Enhancing cancer killing and natural killer cell persistence by targeting NOXA, a predictor of poor patient survival

Natural killer (NK)-cell-based therapeutics have emerged as promising modalities in cancer immunotherapy due to their potent ability to target and kill cancer cells. However, their clinical efficacy is often constrained by the limited <i>in vivo</i> persistence of NK, which hinders sustained therapeutic effects. This study aimed to enhance NK cell survival and functionality by inhibiting apoptosis, thereby boosting the long-term efficacy of NK-cell-mediated therapeutics. Through univariate and multivariate Cox proportional hazards regression analyses, we found that the high expression of Bcl-2-interacting mediator of cell death (<i>BCL2L11</i> [<i>BIM</i>]) and phorbol-12-myristate-13-acetate-induced protein 1 (<i>PMAIP1</i> [<i>NOXA</i>]) in NK cells is associated with poorer survival across various cancer types. Based on potential clinical relevance, we employed CRISPR-Cas9 technology with single-guide RNA to knock out <i>NOXA</i> in NK cells. This modification enhanced the serial cancer-killing efficacy in repeated assays and patient-derived organoid models. In contrast, <i>BIM</i>-knockout (KO) did not confer any additional benefits. Additionally, <i>NOXA</i>-KO NK cells exhibited enhanced post-cryopreservation cytotoxicity, superior metabolic fitness, and extended proliferative capacity. These findings highlight the potential of <i>NOXA</i>-KO NK cells to advance the efficacy of NK-cell-based cancer therapies.