Mode of Induction of Platelet-derived Extracellular Vesicles is a Critical Determinant of Their Phenotype and Function

Overview

Journal Sci Rep

Specialty Science

Date 2020 Oct 23

PMID 33093473

Citations 17

Authors

P M Ferreira

E Bozbas

S D Tannetta

N Alroqaiba

R Zhou

J T B Crawley

J M Gibbins

C I Jones

J Ahnstrom

P Yaqoob

Affiliations

Soon will be listed here.

Abstract

Platelet-derived extracellular vesicles (PDEVs) are the most abundant amongst all types of EVs in the circulation. However, the mechanisms leading to PDEVs release, their role in coagulation and phenotypic composition are poorly understood. PDEVs from washed platelets were generated using different stimuli and were characterised using nanoparticle tracking analysis. Procoagulant properties were evaluated by fluorescence flow cytometry and calibrated automated thrombography. EVs from plasma were isolated and concentrated using a novel protocol involving a combination of size exclusion chromatography and differential centrifugation, which produces pure and concentrated EVs. Agonist stimulation enhanced PDEV release, but did not alter the average size of EVs compared to those produced by unstimulated platelets. Agonist stimulation led to lower negatively-charged phospholipid externalization in PDEVs, which was reflected in the lower procoagulant activity compared to those generated without agonist stimulation. Circulating EVs did not have externalized negatively-charged phospholipids. None of the 4 types of EVs presented tissue factor. The mechanism by which PDEV formation is induced is a critical determinant of its phenotype and function. Importantly, we have developed methods to obtain clean, concentrated and functional EVs derived from platelet-free plasma and washed platelets, which can be used to provide novel insight into their biological functions.

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References

Lechner D, Weltermann A . Circulating tissue factor-exposing microparticles. Thromb Res. 2008; 122 Suppl 1:S47-54. DOI: 10.1016/S0049-3848(08)70019-7. View

Arraud N, Linares R, Tan S, Gounou C, Pasquet J, Mornet S . Extracellular vesicles from blood plasma: determination of their morphology, size, phenotype and concentration. J Thromb Haemost. 2014; 12(5):614-27. DOI: 10.1111/jth.12554. View

Jy W, Horstman L, Arce M, Ahn Y . Clinical significance of platelet microparticles in autoimmune thrombocytopenias. J Lab Clin Med. 1992; 119(4):334-45. View

Hong C, Funk S, Muller L, Boyiadzis M, Whiteside T . Isolation of biologically active and morphologically intact exosomes from plasma of patients with cancer. J Extracell Vesicles. 2016; 5:29289. PMC: 4808740. DOI: 10.3402/jev.v5.29289. View

Morel O, Jesel L, Freyssinet J, Toti F . Cellular mechanisms underlying the formation of circulating microparticles. Arterioscler Thromb Vasc Biol. 2010; 31(1):15-26. DOI: 10.1161/ATVBAHA.109.200956. View

Buzas E, Gyorgy B, Nagy G, Falus A, Gay S . Emerging role of extracellular vesicles in inflammatory diseases. Nat Rev Rheumatol. 2014; 10(6):356-64. DOI: 10.1038/nrrheum.2014.19. View

Tripisciano C, Weiss R, Eichhorn T, Spittler A, Heuser T, Fischer M . Different Potential of Extracellular Vesicles to Support Thrombin Generation: Contributions of Phosphatidylserine, Tissue Factor, and Cellular Origin. Sci Rep. 2017; 7(1):6522. PMC: 5529579. DOI: 10.1038/s41598-017-03262-2. View

Hemker H, Giesen P, AlDieri R, Regnault V, de Smed E, Wagenvoord R . The calibrated automated thrombogram (CAT): a universal routine test for hyper- and hypocoagulability. Pathophysiol Haemost Thromb. 2003; 32(5-6):249-53. DOI: 10.1159/000073575. View

Aatonen M, Ohman T, Nyman T, Laitinen S, Gronholm M, Siljander P . Isolation and characterization of platelet-derived extracellular vesicles. J Extracell Vesicles. 2014; 3. PMC: 4125723. DOI: 10.3402/jev.v3.24692. View

10.

Nordin J, Lee Y, Vader P, Mager I, Johansson H, Heusermann W . Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties. Nanomedicine. 2015; 11(4):879-83. DOI: 10.1016/j.nano.2015.01.003. View

11.

van der Pol E, Boing A, Gool E, Nieuwland R . Recent developments in the nomenclature, presence, isolation, detection and clinical impact of extracellular vesicles. J Thromb Haemost. 2015; 14(1):48-56. DOI: 10.1111/jth.13190. View

12.

Zaldivia M, McFadyen J, Lim B, Wang X, Peter K . Platelet-Derived Microvesicles in Cardiovascular Diseases. Front Cardiovasc Med. 2017; 4:74. PMC: 5702324. DOI: 10.3389/fcvm.2017.00074. View

13.

Larson M, Woodliff J, Hillery C, Kearl T, Zhao M . Phosphatidylethanolamine is externalized at the surface of microparticles. Biochim Biophys Acta. 2012; 1821(12):1501-7. PMC: 3809829. DOI: 10.1016/j.bbalip.2012.08.017. View

14.

Clark S, Thomas C, Hammond V, Aldrovandi M, Wilkinson G, Hart K . Characterization of platelet aminophospholipid externalization reveals fatty acids as molecular determinants that regulate coagulation. Proc Natl Acad Sci U S A. 2013; 110(15):5875-80. PMC: 3625294. DOI: 10.1073/pnas.1222419110. View

15.

Wolf P . The nature and significance of platelet products in human plasma. Br J Haematol. 1967; 13(3):269-88. DOI: 10.1111/j.1365-2141.1967.tb08741.x. View

16.

Berckmans R, Nieuwland R, Boing A, Romijn F, Hack C, Sturk A . Cell-derived microparticles circulate in healthy humans and support low grade thrombin generation. Thromb Haemost. 2001; 85(4):639-46. View

17.

Abid Hussein M, Meesters E, Osmanovic N, Romijn F, Nieuwland R, Sturk A . Antigenic characterization of endothelial cell-derived microparticles and their detection ex vivo. J Thromb Haemost. 2003; 1(11):2434-43. DOI: 10.1046/j.1538-7836.2003.00455.x. View

18.

De Paoli S, Tegegn T, Elhelu O, Strader M, Patel M, Diduch L . Dissecting the biochemical architecture and morphological release pathways of the human platelet extracellular vesiculome. Cell Mol Life Sci. 2018; 75(20):3781-3801. PMC: 11105464. DOI: 10.1007/s00018-018-2771-6. View

19.

Ahnstrom J, Andersson H, Hockey V, Meng Y, McKinnon T, Hamuro T . Identification of functionally important residues in TFPI Kunitz domain 3 required for the enhancement of its activity by protein S. Blood. 2012; 120(25):5059-62. DOI: 10.1182/blood-2012-05-432005. View

20.

Dovizio M, Bruno A, Contursi A, Grande R, Patrignani P . Platelets and extracellular vesicles in cancer: diagnostic and therapeutic implications. Cancer Metastasis Rev. 2018; 37(2-3):455-467. DOI: 10.1007/s10555-018-9730-4. View