title: Evidence of antiangiogenic and antimetastatic activities of the recombinant disintegrin domain of metargidin.
abstract: Metargidin, a transmembrane protein of the adamalysin family, and integrins, e.g., alpha5beta1 and alphav, are preferentially expressed on endothelial cells on angiogenesis. Furthermore, metargidin interacts with these integrins via its disintegrin domain. In this study, recombinant human disintegrin domain (RDD) was produced in Escherichia coli by subcloning its cDNA into the pGEX-2T vector, and the effect of purified RDD on different steps of angiogenesis was evaluated. At concentrations of 2-10 micro g/ml, RDD exhibited inhibitory activities in a variety of in vitro functional assays, including endothelial cell proliferation and adhesion on the integrin substrates fibronectin, vitronectin, and fibrinogen. RDD (10 micro g/ml) totally abrogated endothelial cell migration and blocked most capillary formation in a three-dimensional fibrin gel. To test RDD efficacy in vivo, the RDD gene inserted into pBi vector containing a tetracycline-inducible promoter was electrotransferred into nude mouse muscle. RDD was successfully synthesized by muscle cells in vivo as shown by immunolabeling and Western blotting. In addition, 78% less MDA-MB-231 tumor growth, associated with strong inhibition of tumor angiogenesis, was observed in athymic mice bearing electrotransferred RDD. Moreover, in the presence of RDD, 74% fewer B16F10 melanoma lung metastases were found in C57BL/6 mice. Taken together, these results identified this RDD as a potent intrinsic inhibitor of angiogenesis, tumor growth, and metastasis, making it a promising tool for use in anticancer treatment.
authors.affiliation: INSERM U553, Hôpital Saint-Louis, 1 avenue Claude Vellefaux, 75475 Paris Cedex 10, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Trochon-Joseph
authors.forename: Dominique
authors.initials: D
authors.lastname: Martel-Renoir
authors.forename: Lluis M
authors.initials: LM
authors.lastname: Mir
authors.forename: Annick
authors.initials: A
authors.lastname: Thomaïdis
authors.forename: Paule
authors.initials: P
authors.lastname: Opolon
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Connault
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Christophe
authors.initials: C
authors.lastname: Grenet
authors.forename: Françoise
authors.initials: F
authors.lastname: Fauvel-Lafève
authors.forename: Jeannette
authors.initials: J
authors.lastname: Soria
authors.forename: Chantal
authors.initials: C
authors.lastname: Legrand
authors.forename: Claudine
authors.initials: C
authors.lastname: Soria
authors.forename: Michel
authors.initials: M
authors.lastname: Perricaudet
authors.forename: He
authors.initials: H
authors.lastname: Lu
mesh_terms: D051722:ADAM Proteins; D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D019483:Disintegrins / Q000627:therapeutic use*; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects / Q000473:pathology; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D008175:Lung Neoplasms / Q000098:blood supply / Q000517:prevention & control* / Q000556:secondary; D008546:Melanoma, Experimental / Q000098:blood supply / Q000517:prevention & control* / Q000556:secondary; D008565:Membrane Proteins / Q000627:therapeutic use*; D008666:Metalloendopeptidases / Q000627:therapeutic use*; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D018482:Muscle, Skeletal / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D011994:Recombinant Proteins / Q000627:therapeutic use; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D019483:Disintegrins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D051722:ADAM Proteins; C490902:Adam15 protein, mouse; D008666:Metalloendopeptidases