Induction of Immune-Stimulating Factors and Oncolysis Upon p14ARF Gene Transfer in Melanoma Cell Lines.

Abstract

Together with an anti-tumor immune response, oncolysis using a recombinant viral vector promises to eliminate cancer cells by both gene transfer and host-mediated functions. In this study we explore oncolysis induced by nonreplicating adenoviral vectors used for p14^ARF and interferon-β (hIFNβ) gene transfer in human melanoma cell lines, revealing an unexpected role for p14^ARF in promoting cellular responses predictive of immune stimulation. Oncolysis was confirmed when UACC-62 (p53 wild-type) cells succumbed upon p14^ARF gene transfer in vitro, whereas SK-Mel-29 (p53-mutant) benefitted from its combination with hIFNβ. In the case of UACC-62, in situ gene therapy in nude mice yielded reduced tumor progression in response to the p14^ARF and hIFNβ combination. Potential for immune stimulation was revealed where p14^ARF gene transfer in vitro was sufficient to induce emission of immunogenic cell death factors in UACC-62 and upregulate pro-immune genes, including IRF1, IRF7, IRF9, ISG15, TAP-1, and B2M. In SK-Mel-29, p14^ARF gene transfer induced a subset of these factors. hIFNβ was, as expected, sufficient to induce these immune-stimulating genes in both cell lines. This work is a significant advancement for our melanoma gene therapy strategy because we revealed not only the induction of oncolysis, but also the potential contribution of p14^ARF to immune stimulation.