Mitochondrial NADH-redox inflexibility constrains genomic and epigenetic stability in pluripotent stem cells
regeneration and proliferation. While many cell types tolerate ETC inhibition when pyruvate or aspartate is supplied, pluripotent stem cells (PSCs) enter a reversible paused state even at abundant pyruvate levels. Here, we show that ETC inhibition triggers severe NADH reductive stress in mouse embryonic stem cells (mESCs), driven mainly by threonine dehydrogenase (TDH). TDH-derived NADH establishes a metabolic environment that disfavors cells with compromised mitochondrial function, maintains inhibition of pyruvate dehydrogenase (PDH), and is associated with increased genomic and epigenetic stability at the cellular population level. ETC inhibition similarly induces pausing in early mouse embryos and in human pluripotent stem cells (hPSCs). In hPSCs, combined inhibition of the one-carbon metabolism enzymes serine hydroxymethyltransferase (SHMT1/2) and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) effectively reduced reductive stress and rescued the paused phenotype. Together, these findings support a model in which limited mitochondrial redox adaptability represents a conserved metabolic feature of pluripotent stem cells and in which NADH reductive stress is associated with genomic and epigenetic stability.
Authors
- Miguel A. Esteban (ORCID: https://orcid.org/0000-0002-1426-6809)
- Yifeng Huang
- Runxia Lin
- Mengran Yin
- Yinghua Huang (ORCID: https://orcid.org/0000-0002-2028-8131)
- Ghulam M. Mushraf
- Xueting Xu
- Baoming Qin (ORCID: https://orcid.org/0000-0001-7652-161X)
- Yan Li (ORCID: https://orcid.org/0000-0001-7485-0954)
- Tanveer Ahmed (ORCID: https://orcid.org/0000-0003-0519-2315)
- Peizhi Li (ORCID: https://orcid.org/0009-0009-4966-2754)
- Bushra Mirza
- Hui Zhang (ORCID: https://orcid.org/0000-0002-6966-6957)
- Zeyu Zhang
- Abdul Sammad
- Yin Gao (ORCID: https://orcid.org/0009-0004-4783-6187)
- Qiang Zhang (ORCID: https://orcid.org/0009-0006-9138-1989)
- Liang Ding
- Lulu Wang (ORCID: https://orcid.org/0009-0006-7438-5486)
- Khair Ullah (ORCID: https://orcid.org/0009-0005-6812-5955)
- Dajiang Qin
- Zhenzhu Sun (ORCID: https://orcid.org/0009-0005-0920-782X)
- Xinyu Wu (ORCID: https://orcid.org/0000-0002-5293-5479)
- Yongzhang Pan (ORCID: https://orcid.org/0009-0000-8925-2228)
- Xiangqian Kong
Institutions
- BGI Group (China) (CN)
- Quaid-i-Azam University (PK)
- Xinjiang Medical University (CN)
- Chulalongkorn University (TH)
- University of the Punjab (PK)
- Fifth Affiliated Hospital of Xinjiang Medical University (CN)
- Guangzhou Institutes of Biomedicine and Health (CN)
- Fahd bin Sultan University (SA)
- Second Affiliated Hospital of Guangzhou Medical University (CN)
- Guangzhou Regenerative Medicine and Health Guangdong Laboratory (CN)
- Guangzhou Medical University (CN)
Publication Details
- Journal
- The EMBO Journal
- Published
- 2026-05-27
- DOI
- https://doi.org/10.1038/s44318-026-00784-2
- Primary Topic
- Pluripotent Stem Cells Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- University of Dundee
- Westlake University
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- East China University of Science and Technology
- Bureau of International Cooperation, Chinese Academy of Sciences
- Shenzhen University
- Southern University of Science and Technology
- National Key Research and Development Program of China
- Science and Technology Planning Project of Guangdong Province
- Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences
- High-end Foreign Experts Recruitment Plan of China
- Basic and Applied Basic Research Foundation of Guangdong Province