増殖因子および受容体の蛋白質プロセッシングによる制御システムの解析

• Project Area: Regulation of signal transduction by post-translational modifications and its pathogenic dysregulation
• Project/Area Number: 23117507
• 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: The University of Tokyo
• Principal Investigator: 越川 直彦 東京大学, 医科学研究所, 准教授 (70334282)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2012: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2011: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
• Keywords: EphA / 膜型MMP / プロセシング / がん / シグナル伝達 / HB-EGF / 受容体型チロシンキナーゼ

Research Abstract

申請者はプロテオミクス手法を駆使し、MT1-MMPが細胞膜上で様々な膜蛋白質と複合体を形成し、それらが協働して多様な細胞機能を制御していることを示した。その過程で、受容体型チロシンキナーゼ（EphA2）が膜型MMP（MT1-MMP）と細胞膜上で複合体を形成していること、また、MT1-MMPがEphA2のN末端のフィブロネクチンドメインを切断し、そのリガンド結合ドメインを膜上から遊離させ、EphA2のリガンド結合能を消失させることを見出した。以上から、MT1-MMPによるプロセシングを受けたEphA2断片はEphrin-A1リガンドの存在下においても、リガンド依存的な癌抑制活性を持たないことを新たに見出した。
さらに、これまでにEphA2は種々の悪性癌組織で発現が過剰に亢進していることが報告されているが、種々の癌組織を用いたウエスタンブロット、免疫蛍光組織染色による解析から、EphA2はN末端が切り取られた断片として発現しまた、MT1-MMPと共局在していた。以上の結果は、癌組織中のEphA2はMT1-MMPによるプロセシングによりリガンドに非感受性として、癌化促進に強く寄与すると共に、また、N末端を欠損したEphA2断片が新しい癌治療法を開発するための分子標的になりうる可能性を強く示唆している。

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] Identification of proteins that associate with integrin α2 by proteomic analysis in human fibrosarcoma HT-1080 cells. (2012)
  • Author(s): Uematsu T, Konishi C, Hoshino D, HanX, Tomari T, Egawa N, Takada Y, IsobeT, Seiki M, Koshikawa N.
  • Journal Title: J Cell Physiol. Volume: 227(8) Issue: 8 Pages: 3072-9
  • DOI: 10.1002/jcp.23054
  • Peer Reviewed
• [Journal Article] Establishment and validation of computational model for MT1-MMP dependent ECM degradation and intervention strategies (2012)
  • Author(s): Hoshino, D., Koshikawa, N., Suzuki, T., Quaranta, V., Weaver, A.M., (Seiki, M.), Ichikawa, K.
  • Journal Title: PLoS Comp.Biol. Volume: 8 Issue: 4 Pages: e1002479-e1002479
  • DOI: 10.1371/journal.pcbi.1002479
  • Peer Reviewed
• [Journal Article] Control and inhibition analysis of complex formation processes. (2012)
  • Author(s): Saitou T, Itano K, Hoshino D, Koshikawa N, Seiki M, Ichikawa K, Suzuki T.
  • Journal Title: Theor Biol Med Model. Volume: 3 Issue: 1 Pages: 33-33
  • DOI: 10.1186/1742-4682-9-33
  • Peer Reviewed
• [Journal Article] Detection of the heterogeneous O-glycosylation profile of MT1-MMPexpressed in cancer cells by a simple MALDI-MS method. (2012)
  • Author(s): Shuo T, Koshikawa N, Hoshino D, Minegishi T, Ao-Kondo H, Oyama M, Sekiya S, Iwamoto S, Tanaka K, Seiki M.
  • Journal Title: PLoS One. Volume: 7(8) Issue: 8 Pages: e43751-e43751
  • DOI: 10.1371/journal.pone.0043751
  • Peer Reviewed
• [Journal Article] The phosphoinositide-binding protein ZF21 regulates ECM degradation by invadopodia. (2012)
  • Author(s): Hoshino D, Nagano M, Saitoh A, Koshikawa N, Suzuki T, Seiki M.
  • Journal Title: PLoS One. Volume: 8(1) Issue: 1 Pages: 1-8
  • DOI: 10.1371/journal.pone.0050825
  • Peer Reviewed
• [Journal Article] Turnover of focal adhesions and cancer cell migration. (2012)
  • Author(s): Nagano M, Hoshino D, Koshikawa N, Akizawa T, Seiki M.
  • Journal Title: Int J Cell Biol. Volume: 2012 Pages: 310616-310616
  • DOI: 10.1155/2012/310616
  • Peer Reviewed
• [Journal Article] Mathematical modeling of invadopodia formation (2012)
  • Author(s): T.Saitou, M.Rouzimaimaiti, N.Koshikawa, M.Seiki, K.Ichikawa, T.Suzuki
  • Journal Title: J.Theoretical Biology Volume: 298 Pages: 138-146
  • DOI: 10.1016/j.jtbi.2011.12.018
  • Peer Reviewed
• [Journal Article] Identification of proteins that associate with integrin α2 by proteomic analysis in human fibrosarcoma HT-1080 cells (2012)
  • Author(s): Uematsu T., et al
  • Journal Title: J Cell Physiol Volume: (In press)
  • Peer Reviewed
• [Journal Article] Expression of laminin 5-Y2 chain in cutaneous squamous cell carcinoma and its role in tumour invasion. (2011)
  • Author(s): Hamasaki H, Koga K, Aoki M, Hamasaki M, Koshikawa N, Seiki M, Iwasaki H, Nakayama J, Nabeshima K
  • Journal Title: Br J Cancer. Volume: 6;105(6) Issue: 6 Pages: 824-32
  • DOI: 10.1038/bjc.2011.283
  • Peer Reviewed
• [Journal Article] ZF21 protein, a regulator of the disassembly of focal adhesions and cancer metastasis, contains a novel noncanonical pleckstrin homology domain (2011)
  • Author(s): Nagano M, Hoshino D, Koshiba S, Shuo T Koshikawa N, Tomizawa T, Hayashi F, Tochio N, Harada T, Akizawa T Watanabe S, Handa N, Shirouzu M, Kigawa T, Yokoyama S, Seiki M
  • Journal Title: J Biol Chem. Volume: 286(36) Issue: 36 Pages: 31598-609
  • DOI: 10.1074/jbc.m110.199430
  • Peer Reviewed
• [Journal Article] Membrane-type 4 matrix metalloproteinase (MT4-MMP)modulates water homeostasis in mice. (2011)
  • Author(s): Srichai MB, Colleta H, Gewin L, Matrisian L, Abel TW, Koshikawa N, Seiki M, Pozzi A, Harris RC, Zent R.
  • Journal Title: PLoS One. Volume: 6(2) Issue: 2 Pages: e17099-e17099
  • DOI: 10.1371/journal.pone.0017099
  • Peer Reviewed
• [Journal Article] Cholestatic liver fibrosis and toxin-induced fibrosis are exacerbated in matrix metalloproteinase-2 deficient mice (2011)
  • Author(s): Onozuka I, Sakamoto N, Nakagawa M, Watanabe M, et al.
  • Journal Title: Biochem Biophys Res Commun Volume: 406 Issue: 1 Pages: 134-140
  • DOI: 10.1016/j.bbrc.2011.02.012
  • Peer Reviewed
• [Presentation] Proteolytic activation of HB-EGF by MT1-MMP in ovarian carcinoma cells (2011)
  • Author(s): 越川直彦, 他
  • Organizer: 第70回日本癌学会総会
  • Place of Presentation: 名古屋(招待講演)
  • Year and Date: 2011-10-07
• [Presentation] がん細胞膜上の微小環境-悪性化制御因子としてのMT1-MMP- (2011)
  • Author(s): 越川直彦, 他
  • Organizer: 第20回日本がん転移学会学術総会
  • Place of Presentation: 浜松(招待講演)
  • Year and Date: 2011-06-30
• [Presentation] MT1-MMP is a potent regulator of malignant progression in tumor micro environment (2011)
  • Author(s): Koshikawa, N., Seiki, M.
  • Organizer: Mathematical Methods in Cancer Cell Biology
  • Place of Presentation: Hiroshima, Japan(招待講演)
  • Year and Date: 2011-06-08
• [Presentation] 卵巣がんにおけるMT1-MMPによるHB-EGFの活性化についての研究
  • Author(s): 越川直彦、清木元治
  • Organizer: 第21回 日本がん転移学会学術集会・総会
  • Place of Presentation: 広島
• [Presentation] Membrane type-1 matrix metalloproteinase (MT1-MMP) is a potent regulator of cancer malignant progression through the heparin-binding EGF-like growth factor activation
  • Author(s): Koshikawa, N & Seiki, M
  • Organizer: AACR special meeting, Tumor Invasion and Metastasis
  • Place of Presentation: San Diego, CA USA
• [Presentation] Membrane type-1 matrix metalloproteinase promotes invasive growth signals of cancer cells by cell surface proteolysis
  • Author(s): Koshikawa, N., Seiki, M.
  • Organizer: 第72回日本癌学会総会
  • Place of Presentation: 横浜
  • Invited
• [Remarks]
  • URL: http://www.motoharu-seiki.com/esults/
• [Patent(Industrial Property Rights)] がんの検査方法及び検査用キット (2014)
  • Inventor(s): 越川直彦、清木元治
  • Industrial Property Rights Holder: 越川直彦、清木元治
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-026026
  • Filing Date: 2014-02-13
  • Overseas