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bioRxiv · 10.1101/2025.10.26.684676

A20 restriction of nitric oxide production restores macrophage bioenergetic balance

Abstract

Macrophages play critical roles in regulating host responses to microbial pathogens and other forms of tissue stress and can acquire pro-inflammatory or tissue reparative phenotypes. A20, or TNFAIP3, is a potent regulator of innate immune cell functions and is extensively linked to human inflammatory and autoimmune diseases. We now find that A20 is a powerful regulator of both glycolytic and mitochondrial respiration in macrophages. Differentiated macrophages that are acutely rendered A20 deficient exhibit increased glycolytic activity and markedly decreased mitochondrial respiration after LPS stimulation. These cells are unable to repolarize from an M1-like to M2-like phenotype. Compromised mitochondrial oxygen consumption in A20 deficient macrophages is caused by increased nitric oxide production. Inhibition or genetic ablation of inducible nitric oxide synthase (iNOS) restores mitochondrial oxidative phosphorylation and lactate production in these cells. These metabolic perturbations occur independently of exaggerated cytokine production and despite robust production of IL-10. Therefore, A20 prevents Warburg-like aerobic glycolysis and restores macrophage homeostasis.

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Ma, A., Chan, M.-P., Advincula, R., Achacoso, P., Grossman, E., Nomura, D., Malynn, B.. 2025-10-27. A20 restriction of nitric oxide production restores macrophage bioenergetic balance. https://doi.org/10.1101/2025.10.26.684676

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