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Retrovirally transduced murine T lymphocytes expressing FasL mediate effective killing of prostate cancer cells

Cancer Gene Therapy volume 16, pages 439–452 (2009)Cite this article

Abstract

Adoptively transferred T cells possess anticancer activities partially mediated by T-cell FasL engagement of Fas tumor targets. However, antigen-induced T-cell activation and clonal expansion, which stimulates FasL activity, is often inefficient in tumors. As a gene therapy approach to overcome this obstacle, we have created oncoretroviral vectors to overexpress FasL or non-cleavable FasL (ncFasL) on murine T cells of a diverse T-cell receptor repertoire. Expression of c-FLIP was also engineered to prevent apoptosis of transduced cells. Retroviral transduction of murine T lymphocytes has historically been problematic, and we describe optimized T-cell transduction protocols involving CD3/CD28 co-stimulation of T cells, transduction on ice using concentrated oncoretrovirus, and culture with IL-15. Genetically modified T cells home to established prostate cancer tumors in vivo. Co-stimulated T cells expressing FasL, ncFasL and ncFasL/c-FLIP each mediated cytotoxicity in vitro against RM-1 and LNCaP prostate cancer cells. To evaluate the compatibility of this approach with current prostate cancer therapies, we exposed RM-1, LNCaP, and TRAMP-C1 cells to radiation, mitoxantrone, or docetaxel. Fas and H-2b expression were upregulated by these methods. We have developed a novel FasL-based immuno-gene therapy for prostate cancer that warrants further investigation given the apparent constitutive and inducible Fas pathway expression in this malignancy.

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Acknowledgements

We thank Anton Neschadim and Dr Richard Miller for helpful discussions about transductions and T-cell biology. This work is supported by a grant from the Prostate Cancer Research Foundation of Canada.

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Authors and Affiliations

  1. Department of Medical Biophysics, University of Toronto, Toronto, Canada

    J C Symes & J A Medin

  2. Division of Stem Cell and Developmental Biology, Ontario Cancer Institute, Toronto, Canada

    C Siatskas & J A Medin

  3. Experimental Transplantation and Immunology Branch, National Cancer Institute, NIH, Bethesda, MD, USA

    D H Fowler

  4. Institute of Medical Sciences, University of Toronto, Toronto, Canada

    J A Medin

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  1. J C Symes
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Symes, J., Siatskas, C., Fowler, D. et al. Retrovirally transduced murine T lymphocytes expressing FasL mediate effective killing of prostate cancer cells. Cancer Gene Ther 16, 439–452 (2009). https://doi.org/10.1038/cgt.2008.96

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