title: Efficacy of dendrimer-mediated angiostatin and TIMP-2 gene delivery on inhibition of tumor growth and angiogenesis: in vitro and in vivo studies.
abstract: Gene transfer is an attractive approach to fight cancer by targeting cancer cells or their vasculature. Our study reports the inhibition of tumor growth and angiogenesis by a nonviral method using dendrimers associated with 36-mer anionic oligomers (ON36) for delivering angiostatin (Kringle 1-3) and tissue inhibitor of metalloproteinase (TIMP)-2 genes. The optimal concentrations of dendrimers and ON36 for an efficient green fluorescent protein (GFP) plasmid delivery in endothelial cells (HMEC-1) and cancer cells (MDA-MB-435) were first chosen. Then the efficacy of transfection was determined by testing angiostatin and TIMP-2 secretion by Western blot and the biologic effects were evaluated. Angiostatin gene transfer markedly reduced in vitro (i) HMEC-1 but not MDA-MB-435 proliferation; (ii) HMEC-1 and MDA-MB-435 wound healing reparation; and (iii) capillary tube formation. TIMP-2 gene transfer did not affect cell proliferation but strongly inhibited (i) wound healing of HMEC-1 and MDA-MB-435 cells; and (ii) capillary tube formation. Supernatants of transfected-MDA-MB-435 cells also inhibited the formation of angiogenic networks on Matrigel, indicating a paracrine effect. In vivo, intratumoral angiostatin or TIMP-2 gene delivery using dendrimers associated with ON36 effectively inhibited tumor growth by 71% and 84%, respectively. Combined gene transfer resulted in 96% inhibition of tumor growth. Tumor-associated vascularization was also greatly reduced. These findings provide a basis for the further development of nonviral delivery of genes to fight cancer.
authors.affiliation: Laboratoire de Différenciation Hématopoïétique, Faculté de Médecine et Pharmacie, Rouen, France.
authors.forename: Loïc
authors.initials: L
authors.lastname: Vincent
authors.forename: Julia
authors.initials: J
authors.lastname: Varet
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Pille
authors.forename: Heidi
authors.initials: H
authors.lastname: Bompais
authors.forename: Paule
authors.initials: P
authors.lastname: Opolon
authors.forename: Andrei
authors.initials: A
authors.lastname: Maksimenko
authors.forename: Claude
authors.initials: C
authors.lastname: Malvy
authors.forename: Manouchehr
authors.initials: M
authors.lastname: Mirshahi
authors.forename: He
authors.initials: H
authors.lastname: Lu
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Vannier
authors.forename: Claudine
authors.initials: C
authors.lastname: Soria
authors.forename: Hong
authors.initials: H
authors.lastname: Li
mesh_terms: D043168:Angiostatins; D000818:Animals; D015153:Blotting, Western; D002455:Cell Division; D003094:Collagen / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism; D004338:Drug Combinations; D004727:Endothelium / Q000166:cytology; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000628:therapy; D009389:Neovascularization, Pathologic; D010446:Peptide Fragments / Q000235:genetics* / Q000627:therapeutic use; D010957:Plasmids / Q000378:metabolism; D010958:Plasminogen / Q000235:genetics* / Q000627:therapeutic use; D011509:Proteoglycans / Q000494:pharmacology; D013997:Time Factors; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics* / Q000627:therapeutic use; D014162:Transfection; D014407:Tumor Cells, Cultured; D014945:Wound Healing
chemical_list: D016207:Cytokines; D004338:Drug Combinations; D007797:Laminin; D008164:Luminescent Proteins; D010446:Peptide Fragments; D011509:Proteoglycans; C059485:matrigel; D019716:Tissue Inhibitor of Metalloproteinase-2; D049452:Green Fluorescent Proteins; D043168:Angiostatins; D010958:Plasminogen; D003094:Collagen