Nonredundant Roles of Platelet Glycoprotein VI and Integrin αIIbβ3 in Fibrin-Mediated Microthrombus Formation

Perrella, Gina; Huang, Jingnan; Provenzale, Isabella; Swieringa, Frauke; Heubel‐Moenen, Floor C. J. I.; Farndale, Richard W.; Roest, Mark; Meijden, Paola E. J. van der; Thomas, Mark R.; Ariëns, Robert A. S.; Jandrot‐Perrus, Martine; Watson, Steve P.; Heemskerk, Johan W. M.

doi:10.1161/atvbaha.120.314641

Cited by 24 publications

(37 citation statements)

References 45 publications

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“…GPVI is a key collagen and fibrin receptor on platelets (3-4 k copies per cell), which signals via a cascade of protein tyrosine kinases, leading to strong platelet activation [48,49]. GPVI interaction with subendothelial collagens induces arterial thrombus formation upon vascular injury, as confirmed in multiple in vivo studies with genetically modified mice and in microfluidics studies with mouse or human blood [50,51].…”

Section: Collagen Receptor Glycoprotein VI (Gpvi)mentioning

confidence: 93%

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Huang

Zhang

Solari

et al. 2021

IJMS

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Platelets are small anucleate blood cells that play vital roles in haemostasis and thrombosis, besides other physiological and pathophysiological processes. These roles are tightly regulated by a complex network of signalling pathways. Mass spectrometry-based proteomic techniques are contributing not only to the identification and quantification of new platelet proteins, but also reveal post-translational modifications of these molecules, such as acetylation, glycosylation and phosphorylation. Moreover, target proteomic analysis of platelets can provide molecular biomarkers for genetic aberrations with established or non-established links to platelet dysfunctions. In this report, we review 67 reports regarding platelet proteomic analysis and signalling on a molecular base. Collectively, these provide detailed insight into the: (i) technical developments and limitations of the assessment of platelet (sub)proteomes; (ii) molecular protein changes upon ageing of platelets; (iii) complexity of platelet signalling pathways and functions in response to collagen, rhodocytin, thrombin, thromboxane A2 and ADP; (iv) proteomic effects of endothelial-derived mediators such as prostacyclin and the anti-platelet drug aspirin; and (v) molecular protein changes in platelets from patients with congenital disorders or cardiovascular disease. However, sample sizes are still low and the roles of differentially expressed proteins are often unknown. Based on the practical and technical possibilities and limitations, we provide a perspective for further improvements of the platelet proteomic field.

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Section: Collagen Receptor Glycoprotein VI (Gpvi)mentioning

confidence: 93%

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Huang

Zhang

Solari

et al. 2021

IJMS

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“…Glass coverslips were coated overnight at 4 °C with Horm collagen (50 µg/mL) or plaque homogenate (0.5 mg/mL) before blocking with 10% BSA for 30 min in Hepes buffer (10 mM Hepes, 136 mM NaCl, 2.7 mM KCl, 2 mM MgCl 2 , 5.5 mM glucose, and 0.1% BSA), pH 7.45 [ 7 ]. Citrated whole blood, recalcified with CaCl 2 (7.5 mM) and MgCl 2 (3.75 mM), in the presence of PPACK (40 µM), was perfused for 7 min at 1000 s −1 [ 15 ].…”

Section: Methodsmentioning

confidence: 99%

“…In vitro flow studies have shown that GPVI contributes to the stability of newly formed platelet aggregates on collagen at high shear through use of the blocking anti-GPVI Fab ACT017, which has more recently been named glenzocimab [ 4 ]. Since the activation of GPVI by fibrinogen is also dependent on integrin αIIbβ3, with the interplay of the two receptors driving platelet adhesion and activation [ 7 ]; this suggests that blocking signalling pathways common to both receptors may have a greater antithrombotic effect than blocking GPVI alone.…”

Section: Introductionmentioning

confidence: 99%

Role of Tyrosine Kinase Syk in Thrombus Stabilisation at High Shear

Perrella

Montague

Brown

et al. 2022

IJMS

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Understanding the pathways involved in the formation and stability of the core and shell regions of a platelet-rich arterial thrombus may result in new ways to treat arterial thrombosis. The distinguishing feature between these two regions is the absence of fibrin in the shell which indicates that in vitro flow-based assays over thrombogenic surfaces, in the absence of coagulation, can be used to resemble this region. In this study, we have investigated the contribution of Syk tyrosine kinase in the stability of platelet aggregates (or thrombi) formed on collagen or atherosclerotic plaque homogenate at arterial shear (1000 s−1). We show that post-perfusion of the Syk inhibitor PRT-060318 over preformed thrombi on both surfaces enhances thrombus breakdown and platelet detachment. The resulting loss of thrombus stability led to a reduction in thrombus contractile score which could be detected as early as 3 min after perfusion of the Syk inhibitor. A similar loss of thrombus stability was observed with ticagrelor and indomethacin, inhibitors of platelet adenosine diphosphate (ADP) receptor and thromboxane A2 (TxA2), respectively, and in the presence of the Src inhibitor, dasatinib. In contrast, the Btk inhibitor, ibrutinib, causes only a minor decrease in thrombus contractile score. Weak thrombus breakdown is also seen with the blocking GPVI nanobody, Nb21, which indicates, at best, a minor contribution of collagen to the stability of the platelet aggregate. These results show that Syk regulates thrombus stability in the absence of fibrin in human platelets under flow and provide evidence that this involves pathways additional to activation of GPVI by collagen.

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“…Nonetheless, the genetic absence of GPVI does not abrogate clot retraction and recent studies show that GPVI and αIIbβ3 play complementary non-redundant roles. 89,90 GPVI may also have a physiologic role in inflammatory states, cancer and other acquired states in GT. 19,91 Platelet Proteome Some 40 years ago, McGregor et al 92 studied platelet membrane glycoproteins in GT by associating carbohydrate-specific or protein-specific labeling procedures with high-resolution two-dimensional gel electrophoresis.…”

Section: Gt Platelets and Fibrinmentioning

confidence: 99%

Section: Gt Platelets and Fibrinmentioning

confidence: 99%

Profiling the Genetic and Molecular Characteristics of Glanzmann Thrombasthenia: Can It Guide Current and Future Therapies?

Nurden

2021

JBM

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Glanzmann thrombasthenia (GT) is the most widely studied inherited disease of platelet function. Platelets fail to aggregate due to a defect in platelet-to-platelet attachment. The hemostatic plug fails to form and a moderate to severe bleeding diathesis results. Classically of autosomal recessive inheritance, GT is caused by defects within the ITGA2B and ITGB3 genes that encode the αIIbβ3 integrin expressed at high density on the platelet surface and also in intracellular pools. Activated αIIbβ3 acts as a receptor for fibrinogen and other adhesive proteins that hold platelets together in a thrombus. Over 50 years of careful clinical and biological investigation have provided important advances that have improved not only the quality of life of the patients but which have also contributed to an understanding of how αIIbβ3 functions. Despite major improvements in our knowledge of GT and its genetic causes, extensive biological and clinical variability with respect to the severity and intensity of bleeding remains poorly understood. I now scan the repertoire of ITGA2B and ITGB3 gene defects and highlight the wide genetic and biological heterogeneity within the type II and variant subgroups especially with regard to bleeding, clot retraction, the internal platelet Fg storage pool and the nature of the mutations causing the disease. I underline the continued importance of gene profiling and biological studies and emphasize the multifactorial etiology of the clinical expression of the disease. This is done in a manner to provide guidelines for future studies and future treatments of a disease that has not only aided research on rare diseases but also contributed to advances in antithrombotic therapy.

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Nonredundant Roles of Platelet Glycoprotein VI and Integrin αIIbβ3 in Fibrin-Mediated Microthrombus Formation

Cited by 24 publications

References 45 publications

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Role of Tyrosine Kinase Syk in Thrombus Stabilisation at High Shear

Profiling the Genetic and Molecular Characteristics of Glanzmann Thrombasthenia: Can It Guide Current and Future Therapies?

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