マトリックスメタロプロテアーゼの新規機能解析

• Project/Area Number: 11240206
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Keio University
• Principal Investigator: 岡田 保典 慶應義塾大学, 医学部, 教授 (00115221)
• Co-Investigator(Kenkyū-buntansha): 池田 栄二 慶應義塾大学, 医学部, 講師 (30232177) ; 藤井 豊 福井医科大学, 助教授 (80211522)
• Project Period (FY): 1999 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥123,300,000 (Direct Cost: ¥123,300,000); Fiscal Year 2003: ¥23,400,000 (Direct Cost: ¥23,400,000); Fiscal Year 2002: ¥27,100,000 (Direct Cost: ¥27,100,000); Fiscal Year 2001: ¥22,100,000 (Direct Cost: ¥22,100,000); Fiscal Year 2000: ¥26,100,000 (Direct Cost: ¥26,100,000); Fiscal Year 1999: ¥24,600,000 (Direct Cost: ¥24,600,000)
• Keywords: MMP / ADAM / 細胞外マトリックス / 分解 / 浸潤・転移 / MMPインヒビター / 心筋梗塞 / 増殖因子 / 血管新生 / CTGF / VEGF / Connective tissue growth factor / Vascular endothelial growth factor / プロテオグリカン

Research Abstract

本年度の主な研究成果は、以下の通りである。(1).MT1-MMPのヒトがん細胞浸潤・転移での役割:ヒト唾液腺癌におけるMMPの解析から、粘液類表皮癌ではMT1-MMPによるproMMP-2活性化がリンパ節転移に関与していた。一方、腺癌や腺様嚢胞癌ではTIMP-2の産生増加によりproMMP-2活性化が抑制されていた。ヒト胃癌においても、硬癌ではTIMP-3によりMT1-MMPを介したproMMP-2の活性化抑制がみられ、非硬癌との違いが明らかとなった。(2).MMP-7の機能解折とヒト子宮内膜癌での役割:酵母two-hybrid system法でproMMP-7結合分子としてCD151をクローニングし、その結合によりproMMP-7が活性化されることを明らかにした。また、子宮内膜癌組織において両分子の癌細胞膜上での共存とMMP-7の活性を検出し、組織・細胞レベルでのproMMP-7/CD151系の働きを証明した。これらのデータは、癌細胞浸潤・転移に関与するMMP-7にも、細胞膜表面での活性化機構が存在することを示している。(3).MMP-2の急性心筋梗塞モデルにおける役割:野生型マウスとMMP-2ノックアウトマウスに急性心筋梗塞を作製し、前者にMMP-2選択的インヒビターを投与した。解析の結果、MMP-2ノックアウトマウスとMMP-2インヒビター投与マウスでは、梗塞巣へのマクロファージ遊走抑制により心破裂が予防され、MMP-2インヒビターによる急性心筋梗塞治療の可能性を示した。(4).ADAM分子の機能解析:フィブロネクチンはADAMTS4と細胞膜上で結合し、ADAMTS4活性を阻害することを明らかにした。潜在型ADAM28はMMP-7により活性化され、ADAM28はinsulin-like growth factor-binding protein 3を切断することを証明した。

Research Products

• [Publications] Shiomi T., Okada Y.: "MT1-MMP and MMP-7 in invasion and metastasis of human cancers."Cancer Metastasis Review. 22. 145-152 (2003)
• [Publications] Wada N., Okada Y.(13番目/14名): "Reduced angiogenesis in peritoneal dissemination of gastric cancer through gelatinase inhibition."Clin.Exp.Metastasis. 20. 431-435 (2003)
• [Publications] Mochizuki S., Shimoda M., Shiomi T., Fujii Y., Okada Y.: "ADAM28 is activated by MMP-7 (matrilysin-1) and cleaves insulin-like growth factor binding protein-3."Biochem.Biophys.Res.Commun.. 315. 79-84 (2004)
• [Publications] Kayano K., Okada Y.(8番目/8名): "The activation of proMMP-2 mediated by MT1-MMP in human salivary gland carcinomas : Possible implication of TIMP-2 in the regulation of proMMP-2 activation."J.Pathol.. (In press). (2004)
• [Publications] Inoki I., et al.: "Connective tissue growth factor binds vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis"FASEB. J16. 219-221 (2002)
• [Publications] Aoki T., et al.: "Two-step sandwich enzyme immunoassay using monoclonal antibodies for detection of souble and membrane-associated human membrane type 1-matrix metalloproteina"J.Immunoassay Immunochom.. 23. 49-68 (2002)
• [Publications] Hashimoto G., et al.: "Matrix metalloproteinases cleave connective tissue growth factor and reactivate angiogenic activity of vascular endothelial growth factor 165"J.Biol.Chem.. 277. 36288-36295 (2003)
• [Publications] Enomoto H., et al.: "VEGF isoforms and their receptors are expressed in human osteoarthritic cartilage"Am.J.Pathol.. 162. 171-181 (2003)
• [Publications] Shiomi T., et al.: "Emphysematous changes are caused by degradation of type III collagen in transgenic mice expressing MMP-1"Exp.Lung Res.. 29. 1-15 (2003)
• [Publications] Pucci-Minafra I., et al.: "Qualitative analysis of circulating and tissue forms of gelatinase A(MMP-2) and B(MMP-9) incolon carcinomas by mono-and two-dimensional gelatin zymography"Matrix Biol.. (in press). (2003)
• [Publications] Noda K., et al.: "Production and activation of matrix metalloproteinase-2 in proliferative retinopathy"Invest.Ophthal.Vis.Sci. (in press). (2003)
• [Publications] Foronjy R.F., et al.: "Progressive adult-onset emphysema in transgenic mice expressing human MMP-1 in the lung"Am.J.Physiol.Lung Cell Mole.Biol.. (in press). (2003)
• [Publications] Nakada M., et al.: "The role of matrix metalloproteinases in glioma invasion"Frontiers Bioscience. 8. E261-E269 (2003)
• [Publications] Shiomi T, Okada Y.: "MT1-MMP and MMP-7 in invasion and metastasis of human cancers"Cancer Metastasis Review. (in press). (2003)
• [Publications] Takino T., et al.: "Cleavage metastasis gene product KiSS-1 protein/Metastin by matrix metalloproteinases"Oncogene. (in press). (2003)
• [Publications] Nakada M.et al.: "Roles of membrane-type 1 matrix metalloproteinase and tissue inhibitor of metalloproteinases 2 in invasion and dissemination of human malignant glioma"J.Neurosurg.. 94. 464-473 (2001)
• [Publications] Michizono K.et al.: "Circulating levels of MMP-1, -2, -3, -9 and TIMP-1 are increased in POEMS syndrome"Neurology. 56. 807-810 (2001)
• [Publications] Hashimoto G.et al.: "Inhibition of ADAMTS4(aggrecanase-1) by tissue inhibitors of metalloproteinases(TIMP-1, 2, 3 and 4)"FEBS Lett. 494. 192-195 (2001)
• [Publications] Lemaitre V.et al.: "ApoE knock-out mice expressing human matrix metalloproteinase-1 in macrophages have less advanced atherosclerosis"J.Clin.Invest. 107. 1227-1234 (2001)
• [Publications] Inoki I.et al.: "Connective tissue growth factor binds vascular endothelial growth factor(VEGF) and inhibits VEGF-induced angiogenesis"FASEB J. 16. 219-221 (2002)
• [Publications] Pucci-Minafra I.et al.: "Qualitative analysis of circulating and tissue forms of gelatinase A(MMP-2) and B(MMP-9) in colon carcinomas by mono- and two-dimensional gelatin zymography"Matrix Biol. (in press). (2002)
• [Publications] Okada Y.: "Kelley's Textbook of Rheumatology."W.B.Saunders Company. 55-72 (2001)
• [Publications] Nakamura H. et al.: "Brevican is degraded by matrix metalloproteinases (MMPs) and aggrecanase-1 (ADAMTS4) at the different sites."J.Biol.Chem. 275. 38885-38890 (2000)
• [Publications] Ikeda M. et al: "Expression of vascular endothelial growth factor isoforms and their teceptors Flt-1, KDR and neuropilin-1 in synovial tissues of rheumatoid arthritis."J Pathol. 191. 426-433 (2000)
• [Publications] Yamanaka H. et al: "Expression and tissue localization of membrane-types 1, 2 and 3 matrix : metalloproteinases in rheumatoid arthritis."Lab.Invest.. 80. 677-687 (2000)
• [Publications] Ishida S. et al: "Co-expression of VEGF receptors VEGF-R2 and neuropilin-1 in proliferative diabetic retinopathy."Invest.Ophthalmol.Vis.Sci.. 41. 1649-1656 (2000)
• [Publications] Takizawa M. et al: "Production of tissue inhibitor of metalloproteinases-3 is selectively enhanced by calcium pentosan polysulfate in human rheumatoid synovial fibroblasts."Arthritis Rheum. 43. 812-820 (2000)
• [Publications] Nabeshima K.: "Front-cell-specific expression of membrane-type 1 matrix metalloproteinase and gelatinase A during cohort migration of colon carcinoma cells induced by hepatocyte growth factor/scatter factor"Cancer Res.. 60. 3364-3369 (2000)
• [Publications] Okada Y: "Kelley's Textbook of Rheumatology,"W.B.Saunders Company.. 55-72 (2001)
• [Publications] Okada Y.: "Methods in Molecular Biology"Human Press Inc.. 359-365 (2001)
• [Publications] Nakada M.: "Expression and tissue localization of membrane-types 1, 2, 3 matrix metalloproteinases in human astrocytic tumors"Am. J. Pathol.. 154. 417-428 (1999)
• [Publications] Nakamura H.: "Enhanced production and activation of progelatinase A mediate by membrane-type 1 matrix metalloproteinase in human papillary thyroid carcinomas"Cancer Res.. 59. 467-473 (1999)
• [Publications] Shimada T.: "Characterization of a truncated recombinant from of human membrane type 3 matrix metalloproteinase"Eur. J. Biochem. 262. 907-914 (1999)
• [Publications] Ikeda M.: "Expression of vacular endothelial growth factor isoforms and their teceptors Flt-1, KDR and neuropilin-1 in synovial tissues of rheumatoid arthritis"J Pathol. (in press). (2000)
• [Publications] Yamanaka H.: "Expression and tissue localization of membrane-types 1, 2 and 3 matrix metalloproteinases in rheumatoid arthritis"Lab. Invest. (in press). (2000)
• [Publications] Ishida S.: "Co-expression of VEGF receptors VEGF-R2 and neuropilin-1 in proliferative diabetic retinopathy"Invest. Ophthalmol. Vis. Sci.. (in press). (2000)