AUTHOR=Misztal Tomasz , Golaszewska Agata , Branska-Januszewska Justyna , Marcinczyk Natalia , Chabielska Ewa , Tomasiak Marian , Rusak Tomasz 

TITLE=HAuCl4, Putative General Aquaporins Blocker, Reduces Platelet Spreading, Filopodia Formation, Procoagulant Response, and Thrombus Formation Under Flow

JOURNAL=Frontiers in Physiology

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2020.01025

DOI=10.3389/fphys.2020.01025

ISSN=1664-042X

ABSTRACT=<p><bold>Background</bold>: Recent studies indicate that aquaporin (AQP) water channels have a regulatory function in human platelet secretion and in procoagulant response of murine platelets. However, the engagement of AQPs in morphological changes, procoagulant response, and thrombus formation in human blood has never been investigated.</p><p><bold>Methods</bold>: Confocal microscopy was used to study platelet spreading, filopodia formation, ballooning, and thrombus formation under flow. Flow cytometry was utilized to assess platelet phosphatidylserine (PS) exposure and microparticles shedding. Kinetics of clot formation <italic>in vitro</italic> was evaluated by thromboelastometry. Mouse model of ferric chloride (III) (FeCl<sub>3</sub>)-induced thrombosis was used to investigate thrombus formation <italic>in vivo</italic>.</p><p><bold>Results</bold>: We found that chloroauric(III) acid (HAuCl<sub>4</sub>), a classical AQP inhibitor (10–100 μM), reduced spreading of human platelets on collagen-coated surfaces and inhibited filopodia formation in a fluid phase. Under flow conditions, HAuCl<sub>4</sub> (100 μM) attenuated thrombi growth on collagen, platelet secretion, and PS exposure. Thrombus formation was restored by the addition of exogenous adenosine diphosphate (ADP). Collagen-evoked platelet procoagulant response (evaluated as PS exposure, shedding of microparticles, platelet-dependent thrombin generation, and membrane ballooning) was distinctly reduced by HAuCl<sub>4</sub> (25–200 μM), as well as the dynamics of clot formation. In mouse model of thrombosis, reduction of surface of PS-positive cells within thrombus was observed in the presence of HAuCl<sub>4</sub> (1–10 mg/kg).</p><p><bold>Conclusion</bold>: These results suggest that in human platelets AQPs are crucial for agonist-evoked morphological changes, thrombus formation under flow, and in development of procoagulant response. Antithrombotic effect <italic>in vivo</italic> suggests that nontoxic inhibitors of AQPs may be considered as potential candidates for a novel class of antiplatelet drugs.</p>