From phthalate exposure to venous thrombotic risk: Decoding the DEHP-ROS mediated OTUD1- p53 stabilization loop in endothelium

• Promotion of thrombosis represents a novel toxicological effect of phthalates. • DEHP-ROS-OTUD1-p53 signaling axis is a novel mechanistic link between environmental plasticizer exposure and disordered coagulation. • An L357 point mutation of OTUD1 rescues the endothelial homeostasis disruption caused by DEHP-induced p53 accumulation. • Targeted intervention of endothelial OTUD1, administration of p53 inhibitors or ROS scavengers alleviated DEHP-induced thrombus formation. Phthalates persist in the environment and accumulate in organisms. While epidemiological studies link phthalate (PAE) exposure to increased coagulation risk in humans, the causal relationship and underlying mechanisms remain elusive due to insufficient experimental validation in model systems. This study elucidated the effect of high-dose exposure of di(2-ethylhexyl) phthalate (DEHP) on endothelial redox and coagulation homeostasis. Based on network toxicology, bioinformatics analysis and in vivo and in vitro studies, we found DEHP can induce abnormal coagulation events both in vivo and in vitro. DEHP promotes oxidative stress and a hypercoagulant phenotype via p53 accumulation, which transcriptionally regulates a set of procoagulant genes in endothelial cells (ECs); ROS scavenging suppresses p53 stabilization, alleviating this phenotype. DEHP exposure induces ROS-triggered upregulation of multiple transcription factors, which promote OTUD1 expression. OTUD1 interacts with p53 via its 300–481 region to catalyze K48-dependent de-ubiquitination and consequently stabilizes the p53 protein. Accumulated p53 further promotes ROS production, establishing a detrimental feedback loop. An L357 point mutation of OTUD1 rescues the endothelial homeostasis disruption caused by DEHP-induced p53 accumulation. Targeted intervention of endothelial OTUD1, administration of p53 inhibitors or ROS scavengers alleviated DEHP-induced thrombus formation. Our study reveals a novel mechanism of ROS-OTUD1 signaling in DEHP-induced disruption of coagulant homeostasis—a newly identified toxicological effect. These findings advance the theoretical framework connecting phthalate exposure and cardiovascular pathology, and discover the potential targets in preventing vascular injury and thrombogenesis caused by PAEs.