Transcription factor Zhx2 restricts NK cell maturation and suppresses their antitumor immunity

Abstract

The maturation and functional competence of natural killer (NK) cells is a tightly controlled process that relies on transcription factors (TFs). Here, we identify transcriptional repressor zinc fingers and homeoboxes 2 (Zhx2) as a novel regulator that restricts NK cell maturation and function. Mice with Zhx2 conditional deletion in NK cells (Zhx2Δ/Δ) showed accumulation of matured NK cells. Loss of Zhx2 enhanced NK cell survival and NK cell response to IL-15. Transcriptomic analysis revealed Zeb2, a key TF in NK cell terminal maturation, as a direct downstream target of Zhx2. Therapeutically, transfer of Zhx2-deficient NK cells resulted in inhibition of tumor growth and metastasis in different murine models. Our findings collectively unmask a previously unrecognized role of Zhx2 as a novel negative regulator in NK cell maturation and highlight its therapeutic potential as a promising strategy to enhance NK cell–mediated tumor surveillance.

Optimized protocols for chromatin immunoprecipitation of exogenously expressed epitope-tagged proteins

“…In this protocol, we take Zinc Fingers and Homeoboxes 2 (ZHX2) as an example to perform the ChIP assay to illustrate this process. ZHX2 is a transcriptional factor that plays a critical oncogenic function in multiple cancers, 1 , 3 , 4 , 5 , 6 We first combined site-directed mutagenesis, GATEWAY cloning, and lentivirus infection strategies to construct a HA-tagged ZHX2 res (shRNA #45-resistant) expressed cell line, which could stably express exogenous HA-tagged ZHX2 res proteins. We then used the ZHX2 shRNA #45 to deplete its endogenous gene expression to generate a cell line only express exogenous HA-Tagged ZHX2 res protein.…”

Zinc Fingers and Homeobox Family in Cancer: A Double-Edged Sword

“…The expression levels of ZHX2 in nature killer (NK) cells have shown an important clinical significance [ 65 ]. Kaplan–Meier analysis of overall survival from a TCGA dataset showed that a lower ZHX2 expression level in NK cells was associated with better prognosis in HCC patients.…”

“…Therapeutically, the transfer of ZHX2-deficient NK cells inhibited hepatoma homograft tumor growth and metastasis. These studies reveal a novel regulatory function of ZHX2 in NK cell maturation and its therapeutic potential by enhancing NK cell-mediated cancer surveillance [ 65 ].…”

Zhx2 maintains islet β-cell mass and function by transcriptionally regulating Pax6

“… 13 , 19 However, Zhx2 emerges as a tumor oncogene in clear cell renal cell carcinoma and breast cancer. 12 , 14 , 20 In addition, Zhx2 plays an important role in the regulation of immunocytes and inflammatory diseases, such as restricting NK-cell maturation 21 and promoting macrophage glycolysis and survival in sepsis. 22 , 23 These data suggest that Zhx2 regulates a variety of cellular functions, particularly cell proliferation and inflammation.…”