A gene is required for the high activity of glycolysis and OXPHOS in naïve hESCs

A(gene) depletion led to a marked reduction in the abundance of key glycolytic intermediates, including fructose-6-phosphate (F6P), dihydroxyacetone phosphate (DHAP), glyceraldehyde-3-phosphate (GAP), phosphoenolpyruvate (PEP), pyruvate, and lactate, as well as tricarboxylic acid (TCA) cycle metabolites such as succinate and citric acid. Interestingly, aspartate and glutamine levels were elevated upon A(gene) depletion. To further evaluate the impact of A(gene) on energy metabolism in naïve human embryonic stem cells (hESCs), we measured the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), reflecting mitochondrial respiration and glycolytic activity, respectively. A(gene) depletion significantly impaired basal respiration, maximal respiration, and spare respiratory capacity, and also disrupted glycolytic function. Consistent with these metabolic alterations, cellular NADH levels were significantly decreased in A(gene) -depleted cells. Together, these results uncover a critical role of A(gene) as an essential regulator of the bivalent metabolic state in naïve hESCs, balancing both glycolysis and oxidative phosphorylation.