Efficacy of dendrimer-mediated angiostatin and TIMP-2 gene delivery on inhibition of tumor growth and angiogenesis: in vitro and in vivo studies.
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.
Vincent
Loïc
L
Laboratoire de Différenciation Hématopoïétique, Faculté de Médecine et Pharmacie, Rouen, France.
Varet
Julia
J
Pille
Jean-Yves
JY
Bompais
Heidi
H
Opolon
Paule
P
Maksimenko
Andrei
A
Malvy
Claude
C
Mirshahi
Manouchehr
M
Lu
He
H
Vannier
Jean-Pierre
JP
Soria
Claudine
C
Li
Hong
H
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
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