A Novel Role for Piezo1 in Diabetes-Associated Thrombosis

Abstract

Thrombosis, as the underlying mechanism of coronary heart disease, stroke, and venous thromboembolism, is the leading cause of death. The risk of thrombosis is drastically increased in diabetes. As thrombosis occurs in the blood flow, cells involved undergo dynamic mechanical triggers that can result in downstream cellular responses. One key player of mechano-sensing in blood cells is the Piezo1 channel, which transduces physical stretch to Ca2+ influx in cells. We investigated in the role of Piezo1 in thrombosis and how it is dysregulated in diabetes. We developed a high-throughput imaging-based platform to test Piezo1's function in erythrocytes, leukocytes and platelets from patients’ peripheral blood. The assay utilizes Ca2+ imaging, Piezo1 modulators, and machine-learning-based imaging analysis. To test force-induced Ca2+ entry in vitro, we incorporated microfluidic devices that mimic vessel constrictions. Finally, we used Zebrafish as animal model to assess Piezo1's role in thrombosis in vivo. We found that acute hyperglycemia attenuated Piezo1 activator Yoda1-mediated Ca2+ elevation in erythrocytes by 13.0±3.0% (40 patients, p=0.004). Consistently, we observed in microfluidic devices that hyperglycemia reduced pressure-induced Ca2+ increase in erythrocytes. Conversely, Piezo1-mediated Ca2+ entry in varies blood cells increased in diabetic patients compared to healthy group, suggesting an adaptive regulation of Piezo1 expression in diabetes. To test whether the elevated Piezo1 activity contributes to the prothrombotic state, we tested in WT and Piezo1 knock-down Zebrafish. In vivo clot formation after FeCl3 injury was suppressed in Piezo1 morphants (time to occlusion WT: 64.3±3.5s, Piezo1 morphant: 92.5±4.9s, p=0.0002), suggesting that Piezo1 promotes thrombosis. Overall, we demonstrated that Piezo1's activities in blood cells are modulated in hyperglycemia and diabetes. The upregulated Piezo1 activity in diabetic patients is potentially one of the underlying mechanisms for their increased thrombosis risk, which makes Piezo1 an important diagnostic and therapeutic target.