Role of Kv1.3 Channels in Platelet Functions and Thrombus Formation
Cheng Fan, Xiaofang Yang, Wan Wendy Wang, Jue Wang, Wenzhu Li, Mengyuan Guo, Shiyuan Huang, Zhaohui Wang, Kun Liu
Abstract
Objective: Platelet activation by stimulatory factors leads to an increase in intracellular calcium concentration ([Ca 2+ ] i ), which is essential for almost all platelet functions. Modulation of Ca 2+ influx and [Ca 2+ ] i in platelets has been emerging as a possible strategy for preventing and treating platelet-dependent thrombosis. Voltage-gated potassium 1.3 channels (Kv1.3) are highly expressed in platelets and able to regulate agonist-evoked [Ca 2+ ] i increase. However, the role of Kv1.3 channels in regulating platelet functions and thrombosis has not yet been elucidated. In addition, it is difficult to obtain a specific blocker for this channel, since Kv1.3 shares identical drug-binding sites with other K + channels. Here, we investigate whether specific blockade of Kv1.3 channels by monoclonal antibodies affects platelet functions and thrombosis. Approach and Results: In this study, we produced the anti-Kv1.3 monoclonal antibody 6E12#15, which could specifically recognize both human and mouse Kv1.3 proteins and sufficiently block Kv1.3 channel currents. We found Kv1.3 blockade by 6E12#15 inhibited platelet aggregation, adhesion, and activation upon agonist stimulation. In vivo treatment with 6E12#15 alleviated thrombus formation in a mesenteric arteriole thrombosis mouse model and protected mice from collagen/epinephrine-induced pulmonary thromboembolism. Furthermore, we observed Kv1.3 regulated platelet functions by modulating Ca 2+ influx and [Ca 2+ ] i elevation, and that this is mediated in part by P2X 1 . Interestingly, Kv1.3 −/− mice showed impaired platelet aggregation while displayed no abnormalities in in vivo thrombus formation. This phenomenon was related to more megakaryocytes and platelets produced in Kv1.3 −/− mice compared with wild-type mice. Conclusions: We showed specific inhibition of Kv1.3 by the novel monoclonal antibody 6E12#15 suppressed platelet functions and platelet-dependent thrombosis through modulating platelet [Ca 2+ ] i elevation. These results indicate that Kv1.3 could act as a promising therapeutic target for antiplatelet therapies.