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Enhanced suicide gene effect by adenoviral transduction of a VP22-cytosine deaminase (CD) fusion gene

Gene Therapy volume 8, pages 1654–1664 (2001)Cite this article

Abstract

The low transduction efficiency of viral and nonviral vectors is a major limitation in tumour gene therapy. The HSV-1 tegument protein VP22 has been shown to exhibit a novel intercellular transport property. VP22 wild-type as well as VP22 fusion proteins efficiently spread from the original expressing cell to numerous neighbouring cells, so that protein transport by VP22 chimaeric polypeptides into the surrounding cells offers a possible compensation for the inadequate gene transfer efficiencies. To improve the therapeutic efficacy of the E. coli cytosine deaminase (CD) suicide gene we made use of the VP22 transport property in CD transducing adenoviral (Ad) vectors. C- and N-terminal fusions of CD linked in-frame with VP22 were generated and cloned into recombinant adenoviral vectors. Following in vitro transduction immunofluorescence analysis of Ad-transduced producer cells coplated with naive cells confirmed that the characteristic foci pattern of central producer and adjoining neighbour cells displaying nuclear staining was retained. After transduction of rat hepatoma cells with adenoviral vectors and subsequent incubation with the prodrug 5-FC, we observed enhanced cell cytotoxicity when comparing the CD-VP22 fusion (Ad-CD-VP22) with Ad-vectors expressing the CD gene only (Ad-CD). Thereby employment of Ad-vectors encoding VP22 fusion proteins opens up new possibilities to potentiate the efficiency of suicide gene therapy for the treatment of solid tumours.

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Acknowledgements

We are grateful to Gülüzar Turan, Irina Smirnow and Andrea Schenk for their excellent technical assistance. The plasmids pWZLneoCD and pWZLneoCDglyTK were a kind gift from Kenneth Rogulski, Henry Ford Health System, Detroit, MI, USA. The adenoviral vector Ad-VP22-GFP was a gift from T Harding, Bristol, UK. We thank Transgene SA, Strasbourg, France, for kindly providing us with vectors pTG6600, pTG8347, pTG3602 and the bacteria E. coli BJ5183. This work was supported in part by grants from the Federal Ministry of Education, Science, Research and Technology (Fö. 01KS9602, Fö. 01KV9532), from the Interdisciplinary Clinical Research Center (IZKF) Tübingen, and from the fortüne-program of the Medical Faculty of the Eberhard-Karls-University Tübingen (F.1281127). WAW was supported by a scholarship from the Pinguin Foundation (Henkel KGaA).

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Authors and Affiliations

  1. Internal Medicine I, Medical University Clinic Tübingen, Tübingen, Germany

    WA Wybranietz, CD Groß, M Spiegel, F Graepler, M Bitzer, P Stähler, M Gregor & UM Lauer

  2. Marie Curie Research Institute (MCRI), Oxted, UK

    A Phelan & P O'Hare

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Wybranietz, W., Groß, C., Phelan, A. et al. Enhanced suicide gene effect by adenoviral transduction of a VP22-cytosine deaminase (CD) fusion gene. Gene Ther 8, 1654–1664 (2001). https://doi.org/10.1038/sj.gt.3301564

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