Quantitative Cell Type Specific Immunopeptidome Analysis of Macrophage and Tumor Co-evolution Reveals Therapeutic MHC-I Peptides in Glioblastoma

Abstract Immune checkpoint inhibitors (ICI) have shown impressive performance in treating several types of solid tumors. However, they have been ineffective in glioblastoma (GBM), in part due to the immunosuppressive tumor microenvironment (TME) created by GBM-associated macrophages (GAMs). To uncover MHC-I peptide antigens for targeted immunotherapy, we performed cell type specific immunopeptidome analysis on primary macrophages and GBM tumor cells in a co-culture system to profile MHC-I associated antigen presentation at the tumor-macrophage interface. Co-culturing tumor cells and macrophages induced increased presentation of peptides derived from proteins associated with cytokine signaling pathways on macrophages and from proteins associated with the Rho GTPase pathway on GBM tumor cells. In vivo expression was validated for a cohort of co-culture-induced GAM or GBM associated peptides selected as potential immunotherapy targets, and an mRNA vaccine was developed encoding six peptides from GAMs and GBM tumor cells. Two doses of vaccination generated an antigen specific immune response, significantly delayed GBM tumor growth, and in some cases eradicated tumors. These results demonstrate the translational potential of co-culture induced MHC peptide antigens as therapeutic targets for GBM/GAM targeting vaccines.