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Adenovirus-mediated hypoxia-targeted gene therapy using HSV thymidine kinase and bacterial nitroreductase prodrug-activating genes in vitro and in vivo

Cancer Gene Therapy volume 18, pages 773–784 (2011)Cite this article

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Abstract

Hypoxia is an important factor in tumor growth. It is associated with resistance to conventional anticancer treatments. Gene therapy targeting hypoxic tumor cells therefore has the potential to enhance the efficacy of treatment of solid tumors. Transfection of a panel of tumor cell lines with plasmid constructs containing hypoxia-responsive promoter elements from the genes, vascular endothelial growth factor (VEGF) and erythropoietin, linked to the minimal cytomegalovirus (mCMV) or minimal interleukin-2 (mIL-2) promoters showed optimum hypoxia-inducible luciferase reporter gene expression with five repeats of VEGF hypoxic-response element linked to the mCMV promoter. Adenoviral vectors using this hypoxia-inducible promoter to drive therapeutic transgenes produced hypoxia-specific cell kill of HT1080 and HCT116 cells in the presence of prodrug with both herpes simplex virus thymidine kinase/ganciclovir and nitroreductase (NTR)/CB1954 prodrug-activating systems. Significant cytotoxic effects were also observed in patient-derived human ovarian cancer cells. The NTR/CB1954 system provided more readily controllable transgene expression and so was used for in vivo experiments of human HCT116 xenografts in nude mice. Subjects treated intratumorally with Ad-VEGFmCMV-NTR and intraperitoneal injection of CB1954 demonstrated a statistically significant reduction in tumor growth. Immunohistochemistry of treated xenografts showed a good correlation between transgene expression and hypoxic areas. Further investigation of these hypoxia-inducible adenoviral vectors, alone or in combination with existing modalities of cancer therapy, may aid in the future development of successful Gene-Directed Enzyme Prodrug Therapy systems, which are much needed for targeting solid tumors.

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Abbreviations

CMV:

cytomegalovirus

EPO:

erythropoietin

GCV:

ganciclovir

HREs:

hypoxic-response elements

HSVtk:

herpes simplex virus thymidine kinase

IL-2:

interleukin-2

NTR:

nitroreductase

VEGF:

vascular endothelial growth factor

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Acknowledgements

This work and all authors were supported by Cancer Research UK. We thank Sarah Perry for all the help and advice with the immunohistochemistry and to Luci MacCormac for the primary human ovarian cultures used in this study.

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Authors and Affiliations

  1. Leeds Institute of Molecular Medicine, University of Leeds, St James's University Hospital, Leeds, UK

    T J Harvey, I M Hennig, N Ingram, G D Hall, P J Selby & J D Chester

  2. Institute of Cancer Therapeutics, University of Bradford, Bradford, UK

    S D Shnyder & P A Cooper

  3. Velindre Cancer Centre, Whitchurch, Cardiff, UK

    J D Chester

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  1. T J Harvey
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Correspondence to J D Chester.

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Harvey, T., Hennig, I., Shnyder, S. et al. Adenovirus-mediated hypoxia-targeted gene therapy using HSV thymidine kinase and bacterial nitroreductase prodrug-activating genes in vitro and in vivo. Cancer Gene Ther 18, 773–784 (2011). https://doi.org/10.1038/cgt.2011.43

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