Download PDF
Thromb Haemost 2021; 121(04): 464-476
DOI: 10.1055/s-0040-1718760
Coagulation and Fibrinolysis

Extracellular Histones Inhibit Fibrinolysis through Noncovalent and Covalent Interactions with Fibrin

Matthew Locke 
1   Biotherapeutics Division, National Institute for Biological Standards and Control, South Mimms, Hertfordshire, United Kingdom
,
Colin Longstaff
1   Biotherapeutics Division, National Institute for Biological Standards and Control, South Mimms, Hertfordshire, United Kingdom
› Author Affiliations
› Further Information
Also available at
   Permissions and Reprints

Abstract

Histones released into circulation as neutrophil extracellular traps are causally implicated in the pathogenesis of arterial, venous, and microvascular thrombosis by promoting coagulation and enhancing clot stability. Histones induce structural changes in fibrin rendering it stronger and resistant to fibrinolysis. The current study extends these observations by defining the antifibrinolytic mechanisms of histones in purified, plasma, and whole blood systems. Although histones stimulated plasminogen activation in solution, they inhibited plasmin as competitive substrates. Protection of fibrin from plasmin digestion is enhanced by covalent incorporation of histones into fibrin, catalyzed by activated transglutaminase, coagulation factor FXIII (FXIIIa). All histone subtypes (H1, H2A, H2B, H3, and H4) were crosslinked to fibrin. A distinct, noncovalent mechanism explains histone-accelerated lateral aggregation of fibrin protofibrils, resulting in thicker fibers with higher mass-to-length ratios and in turn hampered fibrinolysis. However, histones were less effective at delaying fibrinolysis in the absence of FXIIIa activity. Therapeutic doses of low-molecular-weight heparin (LMWH) prevented covalent but not noncovalent histone–fibrin interactions and neutralized the effects of histones on fibrinolysis. This suggests an additional antithrombotic mechanism for LMWH beyond anticoagulation. In conclusion, for the first time we report that histones are crosslinked to fibrin by FXIIIa and promote fibrinolytic resistance which can be overcome by FXIIIa inhibitors and histone-binding heparinoids. These findings provide a rationale for targeting the FXIII–histone–fibrin axis to destabilize fibrin and prevent potentially thrombotic fibrin networks.

Keywords

fibrin(ogen) - fibrinolysis - histones - neutrophil extracellular traps - FXIII

Supplementary Material



Publication History

Received: 25 April 2020

Accepted: 15 September 2020

Article published online:
01 November 2020

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Chen R, Kang R, Fan XG, Tang D. Release and activity of histone in diseases. Cell Death Dis 2014; 5: e1370
    CrossrefPubMedSearch in Google Scholar
  • 2 Brinkmann V, Reichard U, Goosmann C. et al. Neutrophil extracellular traps kill bacteria. Science 2004; 303 (5663): 1532-1535
    CrossrefPubMedSearch in Google Scholar
  • 3 Engelmann B, Massberg S. Thrombosis as an intravascular effector of innate immunity. Nat Rev Immunol 2013; 13 (01) 34-45
    CrossrefPubMedSearch in Google Scholar
  • 4 Gould TJ, Lysov Z, Liaw PC. Extracellular DNA and histones: double-edged swords in immunothrombosis. J Thromb Haemost 2015; 13 (Suppl. 01) S82-S91
    CrossrefPubMedSearch in Google Scholar
  • 5 Fuchs TA, Brill A, Duerschmied D. et al. Extracellular DNA traps promote thrombosis. Proc Natl Acad Sci U S A 2010; 107 (36) 15880-15885
    CrossrefPubMedSearch in Google Scholar
  • 6 Thålin C, Hisada Y, Lundström S, Mackman N, Wallén H. Neutrophil extracellular traps: villains and targets in arterial, venous, and cancer-associated thrombosis. Arterioscler Thromb Vasc Biol 2019; 39 (09) 1724-1738
    CrossrefPubMedSearch in Google Scholar
  • 7 Noubouossie DF, Reeves BN, Strahl BD, Key NS. Neutrophils: back in the thrombosis spotlight. Blood 2019; 133 (20) 2186-2197
    CrossrefPubMedSearch in Google Scholar
  • 8 Laridan E, Martinod K, De Meyer SF. Neutrophil extracellular traps in arterial and venous thrombosis. Semin Thromb Hemost 2019; 45 (01) 86-93
    Thieme ConnectPubMedSearch in Google Scholar
  • 9 Wolberg AS. Thrombin generation and fibrin clot structure. Blood Rev 2007; 21 (03) 131-142
    CrossrefPubMedSearch in Google Scholar
  • 10 Longstaff C, Thelwell C, Williams SC, Silva MM, Szabó L, Kolev K. The interplay between tissue plasminogen activator domains and fibrin structures in the regulation of fibrinolysis: kinetic and microscopic studies. Blood 2011; 117 (02) 661-668
    CrossrefPubMedSearch in Google Scholar
  • 11 Gould TJ, Vu TT, Stafford AR. et al. Cell-free DNA modulates clot structure and impairs fibrinolysis in sepsis. Arterioscler Thromb Vasc Biol 2015; 35 (12) 2544-2553
    CrossrefPubMedSearch in Google Scholar
  • 12 Mutch NJ, Engel R, Uitte de Willige S, Philippou H, Ariëns RA. Polyphosphate modifies the fibrin network and down-regulates fibrinolysis by attenuating binding of tPA and plasminogen to fibrin. Blood 2010; 115 (19) 3980-3988
    CrossrefPubMedSearch in Google Scholar
  • 13 Gonias SL, Pasqua JJ, Greenberg C, Pizzo SV. Precipitation of fibrinogen, fibrinogen degradation products and fibrin monomer by histone H3. Thromb Res 1985; 39 (01) 97-116
    CrossrefPubMedSearch in Google Scholar
  • 14 Pemberton AD, Brown JK, Inglis NF. Proteomic identification of interactions between histones and plasma proteins: implications for cytoprotection. Proteomics 2010; 10 (07) 1484-1493
    CrossrefPubMedSearch in Google Scholar
  • 15 Locke M, Francis RJ, Tsaousi E, Longstaff C. Fibrinogen protects neutrophils from the cytotoxic effects of histones and delays neutrophil extracellular trap formation induced by ionomycin. Sci Rep 2020; 10 (01) 11694
    CrossrefPubMedSearch in Google Scholar
  • 16 Kppeć M, Wegrzynowicz Z, Zajdel M, Sawecka J, Szumiel I. Effects of histones and dextran on some properties of fibrin, particularly on its susceptibility to plasmin. Thromb Res 1974; 5 (03) 359-374
    CrossrefPubMedSearch in Google Scholar
  • 17 Longstaff C, Varjú I, Sótonyi P. et al. Mechanical stability and fibrinolytic resistance of clots containing fibrin, DNA, and histones. J Biol Chem 2013; 288 (10) 6946-6956
    CrossrefPubMedSearch in Google Scholar
  • 18 Kalathottukaren MT, Abraham L, Kapopara PR. et al. Alteration of blood clotting and lung damage by protamine are avoided using the heparin and polyphosphate inhibitor UHRA. Blood 2017; 129 (10) 1368-1379
    CrossrefPubMedSearch in Google Scholar
  • 19 Muszbek L, Bereczky Z, Bagoly Z, Komáromi I, Katona É. Factor XIII: a coagulation factor with multiple plasmatic and cellular functions. Physiol Rev 2011; 91 (03) 931-972
    CrossrefPubMedSearch in Google Scholar
  • 20 Byrnes JR, Duval C, Wang Y. et al. Factor XIIIa-dependent retention of red blood cells in clots is mediated by fibrin α-chain crosslinking. Blood 2015; 126 (16) 1940-1948
    CrossrefPubMedSearch in Google Scholar
  • 21 Mutch NJ, Koikkalainen JS, Fraser SR. et al. Model thrombi formed under flow reveal the role of factor XIII-mediated cross-linking in resistance to fibrinolysis. J Thromb Haemost 2010; 8 (09) 2017-2024
    CrossrefPubMedSearch in Google Scholar
  • 22 Rijken DC, Abdul S, Malfliet JJ, Leebeek FW, Uitte de Willige S. Compaction of fibrin clots reveals the antifibrinolytic effect of factor XIII. J Thromb Haemost 2016; 14 (07) 1453-1461
    CrossrefPubMedSearch in Google Scholar
  • 23 Wolberg AS. Fibrinogen and factor XIII: newly recognized roles in venous thrombus formation and composition. Curr Opin Hematol 2018; 25 (05) 358-364
    CrossrefPubMedSearch in Google Scholar
  • 24 Varjú I, Longstaff C, Szabó L. et al. DNA, histones and neutrophil extracellular traps exert anti-fibrinolytic effects in a plasma environment. Thromb Haemost 2015; 113 (06) 1289-1298
    Thieme ConnectPubMedSearch in Google Scholar
  • 25 Longstaff C. subcommittee on fibrinolysis. Development of Shiny app tools to simplify and standardize the analysis of hemostasis assay data: communication from the SSC of the ISTH. J Thromb Haemost 2017; 15 (05) 1044-1046
    CrossrefPubMedSearch in Google Scholar
  • 26 Pieters M, Philippou H, Undas A, de Lange Z, Rijken DC, Mutch NJ. Subcommittee on Factor XIII and Fibrinogen, and the Subcommittee on Fibrinolysis. An international study on the feasibility of a standardized combined plasma clot turbidity and lysis assay: communication from the SSC of the ISTH. J Thromb Haemost 2018; 16 (05) 1007-1012
    CrossrefPubMedSearch in Google Scholar
  • 27 Carr Jr ME, Hermans J. Size and density of fibrin fibers from turbidity. Macromolecules 1978; 11 (01) 46-50
    CrossrefPubMedSearch in Google Scholar
  • 28 Yeromonahos C, Polack B, Caton F. Nanostructure of the fibrin clot. Biophys J 2010; 99 (07) 2018-2027
    CrossrefPubMedSearch in Google Scholar
  • 29 Herren T, Burke TA, Das R, Plow EF. Identification of histone H2B as a regulated plasminogen receptor. Biochemistry 2006; 45 (31) 9463-9474
    CrossrefPubMedSearch in Google Scholar
  • 30 Hethershaw EL, Cilia La Corte AL, Duval C. et al. The effect of blood coagulation factor XIII on fibrin clot structure and fibrinolysis. J Thromb Haemost 2014; 12 (02) 197-205
    CrossrefPubMedSearch in Google Scholar
  • 31 Ryan EA, Mockros LF, Weisel JW, Lorand L. Structural origins of fibrin clot rheology. Biophys J 1999; 77 (05) 2813-2826
    CrossrefPubMedSearch in Google Scholar
  • 32 Longstaff C, Hogwood J, Gray E. et al. Neutralisation of the anti-coagulant effects of heparin by histones in blood plasma and purified systems. Thromb Haemost 2016; 115 (03) 591-599
    Thieme ConnectPubMedSearch in Google Scholar
  • 33 Köhler S, Schmid F, Settanni G. The internal dynamics of fibrinogen and its implications for coagulation and adsorption. PLOS Comput Biol 2015; 11 (09) e1004346
    CrossrefPubMedSearch in Google Scholar
  • 34 Nitzsche R, Köhler J, Kreikemeyer B, Oehmcke-Hecht S. Streptococcus pyogenes escapes killing from extracellular histones through plasminogen binding and activation by streptokinase. J Innate Immun 2016; 8 (06) 589-600
    CrossrefPubMedSearch in Google Scholar
  • 35 Abrams ST, Zhang N, Manson J. et al. Circulating histones are mediators of trauma-associated lung injury. Am J Respir Crit Care Med 2013; 187 (02) 160-169
    CrossrefPubMedSearch in Google Scholar
  • 36 García-Giménez JL, Romá-Mateo C, Carbonell N. et al. A new mass spectrometry-based method for the quantification of histones in plasma from septic shock patients. Sci Rep 2017; 7 (01) 10643
    CrossrefPubMedSearch in Google Scholar
  • 37 Alhamdi Y, Abrams ST, Cheng Z. et al. Circulating histones are major mediators of cardiac injury in patients with sepsis. Crit Care Med 2015; 43 (10) 2094-2103
    CrossrefPubMedSearch in Google Scholar
  • 38 Jiménez-Alcázar M, Napirei M, Panda R. et al. Impaired DNase1-mediated degradation of neutrophil extracellular traps is associated with acute thrombotic microangiopathies. J Thromb Haemost 2015; 13 (05) 732-742
    CrossrefPubMedSearch in Google Scholar
  • 39 Laridan E, Denorme F, Desender L. et al. Neutrophil extracellular traps in ischemic stroke thrombi. Ann Neurol 2017; 82 (02) 223-232
    CrossrefPubMedSearch in Google Scholar
  • 40 Kolaczkowska E, Jenne CN, Surewaard BG. et al. Molecular mechanisms of NET formation and degradation revealed by intravital imaging in the liver vasculature. Nat Commun 2015; 6: 6673
    Search in Google Scholar
  • 41 Albadawi H, Oklu R, Raacke Malley RE. et al. Effect of DNase I treatment and neutrophil depletion on acute limb ischemia-reperfusion injury in mice. J Vasc Surg 2016; 64 (02) 484-493
    CrossrefPubMedSearch in Google Scholar
  • 42 Brill A, Fuchs TA, Savchenko AS. et al. Neutrophil extracellular traps promote deep vein thrombosis in mice. J Thromb Haemost 2012; 10 (01) 136-144
    CrossrefPubMedSearch in Google Scholar
  • 43 Tanka-Salamon A, Kolev K, Machovich R, Komorowicz E. Proteolytic resistance conferred to fibrinogen by von Willebrand factor. Thromb Haemost 2010; 103 (02) 291-298
    Thieme ConnectPubMedSearch in Google Scholar
  • 44 Pasternack R, Büchold C, Jähnig R. et al. Novel inhibitor ZED3197 as potential drug candidate in anticoagulation targeting coagulation FXIIIa (F13a). J Thromb Haemost 2020; 18 (01) 191-200
    CrossrefPubMedSearch in Google Scholar
  • 45 Wang F, Zhang N, Li B. et al. Heparin defends against the toxicity of circulating histones in sepsis. Front Biosci 2015; 20: 1259-1270
    CrossrefPubMedSearch in Google Scholar
  • 46 Wildhagen KC, Wiewel MA, Schultz MJ. et al. Extracellular histone H3 levels are inversely correlated with antithrombin levels and platelet counts and are associated with mortality in sepsis patients. Thromb Res 2015; 136 (03) 542-547
    CrossrefPubMedSearch in Google Scholar
  • 47 Fuchs TA, Bhandari AA, Wagner DD. Histones induce rapid and profound thrombocytopenia in mice. Blood 2011; 118 (13) 3708-3714
    CrossrefPubMedSearch in Google Scholar
  • 48 Semeraro F, Ammollo CT, Morrissey JH. et al. Extracellular histones promote thrombin generation through platelet-dependent mechanisms: involvement of platelet TLR2 and TLR4. Blood 2011; 118 (07) 1952-1961
    CrossrefPubMedSearch in Google Scholar
  • 49 Komorowicz E, Balázs N, Tanka-Salamon A. et al. Biorelevant polyanions stabilize fibrin against mechanical and proteolytic decomposition: effects of polymer size and electric charge. J Mech Behav Biomed Mater 2020; 102: 103459
    CrossrefPubMedSearch in Google Scholar
  • 50 Ammollo CT, Semeraro N, Carratù MR, Colucci M, Semeraro F. Histones differentially modulate the anticoagulant and profibrinolytic activities of heparin, heparin derivatives, and dabigatran. J Pharmacol Exp Ther 2016; 356 (02) 305-313
    CrossrefPubMedSearch in Google Scholar