Acrolein-induced PKM2 modification drives NETosis and glioma progression.

Glioblastoma, the most common and aggressive primary brain tumor, is characterized by profound hypoxia and treatment resistance. Hypoxia drives lipid peroxidation, producing acrolein, a reactive aldehyde that induces DNA damage, mitochondrial dysfunction, and oxidative stress. Neutrophils, through a process known as NETosis, release neutrophil extracellular traps (NETs), which have been linked to tumor progression and a poor prognosis. Here, we identify acrolein as a key regulator of NETosis in glioma. Transcriptomic profiling of peripheral neutrophils from glioma patients revealed upregulated NET-associated pathways, consistent with elevated NET formation and reduced survival. NET levels correlated positively with acrolein accumulation in plasma and tumor tissues. Mechanistically, hypoxia-induced acrolein production in glioma cells promoted NETosis in co-cultured neutrophils, thereby enhancing tumor cell proliferation and migration. Acrolein modified PKM2 at Cys326 and Cys358, inducing its nuclear translocation and co-activation of HIF-1α, which in turn upregulated the expression of IL-6 and IL-8. Pharmacologic activation of PKM2 with TEPP-46 blocked acrolein-induced nuclear PKM2 translocation, reduced NET formation in vitro, and suppressed tumor growth and NET levels in vivo. The acrolein scavenger hydralazine also inhibited NETosis in vitro and both subcutaneous and orthotopic glioma models. These findings define an acrolein-PKM2-NET axis in glioblastoma, highlighting NETosis inhibition as a potential therapeutic approach.