The metalloproteases ADAM17 and ADAM10 regulate endothelial migration and proliferation induced by the mechanoresponsive ion channels Piezo1 and TRPV4

Background Endothelial cells are constantly exposed to mechanical forces generated by blood flow that regulate endothelial homeostasis. Pathological events such as vascular injury or thrombus formation alter these forces, which are sensed by endothelial cells and can trigger repair mechanisms, including inflammation, proliferation and migration. These mechanical changes can be detected by cells through various mechanosensors, particularly ion channels such as Piezo1 and TRPV4. One possible pathway for the translation of mechanical signals into cellular responses involves the downstream activation of membrane-bound proteases of the ADAM family. These proteases cleave transmembrane proteins such as growth factors, adhesion molecules and cytokines, thereby regulating inflammation, proliferation and migration. This study aimed to investigate the link between the ion channels Piezo1/TRPV4 and the metalloproteases ADAM10/17. Methods Primary human umbilical vein endothelial cells (HUVECs) were treated with pharmacological agonists of Piezo1 and TRPV4 or cultured under flow conditions for mechanical stimulation. Pharmacological inhibitors and shRNA-mediated knockdown were used to assess the involvement of specific proteins. ADAM activity was determined indirectly by measuring the release of substrates such as JAM-A or TNFR1 into the culture medium via ELISA or by a reduction in the immunocytochemical staining of VE-cadherin. The effects on cell proliferation and migration were analyzed via live-cell microscopy. Results Mechanical stimulation through flow induced the activation of ADAM10 and ADAM17, which was partially mediated by Piezo1 and TRPV4. This connection was confirmed via the use of specific agonists for both ion channels, which have the potential to independently activate ADAM proteases. Furthermore, Piezo1 but not TRPV4 activation led to ADAM10/17-dependent increases in proliferation and migration. Conclusion Piezo1 and TRPV4 contribute to the mechanical activation of ADAM10/17 in endothelial cells and thus play a role in regulating endothelial functions such as proliferation and migration.