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Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
Humanized and Charge-Optimized CSPG4-Specific CAR-T Cells show Enhanced Efficacy against Head and Neck Squamous Cell
Xiang Xu1,2, Shizhen Qiu3,4, Zhitong Wang1,2
1ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China.
Abstract:
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 (CSPG4Hu) 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, CSPG4Hu 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.
Insights
Researchers developed a new CAR-T cell therapy targeting CSPG4 for head and neck cancer. This approach improves T-cell function and persistence, showing strong antitumor effects in models.
Area of Science:
- Oncology
- Immunotherapy
- Biotechnology
Background:
- Developing CAR-T cell therapy for head and neck squamous cell carcinoma (HNSCC) faces challenges in identifying suitable tumor antigens and CAR designs.
- Chondroitin sulfate proteoglycan 4 (CSPG4) is a promising antigen in HPV-negative HNSCC, linked to tumor proliferation.
Purpose of the Study:
- To validate CSPG4 as a target antigen for HNSCC.
- To engineer enhanced chimeric antigen receptor (CAR)-T cells for improved efficacy and persistence.
- To investigate the impact of CAR design on T-cell metabolism and function.
Main Methods:
- Validated CSPG4 expression and prognostic value in HPV-negative HNSCC.
- Engineered humanized CSPG4 (CSPG4Hu) CAR-T cells by grafting murine scFv 763.74 CDRs onto a human framework.
- Minimized surface positive-charge patches and immunogenic epitopes in the CAR design.
- Utilized transcriptomic and metabolic profiling to assess CAR-T cell state.
- Evaluated CSPG4Hu CAR-T cell persistence and antitumor efficacy in xenograft models.
Main Results:
- CSPG4 was confirmed as a highly expressed, prognostic antigen in HPV-negative HNSCC.
- Engineered CSPG4Hu CAR-T cells exhibited reduced tonic signaling and alleviated exhaustion.
- Biophysical refinement reprogrammed CAR-T cells towards a stem-like state, away from terminal exhaustion.
- CSPG4Hu CAR-T cells demonstrated superior persistence and potent antitumor activity in preclinical models.
Conclusions:
- CSPG4 is a viable target for CAR-T cell therapy in advanced HNSCC.
- A rational engineering framework linking biophysical CAR design to T-cell rejuvenation offers a promising therapeutic strategy.
- CSPG4Hu CAR-T cells represent a potential new treatment for HNSCC.
