biostudies-literatureHuff ALCancer Research UKMayo Foundation for Medical Education and ResearchEuropean Research CouncilNIAID NIH HHSUniversity of MinnesotaMayo ClinicNCI NIH HHSNational Institutes of HealthNIGMS NIH HHS2540-2552https://www.ebi.ac.uk/biostudies/studies/S-EPMC7705043biostudies-literatureUnknown28(12)Enhancing the immunogenicity of tumor-associated antigens would represent a major advance for anti-tumor vaccination strategies. Here, we investigated structure-directed antigen destabilization as a strategy to improve the degradation, immunogenic epitope presentation, and T cell activation against a vesicular stomatitis virus (VSV)-encoded tumor antigen. We used the crystal structure of the model antigen ovalbumin to identify charge-disrupting amino acid mutations that were predicted to decrease the stability of the protein. One mutation, OVA-C12R, significantly reduced the half-life of the protein and was preferentially degraded in a 26-S proteasomal-dependent manner. The destabilized ovalbumin protein exhibited enhanced presentation of the major histocompatibility complex (MHC) class I Molecular therapy : the journal of the American Society of Gene TherapyVesicular Stomatitis Virus Encoding a Destabilized Tumor Antigen Improves Activation of Anti-tumor T Cell Responses.PMC7705043P50 CA108961R01CA108961T32 AI132165R01CA175386P41 GM103311R01 CA175386P41-GM103311Kottke TEvgin LTonne JWongthida PRamirez-Alvarado MShim KGVile RThompson JDriscoll CBSchuelke MHuff ALMer GfalseVesicular Stomatitis Virus Encoding a Destabilized Tumor Antigen Improves Activation of Anti-tumor T Cell Responses.Enhancing the immunogenicity of tumor-associated antigens would represent a major advance for anti-tumor vaccination strategies. Here, we investigated structure-directed antigen destabilization as a strategy to improve the degradation, immunogenic epitope presentation, and T cell activation against a vesicular stomatitis virus (VSV)-encoded tumor antigen. We used the crystal structure of the model antigen ovalbumin to identify charge-disrupting amino acid mutations that were predicted to decrease the stability of the protein. One mutation, OVA-C12R, significantly reduced the half-life of the protein and was preferentially degraded in a 26-S proteasomal-dependent manner. The destabilized ovalbumin protein exhibited enhanced presentation of the major histocompatibility complex (MHC) class I 2020-01-01T00:00:00Z2020 Dec2026-05-07T20:13:09.23Z2022-02-11T13:29:42.069ZS-EPMC77050433287769510.1016/j.ymthe.2020.08.013