PDHA1 Hyperactivation‐Mediated Glucose Metabolism Reprogramming Fuels Neutrophil NETosis and Promotes Acute Lung Injury

Neutrophil extracellular trap (NET) formation, termed NETosis, is a key driver of lung tissue injury in acute lung injury/acute respiratory distress syndrome (ALI/ARDS). However, the precise mechanisms that trigger neutrophil NETosis remain incompletely understood. Here, we observed that neutrophils from both ALI mice and ARDS patients exhibited NETosis alongside a pronounced selective metabolic shift, characterized by enhanced oxidative phosphorylation (OXPHOS) and attenuated glycolytic activity. Phosphoproteomic profiling revealed that this shift was driven by aberrant activation of pyruvate dehydrogenase E1 subunit alpha (PDHA1), resulting from its dephosphorylation at Ser293. Inhibiting PDHA1 activity, using either the inhibitor 3-fluoropyruvate (3-FP) or genetic knockdown, effectively reversed this selective metabolic reprogramming and suppressed NETosis. We further constructed neutrophil-targeted 3-FP-loaded albumin nanoparticles (3-FP-BSA NPs). These nanoparticles were specifically taken up by activated neutrophils, markedly inhibited NETosis, and alleviated lipopolysaccharide-induced ALI in mice. Mechanistically, RNA-seq revealed the enrichment of the AMPK in neutrophils during NETosis. AMPK phosphorylated PDHA1 at Ser314, which disrupted the PDHA1-pyruvate dehydrogenase kinase 1 (PDK1) interaction and inhibited PDHA1 phosphorylation at Ser293. Ultimately, PDHA1 hyperactivation-driven selective metabolic reprogramming precipitated mitochondrial stress and mtROS accumulation, thereby initiating NETosis. This study defines a targetable metabolic pathway in neutrophils and advances a specific PDHA1-nanotherapeutic strategy against ALI/ARDS.