Macrophage-derived exosome miR-146a-5p modulates PNKP/DDOST/JAGN1 complex to regulate NETs formation in...

The progression of atherosclerosis (AS) is closely associated with neutrophil extracellular trap (NET) formation, yet the regulatory mechanisms between macrophages and neutrophils remain unclear.

In this study, we established an AS model using ApoE^-/- mice and isolated exosomes from macrophages stimulated with oxidized low-density lipoprotein (ox-LDL). Multiple techniques, including qRT-PCR, Western blot, co-immunoprecipitation, RIP, and dual-luciferase reporter assays, were employed to analyze the regulatory role of miR-146a-5p on the PNKP/DDOST/JAGN1 complex.

Exosomes derived from ox-LDL-stimulated macrophages exhibited elevated miR-146a-5p expression, which targeted and suppressed polynucleotide kinase 3'-phosphatase (PNKP) expression, thereby reducing PNKP-dolichyl-diphosphooligosaccharide-protein glycosyltransferase non-catalytic subunit (DDOST) interaction and enhancing DDOST phosphorylation. This process activated junctional adhesion molecule-like protein (JAGN1)-dependent NET formation. Inhibition of miR-146a-5p significantly decreased neutrophil reactive oxygen species (ROS) levels, reduced NETs generation, and markedly improved plaque burden and attenuated inflammatory responses in AS mice.

Our study reveals a novel mechanism by which macrophage-derived exosome miR-146a-5p regulates NETs formation through the PNKP/DDOST/JAGN1 axis, providing potential therapeutic strategies for targeted AS treatment.