Thrombin binding to GPIbα induces platelet aggregation and fibrin clot retraction supported by resting αIIbβ3 interaction with polymerized fibrin

Christophe Dubois; Beat Steiner; Nelly Kieffer; Sylvie C. Meyer Reigner

doi:10.1055/s-0037-1613473

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2003; 89(05): 853-865
DOI: 10.1055/s-0037-1613473

Platelets and Blood Cells

Schattauer GmbH

Thrombin binding to GPIbα induces platelet aggregation and fibrin clot retraction supported by resting αIIbβ3 interaction with polymerized fibrin

Christophe Dubois

¹Pharma Division, Discovery Research, F. Hoffmann-La Roche Ltd, Basel, Switzerland

,

Beat Steiner

¹Pharma Division, Discovery Research, F. Hoffmann-La Roche Ltd, Basel, Switzerland

,

Nelly Kieffer

²Laboratoire Franco-Luxembourgeois de Recherche Biomédicale (CNRS-GDRE), University Center, Luxembourg, Luxembourg

,

Sylvie C. Meyer Reigner

¹Pharma Division, Discovery Research, F. Hoffmann-La Roche Ltd, Basel, Switzerland

› Institutsangaben

Abstract

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Summary

We have investigated the mechanisms leading to platelet aggregation following thrombin interaction with the glycoprotein (GP) Ib-IX-V complex. We show that platelets desensitized for the two thrombin receptors, PAR-1 and PAR-4, are still able to aggregate in response to thrombin and that this aggregation can be inhibited by a monoclonal antibody (VM16d) that blocks thrombin binding to GPIbα, or by pretreatment of platelets with Mocarhagin, a protease that specifically cleaves GPIbα. The thrombin/GPIbα-initiated signaling cascade induces platelet shape change through activation of the Rho kinase p160ROCK, independent of calcium mobilization, transient MEK-1 phosphorylation as well as the cleavage of talin through a calcium-independent mechanism. This signaling cascade does not induce the exposure of high affinity αIIbβ3 integrin receptors, nor does it lead to µ-calpain cleavage of filamin or the integrin cytoplasmic tail. In contrast, we provide evidence that binding of thrombin to GPIbα induces fibrin binding to resting αIIbβ3 leading to fibrin-dependent platelet aggregation and clot retraction, that can be selectively inhibited by αIIbβ3 antagonists such as RGDS, the dodecapeptide or lamifiban, as well as by the fibrin polymerization inhibitor GPRP-amide.

Keywords

Platelets - GPIb-IX-V - thrombin - intracellular signaling - aggregation

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Referenzen

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