title: Antitumoral and antimetastatic effect of antiangiogenic plasmids in B16 melanoma: Higher efficiency of the recombinant disintegrin domain of ADAM 15.
abstract: BACKGROUND
  Despite the discovery of novel inhibitors of tumor angiogenesis, protein-based antiangiogenic cancer therapy suffers some limitations that antiangiogenic gene therapy could overcome. We investigated whether intra-tumoral electrotransfer of three angiogenic plasmids could inhibit tumor growth and metastasis.
  
  
  METHODS
  Plasmids encoding recombinant disintegrin domain of ADAM-15 (RDD), thrombospondin 1 (TSP-1), and the soluble isoform of the VEGF receptor 1 (sFlt-1) were injected into B16F10 melanoma-bearing C57BL/6 mice followed by electroporation. Tumor volume was measured daily using a digital caliper. Metastasis was monitored by in vivo bioluminescence after surgical removal of the primary luciferase-encoding B16F10 tumor 5 days after intra-tumoral electrotransfer. Markers of vascularization and cell proliferation were quantified by immunohistochemistry.
  
  
  RESULTS
  Intra-tumoral electrotransfer of the antiangiogenic plasmids induced a significant inhibition of tumor growth, doubling of mean survival time and long-term survivors (∼40% vs 0% in control). When the tumor was removed by surgery after intra-tumoral plasmid electrotransfer, a significant decrease in tumor metastasis was observed leading to long-term tumor-free survival especially after treatment with pRDD plasmid (84% vs 0% in control). Unlike pTSP-1 and psFlt-1, pRDD significantly decreased cell proliferation in B16F10 primary tumors which express αvβ3 and α5β1 integrins. No effect of antiangiogenic plasmid electrotransfer on normal skin blood flow was detected.
  
  
  CONCLUSIONS
  The intra-tumoral electrotransfer of the three antiangiogenic plasmids is a promising method for the treatment of melanoma. The plasmid encoding RDD seems to be particularly effective due to its direct antitumoral activity combined with angiogenesis suppression, and its marked inhibition of metastasis.
authors.affiliation: Université Catholique de Louvain, Louvain Drug Research Institute, Unité de Pharmacie galénique, Brussels, Belgium.
authors.forename: Liévin
authors.initials: L
authors.lastname: Daugimont
authors.forename: Gaelle
authors.initials: G
authors.lastname: Vandermeulen
authors.forename: Florence
authors.initials: F
authors.lastname: Defresne
authors.forename: Caroline
authors.initials: C
authors.lastname: Bouzin
authors.forename: Lluis M
authors.initials: LM
authors.lastname: Mir
authors.forename: Céline
authors.initials: C
authors.lastname: Bouquet
authors.forename: Olivier
authors.initials: O
authors.lastname: Feron
authors.forename: Véronique
authors.initials: V
authors.lastname: Préat
mesh_terms: D000818:Animals; D008297:Male; D051379:Mice; D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism; D020533:Angiogenesis Inhibitors / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use; D019483:Disintegrins / Q000235:genetics; D018274:Electroporation; D015316:Genetic Therapy / Q000379:methods; D008545:Melanoma / Q000235:genetics / Q000556:secondary / Q000628:therapy; D008546:Melanoma, Experimental / Q000235:genetics / Q000628:therapy; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000628:therapy; D010957:Plasmids / Q000627:therapeutic use; D040262:Receptors, Vascular Endothelial Growth Factor / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D019700:Thrombospondin 1 / Q000235:genetics / Q000627:therapeutic use; D014162:Transfection
chemical_list: D051722:ADAM Proteins; C098660:ADAM15 protein, human; D020533:Angiogenesis Inhibitors; D019483:Disintegrins; D008565:Membrane Proteins; D040262:Receptors, Vascular Endothelial Growth Factor; D019700:Thrombospondin 1