Abstract 148: Cardiac Myosin Promotes Thrombin Generation and Attenuates Tissue Plasminogen Activator-induced Plasma Clot Lysis

Abstract

Recently we discovered that skeletal muscle myosin, which is in the same family as cardiac myosin, exerts prothrombotic effects by binding factor Xa and enhancing prothrombin activation in the presence of factor Va. Thus, we tested the influence of cardiac myosin on thrombus formation and fibrinolysis. Studies of the effects of cardiac myosin on thrombogenesis ex vivo using fresh human flowing blood showed that perfusion of blood over cardiac myosin-coated surfaces at 300 s -1 shear rate caused extensive fibrin deposition. Addition of cardiac myosin to blood also promoted the thrombotic responses of human blood flowing over collagen-coated surfaces, evidence of myosin’s thrombogenicity. Further studies showed that cardiac myosin enhanced thrombin generation in whole blood, platelet rich plasma and platelet poor plasma, indicating that myosin promotes thrombin generation in plasma primarily independently of platelets or other blood cell components. In a purified system composed of factor Xa, factor Va, prothrombin and calcium ions, cardiac myosin greatly enhanced prothrombinase activity. Experiments using Gla-domainless factor Xa showed that the Gla domain of factor Xa was not required for cardiac myosin’s prothrombinase enhancement in contrast to phospholipid-enhanced prothrombinase activity which requires that Gla domain. In studies of tissue plasminogen activator (tPA)-induced plasma clot lysis, increasing concentrations of cardiac myosin attenuated tPA-mediated clot lysis. The ability of cardiac myosin to inhibit tPA-induced plasma clot lysis was ablated in the presence of the carboxypeptidase inhibitor from potatoes, an inhibitor of thrombin activatable fibrinolysis inhibitor (TAFI). Clot lysis assays using TAFI-deficient plasma confirmed the requirement for TAFI for the antifibrinolytic action of cardiac myosin. We hypothesize that cardiac myosin-dependent thrombin generation increases TAFI activation and subsequent inhibition of clot lysis. In summary, here we show that cardiac myosin is both procoagulant and anti-fibrinolytic due to its ability to bind factor Xa and strongly promote thrombin generation. This raises new questions about potential procoagulant functions for cardiac myosin in coronary health and disease.