High-throughput elucidation of thrombus formation reveals sources of platelet function variability

Geffen, Johanna P. van; Brouns, Sanne L. N.; Batista, Joana; McKinney, Harriet; Kempster, Carly; Nagy, Magdolna; Sivapalaratnam, Suthesh; Baaten, Constance C. F. M. J.; Bourry, Nikki; Frontini, Mattia; Jurk, Kerstin; Krause, Manuela; Pillitteri, Daniele; Swieringa, Frauke; Verdoold, Remco; Cavill, Rachel; Kuijpers, Marijke J. E.; Ouwehand, Willem H.; Downes, Kate; Heemskerk, Johan W. M.

doi:10.3324/haematol.2018.198853

Cited by 64 publications

(125 citation statements)

References 29 publications

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“…120 The use of computer-based high-throughput testing of platelet function and thrombus formation under flow on extracellular matrix proteins on microchips is full of promise but has been little tested for thrombocytopenias. 121 As highlighted throughout our review, applying NGS to inherited thrombocytopenias is greatly expanding the list of causative genes as well as increasing the number of variants implicated in the classic diseases. The initial success of WES in genotyping gray platelet syndrome and the TAR syndromes led to the BRIDGE-Bleeding and Platelet Disorders (BRIDGE-BPD) project, orchestrated by Professor W. Ouwehand, which combines the unique sequencing and bioinformatics resources of the Sanger Institute in Cambridge (UK).…”

Section: Diagnosismentioning

confidence: 98%

Inherited thrombocytopenias: history, advances and perspectives

Nurden

2020

Haematologica

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In the late 19 th century improvements to the light microscope led to anucleate platelets being visualized in great numbers in human blood. Early pioneers in the field of platelet research included the Canadian William Osler, a Paris hematologist, George Hayem, who performed the first accurate platelet count, and the Italian Giulio Bizzozero. 1 In 1906, James Homer Wright confirmed that platelets were produced by bone marrow megakaryocytes. 2 When, in 1951, Harrington et al. 3 observed purpura in a child of a mother with immune thrombocytopenic purpura, a maternal factor was said to be destroying the platelets. Anti-platelet antibodies were identified as the cause and platelet transfusions, immunosuppressive therapy and splenectomy became standard treatments. Acquired thrombocytopenia, often with defective platelet function, has many causes. For example, it may be an immune response linked to blood transfusions and drugs, a direct consequence of viral or bacterial infections, be the result of other hematologic disorders or be secondary to many major illnesses. However, some thrombocytopenias are inherited and Professors Jean Bernard and Jean-Pierre Soulier in Paris were pioneers in this domain, describing in 1948 what they called in French "dystrophie thrombocytaire hémorragipare congénitale", later renamed the Bernard-Soulier syndrome (BSS). 4 Strikingly, many platelets were enlarged and some were giant. A key to the molecular defect was the platelet deficit of sialic acid, a negatively charged monosaccharide terminating many of the oligosaccharides of platelet glycoproteins (GP) and

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Section: Diagnosismentioning

confidence: 98%

Inherited thrombocytopenias: history, advances and perspectives

Nurden

2020

Haematologica

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“…GP5 (human platelet glycoprotein V) is a part of the Ib-V-IX system of surface glycoproteins that comprise the receptor for von Willebrand factor and mediate the adhesion of platelets to injured vascular surfaces in the arterial circulation (274)(275)(276)(277)(278). GP6 forms a complex with the Fc receptor gamma-chain to initiate the platelet activation signaling cascade upon collagen binding (279)(280)(281)(282)(283)(284) and may be associated with an increased risk of platelet-related disorders including ischemic stroke. PLOD2 (procollagen-lysine,2-oxoglutarate 5-dioxygenase 2) catalyzes the hydroxylation of lysyl residues in collagen-like peptides and COL6A3 (collagen type VI alpha 3 chain) is a general structural matrix protein and is fairly ubiquitously expressed.…”

Section: Myl9mentioning

confidence: 99%

Development of a Bioinformatics Framework for Identification and Validation of Genomic Biomarkers and Key Immunopathology Processes and Controllers in Infectious and Non-infectious Severe Inflammatory Response Syndrome

et al. 2020

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Sepsis is defined as dysregulated host response caused by systemic infection, leading to organ failure. It is a life-threatening condition, often requiring admission to an intensive care unit (ICU). The causative agents and processes involved are multifactorial but are characterized by an overarching inflammatory response, sharing elements in common with severe inflammatory response syndrome (SIRS) of non-infectious origin. Sepsis presents with a range of pathophysiological and genetic features which make clinical differentiation from SIRS very challenging. This may reflect a poor understanding of the key gene inter-activities and/or pathway associations underlying these disease processes. Improved understanding is critical for early differential recognition of sepsis and SIRS and to improve patient management and clinical outcomes. Judicious selection of gene biomarkers suitable for development of diagnostic tests/testing could make differentiation of sepsis and SIRS feasible. Here we describe a methodologic framework for the identification and validation of biomarkers in SIRS, sepsis and septic shock patients, using a 2-tier gene screening, artificial neural network (ANN) data mining technique, using previously published gene expression datasets. Eight key hub markers have been identified which may delineate distinct, core disease processes and which show potential for informing underlying immunological and pathological processes and thus patient stratification and treatment. These do not show sufficient fold change differences between the different disease states to be useful as primary diagnostic biomarkers, but are instrumental in identifying candidate pathways and other associated biomarkers for further exploration.

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“…Images were analysed using Fiji software [32]. Four outcome parameters were assessed: platelet surface area coverage, thrombus morphological score (0-5), thrombus contraction score (0-3) and platelet multilayer score (0-3) as previously explained [31,33]. Scoring was done by visual inspection of brightfield images based on a pre-defined score system.…”

Section: Whole Blood Thrombus Formationmentioning

confidence: 99%

“…Whole blood perfusion experiments were performed on collagen-coated microspots to assess the effects of platelet pretreatment with losartan or honokiol (25-100 µM). Thrombi were analysed for platelet deposition (% surface area coverage) and by visual inspection using a pre-defined score system (morphological, contraction and multilayer score) [33]. Losartan treatment did not affect platelet deposition (Figure 6a multilayer scores (Figure 6d,e) as an indication of lower thrombus integrity and the prpesence of loose platelet aggregates.…”

Section: Losartan and Honokiol Inhibit Thrombus Formationmentioning

confidence: 99%

Comparison of the GPVI inhibitors losartan and honokiol

et al. 2019

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Losartan and honokiol are small molecules which have been described to inhibit aggregation of platelets by collagen. Losartan has been proposed to block clustering of GPVI but not to affect binding of collagen. Honokiol has been reported to bind directly to GPVI but only at a concentration that is three orders of magnitude higher than that needed for inhibition of aggregation. The mechanism of action of both inhibitors is so far unclear. In the present study, we confirm the inhibitory effects of both agents on platelet aggregation by collagen and show that both also block the aggregation induced by the activation of CLEC-2 or the low affinity immune receptor FcγRIIa at similar concentrations. For GPVI and CLEC-2, this inhibition is associated with a reduction in protein tyrosine phosphorylation of multiple proteins including Syk. In contrast, on a collagen surface, spreading of platelets and clustering of GPVI (measured by single molecule localisation microscopy) was not altered by losartan or honokiol. Furthermore, in flow wholeblood, both inhibitors suppressed the formation of multi-layered platelet thrombi at arteriolar shear rates at concentrations that hardly affect collagen-induced platelet aggregation in platelet rich plasma. Together, these results demonstrate that losartan and honokiol have multiple effects on platelets which should be considered in the use of these compounds as anti-platelet agents.

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High-throughput elucidation of thrombus formation reveals sources of platelet function variability

Cited by 64 publications

References 29 publications

Inherited thrombocytopenias: history, advances and perspectives

Inherited thrombocytopenias: history, advances and perspectives

Development of a Bioinformatics Framework for Identification and Validation of Genomic Biomarkers and Key Immunopathology Processes and Controllers in Infectious and Non-infectious Severe Inflammatory Response Syndrome

Comparison of the GPVI inhibitors losartan and honokiol

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