CD3e-targeted circRNA-LNP enables One-Step in vivo CAR-T cell therapy with durable solid tumor remission

This study analyzed the transcriptional changes of CAR-T cells after co-culture and cytotoxic activity against gastric cancer cell lines HGC-27 and AGS using RNA sequencing. Compared with control Mock T cells, CAR-T cells co-cultured with HGC-27 showed upregulation of CXCL10, CCL17, IL26, SPHK1, SFN, TRBV7-7, and TRAJ36, while TP53INP1, KLRB1, KLF10, TRIB1, and PIK3CG were downregulated. In the AGS co-culture group, CAR-T cells exhibited upregulation of CCR8, TRAV26-2, TRBV10-1, TRBJ1-6, TRBJ2-6, TRAJ5, and TRAV15, and downregulation of KRT19, CEACAM5, F3, KRT18, and PIK3R2. These findings highlight distinct gene expression signatures of CAR-T cells in different gastric cancer cell contexts. Overall design: In this study, anti-Claudin18.2 CAR-T cells were generated and co-cultured with two gastric cancer cell lines (HGC-27 and AGS). The experimental groups consisted of CAR-T cells after co-culture with tumor cells, while the control groups consisted of Mock T cells co-cultured with tumor cells. Biological replicates were included for each group. After cytotoxic co-culture assays, T cells were collected for RNA sequencing. Transcriptome sequencing was performed using the Illumina platform. D3-C-A = CAR-T cells co-cultured with AGS D3-M-A = Mock T cells co-cultured with AGS D3-C-H = CAR-T cells co-cultured with HGC-27 D3-M-H = Mock T cells co-cultured with HGC-27