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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

Cancer Gene Therapy volume 10, pages 435–444 (2003)Cite this article

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.

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Acknowledgements

We thank Drs J-C Ehrhart, M. Buckle, and J-R Bertrand for critical reading of the manuscript, Professor J-M Lehn and Dr P Lehn for discussions, and A Rouchès and P Ardouin (Laboratoire d'Expérimentation Animale) for technical support.

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Authors and Affiliations

  1. CNRS-UMR 8532, LBPA, Ecole Normale Supérieure, Cachan, 94235, France

    Phuong-Lan Tran, Martial Hervy & Christian Auclair

  2. Chimie des Interactions Moléculaires, Collège de France, Paris, 75005, France

    Jean-Pierre Vigneron

  3. Laboratoire d'Expérimentation Animale, Institut Gustave Roussy, Villejuif, 94805, France

    David Pericat

  4. Laboratoire de Pathologie Cellulaire, Hôpital Beaujon, Clichy, 92110, France

    Sylvie Dubois, Dominique Cazals & Claude Degott

  5. Division of Hematology/Oncology, Children's Hospital of Los Angeles, 90027, California, USA

    Yves A DeClerck

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  1. Phuong-Lan Tran
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Correspondence to Phuong-Lan Tran.

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Tran, PL., Vigneron, JP., Pericat, D. et al. 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. Cancer Gene Ther 10, 435–444 (2003). https://doi.org/10.1038/sj.cgt.7700592

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