title: Gene therapy for hepatocellular carcinoma using non-viral vectors composed of bis guanidinium-tren-cholesterol and plasmids encoding the tissue inhibitors of metalloproteinases TIMP-2 and TIMP-3.
abstract: Metalloproteinases (MMPs) and their natural inhibitors (TIMPs) contribute to the regulation of tumor microenvironment. Their expressions are deregulated in almost all human cancers. We report a novel approach to gene therapy of hepatocellular carcinoma (HCC), using repeated injections of DNA plasmids encoding the tissue inhibitors of metalloproteinases (TIMPs) TIMP-2 or TIMP-3, and a novel competent formulation of gene transfer based on nontoxic cationic cholesterol derivatives. The new gene delivery system was efficient in demonstrating the antitumor efficiency of TIMP-2 or TIMP-3 in inhibiting tumor growth of human HuH7 HCC cells xenografted into nude mice. We show, for the first time, an in vivo effect of TIMP-3 in delaying HCC tumor growth. No treatment-related toxicity was noted. An inhibition of angiogenesis and tumor necrosis accompanied the inhibitory effects of TIMP-2 or TIMP-3 on tumor expansion and invasion. We also report a bystander effect produced by transfected HuH7 tumor cells mixed with untransfected cells in 1:1 ratio in culture that resulted in killing 98% of cells within 96 h. In addition, the soluble forms of TIMP-2 and TIMP-3 expressed by transfected cells exerted a cytotoxic effect on untransfected HuH7 cell cultures. Taken together, these results demonstrate the potential efficacy of repeated treatment of secreted TIMP-2 and TIMP-3 for the design of nonviral gene therapy for hepatocarcinoma.
authors.affiliation: 1CNRS-UMR 8532, LBPA, Ecole Normale Supérieure, 94235 Cachan, France.
authors.forename: Phuong-Lan
authors.initials: PL
authors.lastname: Tran
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Vigneron
authors.forename: David
authors.initials: D
authors.lastname: Pericat
authors.forename: Sylvie
authors.initials: S
authors.lastname: Dubois
authors.forename: Dominique
authors.initials: D
authors.lastname: Cazals
authors.forename: Martial
authors.initials: M
authors.lastname: Hervy
authors.forename: Yves A
authors.initials: YA
authors.lastname: DeClerck
authors.forename: Claude
authors.initials: C
authors.lastname: Degott
authors.forename: Christian
authors.initials: C
authors.lastname: Auclair
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D006528:Carcinoma, Hepatocellular / Q000628:therapy*; D002412:Cations; D002460:Cell Line; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006146:Guanidines / Q000737:chemistry*; D006801:Humans; D015151:Immunoblotting; D020287:In Situ Nick-End Labeling; D007700:Kinetics; D008113:Liver Neoplasms / Q000628:therapy*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010957:Plasmids / Q000378:metabolism*; D013997:Time Factors; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics*; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics*; D014162:Transfection
chemical_list: C101388:3-((N',N'-diguanidinoethylaminoethyl)carbamoyl)cholesterol; D018952:Antigens, CD34; D002412:Cations; D006146:Guanidines; D019717:Tissue Inhibitor of Metalloproteinase-3; D019716:Tissue Inhibitor of Metalloproteinase-2; D004247:DNA; D002784:Cholesterol; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9