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: Phuong-Lan Tran, Jean-Pierre Vigneron, David Pericat, Sylvie Dubois, Dominique Cazals, Martial Hervy, Yves A DeClerck, Claude Degott, Christian Auclair
• Journal: Cancer gene therapy (Cancer Gene Ther) Vol. 10 Issue 6 Pg. 435-44 (Jun 2003) ISSN: 0929-1903 [Print] England
• PMID: 12768188 (Publication Type: Journal Article)

Chemical References

• 3-((N',N'-diguanidinoethylaminoethyl)carbamoyl)cholesterol
• Antigens, CD34
• Cations
• Guanidines
• Tissue Inhibitor of Metalloproteinase-3
• Tissue Inhibitor of Metalloproteinase-2
• DNA
• Cholesterol
• Matrix Metalloproteinase 2
• Matrix Metalloproteinase 9

Topics

• Animals
• Antigens, CD34 (biosynthesis)
• Carcinoma, Hepatocellular (therapy)
• Cations
• Cell Line
• Cell Line, Tumor
• Cholesterol (analogs & derivatives, chemistry, metabolism)
• DNA (metabolism)
• Female
• Gene Transfer Techniques
• Genetic Therapy (methods)
• Genetic Vectors
• Guanidines (chemistry)
• Humans
• Immunoblotting
• In Situ Nick-End Labeling
• Kinetics
• Liver Neoplasms (therapy)
• Matrix Metalloproteinase 2 (metabolism)
• Matrix Metalloproteinase 9 (metabolism)
• Mice
• Mice, Nude
• Neoplasm Transplantation
• Plasmids (metabolism)
• Time Factors
• Tissue Inhibitor of Metalloproteinase-2 (genetics)
• Tissue Inhibitor of Metalloproteinase-3 (genetics)
• Transfection