Wntシグナルによる細胞増殖、分化、運動の制御機構

• Project/Area Number: 18209011
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Hiroshima University
• Principal Investigator: 菊池 章 Hiroshima University, 大学院・医歯薬学総合研究科, 教授 (10204827)
• Co-Investigator(Kenkyū-buntansha): 岸田 昭世 広島大学, 大学院・医歯薬学総合研究科, 助教授 (50274064)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥49,400,000 (Direct Cost: ¥38,000,000、Indirect Cost: ¥11,400,000); Fiscal Year 2008: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000); Fiscal Year 2007: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000); Fiscal Year 2006: ¥30,420,000 (Direct Cost: ¥23,400,000、Indirect Cost: ¥7,020,000)
• Keywords: Wnt3a / LRP6 / 受容体 / カベオリン / β-カテニン / 細胞極性 / Wnt-3a / Axin / インターナリゼーション

Research Abstract

本年度は最終年度であるので、β-カテニンの安定化の抑制機構について重点的に解析した。Dickkopf1(Dkk1)はWnt/β-カテニン経路の阻害分子であるが、その抑制作用は十分に理解されていない。私共はこれまでに、LRP6がカベオリンの局在する細胞膜上の領域(脂質ラフト画分)に存在し、インターナリーゼーションされることがWntによるβ-カテニンの安定化に重要であることを明らかにしてきた。Dkk1はLRP6を細胞膜上の脂質ラフト画分から非脂質ラフト画分に移動させ、クラスリン依存性にインターナリーゼーションさせることにより、β-カテニン経路を抑制した。さらに、Dkk1は低分子量Gタンパク質Rab5を介してLRP6を早期エンドソームに移行し、Rab11を介して細胞膜にリサイクリングさせた。しかも、リサイクリングしたLRP6は再びWnt3aとDkk1に応答した。一方、Wnt5aもWnt3a依存性のβ-カテニン経路を抑制することが知られており、核内でβ-カテニンと転写因子Tcf4の結合を阻害すると考えられていた。しかし、Wnt5aはWnt3aが作用する受容体Fzに競合して結合する結果、β-カテニン経路を抑制した。したがって、β-カテニン経路を抑制するために複数の経路が存在することが明らかになった。
Wntシグナルによる細胞極性の制御機構を明らかにするために、上皮細胞(MDCK細胞とHPPL細胞)を3次元培養の実験系により解析した。MDCK細胞は肝細胞増殖因子(HGF)刺激により管構造を形成するが、DvlをノックダウンするとHGF依存性の管形成が認められなくなった。また、HPPL細胞が管形成を行う場合に、その伸長先端部の細胞の先進部にDvlとAPCが濃縮した。興味深いことに、DvlとAPCは互いに結合して、Wnt5aはこれを増強した。さらに、DvlとAPCは細胞接着斑のパキシリンとFAKとも複合体を形成した。これらの結果を端緒に、Wntシグナルによる細胞形態や極性の制御に関する分子機構が解明されることが期待される。以上、この3年間でWntシグナルによる細胞増殖、分化、運動に関する成果が得られたと考えている。

Research Products

• [Journal Article] Laminin γ2 mediates Wnt5a-induced invasion of Gastric Cancer Cells. (2009)
  • Author(s): Yamamoto, H., et al
  • Journal Title: Gastroenterology (in press)
  • Peer Reviewed
• [Journal Article] Axin localizes to the centrosome and is involved in microtubule nucleation. (2009)
  • Author(s): Fumoto, K., et al
  • Journal Title: EMBO R. (in press)
  • Peer Reviewed
• [Journal Article] Selective activation of mechanisms of Wnt signaling pathways. (2009)
  • Author(s): Kikuchi, A., et al
  • Journal Title: Trends Cell Biol. 19 Pages: 119-129
  • Peer Reviewed
• [Journal Article] CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. (2009)
  • Author(s): Nishiyama, M., et al
  • Journal Title: Nat. Cell Biol. 11 Pages: 172-182
  • Peer Reviewed
• [Journal Article] IFN-induced transmembrane protein 1 promotes the invasion at the early stage of head and neck cancer progression. (2008)
  • Author(s): Hatano, H., et al
  • Journal Title: Cli. Cancer Res. 14 Pages: 6097-6105
  • Peer Reviewed
• [Journal Article] Polycomb-group complex 1 acts as an E3 ubiquitin ligase for Geminin to sustain hematopoietic stem cell activity. (2008)
  • Author(s): Ohtsubo, M., et al
  • Journal Title: Proc. Natl. Acad. Sci. USA 105 Pages: 10396-10401
  • Peer Reviewed
• [Journal Article] AIP regulates stability of Aurora-A at early mitotic phase coordinately with GSK-3β. (2008)
  • Author(s): Fumoto, K., et al
  • Journal Title: Oncogene 27 Pages: 4478-4487
  • Peer Reviewed
• [Journal Article] IGFBP-4 is an inhibitor of canonical Wnt signaling required for cardiogenesis. (2008)
  • Author(s): Zhu, W., et al
  • Journal Title: Nature 454 Pages: 345-349
  • Peer Reviewed
• [Journal Article] Wnt3a and Dkk1 regulate distinct internalization pathways of LRP6 to tune the activation of β-catenin signaling. (2008)
  • Author(s): Yamamoto, H., et al
  • Journal Title: Dev. Cell 15 Pages: 37-48
  • Peer Reviewed
• [Journal Article] GSK-3β regulates proper mitotic spindle formation in cooperation with a component of the γ-tubulin ring complex, GCP5. (2008)
  • Author(s): Izumi, N., et al.
  • Journal Title: J. Biol. Chem. 283 Pages: 12981-12991
  • Peer Reviewed
• [Journal Article] Tumor formation due to abnormalities in the β-catenin-independent pathway of Wnt signaling. (2008)
  • Author(s): Kikuchi, A., et al
  • Journal Title: Cancer Sci. 99 Pages: 202-208
  • NAID: 10024001624
  • Peer Reviewed
• [Journal Article] Tumor formation due to abnormalities in the β-catenin-independent pathway of Wnt signaling. (2008)
  • Author(s): Kikuchi, A. and Yamamoto, H.
  • Journal Title: Cancer Sci. 99 Pages: 202-208
  • NAID: 10024001624
  • Peer Reviewed
• [Journal Article] GSK-3β regulates proper mitotic spindle formation in cooperation with a component of the γ-tubulin ring complex, GCP5. (2008)
  • Author(s): Izumi, N., et. al.
  • Journal Title: J. Biol. Chem. 283 Pages: 12981-12991
  • Peer Reviewed
• [Journal Article] AIP regulates stability of Aurora-A at early mitotic phase coordinately with GSK-3β. (2008)
  • Author(s): Fumoto, K., et. al.
  • Journal Title: Oncogene (in press)
  • Peer Reviewed
• [Journal Article] IFITM1 promotes the invasion at the early stage of head and neck cancer progression. (2008)
  • Author(s): Hatano, H., et. al.
  • Journal Title: Cli. Cancer Res. (in press)
  • Peer Reviewed
• [Journal Article] Wnt3a and Dkkl regulate internalization of LRP6 differently to tune β-catenin pathway. (2008)
  • Author(s): Yamamoto, H., et. al.
  • Journal Title: Dev. Cell (In press)
  • Peer Reviewed
• [Journal Article] Post-translational palmitoylation and glycosylation of Wnt-5a are necessary for its signaling. (2007)
  • Author(s): Kurayoshi, M., et. al.
  • Journal Title: Biochem. J. 402 Pages: 515-523
  • Peer Reviewed
• [Journal Article] Dvl regulates endo- and exocytotic processes through binding to synaptotagmin. (2007)
  • Author(s): Kishida, S., et. al.
  • Journal Title: Genes Cells 12 Pages: 49-61
  • Peer Reviewed
• [Journal Article] Glycosylation and palmitoylation of Wnt-3a are coupled to produce an active form of Wnt-3a. (2007)
  • Author(s): Komekado, H., et. al.
  • Journal Title: Genes Cells 12 Pages: 521-534
  • Peer Reviewed
• [Journal Article] Wnt5a modulates glycogen synthase kinase 3 to induce phospshorylation of receptor tyrosine kinase Ror2. (2007)
  • Author(s): Yamamoto, H., et. al.
  • Journal Title: Genes Cells 12 Pages: 1215-1223
  • Peer Reviewed
• [Journal Article] Increased expression of h-prune is associated with tumor progression and poor survival in gastric cancer. (2007)
  • Author(s): Oue, N., et. al.
  • Journal Title: Cancer Sci. 98 Pages: 1198-1205
  • Peer Reviewed
• [Journal Article] Sprouty2 and Sprouty4 are essential for embryonic morphogenesis and regulation of FGF signaling. (2007)
  • Author(s): Taniguchi, K., et. al.
  • Journal Title: Biochem. Biophys. Res. Commun. 352 Pages: 896-902
  • Peer Reviewed
• [Journal Article] Multiplicity of the interactions of Wnt proteins and their receptors. (2007)
  • Author(s): Kikuchi, A., et. al.
  • Journal Title: Cell. Signal. 19 Pages: 659-671
  • Peer Reviewed
• [Journal Article] Regulation of Wnt signaling by receptor-mediated endocytosis. (2007)
  • Author(s): Kikuchi, A. and Yamamoto, H.
  • Journal Title: J. Biochem. 141 Pages: 443-451
  • NAID: 10020099687
  • Peer Reviewed
• [Journal Article] The DIX domain of Dishevelled confers Wnt signaling by dynamic polymerization. (2007)
  • Author(s): Schwarz-Romond, T., et. al.
  • Journal Title: Nat. Strut. Mol. Biol. 14 Pages: 484-492
  • Peer Reviewed
• [Journal Article] Non-covalent binding of SUMO protease to SUMO is necessary for enzymatic activities and cell growth. (2007)
  • Author(s): Ihara, M., et. al.
  • Journal Title: J. Biol. Chem. 282 Pages: 16465-16475
  • Peer Reviewed
• [Journal Article] Dvl regulates endo- and exocytotic processes through binding to synaptotagmin. (2007)
  • Author(s): Kishida, S., et al.
  • Journal Title: Genes Cells 12 Pages: 49-61
• [Journal Article] Post-translational palmitoylation and glycosylation of Wnt-5a are necessary for its signaling. (2007)
  • Author(s): Kurayoshi, M., et al.
  • Journal Title: Biochem. J. 402 Pages: 515-523
• [Journal Article] Sprouty2 and Sprouty4 are essential for embryonic morphogenesis and regulation of FGF signaling. (2007)
  • Author(s): Taniguchi, K., et al.
  • Journal Title: Biochem. Biophys. Res. Commun. 352 Pages: 896-902
• [Journal Article] Glycosylation and palmitoylation of Wnt-3a are coupled to produce an active form of Wnt-3a. (2007)
  • Author(s): Komekado, H., et al.
  • Journal Title: Genes Cells (in press)
• [Journal Article] Multiplicity of the interactions of Wnt proteins and their receptors.. (2007)
  • Author(s): Kikuchi, A., et al.
  • Journal Title: Cell. Signal. (in press)
• [Journal Article] Regulation of Wnt signaling by receptor-mediated endocytosis. (2007)
  • Author(s): Kikuchi, A., et al.
  • Journal Title: J. Biochem. (in press)
  • NAID: 10020099687
• [Journal Article] Glycogen synthase kinase-3 And h-prune regulate cell migration by modulating focal adhesions. (2006)
  • Author(s): Kobayashi, T., et al.
  • Journal Title: Mol. Cell. Biol. 26 Pages: 898-911
• [Journal Article] Caveolin is necessary for Wnt-3a-induced internalization of LRP6 and accumulation of β-catenin. (2006)
  • Author(s): Yamamoto, H., et al.
  • Journal Title: Dev. Cell 11 Pages: 213-223
• [Journal Article] GSK-3β-regulated interaction of BICD with dynein is involved in microtubule anchorage at centrosome. (2006)
  • Author(s): Fumoto, K., et al.
  • Journal Title: EMBO J. 25 Pages: 5670-5682
• [Journal Article] Expression of Wnt-5a is correlated with aggressiveness of gastric cancer by stimulating cell migration and invasion. (2006)
  • Author(s): Kurayoshi, M., et al.
  • Journal Title: Cancer Res. 66 Pages: 10439-10448
• [Journal Article] Filopodia formation mediated by receptor tyrosine kinase Ror2 is required for Wnt5a-induced cell migration. (2006)
  • Author(s): Nishita, M., et al.
  • Journal Title: J. Cell Biol. 175 Pages: 555-562
• [Journal Article] Developmemtal stage-specific, biphasic roles of Wnt/β-catenin signaling in cardiomyogenesis and hematopoiesis. (2006)
  • Author(s): Naito, A., et al.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 19812-19817
• [Journal Article] Suppression of STAT3 activity by Duplin, Which is a negative regulator of the Wnt signal. (2006)
  • Author(s): Yamashina, K., et al.
  • Journal Title: J. Biochem. 139 Pages: 305-314
• [Journal Article] Oog1, an oocyte-specific protein, interacts with Ras and Ras-signaling proteins during early embryogenesis. (2006)
  • Author(s): Tsukamoto, S., et al.
  • Journal Title: Biochem. Biophys. Res. Commun. 343 Pages: 1105-1112
• [Journal Article] HTLV-I Tax dysregulates β-catenin signaling. (2006)
  • Author(s): Tomita, M., et al.
  • Journal Title: J. Virol. 80 Pages: 10497-10505
• [Journal Article] Regulation of Wnt signaling by protein-protein interaction and post-translational modifications. (2006)
  • Author(s): Kikuchi, A., et al.
  • Journal Title: Exp. Mol. Med. 38 Pages: 1-10
• [Presentation] Wntシグナルによる細胞運動制御とその異常による癌の悪性化 (2008)
  • Author(s): 菊池章
  • Organizer: 第31回日本分子生物学会年会、第81回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-12
• [Presentation] 3次元培養を用いたDishevelledによる細胞運動・形態形成制御機構の解析 (2008)
  • Author(s): 松本真司
  • Organizer: 第31回日本分子生物学会年会、第81回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-11
• [Presentation] β-カテニン非依存性のWntシグナルによる細胞応答とがん (2008)
  • Author(s): 菊池章
  • Organizer: 第67回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-10-29
• [Presentation] 脂質ラフトにおけるWntシグナルの活性制御 (2008)
  • Author(s): 山本英樹
  • Organizer: 第67回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-10-28
• [Presentation] Wnt5aはlaminin-5γ2の発現を介して胃癌細胞の転移能を促進する (2008)
  • Author(s): 山本英喜
  • Organizer: 第67回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-10-28
• [Presentation] Aggressiveness of cancer induced by abnormality of Wnt signaling. (2007)
  • Author(s): Kikuchi, A.
  • Organizer: The Japan-Korea joint meeting on wnt signaling and human diseases.
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2007-10-26
• [Presentation] Spatial and temporal regulation of Glycogen Synthase kinase-3 and Cellular function. (2007)
  • Author(s): Kikuchi, A.
  • Organizer: 2007 Annual meeting of Korean Society of Medical Biochemistry and Molecular Biology.
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2007-10-25
• [Presentation] Wntシグナルによるがんの悪性化 (2007)
  • Author(s): 菊池章
  • Organizer: 第66回日本癌学会学術総会モーニングレクチャー
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-05
• [Presentation] Regulation of Wnt signaling by receptor-mediated endocytosis. (2007)
  • Author(s): Kikuchi, A.
  • Organizer: Sympojium of Wnt signaling in development and disease.
  • Place of Presentation: Berlin, Germany
  • Year and Date: 2007-09-14
• [Presentation] Expression of Wnt5a is correlated with aggressiveness of gastric and prostate cancer by stimulating cell migration and invasion. (2007)
  • Author(s): Kikuchi, A. and Yamamoto, H.
  • Organizer: Traditional Wnt meeting.
  • Place of Presentation: La Jolla. California, USA
  • Year and Date: 2007-06-22