Neutrophil extracellular trap-induced ferroptosis promotes abdominal aortic aneurysm formation via SLC25A11-mediated depletion of mitochondrial glutathione.

Neutrophil extracellular traps (NETs) induce oxidative stress, which may initiate ferroptosis, an iron-dependent programmed cell death, during abdominal aortic aneurysm (AAA) formation. Mitochondria regulate the progression of ferroptosis, which is characterized by the depletion of mitochondrial glutathione (mitoGSH) levels. However, the mechanisms are poorly understood. This study examined the role of mitoGSH in regulating NET-induced ferroptosis of smooth muscle cells (SMCs) during AAA formation. Concentrations of NET markers were tested in plasma samples. Western blotting and immunofluorescent staining were performed to detect the expression and localization of NET and ferroptosis markers in tissue samples. The role of NETs and SMC ferroptosis during AAA formation was investigated using peptidyl arginine deiminase 4 gene (Padi4) knockout or treatment with a PAD4 inhibitor, ferroptosis inhibitor or activator in an angiotensin II-induced AAA mouse model. The regulatory effect of SLC25A11, a mitochondrial glutathione transporter, on mitoGSH and NET-induced ferroptosis of SMCs was investigated using in vitro and in vivo experiments. Transmission electron microscopy was used to detect mitochondrial damage. Blue native polyacrylamide gel electrophoresis was used to analyze the dimeric and monomeric forms of the protein. Significantly elevated levels of NETosis and ferroptosis markers in aortic tissue samples were observed during AAA formation. Specifically, NETs promoted AAA formation by inducing ferroptosis of SMCs. Subsequently, SLC25A11 was identified as a potential biomarker for evaluating the clinical prognosis of patients with AAA. Furthermore, NETs decreased the stability and dimerization of SLC25A11, leading to the depletion of mitoGSH. This depletion induced the ferroptosis of SMCs and promoted AAA formation. During AAA formation, NETs regulate the stability of the mitochondrial carrier protein SLC25A11, leading to the depletion of mitoGSH and subsequent activation of NET-induced ferroptosis of SMCs. Preventing mitoGSH depletion and ferroptosis in SMCs is a potential strategy for treating AAA. A schematic diagram of main findings. NETs induce iron-mediated cell death in SMCs during AAA formation, accompanied by mitochondrial dysfunction. Mechanistically, NETs modulate the stability of the mitochondrial carrier protein SLC25A11, leading to depletion of mitoGSH in SMCs. The stability of mitochondrial carrier protein SLC25A11 results in depletion of mitoGSH in SMCs, which in turn induces iron dysregulation in SMCs and promotes AAA formation. These findings propose several potential targets for the development of novel therapeutics for AAA treatment, including inhibition of NETs formation, mitigation of mitoGSH depletion, and suppression of SMC ferroptosis. [Display omitted] • During AAA formation, NETs regulate the stability of the mitochondrial carrier protein SLC25A11. • SLC25A11 regulated the depletion of mitochondrial glutathione and activation of NET-induced ferroptosis of SMCs. • Preventing mitoGSH depletion and ferroptosis in SMCs is a potential strategy for treating AAA. [ABSTRACT FROM AUTHOR]