Download PDF
Thromb Haemost 2015; 114(05): 994-1003
DOI: 10.1160/TH14-12-1073
Cellular Haemostasis and Platelets
Schattauer GmbH

HMGB1 binds to activated platelets via the receptor for advanced glycation end products and is present in platelet rich human coronary artery thrombi

Ingo Ahrens
1   Department of Cardiology and Angiology I, Heart Centre, University of Freiburg, Germany
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
,
Yung-Chih Chen   *
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
,
Danijal Topcic   *
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
,
Michael Bode
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
3   Division of Cardiology, University of North Carolina, Chapel Hill, North Carolina, USA
,
David Haenel
1   Department of Cardiology and Angiology I, Heart Centre, University of Freiburg, Germany
,
Christoph E. Hagemeyer
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
,
Hannah Seeba
1   Department of Cardiology and Angiology I, Heart Centre, University of Freiburg, Germany
,
Daniel Duerschmied
1   Department of Cardiology and Angiology I, Heart Centre, University of Freiburg, Germany
,
Nicole Bassler
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
,
Karin A. Jandeleit-Dahm
4   Diabetes Complications Division, Baker IDI, Melbourne, Australia
,
Matthew J. Sweet
5   Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia
,
Alex Agrotis
6   Vascular Biology and Atherosclerosis Laboratory, Baker IDI, Melbourne, Australia
,
Alex Bobik
6   Vascular Biology and Atherosclerosis Laboratory, Baker IDI, Melbourne, Australia
,
Karlheinz Peter
2   Atherothrombosis and Vascular Biology Laboratory, Baker IDI, Melbourne, Australia
› Author Affiliations
Further Information

Publication History

Received: 29 December 2014

Accepted after major revision: 09 June 2015

Publication Date:
06 December 2017 (online)

    Permissions and Reprints

Summary

High mobility group box 1 (HMGB1) acts as both a nuclear protein that regulates gene expression, as well as a pro-inflammatory alarmin that is released from necrotic or activated cells. Recently, HMGB1-expression in human atherosclerotic plaques was identified. Therapeutic blockade of HMGB1 reduced the development of diet-induced atherosclerosis in ApoE knockout mice. Thus, we hypothesised an interaction between HMGB1 and activated platelets. Binding of recombinant HMGB1 to platelets was assessed by flow cytometry. HMGB1 bound to thrombin-activated human platelets (MFI 2.49 vs 25.01, p=0.0079). Blood from wild-type, TLR4 and RAGE knockout mice was used to determine potential HMGB1 receptors on platelets. HMGB1 bound to platelets from wild type C57Bl6 (MFI 2.64 vs 20.3, p< 0.05), and TLR4-/- mice (MFI 2.11 vs 25.65, p< 0.05) but failed to show binding to platelets from RAGE-/- mice (p > 0.05). RAGE expression on human platelets was detected by RT-PCR with mRNA extracted from highly purified platelets and confirmed by Western blot and immunofluorescence microscopy. Platelet activation increased RAGE surface expression (MFI 4.85 vs 6.74, p< 0.05). Expression of HMGB1 in human coronary artery thrombi was demonstrated by immunohistochemistry and revealed high expression levels. Platelets bind HMGB1 upon thrombin-induced activation. Platelet specific expression of RAGE could be detected at the mRNA and protein level and is involved in the binding of HMGB1. Furthermore, platelet activation up-regulates platelet surface expression of RAGE. HMGB1 is highly expressed in platelet-rich human coronary artery thrombi pointing towards a central role for HMGB1 in atherothrombosis, thereby suggesting the possibility of platelet targeted anti-inflammatory therapies for atherothrombosis.

Keywords

HMGB1 - platelets - RAGE - ACS

* Equally contributing authors


 

  • References

  • 1 Scaffidi P, Misteli T, Bianchi ME. Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature 2002; 418: 191-195.
    CrossrefPubMedGoogle Scholar
  • 2 Andersson U, Tracey KJ. HMGB1 is a therapeutic target for sterile inflammation and infection. Annu Rev Immunol 2011; 29: 139-162.
    CrossrefPubMedGoogle Scholar
  • 3 de Souza AW, Westra J, Limburg PC. et al. HMGB1 in vascular diseases: Its role in vascular inflammation and atherosclerosis. Autoimmun Rev 2012; 11: 909-917.
    CrossrefPubMedGoogle Scholar
  • 4 Wang H, Bloom O, Zhang M. et al. HMG-1 as a late mediator of endotoxin lethality in mice. Science 1999; 285: 248-251.
    CrossrefPubMedGoogle Scholar
  • 5 Bianchi ME, Manfredi AA. High-mobility group box 1 (HMGB1) protein at the crossroads between innate and adaptive immunity. Immunol Rev 2007; 220: 35-46.
    CrossrefPubMedGoogle Scholar
  • 6 Libby P, Ridker PM, Hansson GK. Progress and challenges in translating the biology of atherosclerosis. Nature 2011; 473: 317-325.
    CrossrefPubMedGoogle Scholar
  • 7 Inoue K, Kawahara K, Biswas KK. et al. HMGB1 expression by activated vascular smooth muscle cells in advanced human atherosclerosis plaques. Cardiovasc Pathol 2007; 16: 136-143.
    CrossrefPubMedGoogle Scholar
  • 8 Kalinina N, Agrotis A, Antropova Y. et al. Increased expression of the DNA-binding cytokine HMGB1 in human atherosclerotic lesions: role of activated macrophages and cytokines. Arterioscler Thromb Vasc Biol 2004; 24: 2320-2325.
    CrossrefPubMedGoogle Scholar
  • 9 Kanellakis P, Agrotis A, Kyaw TS. et al. High-mobility group box protein 1 neutralisation reduces development of diet-induced atherosclerosis in apolipoprotein e-deficient mice. Arterioscler Thromb Vasc Biol 2011; 31: 313-319.
    CrossrefPubMedGoogle Scholar
  • 10 Huo Y, Schober A, Forlow SB. et al. Circulating activated platelets exacerbate atherosclerosis in mice deficient in apolipoprotein E. Nat Med 2003; 9: 61-67.
    CrossrefPubMedGoogle Scholar
  • 11 Langer HF, Gawaz M. Platelet-vessel wall interactions in atherosclerotic disease. Thromb Haemost 2008; 99: 480-486.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Eisenhardt SU, Habersberger J, Murphy A. et al. Dissociation of pentameric to monomeric C-reactive protein on activated platelets localizes inflammation to atherosclerotic plaques. Circ Res 2009; 105: 128-137.
    CrossrefPubMedGoogle Scholar
  • 13 Schulz C, Massberg S. Platelets in atherosclerosis and thrombosis. Handb Exp Pharmacol 2012; 111-133.
    PubMedGoogle Scholar
  • 14 Duerschmied D, Suidan GL, Demers M. et al. Platelet serotonin promotes the recruitment of neutrophils to sites of acute inflammation in mice. Blood 2013; 121: 1008-1015.
    CrossrefPubMedGoogle Scholar
  • 15 Lievens D, Zernecke A, Seijkens T. et al. Platelet CD40L mediates thrombotic and inflammatory processes in atherosclerosis. Blood 2010; 116: 4317-4327.
    CrossrefPubMedGoogle Scholar
  • 16 Peter K, Bobik A. HMGB1 signals danger in acute coronary syndrome: emergence of a new risk marker for cardiovascular death?. Atherosclerosis 2012; 221: 317-318.
    CrossrefPubMedGoogle Scholar
  • 17 Liu K, Mori S, Takahashi HK. et al. Anti-high mobility group box 1 monoclonal antibody ameliorates brain infarction induced by transient ischemia in rats. FASEB J 2007; 21: 3904-3916.
    CrossrefPubMedGoogle Scholar
  • 18 Birschmann I, Mietner S, Dittrich M. et al. Use of functional highly purified human platelets for the identification of new proteins of the IPP signaling pathway. Thromb Res 2008; 122: 59-68.
    CrossrefPubMedGoogle Scholar
  • 19 Libby P. Inflammation in atherosclerosis. Arterioscler Thromb Vasc Biol 2012; 32: 2045-2051.
    CrossrefPubMedGoogle Scholar
  • 20 Langer HF, Bigalke B, Seizer P. et al. Interaction of platelets and inflammatory endothelium in the development and progression of coronary artery disease. Semin Thromb Hemost 2010; 36: 131-138.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Semple JW, Italiano Jr. JE, Freedman J. Platelets and the immune continuum. Nat Rev Immunol 2011; 11: 264-274.
    CrossrefPubMedGoogle Scholar
  • 22 Duerschmied D, Bode C, Ahrens I. Immune functions of platelets. Thromb Haemost 2014; 112: 678-691.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Maugeri N, Franchini S, Campana L. et al. Circulating platelets as a source of the damage-associated molecular pattern HMGB1 in patients with systemic sclerosis. Autoimmunity 2012; 45: 584-587.
    CrossrefPubMedGoogle Scholar
  • 24 Rouhiainen A, Imai S, Rauvala H. et al. Occurrence of amphoterin (HMG1) as an endogenous protein of human platelets that is exported to the cell surface upon platelet activation. Thromb Haemost 2000; 84: 1087-1094.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Maugeri N, Campana L, Gavina M. 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.
    CrossrefPubMedGoogle Scholar
  • 26 Yan SF, Yan SD, Ramasamy R. et al. Tempering the wrath of RAGE: an emerging therapeutic strategy against diabetic complications, neurodegeneration, and inflammation. Ann Med 2009; 41: 408-422.
    CrossrefPubMedGoogle Scholar
  • 27 Park JS, Gamboni-Robertson F, He Q. et al. High mobility group box 1 protein interacts with multiple Toll-like receptors. Am J Physiol Cell Physiol 2006; 290: C917-924.
    CrossrefPubMedGoogle Scholar
  • 28 Harja E, Bu DX, Hudson BI. et al. Vascular and inflammatory stresses mediate atherosclerosis via RAGE and its ligands in apoE-/- mice. J Clin Invest 2008; 118: 183-194.
    CrossrefPubMedGoogle Scholar
  • 29 Soro-Paavonen A, Watson AM, Li J. et al. Receptor for advanced glycation end products (RAGE) deficiency attenuates the development of atherosclerosis in diabetes. Diabetes 2008; 57: 2461-2469.
    CrossrefPubMedGoogle Scholar
  • 30 Varo N, Libby P, Nuzzo R. et al. Elevated release of sCD40L from platelets of diabetic patients by thrombin, glucose and advanced glycation end products. Diab Vasc Dis Res 2005; 2: 81-87.
    CrossrefPubMedGoogle Scholar
  • 31 Gawlowski T, Stratmann B, Ruetter R. et al. Advanced glycation end products strongly activate platelets. Eur J Nutr 2009; 48: 475-481.
    CrossrefPubMedGoogle Scholar
  • 32 Denis MM, Tolley ND, Bunting M. et al. Escaping the nuclear confines: signal-dependent pre-mRNA splicing in anucleate platelets. Cell 2005; 122: 379-391.
    CrossrefPubMedGoogle Scholar
  • 33 Zimmerman GA, Weyrich AS. Signal-dependent protein synthesis by activated platelets: new pathways to altered phenotype and function. Arterioscler Thromb Vasc Biol 2008; 28: s17-24.
    CrossrefPubMedGoogle Scholar
  • 34 Vazzana N, Guagnano MT, Cuccurullo C. et al. Endogenous secretory RAGE in obese women: association with platelet activation and oxidative stress. J Clin Endocrinol Metab 2012; 97: E1726-1730.
    CrossrefPubMedGoogle Scholar
  • 35 Fukami A, Adachi H, Yamagishi S. et al. Factors associated with serum high mobility group box 1 (HMGB1) levels in a general population. Metabolism 2009; 58: 1688-1693.
    CrossrefPubMedGoogle Scholar
  • 36 Schulz C, Penz S, Hoffmann C. et al. Platelet GPVI binds to collagenous structures in the core region of human atheromatous plaque and is critical for atheroprogression in vivo. Basic Res Cardiol 2008; 103: 356-367.
    CrossrefPubMedGoogle Scholar
  • 37 Seizer P, Gawaz M, May AE. Platelet-monocyte interactions-a dangerous liaison linking thrombosis, inflammation and atherosclerosis. Curr Med Chem 2008; 15: 1976-1980.
    CrossrefPubMedGoogle Scholar
  • 38 Shah PK, Falk E, Badimon JJ. et al. Human monocyte-derived macrophages induce collagen breakdown in fibrous caps of atherosclerotic plaques. Potential role of matrix-degrading metalloproteinases and implications for plaque rupture. Circulation 1995; 92: 1565-1569.
    PubMedGoogle Scholar
  • 39 Martinet W, Schrijvers DM, De Meyer GR. Molecular and cellular mechanisms of macrophage survival in atherosclerosis. Basic Res Cardiol 2012; 107: 297.
    CrossrefPubMedGoogle Scholar
  • 40 Schiraldi M, Raucci A, Munoz LM. et al. HMGB1 promotes recruitment of inflammatory cells to damaged tissues by forming a complex with CXCL12 and signaling via CXCR4. J Exp Med 2012; 209: 551-563.
    CrossrefPubMedGoogle Scholar
  • 41 Zernecke A, Schober A, Bot I. et al. SDF-1alpha/CXCR4 axis is instrumental in neointimal hyperplasia and recruitment of smooth muscle progenitor cells. Circulation Res 2005; 96: 784-791.
    CrossrefPubMedGoogle Scholar
  • 42 Stellos K, Langer H, Daub K. et al. Platelet-derived stromal cell-derived factor-1 regulates adhesion and promotes differentiation of human CD34+ cells to endothelial progenitor cells. Circulation 2008; 117: 206-215.
    CrossrefPubMedGoogle Scholar
  • 43 Stellos K, Bigalke B, Langer H. et al. Expression of stromal-cell-derived factor-1 on circulating platelets is increased in patients with acute coronary syndrome and correlates with the number of CD34+ progenitor cells. Eur Heart J 2009; 30: 584-593.
    CrossrefPubMedGoogle Scholar
  • 44 Stellos K, Bigalke B, Borst O. et al. Circulating platelet-progenitor cell coaggregate formation is increased in patients with acute coronary syndromes and augments recruitment of CD34+ cells in the ischaemic microcirculation. Eur Heart J 2013; 34: 2548-2556.
    CrossrefPubMedGoogle Scholar
  • 45 Schwarz M, Meade G, Stoll P. et al. Conformation-specific blockade of the integrin GPIIb/IIIa: a novel antiplatelet strategy that selectively targets activated platelets. Circ Res 2006; 99: 25-33.
    CrossrefPubMedGoogle Scholar
  • 46 Stoll P, Bassler N, Hagemeyer CE. et al. Targeting ligand-induced binding sites on GPIIb/IIIa via single-chain antibody allows effective anticoagulation without bleeding time prolongation. Arterioscler Thromb Vasc Biol 2007; 27: 1206-1212.
    CrossrefPubMedGoogle Scholar
  • 47 Wang X, Hagemeyer CE, Hohmann JD. et al. Novel single-chain antibody-targeted microbubbles for molecular ultrasound imaging of thrombosis: validation of a unique noninvasive method for rapid and sensitive detection of thrombi and monitoring of success or failure of thrombolysis in mice. Circulation 2012; 125: 3117-3126.
    CrossrefPubMedGoogle Scholar