Glycolysis dependent lactate formation in neutrophils: A metabolic link between NOX-dependent and independent NETosis

Neutrophil extracellular traps (NETs) play a pivotal role in the innate immune defense, as well as in the pathophysiology of various inflammatory disease conditions. Two major types of NETosis have been described - NOX-dependent and independent. The present study was undertaken to assess metabolic requirements of NETs formation by using PMA and A23187 as the inducers of NOX-dependent and NOX-independent NETosis respectively. Both these inducers caused an increase in ECAR, lactate dehydrogenase (LDH) activity, PKM2 dimerization and reduction in pyruvate kinase M2 (PKM2) activity, promoting lactate formation through Warburg effect. Interestingly exogenous treatment with lactate also induced NETs formation in human neutrophils, while inhibition of LDH activity significantly reduced NETosis by both the pathways. Moreover, NETosis and lactate accumulation during LPS induced sepsis in mice was inhibited by sodium oxamate, LDH inhibitor, demonstrating the importance of lactate in an experimental model of NETosis. Present study thus confirms importance of glycolysis in NETosis and also reveals role of lactate in NETs formation. It also reports sharing of the common metabolic pathway by NOX-dependent and independent NETosis. • Glycolysis induced lactate is required for both NOX-dependent and -independent NETosis. • Lactate induced NETs formation, while inhibition of LDH activity significantly reduced NETosis by both the pathways. • NETotic cells exhibit PKM2 dimerization, while enforced tetramerization/activation reduces NETosis.