[Identification of genes regulating human CAR-T cell proliferation by genome-wide CRISPR screening]

In vivo expansion and long-term maintenance of CAR-T cells are considered to be the hallmark of treatment success after CD19 CAR-T cell therapy. Genome-wide CRISPR screening has emerged as a powerful tool for large-scale gene screens. Genome-wide CRISPR screening revealed that CUL5 gene knockout (KO) improved the proliferation of CD19 CAR-T cells. CUL5KO improved not only the proliferation but also the effector function of CAR-T cells. The JAK-STAT pathway was upregulated in CUL5KO CAR-T cells, and CUL5 was associated with the degradation of JAK3 upon activation through IL-2 signaling. CUL5KO CD19 CAR-T cells efficiently suppressed in vivo tumor progression as compared to control CD19 CAR-T cells.