Nicotinamide adenine dinucleotide induces a bivalent metabolism and maintains pluripotency in human embryonic stem cells

Nicotinamide adenine dinucleotide (NAD(+)) and its precursor metabolites are emerging as important regulators of both cell metabolism and cell state. Interestingly, the role of NAD(+) in human embryonic stem cell (hESC) metabolism and the regulation of pluripotent cell state is unresolved. Here we show that NAD(+) simultaneously increases hESC mitochondrial oxidative metabolism and partially suppresses glycolysis and stimulates amino acid turnover, doubling the consumption of glutamine. Concurrent with this metabolic remodeling, NAD(+) increases hESC pluripotent marker expression and proliferation, inhibits BMP4-induced differentiation and reduces global histone 3 lysine 27 trimethylation, plausibly inducing an intermediate naive-to-primed bivalent metabolism and pluripotent state. Furthermore, maintenance of NAD(+) recycling via malate aspartate shuttle activity is identified as an absolute requirement for hESC self-renewal, responsible for 80% of the oxidative capacity of hESC mitochondria. Our findings implicate NAD(+) in the regulation of cell state, suggesting that the hESC pluripotent state is dependent upon cellular NAD(+).