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Hypoxia- and radiation-activated Cre/loxP ‘molecular switch’ vectors for gene therapy of cancer

Gene Therapy volume 13, pages 206–215 (2006)Cite this article

Abstract

Although a significant negative prognostic factor, tumor hypoxia can be exploited for gene therapy. To maximize targeting within the tumor mass, we have developed synthetic gene promoters containing hypoxia-responsive elements (HREs) from the erythropoietin (Epo) gene as well as radiation-responsive CArG elements from the early growth response (Egr) 1 gene. Furthermore, to achieve high and sustained expression of the suicide gene herpes simplex virus thymidine kinase (HSVtk), our gene therapy vectors contain an expression amplification system, or ‘molecular switch’, based on Cre/loxP recombination. In human glioma and breast adenocarcinoma cells exposed to hypoxia and/or radiation, the HRE/CArG promoter rapidly activated Cre recombinase expression leading to selective and sustained HSVtk synthesis. Killing of transfected tumor cells was measured after incubation with the prodrug ganciclovir (GCV; converted by HSVtk into a cytotoxin). In vitro, higher and more selective GCV-mediated toxicity was achieved with the switch vectors, when compared with the same inducible promoters driving HSVtk expression directly. In tumor xenografts implanted in nude mice, the HRE/CArG-switch induced significant growth delay and tumor eradication. In conclusion, hypoxia- and radiation-activated ‘molecular switch’ vectors represent a promising strategy for both targeted and effective gene therapy of solid tumors.

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Acknowledgements

We are very grateful to Drs George Wilson and Michael Borrelli for scientific discussion, and to Dr Yu Wang, Ms Sara Bourne, Mr Evano Piasentin and Ms Jennifer Wright for excellent technical support. This work was supported by The Susan G Komen Breast Cancer Foundation, Wayne State University, Academic Radiation Oncologists and Radiation Oncology Research & Development Center, Detroit, MI, USA.

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Authors and Affiliations

  1. Department of Radiation Oncology, Wayne State University and Karmanos Cancer Institute, Hudson Webber CRC, Detroit, MI, USA

    O Greco, M C Joiner, A Doleh, A D Powell, G G Hillman & S D Scott

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  1. O Greco
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  2. M C Joiner
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  3. A Doleh
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  4. A D Powell
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  5. G G Hillman
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  6. S D Scott
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Correspondence to S D Scott.

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Greco, O., Joiner, M., Doleh, A. et al. Hypoxia- and radiation-activated Cre/loxP ‘molecular switch’ vectors for gene therapy of cancer. Gene Ther 13, 206–215 (2006). https://doi.org/10.1038/sj.gt.3302640

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