mRNA-based precision targeting of neoantigens and tumor-associated antigens in malignant brain tumors

mRNA-based immunotherapeutics have garnered widespread interest for their potential to revolutionize cancer treatment. In this study, we have established a personalized and customizable mRNA-therapeutic approach that effectively targets a plurality of tumor antigens and demonstrated potent anti-tumor response in preclinical brain tumor models. Our platform uniquely addresses the challenge of tumor heterogeneity and low antigen burden, two key deficiencies in targeting classically immunotherapy-resistant CNS malignancies. We created GBM tumor antigen-specific RNA and loaded it into dendritic cells for anti-tumor vaccine. We performed in vivo anti-tumor efficacy study with antigen loaded dendritic cells in combination with adoptive T cell therapy with relevant controls in the KR158 murine GBM model. Post therapy, we isolated the CD45+ve immune cells and CD45 -ve tumor cells from the brain tumor microenvironment and performed scRNA-seq analysis. The samples from each treatment arm were multiplexed together to minimize batch effect. Each treatment arm had 4 replicates for CD45 +ve immune cell subset divided across 3 multiplexed pools and 1 combined replicate for CD45 -ve tumor cells multiplexed into 1 pool.