Role of Kv1.3 Channels in Platelet Functions and Thrombus Formation

Arterioscler Thromb Vasc Biol. 2020 Oct;40(10):2360-2375. doi: 10.1161/ATVBAHA.120.314278. Epub 2020 Aug 13.

Abstract

Objective: Platelet activation by stimulatory factors leads to an increase in intracellular calcium concentration ([Ca2+]i), which is essential for almost all platelet functions. Modulation of Ca2+ influx and [Ca2+]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 [Ca2+]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 Ca2+ influx and [Ca2+]i elevation, and that this is mediated in part by P2X1. 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 [Ca2+]i elevation. These results indicate that Kv1.3 could act as a promising therapeutic target for antiplatelet therapies.

Keywords: Kv1.3; calcium influx; monoclonal antibody; platelets; thrombosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / pharmacology*
  • Blood Coagulation / drug effects
  • Blood Platelets / drug effects*
  • Blood Platelets / metabolism
  • Disease Models, Animal
  • Female
  • Fibrinolytic Agents / pharmacology*
  • HEK293 Cells
  • Humans
  • Kv1.3 Potassium Channel / antagonists & inhibitors*
  • Kv1.3 Potassium Channel / blood
  • Kv1.3 Potassium Channel / deficiency
  • Kv1.3 Potassium Channel / genetics
  • Male
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Platelet Activation / drug effects
  • Platelet Aggregation Inhibitors / pharmacology*
  • Potassium Channel Blockers / pharmacology*
  • Pulmonary Embolism / blood
  • Pulmonary Embolism / genetics
  • Pulmonary Embolism / metabolism
  • Pulmonary Embolism / prevention & control*
  • Signal Transduction
  • Thrombosis / blood
  • Thrombosis / genetics
  • Thrombosis / metabolism
  • Thrombosis / prevention & control*

Substances

  • Antibodies, Monoclonal
  • Fibrinolytic Agents
  • KCNA3 protein, human
  • Kcna3 protein, mouse
  • Kv1.3 Potassium Channel
  • Platelet Aggregation Inhibitors
  • Potassium Channel Blockers