p40-engineered CAR T-cells targeting CD276 enhance anti-tumor efficacy in non-small cell lung cancer

Background: Chimeric antigen receptor (CAR) T-cells have shown remarkable therapeutic efficacy in hematological malignancies; however, effective breakthroughs in solid tumors have not yet been achieved. Objectives: This study aimed to explore the therapeutic potential of p40-engineered CAR T-cells targeting CD276 (p40-H3BBz) for the treatment of non-small cell lung cancer (NSCLC). Methods: A CAR targeting CD276 (H3BBz) was engineered to co-express the p40 subunit via lentiviral transduction. The function of p40-H3BBz was evaluated by measuring cytokine secretion levels and performing luciferase-based cytotoxicity assays in vitro. In vivo efficacy was assessed using tumor-bearing mouse models, and T-cell infiltration was analyzed by immunohistochemistry. Results: The transduction efficiencies of H3BBz and p40-H3BBz were 63.7% and 52.3%, respectively, with both populations predominantly composed of CD4? T-cells. Upon activation, p40-H3BBz exhibited increased IL-23 secretion, while IL-12 levels were comparable to those of H3BBz in vitro. p40-H3BBz also demonstrated enhanced cytotoxicity, degranulation, and increased production of IL-23 and IFN-?. In vivo, p40-H3BBz CAR T-cells showed superior tumor growth inhibition, increased T-cell infiltration, and elevated IL-23 levels within tumors, without inducing significant toxicity in major organs. Conclusion: p40-H3BBz exhibits enhanced cytokine release and cytotoxic activity, representing a promising and safe therapeutic strategy for the treatment of NSCLC.