12-LOX Regulation of ADP Secretion Pathway

Open Access
Author:
Zhu, Junchun
Area of Honors:
Science
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
  • Brittany Anderson, Thesis Supervisor
  • David Scott Witwer, Honors Advisor
  • Nik Tsotakos, Faculty Reader
Keywords:
  • Thrombosis
  • 12-LOX
  • ML355
  • Blood Clotting
  • ADP Secretion
  • Flow Cytometry
  • Aggregometry
Abstract:
Blood clotting is a serious problem, which kills up to 1 in 3 people each year in the United States. Platelets play a vital role in blood clotting, hemostasis, and some other cardiovascular disorders. Platelets are activated following injury or vascular insult and can cause the formation of an occlusive thrombus. Thus, studying platelets is essential to develop new strategies for the prevention of arterial thrombosis. An enzyme found in the platelet and megakaryocyte, 12-Lipoxygenase (12-LOX), was shown to have an important role in platelet activation by a number of research groups. However, the mechanism of how 12-LOX regulates platelet activation needs to be studied further. In this study, we sought to determine the key biochemical steps in platelets that are regulated by 12-LOX, especially whether the adenosine diphosphate (ADP) secretion pathway is involved in 12-LOX signaling. Human platelets were treated with ML355, a selective 12-LOX inhibitor, or the combination of ML355 + ADP receptor inhibitors including P2Y1 and P2Y12 receptor inhibitors (MRS2179 and 2Me-SAMP respectively). The treated platelets will be stimulated with thrombin and assayed for activation using aggregation assays, western blotting, and flow cytometry. Aggregation, granule secretion, and integrin activation data was collected and analyzed. This research could offer a novel target for dual antiplatelet therapy and provide a deeper mechanistic understanding of platelet reactivity and its role in maintaining hemostasis.