Advertisement

The pathway of neutrophil extracellular traps towards atherosclerosis and thrombosis

      Highlights

      • NETs exist in atherosclerotic lesions of both humans and animal models.
      • NETs contribute to atherosclerosis, as well as to arterial and venous thrombosis.
      • The proatherogenic and prothrombotic activities of NETs are expressed through various mechanisms.

      Abstract

      Neutrophil extracellular traps (NETs) are web-like extrusions of genetic material, which are released upon neutrophil activation. NETs consist of a chromatin substructure, onto which a vast array of proteins with various properties is dispersed. NETs production was initially described as an unrecognized defense mechanism of neutrophils, due to their ability to entrap and possibly eliminate a wide range of pathogens. Nevertheless, growing evidence suggests that NETs are implicated in a multitude of pathophysiological conditions, such as autoimmunity, cancer, diabetes mellitus and Alzheimer's disease. Importantly, NETs may also play a decisive role in atherosclerosis and thrombosis. In this context, it has been demonstrated that NETs are present in atherosclerotic lesions of both humans and animal models and are implicated in various mechanisms leading to atherogenesis. Among others, NETs induce oxidative stress and oxidize high-density lipoprotein particles, thus reducing their beneficial cholesterol efflux capacity. NETs also induce endothelial cell dysfunction and apoptosis and promote the generation of anti-double-stranded-DNA autoantibodies. NETs may also play a prothrombotic role, since they form a fibrin-like base for platelet adhesion, activation and aggregation. Furthermore, NETs promote the accumulation of prothrombotic molecules, like von Willebrand factor and fibrinogen, thus significantly contributing to thrombus formation. Notably, there is vast data linking NETs to arterial and venous thrombosis in animal models, as well as in humans. Future large-scale studies should incorporate NETs and their individual components as disease markers, as well as potential therapeutic targets, to reduce atherosclerosis and to prevent thrombosis.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Brinkmann V.
        • et al.
        Neutrophil extracellular traps kill bacteria.
        Science. 2004; 303: 1532-1535
        • Urban C.F.
        • et al.
        Neutrophil extracellular traps contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans.
        PLoS Pathog. 2009; 5 (e1000639)
        • Dwyer M.
        • et al.
        Cystic fibrosis sputum DNA has NETosis characteristics and neutrophil extracellular trap release is regulated by macrophage migration-inhibitory factor.
        J. Innate Immun. 2014; 6: 765-779
        • Welin A.
        • et al.
        The human neutrophil subsets defined by the presence or absence of OLFM4 both transmigrate into tissue in vivo and give rise to distinct NETs in vitro.
        PLoS One. 2013; 8 (e69575)
        • Urban C.F.
        • Nett J.E.
        Neutrophil extracellular traps in fungal infection.
        Semin. Cell Dev. Biol. 2019; 89: 47-57
        • Chang Z.
        • et al.
        The TatD-like DNase of Plasmodium is a virulence factor and a potential malaria vaccine candidate.
        Nat. Commun. 2016; 7: 11537
        • Schonrich G.
        • Raftery M.J.
        Neutrophil extracellular traps go viral.
        Front. Immunol. 2016; 7: 366
        • Gupta S.
        • Kaplan M.J.
        The role of neutrophils and NETosis in autoimmune and renal diseases.
        Nat. Rev. Nephrol. 2016; 12: 402-413
        • Cools-Lartigue J.
        • et al.
        Neutrophil extracellular traps in cancer progression.
        Cell. Mol. Life Sci. 2014; 71: 4179-4194
        • Wong S.L.
        • et al.
        Diabetes primes neutrophils to undergo NETosis, which impairs wound healing.
        Nat. Med. 2015; 21: 815-819
        • Zenaro E.
        • et al.
        Neutrophils promote Alzheimer's disease-like pathology and cognitive decline via LFA-1 integrin.
        Nat. Med. 2015; 21: 880-886
        • Clark S.R.
        • et al.
        Platelet TLR4 activates neutrophil extracellular traps to ensnare bacteria in septic blood.
        Nat. Med. 2007; 13: 463-469
        • Gupta A.K.
        • et al.
        Activated endothelial cells induce neutrophil extracellular traps and are susceptible to NETosis-mediated cell death.
        FEBS Lett. 2010; 584: 3193-3197
        • Saffarzadeh M.
        • et al.
        Neutrophil extracellular traps directly induce epithelial and endothelial cell death: a predominant role of histones.
        PLoS One. 2012; 7 (e32366)
        • Allen C.
        • et al.
        Neutrophil cerebrovascular transmigration triggers rapid neurotoxicity through release of proteases associated with decondensed DNA.
        J. Immunol. 2012; 189: 381-392
        • Doring Y.
        • Soehnlein O.
        • Weber C.
        Neutrophil extracellular traps in atherosclerosis and atherothrombosis.
        Circ. Res. 2017; 120: 736-743
        • Van Avondt K.
        • Maegdefessel L.
        • Soehnlein O.
        Therapeutic targeting of neutrophil extracellular traps in atherogenic inflammation.
        Thromb. Haemostasis. 2019; 119: 542-552
        • Borissoff J.I.
        • 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: 2032-2040
        • Mangold A.
        • et al.
        Coronary neutrophil extracellular trap burden and deoxyribonuclease activity in ST-elevation acute coronary syndrome are predictors of ST-segment resolution and infarct size.
        Circ. Res. 2015; 116: 1182-1192
        • Valles J.
        • et al.
        Neutrophil extracellular traps are increased in patients with acute ischemic stroke: prognostic significance.
        Thromb. Haemostasis. 2017; 117: 1919-1929
        • Fuchs T.A.
        • et al.
        Novel cell death program leads to neutrophil extracellular traps.
        J. Cell Biol. 2007; 176: 231-241
        • Kenny E.F.
        • et al.
        Diverse stimuli engage different neutrophil extracellular trap pathways.
        Elife. 2017; 6
        • Pilsczek F.H.
        • et al.
        A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus.
        J. Immunol. 2010; 185: 7413-7425
        • Yipp B.G.
        • et al.
        Infection-induced NETosis is a dynamic process involving neutrophil multitasking in vivo.
        Nat. Med. 2012; 18: 1386-1393
        • Lood C.
        • et al.
        Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease.
        Nat. Med. 2016; 22: 146-153
        • Douda D.N.
        • et al.
        SK3 channel and mitochondrial ROS mediate NADPH oxidase-independent NETosis induced by calcium influx.
        Proc. Natl. Acad. Sci. U. S. A. 2015; 112: 2817-2822
        • Parker H.
        • et al.
        Myeloperoxidase associated with neutrophil extracellular traps is active and mediates bacterial killing in the presence of hydrogen peroxide.
        J. Leukoc. Biol. 2012; 91: 369-376
        • Papayannopoulos V.
        • et al.
        Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular traps.
        J. Cell Biol. 2010; 191: 677-691
        • Metzler K.D.
        • et al.
        Myeloperoxidase is required for neutrophil extracellular trap formation: implications for innate immunity.
        Blood. 2011; 117: 953-959
        • Metzler K.D.
        • et al.
        A myeloperoxidase-containing complex regulates neutrophil elastase release and actin dynamics during NETosis.
        Cell Rep. 2014; 8: 883-896
        • Neeli I.
        • Khan S.N.
        • Radic M.
        Histone deimination as a response to inflammatory stimuli in neutrophils.
        J. Immunol. 2008; 180: 1895-1902
        • Wang Y.
        • et al.
        Histone hypercitrullination mediates chromatin decondensation and neutrophil extracellular trap formation.
        J. Cell Biol. 2009; 184: 205-213
        • Leshner M.
        • et al.
        PAD4 mediated histone hypercitrullination induces heterochromatin decondensation and chromatin unfolding to form neutrophil extracellular trap-like structures.
        Front. Immunol. 2012; 3: 307
        • Rohrbach A.S.
        • et al.
        Activation of PAD4 in NET formation.
        Front. Immunol. 2012; 3: 360
        • Neeli I.
        • Radic M.
        Opposition between PKC isoforms regulates histone deimination and neutrophil extracellular chromatin release.
        Front. Immunol. 2013; 4: 38
        • Li P.
        • et al.
        PAD4 is essential for antibacterial innate immunity mediated by neutrophil extracellular traps.
        J. Exp. Med. 2010; 207: 1853-1862
        • Remijsen Q.
        • et al.
        Neutrophil extracellular trap cell death requires both autophagy and superoxide generation.
        Cell Res. 2011; 21: 290-304
        • Itakura A.
        • McCarty O.J.
        Pivotal role for the mTOR pathway in the formation of neutrophil extracellular traps via regulation of autophagy.
        Am. J. Physiol. Cell Physiol. 2013; 305: C348-C354
        • Sollberger G.
        • et al.
        Gasdermin D plays a vital role in the generation of neutrophil extracellular traps.
        Sci. Immunol. 2018; 3
        • Chen K.W.
        • et al.
        Noncanonical inflammasome signaling elicits gasdermin D-dependent neutrophil extracellular traps.
        Sci. Immunol. 2018; 3
        • Megens R.T.
        • et al.
        Presence of luminal neutrophil extracellular traps in atherosclerosis.
        Thromb. Haemostasis. 2012; 107: 597-598
        • Awasthi D.
        • et al.
        Oxidized LDL induced extracellular trap formation in human neutrophils via TLR-PKC-IRAK-MAPK and NADPH-oxidase activation.
        Free Radic. Biol. Med. 2016; 93: 190-203
        • Warnatsch A.
        • et al.
        Inflammation. Neutrophil extracellular traps license macrophages for cytokine production in atherosclerosis.
        Science. 2015; 349: 316-320
        • Liu Y.
        • et al.
        Myeloid-specific deletion of peptidylarginine deiminase 4 mitigates atherosclerosis.
        Front. Immunol. 2018; 9: 1680
        • Westerterp M.
        • et al.
        Cholesterol efflux pathways suppress inflammasome activation, NETosis, and atherogenesis.
        Circulation. 2018; 138: 898-912
        • Curcic S.
        • et al.
        Neutrophil effector responses are suppressed by secretory phospholipase A2 modified HDL.
        Biochim. Biophys. Acta. 2015; 1851: 184-193
        • Neumann A.
        • et al.
        Lipid alterations in human blood-derived neutrophils lead to formation of neutrophil extracellular traps.
        Eur. J. Cell Biol. 2014; 93: 347-354
        • Smith C.K.
        • et al.
        Neutrophil extracellular trap-derived enzymes oxidize high-density lipoprotein: an additional proatherogenic mechanism in systemic lupus erythematosus.
        Arthritis Rheum. 2014; 66: 2532-2544
        • Doring Y.
        • et al.
        Auto-antigenic protein-DNA complexes stimulate plasmacytoid dendritic cells to promote atherosclerosis.
        Circulation. 2012; 125: 1673-1683
        • Quillard T.
        • et al.
        TLR2 and neutrophils potentiate endothelial stress, apoptosis and detachment: implications for superficial erosion.
        Eur. Heart J. 2015; 36: 1394-1404
        • Franck G.
        • et al.
        Roles of PAD4 and NETosis in experimental atherosclerosis and arterial injury: implications for superficial erosion.
        Circ. Res. 2018; 123: 33-42
        • Folco E.J.
        • et al.
        Neutrophil extracellular traps induce endothelial cell activation and tissue factor production through interleukin-1alpha and cathepsin G.
        Arterioscler. Thromb. Vasc. Biol. 2018; 38: 1901-1912
        • Wang H.
        • et al.
        Obesity-induced endothelial dysfunction is prevented by neutrophil extracellular trap inhibition.
        Sci. Rep. 2018; 8: 4881
        • Wang Y.
        • et al.
        Mitochondrial oxidative stress promotes atherosclerosis and neutrophil extracellular traps in aged mice.
        Arterioscler. Thromb. Vasc. Biol. 2017; 37: e99-e107
        • Yamamoto K.
        • et al.
        Augmented neutrophil extracellular traps formation promotes atherosclerosis development in socially defeated apoE(-/-) mice.
        Biochem. Biophys. Res. Commun. 2018; 500: 490-496
        • Silvestre-Roig C.
        • et al.
        Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death.
        Nature. 2019; 569: 236-240
        • Oklu R.
        • et al.
        Extracellular traps in lipid-rich lesions of carotid atherosclerotic plaques: implications for lipoprotein retention and lesion progression.
        J. Vasc. Interv. Radiol. 2014; 25: 631-634
        • Range H.
        • et al.
        Periodontal bacteria in human carotid atherothrombosis as a potential trigger for neutrophil activation.
        Atherosclerosis. 2014; 236: 448-455
        • Perez-Sanchez C.
        • et al.
        Diagnostic potential of NETosis-derived products for disease activity, atherosclerosis and therapeutic effectiveness in Rheumatoid Arthritis patients.
        J. Autoimmun. 2017; 82: 31-40
        • Pertiwi K.R.
        • et al.
        Neutrophil extracellular traps participate in all different types of thrombotic and haemorrhagic complications of coronary atherosclerosis.
        Thromb. Haemostasis. 2018; 118: 1078-1087
        • Fuchs T.A.
        • et al.
        Extracellular DNA traps promote thrombosis.
        Proc. Natl. Acad. Sci. U. S. A. 2010; 107: 15880-15885
        • Reyes-Garcia A.M.L.
        • et al.
        Neutrophil extracellular trap components increase the expression of coagulation factors.
        Biomed. Rep. 2019; 10: 195-201
        • Elaskalani O.
        • Abdol Razak N.B.
        • Metharom P.
        Neutrophil extracellular traps induce aggregation of washed human platelets independently of extracellular DNA and histones.
        Cell Commun. Signal. 2018; 16: 24
        • Sambrano G.R.
        • et al.
        Cathepsin G activates protease-activated receptor-4 in human platelets.
        J. Biol. Chem. 2000; 275: 6819-6823
        • Kambas K.
        • et al.
        Autophagy mediates the delivery of thrombogenic tissue factor to neutrophil extracellular traps in human sepsis.
        PLoS One. 2012; 7 (e45427)
        • Stakos D.A.
        • et al.
        Expression of functional tissue factor by neutrophil extracellular traps in culprit artery of acute myocardial infarction.
        Eur. Heart J. 2015; 36: 1405-1414
        • Seif K.
        • et al.
        Neutrophil-mediated proteolysis of thrombospondin-1 promotes platelet adhesion and string formation.
        Thromb. Haemostasis. 2018; 118: 2074-2085
        • Caudrillier A.
        • et al.
        Platelets induce neutrophil extracellular traps in transfusion-related acute lung injury.
        J. Clin. Investig. 2012; 122: 2661-2671
        • Sreeramkumar V.
        • et al.
        Neutrophils scan for activated platelets to initiate inflammation.
        Science. 2014; 346: 1234-1238
        • Maugeri N.
        • et al.
        Activated platelets present high mobility group box 1 to neutrophils, inducing autophagy and promoting the extrusion of neutrophil extracellular traps.
        J. Thromb. Haemost. 2014; 12: 2074-2088
        • Carestia A.
        • et al.
        Mediators and molecular pathways involved in the regulation of neutrophil extracellular trap formation mediated by activated platelets.
        J. Leukoc. Biol. 2016; 99: 153-162
        • Etulain J.
        • et al.
        P-selectin promotes neutrophil extracellular trap formation in mice.
        Blood. 2015; 126: 242-246
        • Panicker S.R.
        • et al.
        Circulating soluble P-selectin must dimerize to promote inflammation and coagulation in mice.
        Blood. 2017; 130: 181-191
        • Erlandsson Harris H.
        • Andersson U.
        Mini-review: the nuclear protein HMGB1 as a proinflammatory mediator.
        Eur. J. Immunol. 2004; 34: 1503-1512
        • Tadie J.M.
        • et al.
        HMGB1 promotes neutrophil extracellular trap formation through interactions with Toll-like receptor 4.
        Am. J. Physiol. Lung Cell Mol. Physiol. 2013; 304: L342-L349
        • Andersson U.
        • Tracey K.J.
        HMGB1 is a therapeutic target for sterile inflammation and infection.
        Annu. Rev. Immunol. 2011; 29: 139-162
        • Moschonas I.
        Platelet-derived microparticles induce the formation of neutrophil extracellular traps.
        Atherosclerosis. 2018; 275 (e106. P1.1.006)
        • Maugeri N.
        • et al.
        Platelet microparticles sustain autophagy-associated activation of neutrophils in systemic sclerosis.
        Sci. Transl. Med. 2018; 10
        • Chrysanthopoulou A.
        • et al.
        Interferon lambda1/IL-29 and inorganic polyphosphate are novel regulators of neutrophil-driven thromboinflammation.
        J. Pathol. 2017; 243: 111-122
        • de Boer O.J.
        • et al.
        Neutrophils, neutrophil extracellular traps and interleukin-17 associate with the organisation of thrombi in acute myocardial infarction.
        Thromb. Haemostasis. 2013; 109: 290-297
        • Pertiwi K.R.
        • et al.
        Extracellular traps derived from macrophages, mast cells, eosinophils and neutrophils are generated in a time-dependent manner during atherothrombosis.
        J. Pathol. 2019; 247: 505-512
        • Riegger J.
        • et al.
        Histopathological evaluation of thrombus in patients presenting with stent thrombosis. A multicenter European study: a report of the prevention of late stent thrombosis by an interdisciplinary global European effort consortium.
        Eur. Heart J. 2016; 37: 1538-1549
        • Kopytek M.
        • et al.
        NETosis is associated with the severity of aortic stenosis: links with inflammation.
        Int. J. Cardiol. 2019; 286: 121-126
        • Yu X.
        • Tan J.
        • Diamond S.L.
        Hemodynamic force triggers rapid NETosis within sterile thrombotic occlusions.
        J. Thromb. Haemost. 2018; 16: 316-329
        • Yu X.
        • Diamond S.L.
        Fibrin modulates shear-induced NETosis in sterile occlusive thrombi formed under haemodynamic flow.
        Thromb. Haemostasis. 2019; 119: 586-593
        • Brill A.
        • et al.
        Neutrophil extracellular traps promote deep vein thrombosis in mice.
        J. Thromb. Haemost. 2012; 10: 136-144
        • von Bruhl M.L.
        • et al.
        Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo.
        J. Exp. Med. 2012; 209: 819-835
        • Martinod K.
        • et al.
        Neutrophil histone modification by peptidylarginine deiminase 4 is critical for deep vein thrombosis in mice.
        Proc. Natl. Acad. Sci. U. S. A. 2013; 110: 8674-8679
        • Martinod K.
        • et al.
        Neutrophil elastase-deficient mice form neutrophil extracellular traps in an experimental model of deep vein thrombosis.
        J. Thromb. Haemost. 2016; 14: 551-558
        • van Montfoort M.L.
        • et al.
        Circulating nucleosomes and neutrophil activation as risk factors for deep vein thrombosis.
        Arterioscler. Thromb. Vasc. Biol. 2013; 33: 147-151
        • Savchenko A.S.
        • et al.
        Neutrophil extracellular traps form predominantly during the organizing stage of human venous thromboembolism development.
        J. Thromb. Haemost. 2014; 12: 860-870
        • Yago T.
        • et al.
        Cooperative PSGL-1 and CXCR2 signaling in neutrophils promotes deep vein thrombosis in mice.
        Blood. 2018; 132: 1426-1437
        • Stark K.
        • et al.
        Disulfide HMGB1 derived from platelets coordinates venous thrombosis in mice.
        Blood. 2016; 128: 2435-2449
        • Dyer M.R.
        • et al.
        Deep vein thrombosis in mice is regulated by platelet HMGB1 through release of neutrophil-extracellular traps and DNA.
        Sci. Rep. 2018; 8: 2068
        • Nakazawa D.
        • et al.
        Activated platelets induce MLKL-driven neutrophil necroptosis and release of neutrophil extracellular traps in venous thrombosis.
        Cell Death Dis. 2018; 4: 6
        • Bertin F.R.
        • et al.
        Natural killer cells induce neutrophil extracellular trap formation in venous thrombosis.
        J. Thromb. Haemost. 2019; 17: 403-414
        • Gollomp K.
        • et al.
        Neutrophil accumulation and NET release contribute to thrombosis in HIT.
        JCI Insight. 2018; 3
        • Li B.
        • et al.
        Neutrophil extracellular traps enhance procoagulant activity in patients with oral squamous cell carcinoma.
        J. Cancer Res. Clin. Oncol. 2019; 145: 1695-1707
        • Hisada Y.
        • et al.
        Neutrophils and Neutrophil Extracellular Traps Enhance Venous Thrombosis in Mice Bearing Human Pancreatic Tumors.
        Haematologica, 2019
        • Jung H.S.
        • et al.
        Cancer cell-induced neutrophil extracellular traps promote both hypercoagulability and cancer progression.
        PLoS One. 2019; 14 (e0216055)
        • Robb C.T.
        • et al.
        Invertebrate extracellular phagocyte traps show that chromatin is an ancient defence weapon.
        Nat. Commun. 2014; 5: 4627
        • Hirsch J.G.
        Bactericidal action of histone.
        J. Exp. Med. 1958; 108: 925-944
        • Kessinger C.W.
        • et al.
        Statins improve the resolution of established murine venous thrombosis: reductions in thrombus burden and vein wall scarring.
        PLoS One. 2015; 10 (e0116621)
        • Totani L.
        • et al.
        Roflumilast inhibits leukocyte-platelet interactions and prevents the prothrombotic functions of polymorphonuclear leukocytes and monocytes.
        J. Thromb. Haemost. 2016; 14: 191-204