Long-term platelet priming after glycoprotein VI stimulation in comparison to Protease-Activating Receptor (PAR) stimulation

Zou, Jinmi; Wu, Jiayu; Roest, Mark; Heemskerk, Johan W. M.

doi:10.1371/journal.pone.0247425

Cited by 10 publications

(8 citation statements)

References 35 publications

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“…A common subsequent event is store-operated Ca 2+ entry ( Mammadova-Bach et al., 2019 ). Although it is understood that the GPCR-induced platelet signaling response is of shorter duration than the GPVI-induced response ( Fernandez et al., 2020 ; Zou et al., 2021 ), this difference has not been examined in detail so far. In the present paper, we hypothesized that detailed analysis of the cytosolic free Ca 2+ dynamics can help in the screening for drugs discriminating between platelet GPCR and ILR agonists.…”

Section: Introductionmentioning

confidence: 99%

Ultra-high-throughput Ca2+ assay in platelets to distinguish ITAM-linked and G-protein-coupled receptor activation

et al. 2022

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Summary Antiplatelet drugs targeting G-protein-coupled receptors (GPCRs), used for the secondary prevention of arterial thrombosis, coincide with an increased bleeding risk. Targeting ITAM-linked receptors, such as the collagen receptor glycoprotein VI (GPVI), is expected to provide a better antithrombotic-hemostatic profile. Here, we developed and characterized an ultra-high-throughput (UHT) method based on intracellular [Ca 2+ ] i increases to differentiate GPVI and GPCR effects on platelets. In 96-, 384-, or 1,536-well formats, Calcium-6-loaded human platelets displayed a slow-prolonged or fast-transient [Ca 2+ ] i increase when stimulated with the GPVI agonist collagen-related peptide or with thrombin and other GPCR agonists, respectively. Semi-automated curve fitting revealed five parameters describing the Ca 2+ responses. Verification of the UHT assay was done with a robustness compound library and clinically relevant platelet inhibitors. Taken together, these results present proof of principle of distinct receptor-type-dependent Ca 2+ signaling curves in platelets, which allow identification of new inhibitors in a UHT way.

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Section: Introductionmentioning

confidence: 99%

Ultra-high-throughput Ca2+ assay in platelets to distinguish ITAM-linked and G-protein-coupled receptor activation

et al. 2022

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“…Our present data suggest that a later, fibrin-dependent role of GPVI is yet smaller than the initial collagen-dependent role. Both GPVI- and PAR-mediated activation can have mutually stimulating effects on platelet activation processes [ 54 ]. It appeared that the ‘memory’ of platelets for GPVI-induced activation is longer than for PAR-induced activation.…”

Section: Discussionmentioning

confidence: 99%

Temporal Roles of Platelet and Coagulation Pathways in Collagen- and Tissue Factor-Induced Thrombus Formation

Navarro

Stegner

Nieswandt

et al. 2021

IJMS

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In hemostasis and thrombosis, the complex process of thrombus formation involves different molecular pathways of platelet and coagulation activation. These pathways are considered as operating together at the same time, but this has not been investigated. The objective of our study was to elucidate the time-dependency of key pathways of thrombus and clot formation, initiated by collagen and tissue factor surfaces, where coagulation is triggered via the extrinsic route. Therefore, we adapted a microfluidics whole-blood assay with the Maastricht flow chamber to acutely block molecular pathways by pharmacological intervention at desired time points. Application of the technique revealed crucial roles of glycoprotein VI (GPVI)-induced platelet signaling via Syk kinase as well as factor VIIa-induced thrombin generation, which were confined to the first minutes of thrombus buildup. A novel anti-GPVI Fab EMF-1 was used for this purpose. In addition, platelet activation with the protease-activating receptors 1/4 (PAR1/4) and integrin αIIbβ3 appeared to be prolongedly active and extended to later stages of thrombus and clot formation. This work thereby revealed a more persistent contribution of thrombin receptor-induced platelet activation than of collagen receptor-induced platelet activation to the thrombotic process.

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“…The importance of PKA in suppressing platelet aggregation activation becomes clear from the fact that the secondary application of iloprost (a prostacyclin analogue) can reverse the integrin activation in response to multiple agonists ( Table 1 ) [ 110 ]. In addition, gain-of-function mutations in the Gsα protein lead to elevated platelet cAMP levels, lower aggregate formation and a bleeding phenotype [ 111 ], whereas loss-of-function mutations leads to an impaired platelet inhibition with iloprost [ 100 ].…”

Section: Platelet Inhibition By Protein Kinases a And Gmentioning

confidence: 99%

Reversible Platelet Integrin αIIbβ3 Activation and Thrombus Instability

Zou

Swieringa

Laat

et al. 2022

IJMS

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Integrin αIIbβ3 activation is essential for platelet aggregation and, accordingly, for hemostasis and arterial thrombosis. The αIIbβ3 integrin is highly expressed on platelets and requires an activation step for binding to fibrinogen, fibrin or von Willebrand factor (VWF). A current model assumes that the process of integrin activation relies on actomyosin force-dependent molecular changes from a bent-closed and extended-closed to an extended-open conformation. In this paper we review the pathways that point to a functional reversibility of platelet αIIbβ3 activation and transient aggregation. Furthermore, we refer to mouse models indicating that genetic defects that lead to reversible platelet aggregation can also cause instable thrombus formation. We discuss the platelet agonists and signaling pathways that lead to a transient binding of ligands to integrin αIIbβ3. Our analysis points to the (autocrine) ADP P2Y1 and P2Y12 receptor signaling via phosphoinositide 3-kinases and Akt as principal pathways linked to reversible integrin activation. Downstream signaling events by protein kinase C, CalDAG-GEFI and Rap1b have not been linked to transient integrin activation. Insight into the functional reversibility of integrin activation pathways will help to better understand the effects of antiplatelet agents.

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Long-term platelet priming after glycoprotein VI stimulation in comparison to Protease-Activating Receptor (PAR) stimulation

Cited by 10 publications

References 35 publications

Ultra-high-throughput Ca2+ assay in platelets to distinguish ITAM-linked and G-protein-coupled receptor activation

Ultra-high-throughput Ca2+ assay in platelets to distinguish ITAM-linked and G-protein-coupled receptor activation

Temporal Roles of Platelet and Coagulation Pathways in Collagen- and Tissue Factor-Induced Thrombus Formation

Reversible Platelet Integrin αIIbβ3 Activation and Thrombus Instability

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