不同启动子的CD276-CAR-T细胞对CD276阳性实体肿瘤细胞杀伤活性

1. Objective: To develop novel CD276 chimeric antigen receptor-modified T cells (CD276-CAR-T) with different promoters within the CAR construct targeting CD276-positive solid tumor cells, and to enhance the persistence of CAR-T cells by incorporating the secretory cytokine IL-2, while screening for promoters that can enhance CAR-T cell function. 2. Methods: A CMV-CAR lentiviral vector（TAA06-AQ-CAR）was constructed based on the CMV promoter and anti-CD276 single-chain antibody sequence. The CMV promoter sequence was replaced with the EF-1/MND/PGK promoter to create EF-1/MND/PGK-CAR（TAA06-BG/BH/BI-CAR）, and TAA06-AQ/BG/BH/BI-CAR-T cells were prepared. Flow cCytotoxic activity of various promoters of CD276-specific CAR T cells against CD276-positive solid tumor cellsytometry was used to assess the in vitro cytotoxic effects of four CAR-T cells against various CD276-positive solid tumor cells (pancreatic cancer cells SW1990 and PANC-1, glioblastoma cells U87, and neuroblastoma SK-N-AS-luc). The antitumor effects of four CAR-T cell therapies were evaluated in vivo using a pancreatic cancer CDX model. 3. Results: Successfully constructed TAA06-AQ/BG/BH/BI-CAR-T cells. Compared to T cells, the four types of CD276-targeting CAR-T cells significantly promote apoptosis in various naturally high CD276-expressing solid tumor cells. All four CAR-T cells exhibited similar in vitro cytotoxic effects; however, TAA06-BH-CAR-T cells demonstrated stronger in vivo toxicity, while TAA06-BG-CAR-T cells showed a distinct safety advantage in vivo. 4. Conclusion: CD276-CAR-T cells can specifically recognize and kill CD276-positive solid tumor cells. TAA06-BG-CAR-T maintains anti-tumor functionality both in vitro and in vivo, while also demonstrating better safety.