TY - JOUR
T1 - High-throughput elucidation of thrombus formation reveals sources of platelet function variability
AU - van Geffen, Ankie
AU - Brouns, Sanne
AU - Batista, Joana
AU - McKinney, Harriet
AU - Kempster, Carly
AU - Nagy, Magdi
AU - Sivapalaratnam, Suthesh
AU - Baaten, Constance
AU - Bourry, Nikki
AU - Frontini, Mattia
AU - Jurk, Kerstin
AU - Krause, Manuela
AU - Pilliteri, Daniele
AU - Swieringa, Frauke
AU - Verdoold, Remco
AU - Cavill, Rachel
AU - Kuijpers, Marijke
AU - Ouwehand, Willem H.
AU - Downes, Kate
AU - Heemskerk, Johan
PY - 2019/5/31
Y1 - 2019/5/31
N2 - In combination with microspotting, whole-blood microfluidics can provide high-throughput information on multiple platelet functions in thrombus formation. Based on assessment of the inter-and intra-subject variability in parameters of microspot-based thrombus formation, we aimed to determine the platelet factors contributing to this variation. Blood samples from 94 genotyped healthy subjects were analyzed for conventional platelet phenotyping: i.e. hematologic parameters, platelet glycoprotein (GP) expression levels and activation markers (24 parameters). Furthermore, platelets were activated by ADP, CRP-XL or TRAP. Parallel samples were investigated for whole-blood thrombus formation (6 microspots, providing 48 parameters of adhesion, aggregation and activation). Microspots triggered platelet activation through GP Ib-V-IX, GPVI, CLEC-2 and integrins. For most thrombus parameters, inter-subject variation was 2-4 times higher than the intra-subject variation. Principal component analyses indicated coherence between the majority of parameters for the GPVI-dependent microspots, partly linked to hematologic parameters, and glycoprotein expression levels. Prediction models identified parameters per microspot that were linked to variation in agonist-induced alpha(IIb)beta(3) activation and secretion. Common sequence variation of GP6 and FCER1G, associated with GPVI-induced alpha(IIb)beta(3) activation and secretion, affected parameters of GPVI-and CLEC-2-dependent thrombus formation. Subsequent analysis of blood samples from patients with Glanzmann thrombasthenia or storage pool disease revealed thrombus signatures of aggregation-dependent parameters that were subject-dependent, but not linked to GPVI activity. Taken together, this high-throughput elucidation of thrombus formation revealed patterns of inter-subject differences in platelet function, which were partly related to GPVI-induced activation and common genetic variance linked to GPVI, but also included a distinct platelet aggregation component.
AB - In combination with microspotting, whole-blood microfluidics can provide high-throughput information on multiple platelet functions in thrombus formation. Based on assessment of the inter-and intra-subject variability in parameters of microspot-based thrombus formation, we aimed to determine the platelet factors contributing to this variation. Blood samples from 94 genotyped healthy subjects were analyzed for conventional platelet phenotyping: i.e. hematologic parameters, platelet glycoprotein (GP) expression levels and activation markers (24 parameters). Furthermore, platelets were activated by ADP, CRP-XL or TRAP. Parallel samples were investigated for whole-blood thrombus formation (6 microspots, providing 48 parameters of adhesion, aggregation and activation). Microspots triggered platelet activation through GP Ib-V-IX, GPVI, CLEC-2 and integrins. For most thrombus parameters, inter-subject variation was 2-4 times higher than the intra-subject variation. Principal component analyses indicated coherence between the majority of parameters for the GPVI-dependent microspots, partly linked to hematologic parameters, and glycoprotein expression levels. Prediction models identified parameters per microspot that were linked to variation in agonist-induced alpha(IIb)beta(3) activation and secretion. Common sequence variation of GP6 and FCER1G, associated with GPVI-induced alpha(IIb)beta(3) activation and secretion, affected parameters of GPVI-and CLEC-2-dependent thrombus formation. Subsequent analysis of blood samples from patients with Glanzmann thrombasthenia or storage pool disease revealed thrombus signatures of aggregation-dependent parameters that were subject-dependent, but not linked to GPVI activity. Taken together, this high-throughput elucidation of thrombus formation revealed patterns of inter-subject differences in platelet function, which were partly related to GPVI-induced activation and common genetic variance linked to GPVI, but also included a distinct platelet aggregation component.
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KW - RESPONSES
KW - DIFFERENCE
KW - ADHESION
U2 - 10.3324/haematol.2018.198853
DO - 10.3324/haematol.2018.198853
M3 - Article
C2 - 30545925
SN - 0390-6078
VL - 104
SP - 1256
EP - 1267
JO - Haematologica-the Hematology Journal
JF - Haematologica-the Hematology Journal
IS - 6
ER -