ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency

Abstract 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 regulat...

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Main Authors: Xudong Ma, Cheng Chen, Xinyu Chen, Songsong Dan, Jianqiong Li, Xiaobing Zhang, Shiqi She, Jianwen Hu, Yan-Wen Zhou, Bo Kang, Ying-Jie Wang, Wenjie Chen
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-97829-z
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author Xudong Ma
Cheng Chen
Xinyu Chen
Songsong Dan
Jianqiong Li
Xiaobing Zhang
Shiqi She
Jianwen Hu
Yan-Wen Zhou
Bo Kang
Ying-Jie Wang
Wenjie Chen
author_facet Xudong Ma
Cheng Chen
Xinyu Chen
Songsong Dan
Jianqiong Li
Xiaobing Zhang
Shiqi She
Jianwen Hu
Yan-Wen Zhou
Bo Kang
Ying-Jie Wang
Wenjie Chen
author_sort Xudong Ma
collection DOAJ
description Abstract 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.
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institution Kabale University
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spelling doaj-art-673b0052d56d473f93eef0f5d221e6f32025-08-20T03:52:20ZengNature PortfolioScientific Reports2045-23222025-05-0115111610.1038/s41598-025-97829-zATR regulates OCT4 phosphorylation and safeguards human naïve pluripotencyXudong Ma0Cheng Chen1Xinyu Chen2Songsong Dan3Jianqiong Li4Xiaobing Zhang5Shiqi She6Jianwen Hu7Yan-Wen Zhou8Bo Kang9Ying-Jie Wang10Wenjie Chen11Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang UniversityShaoxing People’s Hospital; Shaoxing Hospital, Zhejiang University School of MedicineState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityDepartment of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityShanghai Bioprofile Technology Co., Ltd.State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityState Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang UniversityDepartment of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang UniversityAbstract 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.https://doi.org/10.1038/s41598-025-97829-zOCT4ATRNaïve pluripotencyPhosphoproteomicsPost-translational modificationEmbryonic stem cells
spellingShingle Xudong Ma
Cheng Chen
Xinyu Chen
Songsong Dan
Jianqiong Li
Xiaobing Zhang
Shiqi She
Jianwen Hu
Yan-Wen Zhou
Bo Kang
Ying-Jie Wang
Wenjie Chen
ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency
Scientific Reports
OCT4
ATR
Naïve pluripotency
Phosphoproteomics
Post-translational modification
Embryonic stem cells
title ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency
title_full ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency
title_fullStr ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency
title_full_unstemmed ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency
title_short ATR regulates OCT4 phosphorylation and safeguards human naïve pluripotency
title_sort atr regulates oct4 phosphorylation and safeguards human naive pluripotency
topic OCT4
ATR
Naïve pluripotency
Phosphoproteomics
Post-translational modification
Embryonic stem cells
url https://doi.org/10.1038/s41598-025-97829-z
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