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Thromb Haemost 2019; 119(11): 1816-1826
DOI: 10.1055/s-0039-1695008
Blood Cells, Inflammation and Infection
Georg Thieme Verlag KG Stuttgart · New York

Fibrin-VLDL Receptor-Dependent Pathway Promotes Leukocyte Transmigration by Inhibiting Src Kinase Fyn and is a Target for Fibrin β15-42 Peptide

Sergiy Yakovlev
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, United States
2   Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, United States
,
Chunzhang Cao
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, United States
3   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States
,
Rebeca Galisteo
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, United States
,
Li Zhang
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, United States
3   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States
,
Dudley K. Strickland
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, United States
3   Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States
4   Department of Surgery, University of Maryland School of Medicine, Baltimore, Maryland, United States
,
Leonid Medved
1   Center for Vascular and Inflammatory Diseases, University of Maryland School of Medicine, Baltimore, Maryland, United States
2   Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, United States
› Author Affiliations Funding This study was supported by National Institutes of Health grants R01 HL-056051 (L.M.), R35 HL135743 (D.K.S.), R01 NS-082607 (L.Z.), and American Heart Association 18TPA34170550 (L.Z.).
Further Information

Publication History

11 April 2019

02 July 2019

Publication Date:
29 August 2019 (online)

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Abstract

According to the current view, binding of fibrin degradation product E1 fragment to endothelial VE-cadherin promotes transendothelial migration of leukocytes and thereby inflammation, and fibrin-derived β15–42 peptide reduces leukocyte transmigration by competing with E1 for binding to VE-cadherin and, in addition, by signaling through Src kinase Fyn. However, the very low affinity of β15–42 to VE-cadherin raised a question about its ability to inhibit E1–VE-cadherin interaction. Further, our previous study revealed that fibrin promotes leukocyte transmigration through the very-low-density lipoprotein (VLDL) receptor (VLDLR)-dependent pathway and suggested a possible link between the inhibitory properties of β15–42 and this pathway. To test such a link and the proposed inhibitory mechanisms for β15–42, we performed in vitro experiments using surface plasmon resonance, enzyme-linked immunosorbent assay, and leukocyte transendothelial migration assay, and in vivo studies with wild-type and VLDLR-deficient mice using mouse model of peritonitis. The experiments revealed that β15–42 cannot inhibit E1–VE-cadherin interaction at the concentrations used in the previous in vivo studies leaving the proposed Fyn-dependent signaling mechanism as a viable explanation for the inhibitory effect of β15–42. While testing this mechanism, we confirmed that Fyn plays a critical role in controlling fibrin-induced transendothelial migration of leukocytes and found that signaling through the VLDLR-dependent pathway results in inhibition of Fyn, thereby increasing leukocyte transmigration. Furthermore, our in vivo experiments revealed that β15–42 inhibits this pathway, thereby preventing inhibition of Fyn and reducing leukocyte transmigration. Thus, this study clarifies the molecular mechanism underlying the VLDLR-dependent pathway of leukocyte transmigration and reveals that this pathway is a target for β15–42.

Keywords

fibrinogen - fibrin - VLDL receptor - leukocyte transmigration - inflammation
 

  • References

  • 1 Colvin RB, Johnson RA, Mihm Jr MC, Dvorak HF. Role of the clotting system in cell-mediated hypersensitivity. I. Fibrin deposition in delayed skin reactions in man. J Exp Med 1973; 138 (03) 686-698
    CrossrefPubMedGoogle Scholar
  • 2 McRitchie DI, Girotti MJ, Glynn MF, Goldberg JM, Rotstein OD. Effect of systemic fibrinogen depletion on intraabdominal abscess formation. J Lab Clin Med 1991; 118 (01) 48-55
    PubMedGoogle Scholar
  • 3 Languino LR, Plescia J, Duperray A. , et al. Fibrinogen mediates leukocyte adhesion to vascular endothelium through an ICAM-1-dependent pathway. Cell 1993; 73 (07) 1423-1434
    CrossrefPubMedGoogle Scholar
  • 4 Languino LR, Duperray A, Joganic KJ, Fornaro M, Thornton GB, Altieri DC. Regulation of leukocyte-endothelium interaction and leukocyte transendothelial migration by intercellular adhesion molecule 1-fibrinogen recognition. Proc Natl Acad Sci U S A 1995; 92 (05) 1505-1509
    CrossrefPubMedGoogle Scholar
  • 5 Altieri DC. Regulation of leukocyte-endothelium interaction by fibrinogen. Thromb Haemost 1999; 82 (02) 781-786
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Hamaguchi M, Morishita Y, Takahashi I, Ogura M, Takamatsu J, Saito H. FDP D-dimer induces the secretion of interleukin-1, urokinase-type plasminogen activator, and plasminogen activator inhibitor-2 in a human promonocytic leukemia cell line. Blood 1991; 77 (01) 94-100
    CrossrefPubMedGoogle Scholar
  • 7 Robson SC, Shephard EG, Kirsch RE. Fibrin degradation product D-dimer induces the synthesis and release of biologically active IL-1 β, IL-6 and plasminogen activator inhibitors from monocytes in vitro. Br J Haematol 1994; 86 (02) 322-326
    CrossrefPubMedGoogle Scholar
  • 8 Gorlatov S, Medved L. Interaction of fibrin(ogen) with the endothelial cell receptor VE-cadherin: mapping of the receptor-binding site in the NH2-terminal portions of the fibrin β chains. Biochemistry 2002; 41 (12) 4107-4116
    CrossrefPubMedGoogle Scholar
  • 9 Medved L, Weisel JW. ; Fibrinogen and Factor XIII Subcommittee of Scientific Standardization Committee of International Society on Thrombosis and Haemostasis. Recommendations for nomenclature on fibrinogen and fibrin. J Thromb Haemost 2009; 7 (02) 355-359
    CrossrefPubMedGoogle Scholar
  • 10 Skogen WF, Senior RM, Griffin GL, Wilner GD. Fibrinogen-derived peptide Bβ1-42 is a multidomained neutrophil chemoattractant. Blood 1988; 71 (05) 1475-1479
    CrossrefPubMedGoogle Scholar
  • 11 Hamaguchi M, Bunce LA, Sporn LA, Francis CW. Spreading of platelets on fibrin is mediated by the amino terminus of the β chain including peptide β15-42. Blood 1993; 81 (09) 2348-2356
    CrossrefPubMedGoogle Scholar
  • 12 Petzelbauer P, Zacharowski PA, Miyazaki Y. , et al. The fibrin-derived peptide Bβ15-42 protects the myocardium against ischemia-reperfusion injury. Nat Med 2005; 11 (03) 298-304
    CrossrefPubMedGoogle Scholar
  • 13 Zacharowski K, Zacharowski PA, Friedl P. , et al. The effects of the fibrin-derived peptide Bβ15-42 in acute and chronic rodent models of myocardial ischemia-reperfusion. Shock 2007; 27 (06) 631-637
    CrossrefPubMedGoogle Scholar
  • 14 Roesner JP, Petzelbauer P, Koch A. , et al. The fibrin-derived peptide Bβ15-42 is cardioprotective in a pig model of myocardial ischemia-reperfusion injury. Crit Care Med 2007; 35 (07) 1730-1735
    CrossrefPubMedGoogle Scholar
  • 15 Atar D, Petzelbauer P, Schwitter J. , et al; F.I.R.E. Investigators. Effect of intravenous FX06 as an adjunct to primary percutaneous coronary intervention for acute ST-segment elevation myocardial infarction results of the F.I.R.E. (Efficacy of FX06 in the Prevention of Myocardial Reperfusion Injury) trial. J Am Coll Cardiol 2009; 53 (08) 720-729
    CrossrefPubMedGoogle Scholar
  • 16 Hallén J, Petzelbauer P, Schwitter J, Geudelin B, Buser P, Atar D. Impact of time to therapy and presence of collaterals on the efficacy of FX06 in acute ST elevation myocardial infarction: a substudy of the F.I.R.E., the Efficacy of FX06 in the prevention of myocardial reperfusion injury trial. EuroIntervention 2010; 5 (08) 946-952
    CrossrefPubMedGoogle Scholar
  • 17 Roesner JP, Petzelbauer P, Koch A. , et al. Bβ15-42 (FX06) reduces pulmonary, myocardial, liver, and small intestine damage in a pig model of hemorrhagic shock and reperfusion. Crit Care Med 2009; 37 (02) 598-605
    CrossrefPubMedGoogle Scholar
  • 18 Roesner JP, Petzelbauer P, Koch A. , et al. A double blind, single centre, sub-chronic reperfusion trial evaluating FX06 following haemorrhagic shock in pigs. Resuscitation 2009; 80 (02) 264-271
    CrossrefPubMedGoogle Scholar
  • 19 Wiedemann D, Schneeberger S, Friedl P. , et al. The fibrin-derived peptide Bβ15-42 significantly attenuates ischemia-reperfusion injury in a cardiac transplant model. Transplantation 2010; 89 (07) 824-829
    CrossrefPubMedGoogle Scholar
  • 20 Sörensen I, Rong S, Susnik N. , et al. Bβ15-42 attenuates the effect of ischemia-reperfusion injury in renal transplantation. J Am Soc Nephrol 2011; 22 (10) 1887-1896
    CrossrefPubMedGoogle Scholar
  • 21 Krishnamoorthy A, Ajay AK, Hoffmann D. , et al. Fibrinogen β-derived Bβ15-42 peptide protects against kidney ischemia/reperfusion injury. Blood 2011; 118 (07) 1934-1942
    PubMedGoogle Scholar
  • 22 Liu A, Fang H, Yang Y. , et al. The fibrin-derived peptide Bβ15-42 attenuates liver damage in a rat model of liver ischemia/reperfusion injury. Shock 2013; 39 (04) 397-403
    CrossrefPubMedGoogle Scholar
  • 23 Urbschat A, Rupprecht K, Zacharowski K. , et al. Combined peri-ischemic administration of Bβ15-42 in treating ischemia reperfusion injury of the mouse kidney. Microvasc Res 2015; 101: 48-54
    CrossrefPubMedGoogle Scholar
  • 24 Matt U, Warszawska JM, Bauer M. , et al. Bβ15-42 protects against acid-induced acute lung injury and secondary pseudomonas pneumonia in vivo. Am J Respir Crit Care Med 2009; 180 (12) 1208-1217
    CrossrefPubMedGoogle Scholar
  • 25 Gröger M, Pasteiner W, Ignatyev G. , et al. Peptide Bβ15-42 preserves endothelial barrier function in shock. PLoS One 2009; 4 (04) e5391
    CrossrefPubMedGoogle Scholar
  • 26 Bergt S, Gruenewald M, Beltschany C. , et al. The fibrin-derived peptide Bβ15–42 (FX06) ameliorates vascular leakage and improves survival and neurocognitive recovery: implications from two animal models of cardiopulmonary resuscitation. Crit Care Med 2016; 44 (10) e988-e995
    CrossrefPubMedGoogle Scholar
  • 27 Wolf T, Kann G, Becker S. , et al. Severe Ebola virus disease with vascular leakage and multiorgan failure: treatment of a patient in intensive care. Lancet 2015; 385 (9976): 1428-1435
    CrossrefPubMedGoogle Scholar
  • 28 Zacharowski K, Zacharowski P, Reingruber S, Petzelbauer P. Fibrin(ogen) and its fragments in the pathophysiology and treatment of myocardial infarction. J Mol Med (Berl) 2006; 84 (06) 469-477
    PubMedGoogle Scholar
  • 29 Jennewein C, Tran N, Paulus P, Ellinghaus P, Eble JA, Zacharowski K. Novel aspects of fibrin(ogen) fragments during inflammation. Mol Med 2011; 17 (5-6): 568-573
    CrossrefPubMedGoogle Scholar
  • 30 Yakovlev S, Gao Y, Cao C. , et al. Interaction of fibrin with VE-cadherin and anti-inflammatory effect of fibrin-derived fragments. J Thromb Haemost 2011; 9 (09) 1847-1855
    CrossrefPubMedGoogle Scholar
  • 31 Yakovlev S, Mikhailenko I, Cao C, Zhang L, Strickland DK, Medved L. Identification of VLDLR as a novel endothelial cell receptor for fibrin that modulates fibrin-dependent transendothelial migration of leukocytes. Blood 2012; 119 (02) 637-644
    CrossrefPubMedGoogle Scholar
  • 32 Yakovlev S, Medved L. Interaction of fibrin with the very low-density lipoprotein (VLDL) receptor: further characterization and localization of the VLDL receptor-binding site in fibrin βN-domains. Biochemistry 2017; 56 (19) 2518-2528
    CrossrefPubMedGoogle Scholar
  • 33 Bach TL, Barsigian C, Yaen CH, Martinez J. Endothelial cell VE-cadherin functions as a receptor for the β15-42 sequence of fibrin. J Biol Chem 1998; 273 (46) 30719-30728
    CrossrefPubMedGoogle Scholar
  • 34 Yakovlev S, Makogonenko E, Kurochkina N, Nieuwenhuizen W, Ingham K, Medved L. Conversion of fibrinogen to fibrin: mechanism of exposure of tPA- and plasminogen-binding sites. Biochemistry 2000; 39 (51) 15730-15741
    CrossrefPubMedGoogle Scholar
  • 35 Yakovlev S, Gorlatov S, Ingham K, Medved L. Interaction of fibrin(ogen) with heparin: further characterization and localization of the heparin-binding site. Biochemistry 2003; 42 (25) 7709-7716
    CrossrefPubMedGoogle Scholar
  • 36 Yakovlev S, Medved L. Interaction of fibrin(ogen) with the endothelial cell receptor VE-cadherin: localization of the fibrin-binding site within the third extracellular VE-cadherin domain. Biochemistry 2009; 48 (23) 5171-5179
    CrossrefPubMedGoogle Scholar
  • 37 Ruiz J, Kouiavskaia D, Migliorini M. , et al. The apoE isoform binding properties of the VLDL receptor reveal marked differences from LRP and the LDL receptor. J Lipid Res 2005; 46 (08) 1721-1731
    CrossrefPubMedGoogle Scholar
  • 38 Yakovlev S, Belkin AM, Chen L. , et al. Anti-VLDL receptor monoclonal antibodies inhibit fibrin-VLDL receptor interaction and reduce fibrin-dependent leukocyte transmigration. Thromb Haemost 2016; 116 (06) 1122-1130
    Article in Thieme ConnectPubMedGoogle Scholar
  • 39 Kudryk B, Rohoza A, Ahadi M, Chin J, Wiebe ME. Specificity of a monoclonal antibody for the NH2-terminal region of fibrin. Mol Immunol 1984; 21 (01) 89-94
    CrossrefPubMedGoogle Scholar
  • 40 Procyk R, Kudryk B, Callender S, Blombäck B. Accessibility of epitopes on fibrin clots and fibrinogen gels. Blood 1991; 77 (07) 1469-1475
    CrossrefPubMedGoogle Scholar
  • 41 Collins SJ, Ruscetti FW, Gallagher RE, Gallo RC. Terminal differentiation of human promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds. Proc Natl Acad Sci U S A 1978; 75 (05) 2458-2462
    CrossrefPubMedGoogle Scholar
  • 42 Yakovlev S, Medved L. Effect of fibrinogen, fibrin, and fibrin degradation products on transendothelial migration of leukocytes. Thromb Res 2018; 162: 93-100
    CrossrefPubMedGoogle Scholar
  • 43 Holinstat M, Knezevic N, Broman M, Samarel AM, Malik AB, Mehta D. Suppression of RhoA activity by focal adhesion kinase-induced activation of p190RhoGAP: role in regulation of endothelial permeability. J Biol Chem 2006; 281 (04) 2296-2305
    CrossrefPubMedGoogle Scholar