Cationic Zinc Is Required for Factor XII Recruitment and Activation by Stimulated Platelets and for Thrombus Formation in Vivo

J Thromb Haemost. 2020 Jun 16. doi: 10.1111/jth.14964. Online ahead of print.

Abstract

Background: Although divalent zinc (Zn2+ ) is known to bind FXII and affect its sensitivity to autoactivation, little is known about the role of Zn2+ in the binding of FXII to platelets, where FXII activation is thought to occur in vivo, and the function of Zn2+ during thrombus formation following vascular injury remains poorly understood.

Objectives: To evaluate the role of Zn2+ in platelet-dependent FXIIa generation.

Methods: FXII binding to platelets and FXII activation by stimulated platelets were assessed using flow cytometry and a platelet-dependent thrombin generation assay. The mouse cremaster laser injury model was used to evaluate the impact of Zn2+ chelation on thrombus formation in vivo.

Results: Our data demonstrate that stimulated platelets support FXII-dependent thrombin generation and that FXII activation by platelets requires the presence of Zn2+ . By contrast, thrombin generation by stimulated endothelial cells occurred independently of FXII and Zn2+ . Using flow cytometry, we found that FXII-FITC binds to the surfaces of stimulated platelets in a specific and Zn2+ -dependent manner, whereas resting platelets demonstrated minimal binding. Other physiologically-relevant divalent cations are unable to support this interaction. Consistent with these findings, the Zn2+ -specific chelator CaEDTA confers thromboprotection in the mouse cremaster laser injury model without causing increased bleeding. We observed an identical phenotype in FXII null mice tested in the same system.

Conclusions: Our results suggest a novel role for Zn2+ in the binding and activation of FXII at the platelet surface, an interaction that appears crucial to FXII-dependent thrombin generation but dispensable for hemostasis.

Keywords: Contact Pathway; FXII knockout mice; Factor XII; Platelet-dependent thrombin generation; Zinc.