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Characterization of HIV-1 TAT peptide as an enhancer of HSV-TK/GCV cancer gene therapy

Cancer Gene Therapy volume 15, pages 303–314 (2008)Cite this article

Abstract

Cancer suicide gene therapy based on herpes simplex virus type I thymidine kinase (HSV-TK) and ganciclovir (GCV) suffers from the lack of efficacy in clinical use, which is mostly due to low gene-transfer efficiency and absence of bystander effect in tumors. We have previously demonstrated the enhancement of GCV cytotoxicity by fusing the HSV-TK with the cell penetrating peptide from HIV-1 transactivator protein transduction domain (TAT PTD). Despite the earlier promising results, we found that the triple fusion protein HIV-1 transactivator protein transduction domain–thymidine kinase suicide gene–green fluorescent protein marker gene (TAT-TK-GFP) increased GCV cytotoxicity only in 3/12 of different human tumor cell lines. Extended GCV exposure enhanced the cytotoxic effect of HSV-TK/GCV gene therapy, but the difference between TK-GFP and TAT-TK-GFP was not statistically significant. The modest improvement on cell killing mediated by TAT PTD in Chinese hamster ovary cells appeared to be associated with cell-surface heparan sulfate proteoglycan (HSPG) composition. However, TAT-mediated increased cell death did not correlate with the density of cell-surface HSPG expression in different tumor cell lines. In conclusion, although some degree of enhancement by TAT was shown in certain tumor cells in vitro, it is unlikely that TAT peptide linked to a suicide protein could be a useful booster of in vivo gene therapy trials.

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Acknowledgements

This study was financially supported by North Savo Cancer Foundation and Kuopio University Foundation grants (to OR). We thank Marika Ruponen for kindly providing us CHO cells as well as CHO mutant pgsD-677 and pgsB-618 cells. We are greatly thankful to Dr Sari Pesonen (Molecular Cancer Biology Program & Haartman Institute, University of Helsinki, Finland) for kindly providing us the anti-HSPG antibody, Dr Antti Ropponen (Department of Clinical Microbiology, University of Kuopio, Finland) for the phycoerythrin-conjugated anti-mouse antibody and Professor Ilkka Julkunen (National Public Health Institute, Finland) for the anti-MxA antibody. We also thank Professors Juhani Jänne and Leena Alhonen (AI Virtanen Institute, University of Kuopio, Finland) for their valuable advice.

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Author notes

  1. S Lehmusvaara

    Present address: 2Present address: Institute of Medical Technology, University of Tampere, FI-33014 Tampere, Finland,

  2. J Wahlfors

    Present address: 3Present address: Academic Development Unit, University of Tampere, FI-33014 Tampere, Finland,

Authors and Affiliations

  1. Department of Biotechnology and Molecular Medicine, AI Virtanen Institute for Molecular Sciences, University of Kuopio, Kuopio, Finland

    O Rautsi, S Lehmusvaara, A Ketola, A-M Määttä, J Wahlfors & R Pellinen

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Correspondence to O Rautsi.

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Rautsi, O., Lehmusvaara, S., Ketola, A. et al. Characterization of HIV-1 TAT peptide as an enhancer of HSV-TK/GCV cancer gene therapy. Cancer Gene Ther 15, 303–314 (2008). https://doi.org/10.1038/cgt.2008.17

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