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Thromb Haemost 2023; 123(06): 597-612
DOI: 10.1055/s-0043-1761463
Cellular Haemostasis and Platelets

Pharmacological Inhibition of Glycoprotein VI- and Integrin α2β1-Induced Thrombus Formation Modulated by the Collagen Type

Natalie J. Jooss 
1   Department of Biochemistry, Cardiovascular Research Institute, Maastricht University, Maastricht, The Netherlands
2   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
,
Yvonne M.C. Henskens
3   Central Diagnostic Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands
,
Steve P. Watson
2   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
4   Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Midlands, United Kingdom
,
Richard W. Farndale 
5   Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom
6   CambCol Laboratories, Ely, United Kingdom
,
Meinrad P. Gawaz
7   Department of Cardiology and Angiology, University Hospital Tübingen, Eberhard Karls University Tübingen, Tübingen, Germany
,
Martine Jandrot-Perrus
8   UMR_S1148, Laboratory for Vascular Translational Science, INSERM, University Paris Cité, Paris, France
,
Natalie S. Poulter
2   Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom
4   Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Midlands, United Kingdom
,
Johan W. M. Heemskerk 
1   Department of Biochemistry, Cardiovascular Research Institute, Maastricht University, Maastricht, The Netherlands
9   Synapse Research Institute, Maastricht, The Netherlands
› Author Affiliations Funding This work has received funding form the European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No. 766118 (TAPAS). This project was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—project number 374031971-TRR 240 (M.P.G.). N.J.J. is enrolled in a joint PhD program of the Universities of Maastricht and Birmingham (United Kingdom).
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Abstract

Background In secondary cardiovascular disease prevention, treatments blocking platelet-derived secondary mediators pose a risk of bleeding. Pharmacological interference of the interaction of platelets with exposed vascular collagens is an attractive alternative, with clinical trials ongoing. Antagonists of the collagen receptors, glycoprotein VI (GPVI), and integrin α2β1, include recombinant GPVI-Fc dimer construct Revacept, 9O12 mAb based on the GPVI-blocking reagent Glenzocimab, Syk tyrosine-kinase inhibitor PRT-060318, and anti-α2β1 mAb 6F1. No direct comparison has been made of the antithrombic potential of these drugs.

Methods Using a multiparameter whole-blood microfluidic assay, we compared the effects of Revacept, 9O12-Fab, PRT-060318, or 6F1 mAb intervention with vascular collagens and collagen-related substrates with varying dependencies on GPVI and α2β1. To inform on Revacept binding to collagen, we used fluorescent-labelled anti-GPVI nanobody-28.

Results and Conclusion In this first comparison of four inhibitors of platelet–collagen interactions with antithrombotic potential, we find that at arterial shear rate: (1) the thrombus-inhibiting effect of Revacept was restricted to highly GPVI-activating surfaces; (2) 9O12-Fab consistently but partly inhibited thrombus size on all surfaces; (3) effects of GPVI-directed interventions were surpassed by Syk inhibition; and (4) α2β1-directed intervention with 6F1 mAb was strongest for collagens where Revacept and 9O12-Fab were limitedly effective. Our data hence reveal a distinct pharmacological profile for GPVI-binding competition (Revacept), GPVI receptor blockage (9O12-Fab), GPVI signaling (PRT-060318), and α2β1 blockage (6F1 mAb) in flow-dependent thrombus formation, depending on the platelet-activating potential of the collagen substrate. This work thus points to additive antithrombotic action mechanisms of the investigated drugs.

Keywords

collagen - glycoprotein VI - integrin α2β1 - protein tyrosine kinase - thrombus

Authors' Contributions

N.J.J. designed and performed experiments, analyzed data, prepared figures, and wrote the manuscript. M.P.G. provided Revacept and revised the manuscript. M.J.-P. supplied 9O12-Fab and revised the manuscript. Y.M.C.H., N.S.P., and S.P.W. contributed by funding and supervision, and revised the manuscript. J.W.M.H. designed experiments, provided supervision and funding, and wrote the manuscript. All authors have read and approved the manuscript.


Supplementary Material



Publication History

Received: 22 May 2022

Accepted: 20 December 2022

Article published online:
17 February 2023

© 2023. Thieme. All rights reserved.

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