CD73 disruption with tumor-specific CRISPR/Cas9 nanoparticles reduces adenosine-mediated immunosuppression and amplifies response to chemotherapy.

The therapeutic efficacy of cancer immunotherapy is impaired by poor tumor immunogenicity and the immunosuppressive tumor microenvironment (TME), partially driven by reprogrammed adenosine metabolism in the tumor. Here, we develop degradable CRISPR/Cas9 nanoparticles to durably and precisely downregulate CD73 expression across multiple cell subsets within the tumor microenvironment, including tumor cells, endothelial cells, cancer-associated fibroblasts, and tumor-infiltrating immune cells, thereby effectively disrupting adenosine metabolism. In combination with immunogenic cell death induced by the co-delivered chemotherapeutic drug doxorubicin, this strategy synergistically potentiates tumor immunogenicity and reprograms the TME into an immunostimulatory state. This immune remodeling contributes to tumor control, prolonging median survival in preclinical models of melanoma and triple negative breast cancer in female mice. This study provides a strategy for precise and durable regulation of cancer immunometabolism to improve therapeutic effects of chemo-immunotherapy for synergistic cancer therapy.