Degradable CRISPR/Cas9 nanosystem activated by NIR-II light targets genome editing of PD-L1 and metabolic modulation for enhanced antitumor immunity

The disruption of the programmed cell death ligand-1 (PD-L1) on tumor cell membranes holds promise to augment T cell-mediated antitumor immunity. However, the permanent prevention of constant PD-L1 expression by Cas9-mediated editing with spatiotemporal specificity and modulation of immunosuppressive tumor microenvironment (TME) remains challenging. Herein, degradable photothermal nanocarriers (RPP) composed of rough polydopamine nanoparticles and cationic polymers are developed to achieve second near-infrared (NIR-II) light activated CRISPR/Cas9 targeting PD-L1 and resveratrol (Res)-mediated inhibition of tumor glycolysis. The co-delivery of CRISPR/Cas9 plasmids driven by a heat-inducible promoter (pHCP) and natural Res can simultaneously realize permanent genome editing of PD-L1 and regulation of glucose metabolism, thereby reprogramming the immunosuppressive TME. Under NIR-II light irradiation, the photothermal effect of RPP not only drives the transcription of single-guide RNA targeting PD-L1 and Cas9, but also triggers the release of Res and induces immunogenic cell death (ICD) of tumor cells, which contribute to the activation of antitumor immune responses. In addition, the intrinsic immunomodulatory effects of the RPP nanocarriers to enhance immune response are exploited. The proposed RPP-mediated therapeutic modality realizes permanent blockade of PD-L1 and remodeling of the immunosuppressive TME. The current work proposes a facile strategy for CRISPR/Cas9-mediated permanent genome editing of PD-L1, ICD induction, and Res regulation of glucose metabolism, which holds great promise in cancer immunotherapy.