Sourca datasets (numerical values) used to construct all graphs in the publication

Abstract
The safe and effective application of human pluripotent stem cells (hPSCs) in research and regenerative medicine requires precise control over pluripotency and cell fate. Pluripotency is characterized by high levels of histone acetylation and aerobic glycolysis, while differentiation is associated with metabolic shifts and reduced histone acetylation. These transitions are driven, in part, by the availability of metabolic substrates that influence epigenetic regulation. A central enzyme in this process is pyruvate dehydrogenase (PDH), which converts glycolytic pyruvate into acetyl coenzyme A (Ac-CoA), the essential donor for histone acetylation.
Here, we investigate how PDH activity regulates histone acetylation and pluripotency maintenance under physiologically relevant oxygen conditions (5% and 21% O₂), in response to FGF2 signaling and changes in reactive oxygen species (ROS). We show that active PDH promotes global histone H3 acetylation and upregulates the expression of the key pluripotency factor NANOG, specifically under 5% O₂. Mechanistically, we identify a novel FGF2–MEK1/2–ERK1/2–ROS axis that modulates PDH activity via redox-dependent regulation. Notably, this effect is oxygen-sensitive and absent at atmospheric oxygen levels (21% O₂).
Our findings position PDH as a redox-sensitive metabolic switch that connects energy metabolism with epigenetic control of pluripotency by regulating Ac-CoA availability. This work highlights the critical role of oxygen tension, ROS homeostasis, and growth factor signaling in shaping the metabolic–epigenetic landscape of hPSCs, with implications for optimizing stem cell culture and differentiation protocols.