NETosis in Acute Thrombotic Disorders

 

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Abstract

The release of extracellular traps by neutrophils (NETs) represents a novel active mechanism of cell death that has been recently implicated in the pathogenesis of thrombotic disorders. The aim of this study was to investigate the generation of NETs in different groups of patients with acute thrombotic events (ATEs) and to establish whether NETs markers can predict the risk of new cardiovascular events. We performed a case–control study of patients with ATE, including acute coronary syndrome (n = 60), cerebrovascular accident (n = 50), and venous thromboembolism (n = 55). Control subjects (n = 70) were identified among patients admitted for acute chest pain and in which a diagnosis of ATE was excluded. Serum levels of NET markers and neutrophil activation, such as myeloperoxidase (MPO)-DNA complexes, neutrophil gelatinase-associated lipocalin, polymorphonuclear neutrophil elastase, lactoferrin, and MPO, were measured in each patient. We found that circulating levels of MPO-DNA complexes were significantly increased in patients with ATE (p < 0.001) compared with controls and that this association remained significant even after fully adjustment for traditional risk factors (p = 0.001). A receiver operating characteristics analysis of circulating MPO-DNA complexes in discriminating between controls and patients with ATE showed a significant area under the curve of 0.76 (95% confidence interval: 0.69–0.82). After a median follow-up of 40.7 (± 13.8) months, 24 out of the 165 patients with ATE presented a new cardiovascular event and 18 patients died. None of the markers under investigation influenced survival or the incidence of new cardiovascular events. In conclusion, we found that increase of markers of NETosis can be observed in acute thrombotic conditions, occurring both on the arterial and venous site. Nevertheless, the level of neutrophil markers measured during the ATE is not predictive of future risk of mortality and cardiovascular events.

Authors' Contributions

A.B., E.F., and M.R. conceived the study and designed the protocol. A.B., E.F., and M.D. performed the laboratory tests. L.T., R.B., C.N., M.T., F.C., and R.S. collected the blood samples and recorded the patients' data. E.F. and L.V. performed statistical analysis. A.B., E.F., and C.F. wrote the paper. C.A. and P.P. supervised the project. All authors critically reviewed and approved the final version of the manuscript.

Publication History

Article published online:
12 June 2023

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• References

• 1 Naess IA, Christiansen SC, Romundstad P, Cannegieter SC, Rosendaal FR, Hammerstrøm J. Incidence and mortality of venous thrombosis: a population-based study. J Thromb Haemost 2007; 5 (04) 692-699
  CrossrefPubMedGoogle Scholar
• 2 Brinkmann V, Reichard U, Goosmann C. et al. Neutrophil extracellular traps kill bacteria. Science 2004; 303 (5663): 1532-1535
  CrossrefPubMedGoogle Scholar
• 3 Laridan E, Martinod K, De Meyer SF. Neutrophil extracellular traps in arterial and venous thrombosis. Semin Thromb Hemost 2019; 45 (01) 86-93
  Article in Thieme ConnectPubMedGoogle Scholar
• 4 Schumski A, Ortega-Gómez A, Wichapong K. et al. Endotoxinemia accelerates atherosclerosis through electrostatic charge-mediated monocyte adhesion. Circulation 2021; 143 (03) 254-266
  CrossrefPubMedGoogle Scholar
• 5 Franck G, Mawson TL, Folco EJ. et al. Roles of PAD4 and NETosis in experimental atherosclerosis and arterial injury implications for superficial erosion. Circ Res 2018; 123 (01) 33-42
  CrossrefPubMedGoogle Scholar
• 6 Donkel SJ, Wolters FJ, Ikram MA, de Maat MPM. Circulating myeloperoxidase (MPO)-DNA complexes as marker for neutrophil extracellular traps (NETs) levels and the association with cardiovascular risk factors in the general population. PLoS One 2021; 16 (8 August): 1-13
  CrossrefPubMedGoogle Scholar
• 7 Borissoff JI, Joosen IA, Versteylen MO. et al. Elevated levels of circulating DNA and chromatin are independently associated with severe coronary atherosclerosis and a prothrombotic state. Arterioscler Thromb Vasc Biol 2013; 33 (08) 2032-2040
  CrossrefPubMedGoogle Scholar
• 8 An Z, Li J, Yu J. et al. Neutrophil extracellular traps induced by IL-8 aggravate atherosclerosis via activation NF-κB signaling in macrophages. Cell Cycle 2019; 18 (21) 2928-2938
  CrossrefPubMedGoogle Scholar
• 9 Wang W, Peng W, Ning X. Increased levels of neutrophil extracellular trap remnants in the serum of patients with rheumatoid arthritis. Int J Rheum Dis 2018; 21 (02) 415-421
  CrossrefPubMedGoogle Scholar
• 10 Masuda S, Nakazawa D, Shida H. et al. NETosis markers: quest for specific, objective, and quantitative markers. Clin Chim Acta 2016; 459: 89-93
  CrossrefPubMedGoogle Scholar
• 11 Vallés J, Lago A, Santos MT. et al. Neutrophil extracellular traps are increased in patients with acute ischemic stroke: prognostic significance. Thromb Haemost 2017; 117 (10) 1919-1929
  Article in Thieme ConnectPubMedGoogle Scholar
• 12 Savchenko AS, Martinod K, Seidman MA. et al. Neutrophil extracellular traps form predominantly during the organizing stage of human venous thromboembolism development. J Thromb Haemost 2014; 12 (06) 860-870
  CrossrefPubMedGoogle Scholar
• 13 Halpern EJ. Triple-rule-out CT angiography for evaluation of acute chest pain and possible acute coronary syndrome. Radiology 2009; 252 (02) 332-345
  CrossrefPubMedGoogle Scholar
• 14 Kano H, Aminul Huq M, Tsuda M, Noguchi H, Takeyama N. Sandwich ELISA for circulating myeloperoxidase- and neutrophil elastase-DNA complexes released from neutrophil extracellular traps. Adv Tech Biol Med 2017; 05 (01) 1-3
  CrossrefPubMedGoogle Scholar
• 15 Silvain J, Collet JP, Nagaswami C. et al. Composition of coronary thrombus in acute myocardial infarction. J Am Coll Cardiol 2011; 57 (12) 1359-1367
  CrossrefPubMedGoogle Scholar
• 16 Helseth R, Shetelig C, Andersen GØ. et al. Neutrophil extracellular trap components associate with infarct size, ventricular function, and clinical outcome in STEMI. Mediators Inflamm 2019; 2019: 7816491
  CrossrefPubMedGoogle Scholar
• 17 Li T, Peng R, Wang F. et al. Lysophosphatidic acid promotes thrombus stability by inducing rapid formation of neutrophil extracellular traps: a new mechanism of thrombosis. J Thromb Haemost 2020; 18 (08) 1952-1964
  CrossrefPubMedGoogle Scholar
• 18 Wan W, Liu H, Long Y. et al. The association between circulating neutrophil extracellular trap related biomarkers and retinal vein occlusion incidence: a case-control pilot study. Exp Eye Res 2021; 210 (June): 108702
  CrossrefPubMedGoogle Scholar
• 19 Sivalingam Z, Larsen SB, Grove EL, Hvas AM, Kristensen SD, Magnusson NE. Neutrophil gelatinase-associated lipocalin as a risk marker in cardiovascular disease. Clin Chem Lab Med 2017; 56 (01) 5-18
  CrossrefPubMedGoogle Scholar
• 20 Lupu L, Rozenfeld KL, Zahler D. et al. Detection of renal injury following primary coronary intervention among ST-segment elevation myocardial infarction patients: doubling the incidence using neutrophil gelatinase-associated lipocalin as a renal biomarker. J Clin Med 2021; 10 (10) 4-11
  CrossrefPubMedGoogle Scholar
• 21 Zahler D, Merdler I, Banai A. et al. Predictive value of elevated neutrophil gelatinase-associated lipocalin (NGAL) levels for assessment of cardio-renal interactions among ST-segment elevation myocardial infarction patients. J Clin Med 2022; 11 (08) 2162
  CrossrefPubMedGoogle Scholar
• 22 Sivalingam Z, Erik Magnusson N, Grove EL, Hvas AM, Dalby Kristensen S, Bøjet Larsen S. Neutrophil gelatinase-associated lipocalin (NGAL) and cardiovascular events in patients with stable coronary artery disease. Scand J Clin Lab Invest 2018; 78 (06) 470-476
  CrossrefPubMedGoogle Scholar

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