Phenotyping of human platelets in response to platelet agonists and inhibitors using multiparameter flow cytometry and unbiased high-dimensional analysis
Year: 2025
Type: article
Abstract: Background: Despite increasing evidence of functionally distinct platelet subpopulations, the role of receptor remodeling in this process is unclear. Objectives: To develop a high-dimensional multiparameter flow cytometry panel for platelet phenotyping and subpopulation analysis. Methods: Using conventional flow cytometry, we assessed the expression of 9 platelet surface receptors in whole blood (WB) samples from healthy adults in response to different platelet activators and inhibitors. Platelet subpopulations were identified by Full Annotation Using Shape-constrained Trees (FAUST). Results: (defined by PAC1 antibody binding) and expression of CD62P, CD40L, CD63, CD42b, CD147, CXCR4, CD32a, and CD36. After WB stimulation with adenosine diphosphate, thrombin receptor-activating peptide-6, or convulxin, FAUST identified 8 platelet subpopulations that were defined by differential levels of PAC1, CD62P, and CD63. To determine how inhibitors affect platelet subpopulations, WB was pretreated with prostacyclin, dasatinib, vorapaxar, or ticagrelor prior to stimulation with adenosine diphosphate, thrombin receptor-activating peptide-6, or convulxin. Again, FAUST identified 8 distinct platelet subpopulations, which were defined by differential levels of PAC1, CD40L, and CD62P, showing that under conditions of platelet inhibition, CD40L expression is important for subpopulation annotation. Conclusion: Platelet subpopulations are dynamic and undergo remodeling in response to different agonists and inhibitors. This high-dimensional single-cell flow cytometry assay is a practicable approach to evaluate the effects of agonists and inhibitors on platelet receptor expression and platelet subpopulations.
Authors: Daisie M. Yates, Benjamin E.J. Spurgeon, Lih T. Cheah, James I. Robinson, Ann W. Morgan, Khalid M. Naseem
Language: English
Subfield: Hematology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,250
Awards: Leeds 1st intake 2021 – 4-year PhD Studentship (5th) Scheme: Ms Marcella Conning-Rowland; Mr Michael Murray; Ms Jessica Wan, G1114017013, G1216683414, NIHR Senior Investigator Award, MICA: Treatment According to Response in Giant cEll arTeritis (TARGET), G6697820746, Dysregulation of platelet phosphodiesterase 3A isoforms promote platelet hyperactivity in atherothrombotic disease
OpenAlex ID: W4414477198