Glutathione S‐Transferase Omega 1 Promotes Macrophage Inflammation‐Associated Coagulopathy in Sepsis by S‐Glutathionylating ANXA5 to Enhance Its Ubiquitination

Sepsis-associated coagulopathy contributes to organ dysfunction and mortality. Macrophage-mediated inflammation disrupts anticoagulant homeostasis, but the role of glutathione S-transferase omega 1 (GSTO1) in this process remains unclear. Clinical samples from sepsis patients were analyzed for correlations among GSTO1, ANXA5, and coagulation parameters were evaluated using Spearman rank correlation analysis. In vitro, loss- and gain-of-function approaches were employed in macrophages to examine the regulatory axis among GSTO1, ANXA5, and HUWE1. Protein interactions, post-translational modifications (S-glutathionylation and ubiquitination), and protein stability were assessed using co-immunoprecipitation, Western blotting, and cycloheximide chase assays. Macrophage polarization and tissue factor activity were detected with a commercial assay kit and flow cytometry. Coagulation function was assessed by incubating macrophage-conditioned media with human plasma and measuring APTT, PT, and TT. In vivo, myeloid-specific GSTO1 knockout mice and pharmacological GSTO1 inhibition were utilized to dissect the function of GSTO1 in a cecal ligation and puncture (CLP)-induced septic mouse model. The results indicated that upregulation of GSTO1 was associated with coagulation abnormalities in sepsis, where it promoted macrophage-mediated coagulopathy. GSTO1 reduced ANXA5 protein stability by facilitating its S-glutathionylation at Cys314 in a catalytic activity-dependent manner. Additionally, GSTO1 promoted HUWE1-dependent ubiquitination and subsequent degradation of ANXA5. ANXA5 overexpression rescued the loss of anticoagulant function caused by GSTO1-induced inflammation in macrophages. In vivo, knockout or inhibition of GSTO1 reduced the incidence of coagulopathy in a CLP-induced septic mouse model. GSTO1 facilitated ANXA5 degradation via S-glutathionylation and HUWE1-mediated ubiquitination, promoting macrophage inflammation and anticoagulation impairment, identifying GSTO1 as a candidate for translational exploration in sepsis-associated coagulopathy.