CELL INTELLIGENCE · 肿瘤细胞治疗研究

Major challenges of developing CAR T cell therapy for head and neck squamous cell carcinoma (HNSCC) include identifying a robust tumor antigen and a suitable CAR design. Here, we validate chondroitin sulfate proteoglycan 4 (CSPG4) as a highly expressed, prognostic antigen in HPV negative HNSCC that drives tumor proliferation. By grafting the murine single-chain variable fragment (scFv) 763.74 complementarity determining regions (CDRs) onto a human antibody framework engineered to minimize surface positive charge patches and immunogenic epitopes, we generated humanized CSPG4 (CSPG4 Hu ) CAR T cells with reduced tonic signaling and alleviated exhaustion. Transcriptomic and metabolic profiling reveal that this biophysical refinement reprograms CAR-T cells away from a glycolytic, terminal exhaustion state toward a PI3K/Akt-driven stem-like state. Consequently, CSPG4 Hu CAR-T cells demonstrate superior persistence and potent antitumor efficacy across systemic xenograft and patient-derived xenograft models. Our study establishes a rational engineering framework that links biophysical CAR design to transcriptomic and metabolic rejuvenation, offering a promising therapeutic candidate for advanced HNSCC.