iPSCs-derived NK cells with site-specific integration of CAR19 and IL24 at the multi-copy rDNA locus enhanced antitumor activity and proliferation

The generation of CAR-NK cells using induced pluripotent stem cells (iPSCs) has emerged as one of the paradigms for manufacturing off-the-shelf universal immunotherapy.iPSCs here were site-specifically integrated at the ribosomal DNA (rDNA) locus with interleukin 24 (IL24) and CD19-specific chimeric antigen receptor (CAR19), and successfully differentiated into iPSCs-derived NK (iNK) cells followed by expansion using magnetic beads in vitro. To explore the anti-tumor phenotype of CAR-loaded iNK cells and the likely role of IL24, RNA from four iNK cells was isolated, sequenced, and gene expression of the cell populations were compared. To explore the molecular mechanism underlying the anti-tumor phenotype of CAR-loaded iNK cells and the potential impact of IL24 on CAR-loaded iNK cells, iPSCs were site-specifically integrated at the ribosomal DNA (rDNA) locus with interleukin 24 (IL24) and CD19-specific chimeric antigen receptor (CAR19), and successfully differentiated into iPSCs-derived NK (iNK) cells followed by expansion using magnetic beads in vitro. We then performed gene expression profile analysis using data obtained from RNA-seq of four iNK cells populations. Comparative gene expression profile analysis using data obtained from RNA-seq of the four iNK cells (CAR19-IL24-iNK, CAR19-iNK cells, LV-CAR19-iNK cells and iNK cells).