Prospective, international, multisite comparison of platelet isolation techniques for genome-wide transcriptomics: communication from the SSC of the ISTH.
| Autor: | Banerjee M; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA., Rowley JW; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA; Department of Internal Medicine, University of Utah Health, Salt Lake City, Utah, USA., Stubben CJ; Bioinformatics Shared Resource, Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah, USA., Tolley ND; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA., Freson K; Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, Katholeike Universiteit (KU) Leuven, Leuven, Belgium., Nelson B; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA., Nagy B Jr; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary., Fejes Z; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary., Blair AM; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA., Turro E; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA., Gresele P; Department of Medicine and Surgery, University of Perugia, Perugia, Italy., Taranta GC; Department of Medicine and Surgery, University of Perugia, Perugia, Italy., Bury L; Department of Medicine and Surgery, University of Perugia, Perugia, Italy., Falcinelli E; Department of Medicine and Surgery, University of Perugia, Perugia, Italy., Lordkipanidzé M; Research Center, Montreal Heart Institute, Montreal, Quebec, Canada; Faculty of Pharmacy, Université de Montréal, Montreal, Quebec, Canada., Alessi MC; Cardiovascular and Nutrition Centre (C2VN), Aix Marseille University, Institut National de la Sante et de la Recherche Medicale (INSERM), National Research Institute for Agriculture, Food and Environment (INRAE), Marseille, France., Johnson AD; Population Sciences Branch, Division of Intramural Research, National Heart, Lung and Blood Institute, Framingham, Massachusetts, USA; The Framingham Heart Study, Framingham, Massachusetts, USA., Bakchoul T; Department of Transfusion Medicine, Medical Faculty of Tubingen, University of Tubingen, Tubingen, Germany., Ramstrom S; Cardiovascular Research Centre, School of Medical Sciences, Faculty of Medicine and Health, Örebro University, Örebro, Sweden., Frontini M; Department of Clinical and Biomedical Sciences, University of Exeter Medical School, Faculty of Health and Life Sciences, Exeter, United Kingdom., Camera M; Unit of Cell and Molecular Biology in Cardiovascular Diseases, Centro Cardiologico Monzino, Instituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Milan, Italy; Department of Pharmaceutical Sciences, Università Degli Studi Di Milano, Milan, Italy., Brambilla M; Unit of Cell and Molecular Biology in Cardiovascular Diseases, Centro Cardiologico Monzino, Instituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Milan, Italy., Campbell RA; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA; Department of Internal Medicine, University of Utah Health, Salt Lake City, Utah, USA., Rondina MT; University of Utah Molecular Medicine Program, Eccles Institute of Human Genetics, Salt Lake City, Utah, USA; Department of Internal Medicine, University of Utah Health, Salt Lake City, Utah, USA; George E. Wahlen Veterans Affairs Medical Center & Geriatric Research Education and Clinical Center (GRECC), Salt Lake City, Utah, USA. Electronic address: matthew.rondina@hsc.utah.edu. |
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| Jazyk: | angličtina |
| Zdroj: | Journal of thrombosis and haemostasis : JTH [J Thromb Haemost] 2024 Oct; Vol. 22 (10), pp. 2922-2934. Date of Electronic Publication: 2024 Jul 03. |
| DOI: | 10.1016/j.jtha.2024.06.017 |
| Abstrakt: | Genome-wide platelet transcriptomics is increasingly used to uncover new aspects of platelet biology and as a diagnostic and prognostic tool. Nevertheless, platelet isolation methods for transcriptomic studies are not standardized, introducing challenges for cross-study comparisons, data integration, and replication. In this prospective multicenter study, called "Standardizing Platelet Transcriptomics for Discovery, Diagnostics, and Therapeutics in the Thrombosis and Hemostasis Community (STRIDE)" by the International Society on Thrombosis and Haemostasis Scientific and Standardization Committees, we assessed how 3 of the most commonly used platelet isolation protocols influence metrics from next-generation bulk RNA sequencing and functional assays. Compared with washing alone, more stringent removal of leukocytes by anti-CD45 beads or PALL filters resulted in a sufficient quantity of RNA for next-generation sequencing and similar quality of RNA sequencing metrics. Importantly, stringent removal of leukocytes resulted in the lower relative expression of known leukocyte-specific genes and the higher relative expression of known platelet-specific genes. The results were consistent across enrolling sites, suggesting that the techniques are transferrable and reproducible. Moreover, all 3 isolation techniques did not influence basal platelet reactivity, but agonist-induced integrin αIIbβ Competing Interests: Declaration of competing interests M.T.R. reports patents pending or issued on using platelet transcriptomics. All other authors have no conflicts to declare. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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