RNA-seq of B7H3_CAR_T cells loaded with oncolytic virus VSVΔ51

Oncolytic viruses (OVs) have been evaluated as an additional method to increase the efficacy of chimeric antigen receptor (CAR)-T cells in treating solid tumors. Here, we designed a CAR moiety by inserting the CR2 and CR3 domains of low-density lipoprotein receptor, enabling specifically and significantly surface-loading of vesicular stomatitis virus (VSV) onto CAR-T cells. We used RNA sequence to study the global gene expression and identified differentially expressed genes in B7H3-CAR-T cells and CR2/3-B7H3-CAR-T cells loaded with loading VSVΔ51, aiming to identify the changed pathways that are influenced by the cross-connection between VSV-G protein and CR2/3-CAR moiety. For CAR-T cell construction, the activated CD3 positive human T lymphocytes were transduced with lentivirus carrying the B7H3-CAR and CR2/3-B7H3-CAR. To preload VSVΔ51, B7H3-CAR-T or CR2/3-B7H3-CAR-T cells were co-incubated with VSVΔ51 (MOI=10) on ice for 1 hour. Then the CAR-T cells were incubated in 37℃ for 3 hours. The samples used in the RNA sequence analysis were in triplicates.