GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE310nnn/GSE310988/primaryOK200TranscriptomicsHomo sapiensExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE310988GEOGSEfalseEZH2-Mediated Epigenetic Regulation of NK Cell Expansion, Survival, and Effector Function [RNA-seq]NK cell expansion and preparation is the key step for natural killer (NK) cell-based immunotherapy. During this process, we observed with great precision that all ex vivo expanded NK cells upregulated the expression of the epigenetic regulator EZH2 (enhancer Zeste homolog 2). EZH2 acts canonically as the catalytic core of the Polycomb Repressive Complex 2 (PRC2), mediating H3K27 trimethylation and transcriptional repression. While non-canonically, it also functions as a coactivator to promote gene expression. However, the regulatory role and function of EZH2 in NK cell expansion remained unclear. Here, by establishing a PBNK-derived NK cell expansion system, combining with the mRNA-seq and CUT&RUN-seq, we deeply characterize the dynamics of EZH2 and H3K27me3 binding, along with gene expression changes, between resting and expanded NK cells, and to infer the regulatory mechanisms employed by EZH2. We treated ExNK cells with different type of EZH2 inhibitor to test EZH2 function in ExNK cells. Using the EZH2 degrader MS177 as well as the catalytic inhibitors GSK126 and Tazemetostat, we demonstrated that the non-canonical functions of EZH2 are critical for NK cell activation, expansion, and functional enhancement in vitro, while its canonical methyltransferase activity also plays an indispensable role in maintaining NK cell fitness. Our study is the first to reveal the functional role of EZH2-mediated epigenetic regulation in expanded human NK cells, providing key theoretical basis and core indicators for the clinical preparation of sufficient quantities of functional NK cells.2026/08/31GSE310988GSM9314705GSM9314716GSM9314706GSM9314717GSM9314703GSM9314714GSM9314715GSM9314704GSM9314712GSM9314702GSM9314713GSM9314721GSM9314710GSM9314711GSM9314722GSM9314720GSM9314709GSM9314718GSM9314707GSM9314708GSM93147193428429480310988Homo sapiens