biostudies-literaturePereira MSFNationwide Children's HospitalNIH HHS705https://www.ebi.ac.uk/biostudies/studies/S-EPMC9913837biostudies-literatureUnknown15(3)Loss of cytotoxicity and defective metabolism are linked to glycogen synthase kinase 3 beta (GSK3β) overexpression in natural killer (NK) cells from patients with acute myeloid leukemia or from healthy donors after expansion ex vivo with IL-15. Drug inhibition of GSK3β in these NK cells improves their maturation and cytotoxic activity, but the mechanisms of GSK3β-mediated dysfunction have not been well studied. Here, we show that expansion of NK cells with feeder cells expressing membrane-bound IL-21 maintained normal GSK3β levels, allowing us to study GSK3β function using CRISPR gene editing. We deleted <i>GSK3B</i> and expanded paired-donor knockout and wild-type (WT) NK cells and then assessed transcriptional and functional alterations induced by loss of GSK3β. Surprisingly, our data shCancersDeletion of Glycogen Synthase Kinase 3 Beta Reprograms NK Cell Metabolism.PMC9913837AWD00001042U54-CA232561Maguire CNaeimi Kararoudi MMundy-Bosse BLSorathia KThakkar ALee DACollins PLPereira MSFSezgin YfalseDeletion of Glycogen Synthase Kinase 3 Beta Reprograms NK Cell Metabolism.Loss of cytotoxicity and defective metabolism are linked to glycogen synthase kinase 3 beta (GSK3β) overexpression in natural killer (NK) cells from patients with acute myeloid leukemia or from healthy donors after expansion ex vivo with IL-15. Drug inhibition of GSK3β in these NK cells improves their maturation and cytotoxic activity, but the mechanisms of GSK3β-mediated dysfunction have not been well studied. Here, we show that expansion of NK cells with feeder cells expressing membrane-bound IL-21 maintained normal GSK3β levels, allowing us to study GSK3β function using CRISPR gene editing. We deleted <i>GSK3B</i> and expanded paired-donor knockout and wild-type (WT) NK cells and then assessed transcriptional and functional alterations induced by loss of GSK3β. Surprisingly, our data sh2023-01-01T00:00:00Z2023 Jan2025-04-25T19:25:01.9Z2025-04-06T07:57:57.024ZS-EPMC99138373676566310.3390/cancers15030705