Engineering next-generation CAR-NK cells for hematological malignancies: genome editing, synthetic biology, and...

Chimeric antigen receptor (CAR)-based immunotherapy has transformed cancer treatment by enabling engineered immune cells to selectively recognize and eliminate malignant targets, with CAR-T cells establishing the first major clinical success in this field. However, severe associated toxicities, logistical complexities of autologous manufacturing, and high rates of antigen-loss relapse have accelerated the development of alternative effector platforms such as CAR-NK cells. CAR-NK cells combine antigen-directed cytotoxicity with the innate antitumor functions of natural killer cells, offering a potentially safer and more readily available off-the-shelf therapeutic strategy. Nevertheless, the clinical efficacy of CAR-NK therapy is constrained by key biological and translational hurdles: brief in vivo persistence, inadequate trafficking and solid tumor infiltration, severe susceptibility to tumor microenvironment-mediated immunosuppression, and antigen escape driven by tumor heterogeneity. These biological constraints are compounded by technical bottlenecks, including cell source variability, limited large-scale expansion capacity, ex vivo functional exhaustion, and an absence of standardized manufacturing protocols. To address these multi-faceted barriers, recent advances in genome engineering, systems biology, and computational modeling provide promising routes forward. Given that prolonged exposure to immunosuppressive niches induces functional impairment, genome-editing strategies targeting inhibitory checkpoints and persistence programs are crucial; however, enhancing persistence requires stringent safeguards to mitigate toxicity risks from constitutive signaling or genomic instability. Thus, engineering next-generation CAR-NK cells necessitates balancing extended longevity, potent cytotoxicity, and safety. Multiomics approaches facilitate deeper characterization of CAR-NK functional states and tumor-immune interactions, whereas artificial intelligence-guided engineering integrates high-dimensional biological and manufacturing data to optimize receptor design, predict therapeutic performance, and accelerate translational workflows. Collectively, these innovations support a paradigm shift toward programmable CAR-NK cells that can dynamically sense, adapt, and respond to tumor evolution, driving the transition toward autonomous, highly scalable, and precisely controllable cellular immunotherapies in precision oncology.