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Fibrin exposure triggers αIIbβ3-independent platelet aggregate formation, ADAM10 activity and glycoprotein VI shedding in a charge-dependent manner

Montague, Samantha; Hicks, Sarah; Lee, Christine; Coupland, Lucy; Parish, Christopher; Lee, W M Steve; Andrews, Robert; Gardiner, Elizabeth

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Background Collagen and fibrin engagement and activation of glycoprotein (GP) VI induces proteolytic cleavage of the GPVI ectodomain generating shed soluble GPVI (sGPVI). Collagen‐mediated GPVI shedding requires intracellular signalling to release the sGPVI, mediated by A Disintegrin And Metalloproteinase 10 (ADAM10); however, the precise mechanism by which fibrin induces GPVI shedding remains elusive. Plasma sGPVI levels are elevated in patients with coagulopathies, sepsis, or inflammation...[Show more]

dc.contributor.authorMontague, Samantha
dc.contributor.authorHicks, Sarah
dc.contributor.authorLee, Christine
dc.contributor.authorCoupland, Lucy
dc.contributor.authorParish, Christopher
dc.contributor.authorLee, W M Steve
dc.contributor.authorAndrews, Robert
dc.contributor.authorGardiner, Elizabeth
dc.date.accessioned2021-01-12T03:30:03Z
dc.identifier.issn1538-7933
dc.identifier.urihttp://hdl.handle.net/1885/219290
dc.description.abstractBackground Collagen and fibrin engagement and activation of glycoprotein (GP) VI induces proteolytic cleavage of the GPVI ectodomain generating shed soluble GPVI (sGPVI). Collagen‐mediated GPVI shedding requires intracellular signalling to release the sGPVI, mediated by A Disintegrin And Metalloproteinase 10 (ADAM10); however, the precise mechanism by which fibrin induces GPVI shedding remains elusive. Plasma sGPVI levels are elevated in patients with coagulopathies, sepsis, or inflammation and can predict onset of sepsis and sepsis‐related mortality; therefore, it is clinically important to understand the mechanisms of GPVI shedding under conditions of minimal collagen exposure. Objectives Our aim was to characterize mechanisms by which fibrin‐GPVI interactions trigger GPVI shedding. Methods Platelet aggregometry, sGPVI ELISA, and an ADAM10 fluorescence resonance energy transfer assay were used to measure fibrin‐mediated platelet responses. Results Fibrin induced αIIbβ3‐independent washed platelet aggregate formation, GPVI shedding, and increased ADAM10 activity, all of which were insensitive to pre‐treatment with inhibitors of Src family kinases but were divalent cation‐ and metalloproteinase‐dependent. In contrast, treatment of washed platelets with other GPVI ligands, collagen, and collagen‐related peptide caused αIIbβ3‐dependent platelet aggregation and GPVI release but did not increase constitutive ADAM10 activity. Conclusions Fibrin engages GPVI in a manner that differs from other GPVI ligands. Inclusion of polyanionic molecules disrupted fibrin‐induced platelet aggregate formation and sGPVI release, suggesting that electrostatic charge may play a role in fibrin/GPVI engagement. It may be feasible to exploit this property and specifically disrupt GPVI/fibrin interactions whilst sparing GPVI/collagen engagement.Fibrin engages GPVI in a manner that differs from other GPVI ligands. Inclusion of polyanionic molecules disrupted fibrin‐induced platelet aggregate formation and sGPVI release, suggesting that electrostatic charge may play a role in fibrin/GPVI engagement. It may be feasible to exploit this property and specifically disrupt GPVI/fibrin interactions whilst sparing GPVI/collagen engagement.
dc.description.sponsorshipNational Health and Medical Research Council of Australia; Australian Research Council; THANZ Science and Education Research Grant
dc.format.mimetypeapplication/pdf
dc.language.isoen_AU
dc.publisherWiley-Blackwell Publishing Ltd
dc.rights© 2020 International Society on Thrombosis and Haemostasis
dc.sourceJournal of Thrombosis and Haemostasis
dc.subjectfibrin
dc.subjectGPVI
dc.subjectADAM10
dc.subjectreceptor shedding
dc.subjectthrombosis
dc.titleFibrin exposure triggers αIIbβ3-independent platelet aggregate formation, ADAM10 activity and glycoprotein VI shedding in a charge-dependent manner
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume18
dc.date.issued2020
local.identifier.absfor111202 - Cancer Diagnosis
local.identifier.ariespublicationa383154xPUB11350
local.publisher.urlhttps://www.wiley.com/en-gb
local.type.statusPublished Version
local.contributor.affiliationMontague, Samantha, College of Health and Medicine, ANU
local.contributor.affiliationHicks, Sarah, College of Health and Medicine, ANU
local.contributor.affiliationLee, Christine, College of Health and Medicine, ANU
local.contributor.affiliationCoupland, Lucy, College of Health and Medicine, ANU
local.contributor.affiliationParish, Christopher, College of Health and Medicine, ANU
local.contributor.affiliationLee, Steve, College of Engineering and Computer Science, ANU
local.contributor.affiliationAndrews, Robert, College of Health and Medicine, ANU
local.contributor.affiliationGardiner, Elizabeth, College of Health and Medicine, ANU
local.description.embargo2099-12-31
local.bibliographicCitation.issue6
local.bibliographicCitation.startpage1447
local.bibliographicCitation.lastpage1458
local.identifier.doi10.1111/jth.14797
local.identifier.absseo920103 - Cardiovascular System and Diseases
dc.date.updated2020-11-02T04:17:16Z
CollectionsANU Research Publications

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