CLEC-2 Supports Platelet Aggregation in Mouse but not Human Blood at Arterial Shear

Abstract

C-type lectin-like receptor 2 (CLEC-2) is highly expressed on platelets and a subpopulation of myeloid cells, and is critical in lymphatic development. CLEC-2 has been shown to support thrombus formation at sites of inflammation, but to have a minor/negligible role in hemostasis. This identifies CLEC-2 as a promising therapeutic target in thromboinflammatory disorders, without hemostatic detriment. We utilized a GPIbα-Cre recombinase mouse for more restricted deletion of platelet-CLEC-2 than the previously used PF4-Cre mouse. clec1b^fl/flGPIbα-Cre⁺ mice are born at a Mendelian ratio, with a mild reduction in platelet count, and present with reduced thrombus size post-FeCl₃-induced thrombosis, compared to littermates. Antibody-mediated depletion of platelet count in C57BL/6 mice, to match clec1b^fl/flGPIbα-Cre⁺ mice, revealed that the reduced thrombus size post-FeCl₃-injury was due to the loss of CLEC-2, and not mild thrombocytopenia. Similarly, clec1b^fl/flGPIbα-Cre⁺ mouse blood replenished with CLEC-2-deficient platelets ex vivo to match littermates had reduced aggregate formation when perfused over collagen at arterial flow rates. In contrast, platelet-rich thrombi formed following perfusion of human blood under flow conditions over collagen types I or III, atherosclerotic plaque, or inflammatory endothelial cells were unaltered in the presence of CLEC-2-blocking antibody, AYP1, or recombinant CLEC-2-Fc. The reduction in platelet aggregation observed in clec1b^fl/flGPIbα-Cre⁺ mice during arterial thrombosis is mediated by the loss of CLEC-2 on mouse platelets. In contrast, CLEC-2 does not support thrombus generation on collagen, atherosclerotic plaque, or inflamed endothelial cells in human at arterial shear.

Keywords

CLEC-2 - thrombosis - platelets - AYP1 - thromboinflammation

Author Contributions

J.H.B. designed and performed research, collected, analyzed and interpreted data, and wrote the manuscript; C.W.S.., N.J.J., and H.C.B. performed experiments and analyzed data; Y.D. and M.R.T. produced reagents; S.J.M., N.S.P., and J.R. interpreted data; S.P.W. designed research and interpreted data. All authors edited and approved the final version of the manuscript.

Publikationsverlauf

Eingereicht: 05. Januar 2022

Angenommen: 01. Juni 2022

Accepted Manuscript online:
11. Juli 2022

Artikel online veröffentlicht:
18. Oktober 2022

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