Abstract Body: Alterations in the composition of extracellular vesicles (EV) offer promising insights into identifying critical molecules as potential biomarkers for early diagnostic or novel therapeutic targets in the treatment of hypertension. In this study, we investigated the protein composition of circulating EV from DOCA+Salt hypertensive mice (fold change <0.71 or >2.3; P<0.05). Additionally, Ingenuity Pathway Analysis highlighted 257 altered pathways in male mice, with only 17 pathways specific to this sex. Among the 235 independent proteins identified, 132 demonstrated differential expression in hypertensive males in comparison to hypertensive female mice. Additionally, 72 proteins were exclusively detected in hypertensive male mice, and 4 were common across the 6 major pathways associated with hypertension, notably Gnai2 which was uniquely expressed in hypertensive male mice. Diabetic cardiomyopathy emerged as one of the top ten signaling pathways based on the enrichment scores for each pathway. Furthermore, our findings suggest that the Renin-Angiotensin System is significantly overexpressed in males compared to females (q-values: 1.4E-05 vs. 5.0E-4). This was evidenced by the upregulation of Leucyl/cystinyl aminopeptidase (IRAP) and aminopeptidase N which are involved in Ang-IV formation and signaling, respectively. Among other differentially expressed proteins, cyclin B1 (CCNB1) was upregulated, consistent with previous studies showing an increase in CCNB1 in rats with pulmonary arterial hypertension and our recent findings showing a significant decrease in miR-410-3p in plasma samples of hypertensive mice, a miRNA known to target CCNB1. These results support a negative correlation between miR-410-3p and CCNB1 in hypertension. Conversely, we noted a significant decrease in laminin levels, which are known to regulate blood pressure by modulating the expression of contractile proteins. Surprisingly, while it was observed that the hypertrophic cardiomyopathy pathway was significantly elevated, this increase was more prominent in females than males. Together, the Ingenuity Pathway Analysis revealed enrichment in proteins associated with diabetic cardiomyopathy signaling, the complement system, and NADH-ubiquinone oxidoreductase in extracellular vesicles from hypertensive mice. Further exploration of these alterations may enhance our understanding of salt sensitive hypertension.