Download PDF
Thromb Haemost 2014; 112(01): 128-136
DOI: 10.1160/TH13-11-0977
Platelets and Blood Cells
Schattauer GmbH

Smoking alters circulating plasma microvesicle pattern and microRNA signatures

Sigrun Badrnya
1   Institute of Physiology, Centre for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
,
Roland Baumgartner
1   Institute of Physiology, Centre for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
,
Alice Assinger
1   Institute of Physiology, Centre for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria
› Author Affiliations Financial support: This work was supported by a grant of the Anniversary Foundation for Higher Education of Vienna [Hochschuljubiläumsstiftung, H-3103/2011] and the Austrian Science Fund [FWF P-24978-B13].
Further Information

Publication History

Received: 26 November 2013

Accepted after major revision: 07 February 2014

Publication Date:
01 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

Circulating plasma microvesicles (PMVs) and their microRNA content are involved in the development of atherosclerosis and could serve as biomarkers for cardiovascular disease (CVD) progression. However, little is known on how smoking influences the levels of PMVs and microRNA signatures in vivo. Therefore, we aimed to investigate the effects of smoking on circulating PMV levels and CVD-related PMV-derived microRNAs in young, healthy smokers. Twenty young (10 female, 10 male; 25 ± 4 years) healthy smokers (16 ± 6 cigarettes per day for 8 ± 4 years) and age- and sex-matched controls were included in this study. While complete blood count revealed no differences between both groups, smoking significantly enhanced intracellular reactive oxygen species in platelets and leukocytes as well as platelet-leukocyte aggregate formation. Total circulating PMV counts were significantly reduced in smokers, which could be attributed to decreased platelet-derived PMVs. While the number of endothelial PMVs remained unaffected, smoking propagated circulating leukocyte-derived PMVs. Despite reduced total PMVs, PMV-derived microRNA-profiling of six smoker/control pairs revealed a decrease of only a single microRNA, the major platelet-derived microRNA miR-223. Conversely, miR-29b, a microRNA associated with aortic aneurysm and fibrosis, and RNU6–2, a commonly used reference-RNA, were significantly up-regulated. Smoking leads to alterations in the circulating PMV profile and changes in the PMV-derived microRNA signature already in young, healthy adults. These changes may contribute to the development of smoking-related cardiovascular pathologies. Moreover, these smoking-related changes have to be considered when microRNA or PMV profiles are used as disease-specific biomarkers.

Keywords

Blood cells - cardiovascular diseases - cell-derived microvesicles - microRNA - smoking
 

  • References

  • 1 Burger D, Schock S, Thompson CS. et al. Microparticles: biomarkers and beyond. Clin Sci 2013; 124: 423-441.
    CrossrefPubMedGoogle Scholar
  • 2 Hunter MP, Ismail N, Zhang X. et al. Detection of microRNA expression in human peripheral blood microvesicles. PLoS One 2008; 03: e3694.
    CrossrefPubMedGoogle Scholar
  • 3 Zampetaki A, Willeit P, Drozdov I. et al. Profiling of circulating microRNAs: from single biomarkers to re-wired networks. Cardiovasc Res 2012; 93: 555-562.
    CrossrefPubMedGoogle Scholar
  • 4 Diehl P, Fricke A, Sander L. et al. Microparticles: major transport vehicles for distinct microRNAs in circulation. Cardiovasc Res 2012; 93: 633-644.
    CrossrefPubMedGoogle Scholar
  • 5 Ambrose JA, Barua RS. The pathophysiology of cigarette smoking and cardiovascular disease: an update. J Am Coll Cardiol 2004; 43: 1731-1737.
    CrossrefPubMedGoogle Scholar
  • 6 Csordas A, Bernhard D. The biology behind the atherothrombotic effects of cigarette smoke. Nat Rev Cardiol 2013; 10: 219-230.
    CrossrefPubMedGoogle Scholar
  • 7 Li M, Yu D, Williams KJ. et al. Tobacco smoke induces the generation of procoagulant microvesicles from human monocytes/macrophages. Arterioscler Thromb Vasc Biol 2010; 30: 1818-1824.
    CrossrefPubMedGoogle Scholar
  • 8 Grant R, Ansa-Addo E, Stratton D. et al. A filtration-based protocol to isolate human plasma membrane-derived vesicles and exosomes from blood plasma. J Immunol Methods 2011; 371: 143-151.
    CrossrefPubMedGoogle Scholar
  • 9 Casey RG, Joyce M, Roche-Nagle G. et al. Young male smokers have altered platelets and endothelium that precedes atherosclerosis. J Surg Res 2004; 116: 227-233.
    CrossrefPubMedGoogle Scholar
  • 10 Assinger A, Laky M, Schabbauer G. et al. Efficient phagocytosis of periodontopathogens by neutrophils requires plasma factors, platelets and TLR2. J Thromb Haemost 2011; 09: 799-809.
    CrossrefPubMedGoogle Scholar
  • 11 Mobarrez F, Antovic J, Egberg N. et al. A multicolor flow cytometric assay for measurement of platelet-derived microparticles. Thromb Res 2010; 125: e110-e116.
    CrossrefPubMedGoogle Scholar
  • 12 Robert S, Poncelet P, Lacroix R. et al. Standardization of platelet-derived microparticle counting using calibrated beads and a Cytomics FC500 routine flow cytometer: a first step towards multicenter studies?. J Thromb Haemost 2009; 07: 190-197.
    CrossrefPubMedGoogle Scholar
  • 13 Biasucci LM, Porto I, Di VL. et al. Differences in microparticle release in patients with acute coronary syndrome and stable angina. Circ J 2012; 76: 2174-2182.
    CrossrefPubMedGoogle Scholar
  • 14 van Ierssel SH, Van Craenenbroeck EM, Conraads VM. et al. Flow cytometric detection of endothelial microparticles (EMP): effects of centrifugation and storage alter with the phenotype studied. Thromb Res 2010; 125: 332-339.
    CrossrefPubMedGoogle Scholar
  • 15 Huang X, Liang M, Dittmar R. et al. Extracellular MicroRNAs in Urologic Malignancies: Chances and Challenges. Int J Mol Sci 2013; 14: 14785-14799.
    CrossrefPubMedGoogle Scholar
  • 16 Yuana Y, Bertina RM, Osanto S. Pre-analytical and analytical issues in the analysis of blood microparticles. Thromb Haemost 2011; 105: 396-408.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Mobarrez F, He S, Broijersen A. et al. Atorvastatin reduces thrombin generation and expression of tissue factor, P-selectin and GPIIIa on platelet-derived micro-particles in patients with peripheral arterial occlusive disease. Thromb Haemost 2011; 106: 344-352.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Michelson AD, Barnard MR, Krueger LA. et al. Circulating monocyte-platelet aggregates are a more sensitive marker of in vivo platelet activation than platelet surface P-selectin: studies in baboons, human coronary intervention, and human acute myocardial infarction. Circulation 2001; 104: 1533-1537.
    CrossrefPubMedGoogle Scholar
  • 19 Furman MI, Barnard MR, Krueger LA. et al. Circulating monocyte-platelet aggregates are an early marker of acute myocardial infarction. J Am Coll Cardiol 2001; 38: 1002-106.
    CrossrefPubMedGoogle Scholar
  • 20 Horstman LL, Jy W, Jimenez JJ. et al. New horizons in the analysis of circulating cell-derived microparticles. Keio J Med 2004; 53: 210-230.
    CrossrefPubMedGoogle Scholar
  • 21 Connor DE, Exner T, Ma DD. et al. The majority of circulating platelet-derived microparticles fail to bind annexin V, lack phospholipid-dependent procoagulant activity and demonstrate greater expression of glycoprotein Ib. Thromb Haemost 2010; 103: 1044-1052.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Wright SC, Zhong J, Zheng H. et al. Nicotine inhibition of apoptosis suggests a role in tumor promotion. FASEB J 1993; 07: 1045-1051.
    CrossrefPubMedGoogle Scholar
  • 23 Flaumenhaft R, Dilks JR, Richardson J. et al. Megakaryocyte-derived microparticles: direct visualization and distinction from platelet-derived microparticles. Blood 2009; 113: 1112-1121.
    CrossrefPubMedGoogle Scholar
  • 24 Italiano Jr JE, Mairuhu AT, Flaumenhaft R. Clinical relevance of microparticles from platelets and megakaryocytes. Curr Opin Hematol 2010; 17: 578-584.
    CrossrefPubMedGoogle Scholar
  • 25 Gordon C, Gudi K, Krause A. et al. Circulating endothelial microparticles as a measure of early lung destruction in cigarette smokers. Am J Respir Crit Care Med 2011; 184: 224-232.
    CrossrefPubMedGoogle Scholar
  • 26 Chironi G, Simon A, Hugel B. et al. Circulating leukocyte-derived microparticles predict subclinical atherosclerosis burden in asymptomatic subjects. Arterioscler Thromb Vasc Biol 2006; 26: 2775-2780.
    CrossrefPubMedGoogle Scholar
  • 27 Angelillo-Scherrer A. Leukocyte-derived microparticles in vascular homeostasis. Circ Res 2012; 110: 356-369.
    CrossrefPubMedGoogle Scholar
  • 28 Mallat Z, Hugel B, Ohan J. et al. Shed membrane microparticles with procoagulant potential in human atherosclerotic plaques: a role for apoptosis in plaque thrombogenicity. Circulation 1999; 99: 348-353.
    CrossrefPubMedGoogle Scholar
  • 29 Matetzky S, Tani S, Kangavari S. et al. Smoking increases tissue factor expression in atherosclerotic plaques: implications for plaque thrombogenicity. Circulation 2000; 102: 602-604.
    CrossrefPubMedGoogle Scholar
  • 30 Sambola A, Osende J, Hathcock J. et al. Role of risk factors in the modulation of tissue factor activity and blood thrombogenicity. Circulation 2003; 107: 973-977.
    CrossrefPubMedGoogle Scholar
  • 31 Winckers K, ten Cate H, Hackeng TM. The role of tissue factor pathway inhibitor in atherosclerosis and arterial thrombosis. Blood Rev 2013; 27: 119-132.
    CrossrefPubMedGoogle Scholar
  • 32 Zampetaki A, Willeit P, Tilling L. et al. Prospective study on circulating MicroR-NAs and risk of myocardial infarction. J Am Coll Cardiol 2012; 60: 290-299.
    CrossrefPubMedGoogle Scholar
  • 33 Willeit P, Zampetaki A, Dudek K. et al. Circulating microRNAs as novel biomarkers for platelet activation. Circ Res 2013; 112: 595-600.
    CrossrefPubMedGoogle Scholar
  • 34 Shi R, Ge L, Zhou X. et al. Decreased platelet miR-223 expression is associated with high on-clopidogrel platelet reactivity. Thromb Res 2013; 131: 508-13.
    CrossrefPubMedGoogle Scholar
  • 35 Shanker G, Kontos JL, Eckman DM. et al. Nicotine upregulates the expression of P2Y12 on vascular cells and megakaryoblasts. J Thromb Thrombolysis 2006; 22: 213-220.
    CrossrefPubMedGoogle Scholar
  • 36 Laffont B, Corduan A, Ple H. et al. Activated platelets can deliver mRNA regulatory Ago2·microRNA complexes to endothelial cells via microparticles. Blood 2013; 122: 253-261.
    CrossrefPubMedGoogle Scholar
  • 37 Mittelbrunn M, Gutierrez-Vazquez C, Villarroya-Beltri C. et al. Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat Commun 2011; 02: 282.
    CrossrefPubMedGoogle Scholar
  • 38 Raitoharju E, Lyytikainen LP, Levula M. et al. miR-21, miR-210, miR-34a, and miR-146a/b are up-regulated in human atherosclerotic plaques in the Tampere Vascular Study. Atherosclerosis 2011; 219: 211-217.
    CrossrefPubMedGoogle Scholar
  • 39 Graff JW, Dickson AM, Clay G. et al. Identifying functional microRNAs in macrophages with polarized phenotypes. J Biol Chem 2012; 287: 21816-21825.
    CrossrefPubMedGoogle Scholar
  • 40 Boon RA, Seeger T, Heydt S. et al. MicroRNA-29 in aortic dilation: implications for aneurysm formation. Circ Res 2011; 109: 1115-1119.
    CrossrefPubMedGoogle Scholar
  • 41 Merk DR, Chin JT, Dake BA. et al. miR-29b participates in early aneurysm development in Marfan syndrome. Circ Res 2012; 110: 312-324.
    CrossrefPubMedGoogle Scholar
  • 42 Chen KC, Wang YS, Hu CY. et al. OxLDL up-regulates microRNA-29b, leading to epigenetic modifications of MMP-2/MMP-9 genes: a novel mechanism for cardiovascular diseases. FASEB J 2011; 25: 1718-1728.
    CrossrefPubMedGoogle Scholar
  • 43 Maegdefessel L, Azuma J, Toh R. et al. Inhibition of microRNA-29b reduces murine abdominal aortic aneurysm development. J Clin Invest 2012; 122: 497-506.
    CrossrefPubMedGoogle Scholar
  • 44 Yongxin S, Wenjun D, Qiang W. et al. Heavy smoking before coronary surgical procedures affects the native matrix metalloproteinase-2 and matrix metallo-proteinase-9 gene expression in saphenous vein conduits. Ann Thorac Surg 2013; 95: 55-61.
    CrossrefPubMedGoogle Scholar
  • 45 Martell EA. Radioactivity of tobacco trichomes and insoluble cigarette smoke particles. Nature 1974; 249: 215-217.
    CrossrefPubMedGoogle Scholar
  • 46 Li G, Zhao J, Peng X. et al. The mechanism involved in the loss of PTEN expression in NSCLC tumor cells. Biochem Biophys Res Commun 2012; 418: 547-52.
    CrossrefPubMedGoogle Scholar
  • 47 Wang K, Yuan Y, Cho JH. et al. Comparing the MicroRNA spectrum between serum and plasma. PLoS One 2012; 07: e41561.
    CrossrefPubMedGoogle Scholar
  • 48 Corsten MF, Dennert R, Jochems S. et al. Circulating MicroRNA-208b and MicroRNA-499 reflect myocardial damage in cardiovascular disease. Circ Car-diovasc Genet 2010; 03: 499-506.
    CrossrefPubMedGoogle Scholar
  • 49 Zernecke A, Bidzhekov K, Noels H. et al. Delivery of microRNA-126 by apoptotic bodies induces CXCL12-dependent vascular protection. Sci Signal. 2009 02. ra81.
    PubMedGoogle Scholar
  • 50 Finn NA, Eapen D, Manocha P. et al. Coronary heart disease alters intercellular communication by modifying microparticle-mediated microRNA transport. FEBS Lett 2013; 587: 3456-3463.
    CrossrefPubMedGoogle Scholar