal Title

      Biochem Biophys Res Commun.

      Volume: 443 Pages: 840-6

    • DOI

      10.1016/j.bbrc.2013.12.061

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Lineage-specific expression of bestrophin-2 and bestrophin-4 in human intestinal epithelial cells2013

    • Author(s)
      Ito G, Okamoto R, Nakamura T, Watanabe M et al
    • Journal Title

      PLoS One.

      Volume: 8

    • DOI

      10.1371/journal.pone.0079693

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] IL-7 promotes long-term in vitro survival of unique long-lived memory subset generated from mucosal effector memory CD4+ T cells in chronic colitis mice2013

    • Author(s)
      Takahara M, Nemoto Y, Oshima S, Matsuzawa Y, Kanai T, Okamoto R, Tsuchiya K, Nakamura T, Yamamoto K, Watanabe M.
    • Journal Title

      Immunol Lett

      Volume: 156 Pages: 82-93

    • DOI

      10.1016/j.imlet.2013.09.001

    • NAID

      120005468269

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Th1/Th17-mediated interstitial pneumonia in chronic colitis mice independent of intestinal microbiota2013

    • Author(s)
      Nemoto Y, Watanabe M et al
    • Journal Title

      J Immunol

      Volume: 190 Pages: 6616-25

    • DOI

      10.4049/jimmunol.1202930

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 【特集:機能性消化管障害(FGID):診断と治療の進歩】I. FGIDの基本的知識 4. 感染症や腸内細菌の意義(免疫学的異常について).2013

    • Author(s)
      土屋輝一郎
    • Journal Title

      日本内科学会雑誌

      Volume: 102 Pages: 25-31

    • Related Report
      2013 Annual Research Report
  • [Journal Article] 消化管幹細胞と消化器がん幹細胞2013

    • Author(s)
      土屋輝一郎
    • Journal Title

      日本内科学会雑誌

      Volume: 102 Pages: 2273-2278

    • NAID

      10031202663

    • Related Report
      2013 Annual Research Report
  • [Presentation] The Acquisition of Cancer Stemness by Atoh1 in Colitis Associated Cancer2015

    • Author(s)
      Kiichiro Tsuchiya, Keita Fukushima, Mamoru Watanabe
    • Organizer
      8th Annual World Congress of Regenerative Medicine & Stem Cells
    • Place of Presentation
      釜山(韓国)
    • Year and Date
      2015-03-21
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] Innate immune spiral of intestinal epithelial cells by the long-term inflammation.2014

    • Author(s)
      Tsuchiya K, Hibiya S, Watanabe M
    • Organizer
      The 73rd Annual Meeting of the Japanese Cancer Association.
    • Place of Presentation
      パシフィコ横浜(横浜)
    • Year and Date
      2014-09-25
    • Related Report
      2014 Annual Research Report
  • [Presentation] Molecular mechanism in the pathogenesis of the colitis-associated colorectal cancer.2014

    • Author(s)
      Tsuchiya K
    • Organizer
      The 32th Annual Meeting of Japanese Society for Cytoprotection & Cytobiology
    • Place of Presentation
      Kyoto, Japan
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Live imaging of single cell reveals single stem cell dynamics in an organoid derived from murine small intestine.2013

    • Author(s)
      Horita N, Tsuchiya K, Hibiya S, Fukushima K, Kano Y, Xiu Zheng, Okamoto R, Nakamura T , Watanabe M
    • Organizer
      DDW2013
    • Place of Presentation
      Orland,USA
    • Related Report
      2013 Annual Research Report
  • [Presentation] Stabilization of Atoh1 protein by TNF-α in colitic cancer might acquire the cancer stemness.DDW20132013

    • Author(s)
      Tsuchiya K, Fukushima K, Kano Y, Hibiya S, Horita N, Xiu Zheng, Okamoto R,Nakamura T, Watanabe M
    • Organizer
      DDW2013
    • Place of Presentation
      Orland,USA
    • Related Report
      2013 Annual Research Report
  • [Presentation] Atoh1 protein stabilization acquired cancer stem cell phenotype and niche in colon cancer2013

    • Author(s)
      Kano Y, Tsuchiya K, Okamoto R, Nakamura T: Watanabe M
    • Organizer
      GI Research Academy 2013
    • Place of Presentation
      Kyoto, Japan
    • Related Report
      2013 Annual Research Report
  • [Presentation] Atoh1proteinstabilizationacquiredcancerstemcell phenotype and niche in colon cancer.2013

    • Author(s)
      Kano Y, Tsuchiya K, Watanabe M
    • Organizer
      第72回日本癌学会学術集会
    • Place of Presentation
      横浜
    • Related Report
      2013 Annual Research Report
  • [Presentation] Cancer Stemness In Mucinous Colon Cancer.2013

    • Author(s)
      Tsuchiya K, Kano Y, Watanabe M
    • Organizer
      BIT's 6th Annual World Congress of Regenerative Medicine & Stem Cells-2013.
    • Place of Presentation
      大連、中国
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 消化管幹細胞と消化器癌幹細胞2013

    • Author(s)
      土屋輝一郎
    • Organizer
      第110回 日本内科学会講演会.
    • Place of Presentation
      東京
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 全小腸マッピング生検検体を用いた網羅的遺伝子発現解析によるクローン病特異的遺伝子の同定2013

    • Author(s)
      堀田伸勝、土屋輝一郎、渡辺 守
    • Organizer
      第85回 日本消化器内視鏡学会総会
    • Place of Presentation
      京都
    • Related Report
      2013 Annual Research Report
  • [Presentation] 炎症関連大腸がんにおける悪性形質獲得機構解析2013

    • Author(s)
      土屋輝一郎
    • 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
      東京
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Atoh1発現大腸癌における幹細胞形質獲得とニッチ形成2013

    • Author(s)
      加納嘉人、土屋輝一郎、渡辺 守
    • Organizer
      JDDW2013
    • Place of Presentation
      東京
    • Related Report
      2013 Annual Research Report
  • [Presentation] 白血球除去療法(LCAP)による潰瘍性大腸炎の治療戦略2013

    • Author(s)
      土屋輝一郎
    • Organizer
      JDDW2013
    • Place of Presentation
      東京
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 全小腸マッピング生検によるクローン病病態解析2013

    • Author(s)
      土屋輝一郎、堀田伸勝、林 亮平、日比谷秀爾、福島啓太、加納嘉人、渡辺 守
    • Organizer
      第51回 小腸研究会
    • Place of Presentation
      名古屋
    • Related Report
      2013 Annual Research Report

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Published: 2013-05-15   Modified: 2019-07-29  

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