Surgery increases the risk of thrombosis but prevention and management of thromboembolic disease in the perioperative period is complicated by the risk of hemorrhage. Neutrophils (PMNs) play a vital role in wound healing, but their contribution to perioperative thrombosis and implications for thromboprophylaxis are unclear. Adherence of PMNs to wounds and to the vasculature generates a unique localized sequestrum enriched in enzymes and antimicrobial peptides protected from plasma inhibitors that contribute to innate immunity in part through the orderly formation and dissolution of fibrin. We posit that post-operative and persistent inflammation disrupt this balance, predisposing to thrombosis by promoting formation and persistence of fibrin. We have previously observed that alpha-defensins (?-def), antimicrobial peptides that constitute 5% of total human PMN protein that are released upon activation, promote clotting and inhibit fibrinolysis. The absence of ?-def in murine PMNs has hindered a better understanding of how these peptides contribute to thromboembolic disease in the perioperative setting. Using a novel transgenic mouse that expresses PMN ?-def (Def++), we show that ?-def circulates in a complex with fibrinogen and promotes polymerization and retraction of fibrin in vitro, deposits in the vasculature, induces occlusive arterial and venous thrombosis, and inhibits lysis of pulmonary emboli in vivo. These pathogenic properties can be prevented and possibly reversed by preventing ?-def release using colchicine. We now propose an integrated approach to understanding the mechanism and implications of PMN ?-def on perioperative thrombosis by examining: a) The biophysical effects of ?-def on clot formation and structure~ b) The mechanism of accelerated thrombosis and impaired fibrinolysis in Def++ mice and the salutary effect of blocking ?-def release~ and c) The utility of ?-def gene and protein expression as biomarkers of risk for venous thrombosis and post-thrombotic syndrome. These studies will provide insight into an unappreciated contribution of PMN ?-def to arterial and venous thrombosis, identify novel genetic and protein biomarkers of patients at risk for surgery related thrombosis and provide evidence for the use of a safe and effective intervention to prevent thrombosis in the perioperative period.
There is a clear link between infection and inflammation with venous and arterial thrombosis, but the mediators and implications for therapy are uncertain. We have shown that alpha-defensins, peptides released from neutrophils, are prothrombotic and anti-fibrinolytic and that these properties are attenuated with colchicine, a drug used in clinical medicine for decades. This project will examine the mechanisms by which alpha-defensins promote thrombosis, determine if plasma levels identify patients at risk for venous thromboembolism and help identify a novel approach to anti-thrombotic therapy without bleeding risk.
|Pillai, Vikram G; Bao, Jialing; Zander, Catherine B et al. (2016) Human neutrophil peptides inhibit cleavage of von Willebrand factor by ADAMTS13: a potential link of inflammation to TTP. Blood 128:110-9|
|Abu-Fanne, Rami; Maraga, Emad; Abd-Elrahman, Ihab et al. (2016) ?-Defensins Induce a Post-translational Modification of Low Density Lipoprotein (LDL) That Promotes Atherosclerosis at Normal Levels of Plasma Cholesterol. J Biol Chem 291:2777-86|