SLC25A51 is a mitochondrial NAD+ transporter that selectively imports oxidized NAD+ into the mitochondrial matrix and is essential for sustaining cell respiration. The study found that elevated expression of SLC25A51 correlated with poorer outcomes in patients with Acute Myeloid Leukemia (AML). Depletion of SLC25A51 led to increased apoptosis and prolonged survival in orthotopic xenograft models of AML. Metabolic flux analyses indicated that SLC25A51 depletion shunted flux away from mitochondrial oxidative pathways without increasing glycolytic flux. Combining SLC25A51 depletion with 5-Azacytidine treatment further limited the expansion of AML cells in vivo. These findings suggest that SLC25A51 plays a critical role in AML by decoupling the mitochondrial NAD+/NADH ratio to support oxidative reactions and promote proliferation. SLC25A51 represents a potential therapeutic target for refractory AML.SLC25A51 is a mitochondrial NAD+ transporter that selectively imports oxidized NAD+ into the mitochondrial matrix and is essential for sustaining cell respiration. The study found that elevated expression of SLC25A51 correlated with poorer outcomes in patients with Acute Myeloid Leukemia (AML). Depletion of SLC25A51 led to increased apoptosis and prolonged survival in orthotopic xenograft models of AML. Metabolic flux analyses indicated that SLC25A51 depletion shunted flux away from mitochondrial oxidative pathways without increasing glycolytic flux. Combining SLC25A51 depletion with 5-Azacytidine treatment further limited the expansion of AML cells in vivo. These findings suggest that SLC25A51 plays a critical role in AML by decoupling the mitochondrial NAD+/NADH ratio to support oxidative reactions and promote proliferation. SLC25A51 represents a potential therapeutic target for refractory AML.