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Transcellular transfer of active HSV-1 thymidine kinase mediated by an 11-amino-acid peptide from HIV-1 Tat

Cancer Gene Therapy volume 10, pages 64–74 (2003)Cite this article

Abstract

Suicide gene therapy using herpes simplex virus type-1 (HSV-1) thymidine kinase (TK) is a widely exploited approach for gene therapy of cancer and other hyperproliferative disorders. Despite its popularity, clinical success has been so far hampered mostly by the relative inefficiency of TK gene transfer and its limited bystander effect. Here we report that fusion of TK to an 11-amino-acid peptide from the basic domain of the HIV-1 Tat protein (Tat11) imparts cell membrane translocating ability to the enzyme and significantly increases its cytotoxic efficacy. In cells expressing Tat11-TK, this protein is found extracellularly, associated with cell surface heparan sulfate proteoglycans, and is released into the cell culture medium. Based on its interaction with HSPGs, the protein is then internalized by neighboring, nonexpressing cells, which become susceptible to cell death when treated with the nucleoside analogue acyclovir. As a consequence, co-cultures of wild-type cells with cells expressing Tat11-TK show increased sensitivity to ACV through a mechanism involving apoptosis. Modification of TK by fusion with Tat11 might constitute an important step for the optimization of TK suicide gene strategy for gene therapy of cellular proliferation.

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Acknowledgements

This work was supported by Grants from the National Research Program on AIDS of the Istituto Superiore di Sanità (Rome, Italy) and from the Ministero dell'Istruzione, Universitá e Ricerca. We are grateful to M Janicot for the anti–TK antibody, and to B Boziglav and ME Lopez for excellent technical assistance.

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Authors and Affiliations

  1. Molecular Medicine Laboratory, International Center for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy

    Ennio Tasciotti, Monica Zoppè & Mauro Giacca

  2. Laboratorio di Biologia Molecolare and NEST/INFM, Scuola Normale Superiore, Pisa, Italy

    Ennio Tasciotti & Mauro Giacca

  3. Laboratorio di Terapia Genica e Molecolare, Istituto di Fisiologia Clinica, CNR, Pisa, Italy

    Monica Zoppè

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  1. Ennio Tasciotti
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Correspondence to Monica Zoppè.

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Tasciotti, E., Zoppè, M. & Giacca, M. Transcellular transfer of active HSV-1 thymidine kinase mediated by an 11-amino-acid peptide from HIV-1 Tat. Cancer Gene Ther 10, 64–74 (2003). https://doi.org/10.1038/sj.cgt.7700526

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