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