Generation of murine tumor cell lines to assess in vivo anti-tumor immune responses to melanoma.

Abstract

Abstract Mouse models of cancer remain the preferred means to assess the efficacy of anti-cancer therapies, and several models have been used to address the effect of cancer drugs or in the discovery of biomarkers. The Yale University Mutant melanoma lines, YUMM1.5 and YUMMER1.7 murine tumor models were developed from the BrafV600E/Pten/−/−Cdkn2a−/− engineered mouse model to more closely reflect poorly immunogenic and highly mutated melanomas, respectively, that are driven by human disease-relevant mutations; however, the dominant T cell-recognized epitopes specific for these tumors remains unknown. We therefore sought to engineer OVA-expressing versions of the YUMM1.5 and YUMMER1.7 tumor lines, hereafter referred to as YUMM1.5-OVA and YUMMER1.7-OVA. Using stable transfection to insert a plasmid expressing full-length secretory ovalbumin, we have generated polyclonal and monoclonal cell lines that generate an in vivo OVA-specific response as validated by MHC class I tetramer staining and cytokine production following OVA peptide stimulation. Together, these two new models for melanoma tumors will allow us to assess intrinsic and extrinsic mechanisms regulating the generation of effective and recovery of exhausted T cell responses to melanoma in vivo.