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Retrovirally transduced murine T lymphocytes expressing FasL mediate effective killing of prostate cancer cells
J C Symes1, C Siatskas, D H Fowler
1Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
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-2(b) 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.
Insights
This study developed a novel gene therapy for prostate cancer using T cells engineered to express FasL. These modified T cells effectively kill cancer cells, offering a promising new treatment strategy.
Area of Science:
- Immunology
- Cancer Biology
- Gene Therapy
Background:
- Adoptively transferred T cells fight cancer via FasL-Fas interactions.
- Tumor microenvironments often hinder T-cell activation and FasL efficacy.
- Existing T-cell therapies face challenges with in vivo activation and expansion.
Purpose of the Study:
- To engineer T cells for enhanced anticancer activity against prostate cancer.
- To overcome limitations in T-cell activation and FasL function within tumors.
- To develop a novel FasL-based immuno-gene therapy for prostate cancer.
Main Methods:
- Created oncoretroviral vectors to overexpress FasL or non-cleavable FasL (ncFasL) on murine T cells.
- Engineered T cells for c-FLIP expression to prevent apoptosis of transduced cells.
- Optimized T-cell transduction protocols using CD3/CD28 co-stimulation, low-temperature transduction, and IL-15.
Main Results:
- Genetically modified T cells demonstrated homing to established prostate tumors in vivo.
- Engineered T cells exhibited potent in vitro cytotoxicity against prostate cancer cell lines (RM-1, LNCaP).
- Standard prostate cancer treatments (radiation, chemotherapy) upregulated Fas and H-2b expression on cancer cells, suggesting synergistic potential.
Conclusions:
- Developed a novel FasL-based immuno-gene therapy for prostate cancer.
- Demonstrated efficacy of engineered T cells in targeting and killing prostate cancer cells.
- Highlighted the potential of this approach due to constitutive and inducible Fas pathway expression in prostate cancer.
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