423 An omega-6-derived eicosanoid negatively regulates platelet reactivity of cardiovascular patients at increased risk for thrombosis

Abstract

OBJECTIVES/GOALS: This study aimed to investigate the mechanistic effects of the omega-6-derived eicosanoid 12-HETrE on platelets of cardiovascular patients at risk for a recurrent cardiovascular event triggered by thrombosis. 12-HETrE negatively regulates platelet reactivity through binding to the prostacyclin receptor in platelets. METHODS/STUDY POPULATION: Healthy donors were recruited from the Ann Arbor community and the University of Michigan Medicine Center. Cardiovascular patients with elevated cardiovascular risk were recruited from the Cardiac Catheterization Laboratory at Michigan Medicine Hospital. All subjects were recruited under study protocols approved by the University of Michigan IRB. Healthy donors were matched with the cardiovascular patients regarding age, sex, race, and BMI. Whole blood was collected via venipuncture into vacutainers containing sodium citrate. Platelets were isolated via serial centrifugation and treated ex vivo with vehicle control, 12-HETrE, or Iloprost. RESULTS/ANTICIPATED RESULTS: Based on our previous studies, we chose to treat platelets ex vivo with 25uM of 12-HETrE for 10 minutes. Using platelets of healthy donors, we have shown that treatment with 25uM of 12-HETrE for 10 minutes inhibited platelet aggregation and activation, and activated protein kinase A, suggesting activation of the prostacyclin receptor. We conducted a preliminary study to demonstrate that ex vivo treatment of 12-HETrE regulated signaling pathways in platelets such as cell-to-cell interaction, platelet activation and cytoskeleton rearrangements. In this study, we have demonstrated that treatment with 12-HETrE regulated receptors and intraplatelet proteins in platelets of cardiovascular patients. Furthermore, these proteins are involved in critical pathways in the platelet. DISCUSSION/SIGNIFICANCE: Dual antiplatelet therapy has significantly decreased mortality due to thrombotic events. However, cardiovascular events triggered by thrombosis persist as the leading cause of death in the US. This study may uncover key regulators to be targeted for the long-term goal of providing additional protection to reduce future incidence of thrombosis.