High-throughput platelet spreading analysis: a tool for the diagnosis of platelet-based bleeding disorders

Khan, Abdullah O.; Maclachlan, Annabel; Lowe, Gillian; Nicolson, Phillip L.R.; Ghaithi, Rashid Al; Thomas, Steven G.; Watson, Steve P.; Pike, Jeremy; Morgan, Neil V.

doi:10.3324/haematol.2019.225912

Cited by 22 publications

(21 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…32,117 The evaluation of platelet spreading on single protein substrates provides more information. 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.…”

Section: Diagnosismentioning

confidence: 99%

See 1 more Smart Citation

Inherited thrombocytopenias: history, advances and perspectives

Nurden

2020

Haematologica

View full text Add to dashboard Cite

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

show abstract

Section: Diagnosismentioning

confidence: 99%

Section: Diagnosismentioning

confidence: 99%

Inherited thrombocytopenias: history, advances and perspectives

Nurden

2020

Haematologica

View full text Add to dashboard Cite

show abstract

“…Peripheral blood was collected from patients and platelet phenotyping using lumi-aggregometry or flow cytometry was performed on platelet-rich plasma, as previously described. 6,7 Our study cohort consisted of 129 patients with a strong history of bleeding and suspected of having a platelet function disorder of unknown cause as previously described. 1,2 The ISTH Bleeding Assessment Tool (BAT) showed a mean score of 9.825 overall (range 2-23).…”

Section: Research Ethics Service By the Research Ethics Committee Of mentioning

confidence: 99%

Novel gene variants in patients with platelet‐based bleeding using combined exome sequencing and RNAseq murine expression data

Khan

Stapley

Pike

et al. 2021

Journal of Thrombosis and Haemostasis

Self Cite

View full text Add to dashboard Cite

Identifying genetic variants in platelet disorders is challenging due to its heterogenous nature. We combine WES, RNAseq, and python‐based bioinformatics to identify novel gene variants. We find novel candidates in patient data by cross‐referencing against a murine RNAseq model of thrombopoiesis. This innovative combined bioinformatic approach provides novel data for future research in the field. Abstract BackgroundThe UK Genotyping and Phenotyping of Platelets study has recruited and analyzed 129 patients with suspected heritable bleeding. Previously, 55 individuals had a definitive genetic diagnosis based on whole exome sequencing (WES) and platelet morphological and functional testing. A significant challenge in this field is defining filtering criteria to identify the most likely candidate mutations for diagnosis and further study. ObjectiveIdentify candidate gene mutations for the remaining 74 patients with platelet‐based bleeding with unknown genetic cause, forming the basis of future re‐recruitment and further functional testing and assessment. MethodsUsing python‐based data frame indexing, we first identify and filter all novel and rare variants using a panel of 116 genes known to cause bleeding across the full cohort of WES data. This identified new variants not previously reported in this cohort. We then index the remaining patients, with rare or novel variants in known bleeding genes against a murine RNA sequencing dataset that models proplatelet‐forming megakaryocytes. ResultsFiltering against known genes identified candidate variants in 59 individuals, including novel variants in several known genes. In the remaining cohort of “unknown” patients, indexing against differentially expressed genes revealed candidate gene variants in several novel unreported genes, focusing on 14 patients with a severe clinical presentation. ConclusionsWe identified candidate mutations in a cohort of patients with no previous genetic diagnosis. This work involves innovative coupling of RNA sequencing and WES to identify candidate variants forming the basis of future study in a significant number of undiagnosed patients.

show abstract

“…These syndromes are challenging to detect or evaluate using conventional platelet function testing due to associated thrombocytopenia (low platelet count). Of note, platelet spreading assays, which are not affected by low platelet count, can help define bleeding phenotypes in patient samples that are negative for an aggregation defect (85). The combination of biological optical microimaging with genomic information has opened up new avenues to test and evaluate these rare conditions that are not limited by low platelet counts but are still constrained by the limits of optical diffraction (86,87).…”

Section: Micro-imaging Platelets In a Dishmentioning

confidence: 99%

Imaging Platelet Processes and Function—Current and Emerging Approaches for Imaging in vitro and in vivo

et al. 2020

View full text Add to dashboard Cite

Platelets are small anucleate cells that are essential for many biological processes including hemostasis, thrombosis, inflammation, innate immunity, tumor metastasis, and wound healing. Platelets circulate in the blood and in order to perform all of their biological roles, platelets must be able to arrest their movement at an appropriate site and time. Our knowledge of how platelets achieve this has expanded as our ability to visualize and quantify discreet platelet events has improved. Platelets are exquisitely sensitive to changes in blood flow parameters and so the visualization of rapid intricate platelet processes under conditions found in flowing blood provides a substantial challenge to the platelet imaging field. The platelet's size (∼2 µm), rapid activation (milliseconds), and unsuitability for genetic manipulation, means that appropriate imaging tools are limited. However, with the application of modern imaging systems to study platelet function, our understanding of molecular events mediating platelet adhesion from a single-cell perspective, to platelet recruitment and activation, leading to thrombus (clot) formation has expanded dramatically. This review will discuss current platelet imaging techniques in vitro and in vivo, describing how the advancements in imaging have helped answer/expand on platelet biology with a particular focus on hemostasis. We will focus on platelet aggregation and thrombus formation, and how platelet imaging has enhanced our understanding of key events, highlighting the knowledge gained through the application of imaging modalities to experimental models in vitro and in vivo. Furthermore, we will review the limitations of current imaging techniques, and questions in thrombosis research that remain to be addressed. Finally, we will speculate how the same imaging advancements might be applied to the imaging of other vascular cell biological functions and visualization of dynamic cell-cell interactions.

show abstract

High-throughput platelet spreading analysis: a tool for the diagnosis of platelet-based bleeding disorders

Cited by 22 publications

References 15 publications

Inherited thrombocytopenias: history, advances and perspectives

Inherited thrombocytopenias: history, advances and perspectives

Novel gene variants in patients with platelet‐based bleeding using combined exome sequencing and RNAseq murine expression data

Imaging Platelet Processes and Function—Current and Emerging Approaches for Imaging in vitro and in vivo

Contact Info

Product

Resources

About