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Suicide gene therapy of sarcoma cell lines using recombinant adeno-associated virus 2 vectors

Abstract

Soft-tissue sarcomas are mesenchymal tumors that respond poorly to systemic chemotherapy. Suicide gene therapy may be an alternative treatment strategy. Here we show a high susceptibility of human sarcoma cell lines for recombinant adeno-associated virus 2 (rAAV-2) suicide vectors: connective tissue sarcoma (HS-1), fibrosarcoma (HT-1080), Ewing sarcoma (RD-ES), Askin tumor (SK-N-MC), rhabdomyosarcoma (A-204) and soft-tissue sarcoma (WSKL-1). Several vectors containing the thymidine kinase (TK) gene under the control of either the cytomegalovirus promoter or the elongation-factor 1 alpha (EF1α) promoter were cloned and tested. Higher expression levels of the transgene were observed in the sarcoma lines when using the EF1α-suicide gene-containing vectors. A complete eradication of rAAV-2-EF1α-TK/eGFP (TK/enhanced green fluorescent protein fusion gene)-transduced tumor cells was shown following exposure to ganciclovir (2.5 μg/ml) in vitro, while at this dose level >90% of mock-transduced tumor cells survived. Xenotransplantation tumor models (intraperitoneal, subcutaneous) for the human sarcoma cell line HS-1 were established in nonobese diabetic/severe-combined immunodeficient mice. Mice transplanted with rAAV-2-EF1α-TK/eGFP-transduced and ganciclovir-exposed tumor cells survived >5 months while in the nontransduced group all mice had died approximately 1 month after inoculation. These data hold promise for further development of rAAV-2-based suicide gene therapy of sarcomas.

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Acknowledgements

We are grateful to Sigrid Heil, Hans-Jürgen Engel, Bernhard Berkus and the staff of the German Cancer Research Center animal facility for their technical support (German Cancer Research Center, Heidelberg, Germany). This work was supported in part by a grant of the Joachim-Kuhlmann-Stiftung (Essen, Germany). We are grateful to Dr. J.A. Kleinschmidt (German Cancer Research Center, Heidelberg) for providing the plasmid p.DG and for providing the plasmid PHR-HIV-EF1α. Prof. Dr. D. Trono (University of Geneva)

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Correspondence to Stefan Fruehauf.

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Veldwijk, M., Berlinghoff, S., Laufs, S. et al. Suicide gene therapy of sarcoma cell lines using recombinant adeno-associated virus 2 vectors. Cancer Gene Ther 11, 577–584 (2004). https://doi.org/10.1038/sj.cgt.7700718

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