Genetically modified extracellular vesicles loaded with activated gasdermin D potentially inhibit prostate-specific membrane antigen-positive prostate carcinoma growth and enhance immunotherapy
Ke Gao, Wenjin Xi, Jianxin Ni, Jun Jiang, Yonghua Lei, Li Li, Jie Chu, Ruixiao Li, Yongpan An, Yanan Ouyang, Rujuan Su, Rui Zhang, Guojun Wu
Abstract
Prostate cancer (PCa) is associated with poor immunogenicity and lymphocytic infiltration, and immunotherapy effective against PCa remains unavailable. Pyroptosis, a novel immunotherapeutic modality for cancer, promotes systemic immune responses leading to immunogenic cell death in solid tumors. This paper describes the preparation and analysis of PSMA scFv -EV N-GSDMD ; this genetically engineered recombinant extracellular vesicle (EV) expresses a single-chain variable antibody fragment (scFv) with high affinity for prostate-specific membrane antigen (PSMA) on their surfaces and is loaded with the N-terminal domain of gasdermin D (GSDMD). Both in vitro and in vivo, PSMA scFv -EV N-GSDMD effectively targeted PSMA-positive PCa cells and induced pyroptosis through the carrier properties of EVs and the specificity of PSMA scFv . In the 22RV1 and PSMA-transfected RM-1-inoculated PCa mouse models, PSMA scFv -EV N-GSDMD efficiently inhibited tumor growth and promoted tumor immune responses. In conclusion, PSMA scFv -EV N-GSDMD can convert the immunosuppressive “cold” tumor microenvironment of PCa into an immunogenic “hot” tumor microenvironment. The multifunctional HEK-293T EV-based targeted delivery system (PSMA scFv -EV N-GSDMD ) loaded N-GSDMD is used to inhibit prostate cancer growth and activate antitumor immunity. • A novel engineered PSMA-targeted extracellular vesicle delivery system "PSMA scFv -EV N-GSDMD " was successfully constructed. • The drug delivery system can exert effective antitumor effects in vitro by delivering the pyroptosis molecule N-GSDMD. • PSMA scFv -EV N-GSDMD improves the antitumor immune microenvironment by promoting pyroptosis in PCa. • PSMA scFv -EV N-GSDMD exerts synergistic antitumor effects of immunotherapy in vivo.