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

Navarro, Stefano; Stegner, David; Nieswandt, Bernhard; Heemskerk, Johan W. M.; Kuijpers, Marijke J. E.

doi:10.3390/ijms23010358

Cited by 23 publications

(18 citation statements)

References 62 publications

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“…Next, we assessed whether JAQ1 binding would be affected by the pre-incubation of human platelets with different, established anti-huGPVI monoclonal antibodies (mAbs). Indeed, JAQ1-FITC binding was reduced by ~66% after pre-incubation with the function-blocking anti-huGPVI mAb EMF-1 [ 30 ], but only partially by the non-function-blocking EMF-2 (~26%) ( Figure 1 B). Subsequently, we tested whether JAQ1 recognizes huGPVI in a Western blot analysis of human platelet lysate.…”

Section: Resultsmentioning

confidence: 99%

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Targeting of a Conserved Epitope in Mouse and Human GPVI Differently Affects Receptor Function

Navarro

Starke

Heemskerk

et al. 2022

IJMS

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Glycoprotein (GP) VI is the major platelet collagen receptor and a promising anti-thrombotic target. This was first demonstrated in mice using the rat monoclonal antibody JAQ1, which completely blocks the Collagen-Related Peptide (CRP)-binding site on mouse GPVI and efficiently inhibits mouse platelet adhesion, activation and aggregation on collagen. Here, we show for the first time that JAQ1 cross-reacts with human GPVI (huGPVI), but not with GPVI in other tested species, including rat, rabbit, guinea pig, swine, and dog. We further demonstrate that JAQ1 differently modulates mouse and human GPVI function. Similar to its effects on mouse GPVI (mGPVI), JAQ1 inhibits CRP-induced activation in human platelets, whereas, in stark contrast to mouse GPVI, it does not inhibit the adhesion, activation or aggregate formation of human platelets on collagen, but causes instead an increased response. This effect was also seen with platelets from newly generated human GPVI knockin mice (hGP6tg/tg). These results indicate that the binding of JAQ1 to a structurally conserved epitope in GPVI differently affects its function in human and mouse platelets.

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Section: Resultsmentioning

confidence: 99%

“…The heparin was purchased from Ratiopharm (Ulm, Germany). The JAQ1 [ 24 ], EMF-1 [ 30 ], EMF-2, JON/A [ 42 , 43 ] and WUG 1.9 were generated in house. The IV.3 antibody was purchased from Biozol (Eching, Germany).…”

Section: Methodsmentioning

confidence: 99%

Targeting of a Conserved Epitope in Mouse and Human GPVI Differently Affects Receptor Function

Navarro

Starke

Heemskerk

et al. 2022

IJMS

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“…In hemostasis and thrombosis, the generation of thrombin is in part triggered by vascular-exposed tissue factor. Kinetic studies with flowed blood have shown that the role of tissue factor in platelet aggregation and thrombus formation is only short-term [ 44 ]. Interestingly, one report states that this role of tissue factor can depend on factor VII-activating protein (FSAP, gene HAPP2 ).…”

Section: Thrombin Par1 and Par4 Receptor Stimulationmentioning

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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“…Recent multiparameter microfluidics assays have provided first understanding of the molecular mechanisms, implicated in whole-blood thrombus formation involving GPVI [ 10 , 11 , 12 ]. The heterogenous buildup of arterial thrombi takes place by a series of consecutive platelet activation events, i.e., flow-dependent platelet adhesion, integrin αIIbβ3 activation, granular secretion, platelet aggregation, and thrombus contraction and stabilization [ 8 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

Roles of Focal Adhesion Kinase PTK2 and Integrin αIIbβ3 Signaling in Collagen- and GPVI-Dependent Thrombus Formation under Shear

Huang

Jooss

Fernández

et al. 2022

IJMS

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Glycoprotein (GP)VI and integrin αIIbβ3 are key signaling receptors in collagen-dependent platelet aggregation and in arterial thrombus formation under shear. The multiple downstream signaling pathways are still poorly understood. Here, we focused on disclosing the integrin-dependent roles of focal adhesion kinase (protein tyrosine kinase 2, PTK2), the shear-dependent collagen receptor GPR56 (ADGRG1 gene), and calcium and integrin-binding protein 1 (CIB1). We designed and synthetized peptides that interfered with integrin αIIb binding (pCIB and pCIBm) or mimicked the activation of GPR56 (pGRP). The results show that the combination of pGRP with PTK2 inhibition or of pGRP with pCIB > pCIBm in additive ways suppressed collagen- and GPVI-dependent platelet activation, thrombus buildup, and contraction. Microscopic thrombus formation was assessed by eight parameters (with script descriptions enclosed). The suppressive rather than activating effects of pGRP were confined to blood flow at a high shear rate. Blockage of PTK2 or interference of CIB1 no more than slightly affected thrombus formation at a low shear rate. Peptides did not influence GPVI-induced aggregation and Ca2+ signaling in the absence of shear. Together, these data reveal a shear-dependent signaling axis of PTK2, integrin αIIbβ3, and CIB1 in collagen- and GPVI-dependent thrombus formation, which is modulated by GPR56 and exclusively at high shear. This work thereby supports the role of PTK2 in integrin αIIbβ3 activation and signaling.

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

Cited by 23 publications

References 62 publications

Targeting of a Conserved Epitope in Mouse and Human GPVI Differently Affects Receptor Function

Targeting of a Conserved Epitope in Mouse and Human GPVI Differently Affects Receptor Function

Reversible Platelet Integrin αIIbβ3 Activation and Thrombus Instability

Roles of Focal Adhesion Kinase PTK2 and Integrin αIIbβ3 Signaling in Collagen- and GPVI-Dependent Thrombus Formation under Shear

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