The endothelial protein C receptor (EPCR) binds to protein C (PC) and increases the rate of activated protein C (APC) generation by the thrombin-thrombomodulin (TM) complex. APC exerts anticoagulant, anti-inflammatory, and cytoprotective effects, which are EPCR-dependent. The thrombin-TM complex is also a potent activator of thrombin activable fibrinolysis inhibitor (TAFI), leading to impaired clot lysis. Mutations and polymorphisms identified in the EPCR gene, which can affect the efficiency of PC activation, have been associated with an increased risk of thrombosis. In this thesis we investigate the impact of impaired PC binding to EPCR on coagulation, inflammation, and fibrinolysis using novel in vitro and in vivo models. Using a murine model that harbours a variant of EPCR that does not bind PC (R84A), we demonstrate that upon thrombotic challenge, there is an increase in thrombin generation and fibrin deposition in the lungs. Upon inflammatory challenge, impaired PC/EPCR interactions also result in increased thrombin generation and increased neutrophil infiltration into the lungs. Using cells that express TM and a human variant of EPCR that does not bind PC (R96C), we demonstrate that clot lysis is delayed in normal plasma independent of TAFI activation, suggesting PC and TAFI do not compete for activation by the thrombin-TM complex. In contrast, delayed clot lysis in plasma deficient of PC is a result of greater TAFI activation by the thrombin-TM complex. Taken together, impairment of the PC pathway contributes to thrombosis through pro-coagulant, pro-inflammatory and anti-fibrinolytic mechanisms. Interestingly, mice with EPCR variant R84A, develop bone marrow failure and splenomegaly, revealing a novel role for EPCR in the bone marrow. Taken together, PC/EPCR interactions regulate the coagulation, inflammation, and fibrinolytic pathways, which may have a significant impact on maintaining hematopoietic homeostasis. / Thesis / Doctor of Philosophy (PhD) / Under normal conditions, blood is maintained in a fluid state. Upon injury or infection, the blood begins to form a clot to prevent bleeding. Once bleeding has stopped the clot is dissolved and blood regains its fluid state. The formation of a blood clot is a serious and potentially life threatening disease. A blood clot formed inside a blood vessel can block the flow of blood through the circulation, leading to organ damage. Approximately 50% of blood clots are caused by known genetic or environmental factors, leaving 50% of blood clots caused by unknown factors. In this thesis we investigate the unknown factors that contribute to blood clotting. In patients who have experienced blood clots with no known cause, we have identified genetic mutations in a blood vessel wall protein, known as the endothelial protein C receptor (EPCR) that renders it non-functional. We demonstrate both in vitro and in vivo that non-functional EPCR not only leads to the formation of a blood clot but also delays the removal of the blood clot. Our in vivo studies have also revealed a previously unknown role for EPCR in the bone marrow, likely through its effects on blood coagulation. Taken together, loss of EPCR function contributes to the development of clot formation and likely impacts other organ systems.
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/19898 |
| Date | January 2016 |
| Creators | Pepler, Laura |
| Contributors | Liaw, Patricia, Medical Sciences (Blood and Cardiovascular) |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
| Type | Thesis |
Page generated in 0.002 seconds