Site-specific neoepitope induction by RNA editing reprograms tumor immunogenicity.

Introduction: Neoantigens derived from somatic mutations exhibit high immunogenicity and specificity. However, their stochastic nature necessitates personalized vaccine design, and cancers with low tumor mutational burden often have insufficient neoantigen availability, limiting their clinical applicability. Here, we introduce RNA Editing-Induced Site-specific Neoantigen (REISN) as a strategy to remodel tumor immunogenicity by targeting predefined epitopes in widely expressed cancer/testis and embryonic antigens, including murine P1A and human AFP. Methods: We leveraged an MS2 coat protein (MCP)-ADAR-based A-to-I RNA editing system to selectively modify T cell receptor (TCR)-contact residues in major histocompatibility complex class I (MHC-I)-presented peptides derived from P1A and AFP. We further evaluated the immunogenicity and antitumor efficacy of lipid nanoparticle-formulated P1A-REISN circular RNA vaccination in immunocompetent BALB/c mice bearing tumors with P1A-REISN induction. Results: Site-directed RNA editing generated REISN-modified epitopes capable of activating REISN-specific naïve T cells. Vaccination with lipid nanoparticle-formulated P1A-REISN circular RNA elicited potent antigen-specific CD8+ T cell responses and suppressed tumor growth in immunocompetent BALB/c mice. Discussion: These findings provide proof-of-concept that site-directed RNA editing can be used to reprogram tumor immunogenicity by generating defined neoantigen-like epitopes from shared tumor-associated antigens. This strategy may expand neoantigen-based immunotherapy beyond tumors with high mutational burden and warrants further investigation for clinical translation.