Ma Xudong, Chen Cheng, Chen Xinyu, Dan Songsong, Li Jianqiong, Zhang Xiaobing, She Shiqi, Hu Jianwen, Zhou Yan-Wen, Kang Bo, Wang Ying-Jie, Chen Wenjie
Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, 310016, Zhejiang, China.
Shaoxing People's Hospital; Shaoxing Hospital, Zhejiang University School of Medicine, Shaoxing, 312000, Zhejiang, China.
Sci Rep. 2025 May 1;15(1):15274. doi: 10.1038/s41598-025-97829-z.
Under specific conditions, cultured human embryonic stem cells (hESCs) corresponding to primed post-implantation epiblasts can be converted back to a 'naïve pluripotency' state that resembles the pre-implantation epiblasts. The core pluripotency factor OCT4 is known to be crucial in regulating different states of pluripotency, but its potential regulatory role in human naïve pluripotency remains unexplored. In this study, we systematically mapped out phosphorylation sites in OCT4 protein that are differentially phosphorylated between two states of pluripotency, and further identified ATR as a key kinase that phosphorylated OCT4 in naïve but not primed hESCs. The kinase activity levels of ATR in naïve hESCs were higher than those in primed hESCs. Ablating cellular ATR activity significantly halted the induction of naïve hESCs from their primed counterparts, and increased early apoptotic death of naïve hESCs upon UV and CPT treatment. Thus, our work reveals the importance of ATR activity in safeguarding human naïve pluripotency, and implicates a potential association of OCT4 phosphorylation, DNA damage sensing and repairing system in regulating different states of pluripotency during early development.
在特定条件下，与着床后原始态上胚层相对应的培养人胚胎干细胞（hESCs）可以转化回类似于着床前上胚层的“幼稚多能性”状态。核心多能性因子OCT4在调节多能性的不同状态中起着关键作用，但其在人类幼稚多能性中的潜在调节作用仍未被探索。在本研究中，我们系统地绘制了OCT4蛋白在两种多能性状态之间差异磷酸化的磷酸化位点，并进一步确定ATR是在幼稚而非原始态hESCs中磷酸化OCT4的关键激酶。幼稚hESCs中ATR的激酶活性水平高于原始态hESCs。消除细胞ATR活性显著阻止了从原始态hESCs诱导产生幼稚hESCs，并增加了幼稚hESCs在紫外线和喜树碱处理后的早期凋亡死亡。因此，我们的工作揭示了ATR活性在维护人类幼稚多能性中的重要性，并暗示了OCT4磷酸化、DNA损伤感知和修复系统在早期发育过程中调节多能性不同状态的潜在关联。
