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
N1 - Funding Information:
Support was obtained from the Cardiovascular Center (HVC), Maastricht University Medical Center, the Center for Translational Molecular Medicine (Incoag/Mikrobat), Interreg Euregio Meuse-Rhin (Polyvalve), the National Institute for Health Research (NIHR) Cambridge Biomedical Research Centre and the National Health Service Blood and Transplant (NHSBT). We gratefully acknowledge the participation of all NIHR BioResource volunteers, and thank the NIHR BioResource center and staff for their contribution. We also thank the National Institute for Health Research and NHS Blood and Transplant for funding support. KD is supported as a HSST trainee by Health Education England. Funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
© 2019 Ferrata Storti Foundation.
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.
KW - ADHESION
KW - DIFFERENCE
KW - DISORDERS
KW - RESPONSES
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 -