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CC BY-NC-ND 4.0 · Thromb Haemost 2022; 122(12): 1988-2000
DOI: 10.1055/a-1896-6992
Cellular Haemostasis and Platelets

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

Joshua H. Bourne 
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
,
Christopher W. Smith
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
,
Natalie J. Jooss
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
2   Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, The Netherlands
,
Ying Di
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
,
Helena C. Brown
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
3   Institute of Experimental Biomedicine I, University Hospital and Rudolf Virchow Center for Integrative and Translational Bioimaging, University of Würzburg, Würzburg, Germany.
,
Samantha J. Montague
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
,
Mark R. Thomas
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
4   UHB and SWBH NHS Trusts, Birmingham, United Kingdom
,
Natalie S. Poulter
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
5   Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, The Midlands, United Kingdom
,
Julie Rayes
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
5   Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, The Midlands, United Kingdom
,
Steve P. Watson
1   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
5   Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, The Midlands, United Kingdom
› Author Affiliations Funding This work was supported by a college-funded PhD studentship (University of Birmingham), N.J.J. and H.C.B. are funded by a European Union's Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement (766118). S.J.M. is supported by the BHF Accelerator Award (AA/18/2/34218). J.R. is supported by a BHF Intermediate Basic Science Fellowship Application (FS/IBSRF/20/25039). S.P.W. holds a BHF Chair (CH/03/003).
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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. clec1bfl/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 clec1bfl/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, clec1bfl/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 clec1bfl/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.


Supplementary Material



Publication History

Received: 05 January 2022

Accepted: 01 June 2022

Accepted Manuscript online:
11 July 2022

Article published online:
18 October 2022

© 2022. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

Comment to this article:

Of Mice and Man: The Unwinding of CLEC-2 as an Antithrombotic Target?Thromb Haemost 2022; 122(12): 1963-1965
DOI: 10.1055/a-1938-1380

 

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