Abstract
Herpes simplex virus type 1-thymidine kinase (HSV1-TK) in combination with ganciclovir is an efficient and widely used strategy in brain tumour gene therapy. Recently, we have shown effective inhibition of glioma growth in a syngeneic rat model using recombinant adenoviruses expressing the full-length HSV1-TK and an N-terminus truncated variant, HSV1-ΔTK in the presence of ganciclovir. We also showed active chronic brain inflammation in the long-term survivors (3 months) treated with HSV1-TK plus GCV. Furthermore, our results indicated loss of myelinated fibres, oedema and indices of ongoing axonal degeneration. In this study, we assessed the cytotoxicity of both HSV1-TK variants in the presence or absence of ganciclovir, in primary cultures of neurones and glia, and in the rat brain in vivo. Our results indicate that, at viral doses where tumour cells are sensitive to the enzyme/prodrug system, (1) there is no major cytotoxicity for either neurones or glial cells grown in primary cultures, (2) on its own the full-length HSV1-TK is more cytotoxic than its truncated version HSV1-ΔTK for a population of non-neuronal and non-glial cells within neocortical primary cultures, and (3) in vivo, when delivered into the striatum, RAds encoding HSV1-TK are more cytotoxic than RAds encoding HSV1-ΔTK, after administration of ganciclovir. The effectiveness of HSV1-ΔTK in preventing brain tumour growth in vivo, combined with its reduced cytotoxicity, both in vivo and in primary cultures of CNS cells, could represent an advantage for treatment of brain tumours using gene therapy.
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Acknowledgements
This work was supported by a project grant (SP2332/0101) from the Cancer Research Campaign (UK) (CRC), the BBSRC (34/T08236) and EU-Biomed programmes (Contract No BMH4-CT96–1436; B104-CT98–0297; BMH4-CT98–3277) to MGC and PRL. We would also like to acknowledge the support which our laboratory receives from Action Research, The Welcome Trust, The MRC, The British Heart Foundation, The Royal Society, The Parkinson's Disease Society, REMEDI, The Sir Halley Stewart Trust, The Lister Institute for Preventive Medicine, and the Faculty of Medicine Bequest Fund. We thank Prof AM Heagerty for his support and encouragement. The skilful secretarial of Mrs Ros Poulton is very gratefully acknowledged. We also wish to thank Roche Products Ltd for their generous gift of Cymevene (GCV) for this study and Michel Janicot from Rhone Poulenc Rorer, France for kindly providing the anti-TK antibodies. TDS is a Research Training Fellow supported by Action Research, AEM was a fellow of CONICET (Argentina), and PRL is a Research Fellow of the Lister Institute of Preventive Medicine.
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Cowsill, C., Southgate, T., Morrissey, G. et al. Central nervous system toxicity of two adenoviral vectors encoding variants of the herpes simplex virus type 1 thymidine kinase: reduced cytotoxicity of a truncated HSV1-TK. Gene Ther 7, 679–685 (2000). https://doi.org/10.1038/sj.gt.3301147
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DOI: https://doi.org/10.1038/sj.gt.3301147
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