title: The low-frequency allele of the platelet collagen signaling receptor glycoprotein VI is associated with reduced functional responses and expression.
abstract: Interaction of platelets with collagen under conditions of blood flow is a multi-step process with tethering via glycoprotein IbIXV (GPIbIXV) over von Willebrand factor, adhesion by direct interaction with the integrin GPIaIIa, and signaling via GPVI. GPVI can be specifically agonized by cross-linked collagen-related peptide (CRP-XL), which results in a signaling cascade very similar to that evoked by native collagen. The GPVI gene has 2 common alleles that differ by 3 replacements in the glycosylated stem and 2 in the cytoplasmic domain. We used CRP-XL to elucidate the variation in responses observed in platelet function in different individuals. We observed a 3-fold difference in the response to CRP-XL in platelet aggregation when comparing platelets from 10 high-frequency allele homozygotes with 8 low-frequency ones (2-way analysis of variance [ANOVA], P <.0001). The difference in functional responses was reflected in fibrinogen binding and in downstream signaling events as measured by tyrosine phosphorylation, the expression of P-selectin, and the binding of annexin V and the generation of thrombin on the platelet surface (2-way ANOVA, P <.001). Platelets homozygous for the low-frequency allele tended to be less able to form a thrombus on a collagen surface in flowing whole blood or in the platelet function analyzer-100 (t test, P =.065 and P =.061, respectively). The functional difference was correlated to a difference in total and membrane-expressed GPVI measured by monoclonal and polyclonal antibodies. This study demonstrates for the first time that platelet function may be altered by allelic differences in GPVI.
authors.affiliation: Department of Haematology, University of Cambridge, United Kingdom.
authors.forename: Lotta
authors.initials: L
authors.lastname: Joutsi-Korhonen
authors.forename: Peter A
authors.initials: PA
authors.lastname: Smethurst
authors.forename: Angela
authors.initials: A
authors.lastname: Rankin
authors.forename: Elaine
authors.initials: E
authors.lastname: Gray
authors.forename: Martin
authors.initials: M
authors.lastname: IJsseldijk
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Onley
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Watkins
authors.forename: Lorna M
authors.initials: LM
authors.lastname: Williamson
authors.forename: Alison H
authors.initials: AH
authors.lastname: Goodall
authors.forename: Philip G
authors.initials: PG
authors.lastname: de Groot
authors.forename: Richard W
authors.initials: RW
authors.lastname: Farndale
authors.forename: Willem H
authors.initials: WH
authors.lastname: Ouwehand
mesh_terms: D000328:Adult; D002352:Carrier Proteins / Q000494:pharmacology; D003094:Collagen / Q000494:pharmacology; D005260:Female; D005787:Gene Frequency; D006720:Homozygote; D006801:Humans; D008297:Male; D008875:Middle Aged; D010455:Peptides; D010766:Phosphorylation / Q000187:drug effects; D015539:Platelet Activation / Q000187:drug effects; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000235:genetics* / Q000502:physiology*; D020641:Polymorphism, Single Nucleotide; D038981:Receptors, Collagen; D013917:Thrombin / Q000096:biosynthesis / Q000187:drug effects
chemical_list: D002352:Carrier Proteins; D010455:Peptides; D010980:Platelet Membrane Glycoproteins; D038981:Receptors, Collagen; C435908:collagen-related peptide; C098341:platelet membrane glycoprotein VI; D003094:Collagen; D013917:Thrombin