A hierarchical self-adjuvanted nanoCRISPR-based vaccine restores endogenous immune recognition and surveillance to amplify adaptive immune responses.

Tumor vaccines are considered a promising approach in immunotherapy, designed to boost the immune system's capacity to identify tumor-associated antigens and subsequently trigger immune responses against tumors. However, the inherent genetic instability of tumor cells frequently results in decreased expression or loss of antigen and/or major histocompatibility complex (MHC) expression and upregulation of immune checkpoint molecule PD-L1, thus evading endogenous immune recognition and surveillance. Herein, we developed a hierarchical self-adjuvanted nanoCRISPR-based vaccine (HEDERA) loaded with LSD1/PD-L1 dual-editing CRISPR/Cas9 system, seeking to reinstate the endogenous immune detection and monitoring mechanisms to enhance adaptive immune reactions. Knockdown of LSD1 increases the presence of tumor-specific antigens and major histocompatibility complex class I molecules on the surface of cancer cells, thereby restoring immune recognition. Simultaneously, silencing PD-L1 alleviates the "exhaustion" of T cells and reactivates their cytotoxic activity. Moreover, LSD1 knockdown activates the type I interferon pathway to induce a self-adjuvant effect that enhances innate immune responses and thereby strengthens T cell-mediated adaptive immunity. This dual strategy achieves unprecedented efficacy, with 90% primary tumor inhibition, and demonstrates an 87.3% and 90.6% inhibition rate for post-surgical metastatic and recurrent tumors, respectively. Overall, HEDERA overcomes the single-action constraint of traditional tumor vaccines, and avoids combined medication-related poor patient compliance, delivering a more efficient, convenient integrated tumor immunotherapy solution.