Guo Dan, Du Zeling, Liu Youqi, Lin Meiqi, Lu Yue, Hardikar Swanand, Xue Yanna, Zhang Jinghong, Chen Taiping, Dan Jiameng
State Key Laboratory of Primate Biomedical Research, Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming, Yunnan 650500, China.
Yunnan Key Laboratory of Primate Biomedical Research, Kunming, Yunnan 650500, China.
Nucleic Acids Res. 2025 Apr 10;53(7). doi: 10.1093/nar/gkaf302.
Two-cell-like cells (2CLCs), a rare population (∼0.5%) in mouse embryonic stem cell (mESC) cultures, are in a transient totipotent-like state resembling that of 2C-stage embryos, and their discovery and characterization have greatly facilitated the study of early developmental events, such as zygotic genome activation. However, the molecular determinants governing 2C-like reprogramming remain to be elucidated. Here, we show that ZBTB24, CDCA7, and HELLS, components of a molecular pathway that is involved in the pathogenesis of immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome, function as negative regulators of 2C-like reprogramming by maintaining DNA methylation of the Dux cluster, a master inducer of the 2C-like state. Disruption of the ZBTB24-CDCA7-HELLS axis results in Dux hypomethylation and derepression, leading to dramatic upregulation of 2C-specific genes, which can be reversed by site-specific re-methylation in the Dux promoter. We also provide evidence that CDCA7 is enriched at the Dux cluster and recruits the CDCA7-HELLS chromatin remodeling complex to constitutive heterochromatin. Our study uncovers a key role for the ZBTB24-CDCA7-HELLS axis in safeguarding the mESC state by suppressing the 2C-like reprogramming.
双细胞样细胞（2CLCs）是小鼠胚胎干细胞（mESC）培养物中一种罕见的细胞群体（约0.5%），处于类似二细胞期胚胎的短暂全能样状态，它们的发现和表征极大地促进了对早期发育事件的研究，如合子基因组激活。然而，控制2C样重编程的分子决定因素仍有待阐明。在这里，我们表明，ZBTB24、CDCA7和HELLS是参与免疫缺陷、着丝粒不稳定和面部异常（ICF）综合征发病机制的分子途径的组成部分，它们通过维持2C样状态的主要诱导因子Dux簇DNA甲基化，作为2C样重编程的负调节因子。ZBTB24 - CDCA7 - HELLS轴的破坏导致Dux去甲基化和去抑制，从而导致2C特异性基因的显著上调，这可以通过Dux启动子中的位点特异性重新甲基化来逆转。我们还提供证据表明，CDCA7在Dux簇处富集，并将CDCA7 - HELLS染色质重塑复合物招募到组成型异染色质。我们的研究揭示了ZBTB24 - CDCA7 - HELLS轴在通过抑制2C样重编程来维持mESC状态方面的关键作用。
