Genetically stable multi-gene edited iPSCs-derived NK cells for enhanced cancer immunotherapy.

Chimeric antigen receptor (CAR)-T cell treatment is an innovative drug with excellent therapeutic effects against B cell blood cancer. However, multiple side effects and ultra-high treatment costs must be overcome. Off-the-shelf CAR natural killer (NK) cells can be a good alternative to patient-specific CAR-T cells. The purpose of this study was to combine cellular reprogramming, gene editing, and differentiation technologies to produce full-off-the-shelf NK cells and to verify their efficacy and safety. Genetically stable universal and potent CAR (upCAR)-induced pluripotent stem cells (iPSCs) showed biallelic insertions and deletions in the coding sequence and no off-target effects. upCAR-NK cells showed a very high differentiation yield and in vitro proliferation, and freezing/thawing was possible. In addition, upCAR-NK cells secrete interferon-γ when they meet cancer cells, showing cytotoxic effects in vitro and in vivo. upCAR-NK cells show no obvious toxicity in vivo. In conclusion, this study developed genetically stable upCAR-iPSCs and upCAR-NK cell platform technologies that are less likely to have side effects and can be more economically developed for B cell blood cancer than CAR-T cells. In the future, this technology could be useful in developing a full-off-the-shelf CAR-NK cells anti-cancer immune cell therapy with low side effects, high efficacy, and a low price.