• Deletion of pyruvate dehydrogenase kinases 2 and 4 (PDK2/4) reduces susceptibility to deep vein thrombosis by limiting NETosis.

  • PDK2/4 potentiates NETosis by increasing intracellular [Ca2+] concentration, Erk1/2 phosphorylation, and aerobic glycolysis.

Neutrophils contribute to deep vein thrombosis (DVT) by releasing prothrombotic neutrophil extracellular traps (NETs). NETs formation (known as NETosis) is an energy-intensive process that requires an increased rate of aerobic glycolysis. The metabolic enzymes pyruvate dehydrogenase kinases (PDKs) inhibit the pyruvate dehydrogenase (PDH) complex to divert the pyruvate flux from oxidative phosphorylation towards aerobic glycolysis. Herein, we identified that the combined deletion of PDK2 and PDK4 (PDK2/4-/-) renders mice less susceptible to DVT (measured by thrombus incidence, weight, and length) in the inferior vena cava (IVC)-stenosis model at day 2 post-surgery. Compared to wild-type (WT) mice, the venous thrombus obtained from PDK2/4-/- mice exhibited reduced citrullinated histone content, a known marker of NETs. In line with in vivo observations, phorbol 12-myristate 13-acetate (PMA)-stimulated PDK2/4-/- neutrophils displayed reduced NETosis and secretion of cathepsin G & elastase compared to PMA-stimulated WT neutrophils. The formation of platelet aggregates mediated by PMA-stimulated PDK2/4-/- neutrophils were significantly reduced compared to PMA-stimulated WT neutrophils. Finally, PDK2/4-/- neutrophils exhibited reduced levels of intracellular Ca2+ concentration, Erk1/2 phosphorylation, and glycoPER (a measure of aerobic glycolysis), known to facilitate NETosis. Together, these findings elucidate for the first time the fundamental role of PDK2/4 in regulating NETosis and acute DVT.

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