Pharmacologic inhibition of TMEM16 with benzbromarone or genetic deletion of TMEM16E significantly reduced vessel-wall phosphatidylserine externalization and fibrin formation during thrombosis.
TMEM16E and TMEM16F regulate endothelial cell phosphatidylserine externalization, and their inhibition reduces vessel-wall-dependent thrombosis without increasing bleeding in preclinical models.
p-value: p=<0.001
Endothelial cells (ECs) normally form an anticoagulant surface under physiological conditions, but switch to support coagulation following pathogenic stimuli. This switch promotes thrombotic cardiovascular disease. To generate thrombin at physiologic rates, coagulation proteins assemble on a membrane containing anionic phospholipid, most notably phosphatidylserine (PS). PS can be rapidly externalized to the outer cell membrane leaflet by phospholipid "scramblases," such as TMEM16F. TMEM16F-dependent PS externalization is well characterized in platelets. In contrast, how ECs externalize phospholipids to support coagulation is not understood. We employed a focused genetic screen to evaluate the contribution of transmembrane phospholipid transport on EC procoagulant activity. We identified 2 TMEM16 family members, TMEM16F and its closest paralog, TMEM16E, which were both required to support coagulation on ECs via PS externalization. Applying an intravital laser-injury model of thrombosis, we observed, unexpectedly, that PS externalization was concentrated at the vessel wall, not on platelets. TMEM16E-null mice demonstrated reduced vessel-wall-dependent fibrin formation. The TMEM16 inhibitor benzbromarone prevented PS externalization and EC procoagulant activity and protected mice from thrombosis without increasing bleeding following tail transection. These findings indicate the activated endothelial surface is a source of procoagulant phospholipid contributing to thrombus formation. TMEM16 phospholipid scramblases may be a therapeutic target for thrombotic cardiovascular disease.
Schmaier et al. (2023) studied Thrombosis. TMEM16 inhibitors (benzbromarone) or TMEM16E genetic deletion vs. Vehicle or wild-type littermates was evaluated on Fibrin formation following laser injury (median AUC) (p=<0.001). Pharmacologic inhibition of TMEM16 with benzbromarone or genetic deletion of TMEM16E significantly reduced vessel-wall phosphatidylserine externalization and fibrin formation during thrombosis.