Download PDF
Thromb Haemost 2018; 118(01): 042-053
DOI: 10.1160/TH17-02-0132
Coagulation and Fibrinolysis
Schattauer GmbH Stuttgart

Amino-Fucoidan as a Vector for rtPA-Induced Fibrinolysis in Experimental Thrombotic Events

Radouane Ghebouli   *
,
Stephane Loyau   *
,
Murielle Maire
,
Pierre Saboural
,
Jean-Philippe Collet
,
Martine Jandrot-Perrus
,
Didier Letourneur
,
Frédéric Chaubet
,
Jean-Baptiste Michel
Further Information

Publication History

24 February 2017

22 August 2017

Publication Date:
05 January 2018 (online)

Also available at
    Permissions and Reprints

Abstract

Acute ischaemic stroke, myocardial infarction and pulmonary embolism are the main causes of mortality and morbidity worldwide. Thrombolysis by intravenous injection of recombinant tissue plasminogen activator (rtPA) remains the most common non-interventional treatment to recanalize occluded vessels. However, this procedure is limited by significant drawbacks, including high doses and bleeding complications. Recent studies showed that fucoidan targets the intraluminal thrombus in vivo. We have developed a chimaera covalently linking fucoidan, able to target platelets within the thrombus, to dilysine, able to non-covalently bind rtPA. We hypothesize that this construct should vectorize rtPA to the thrombus, thus increasing its fibrinolytic efficacy and avoiding its deleterious effects. In vitro, rtPA mixed to dilysine fucoidan (DLF) shows a greater fibrinolytic effect than rtPA alone, both on platelet-rich thrombus and in whole blood. In vivo, occluded mesenteric vessels, carotid artery and vena cava were more efficiently recanalized by DLF complexed to rtPA than by rtPA alone. This study thus provides evidence that DLF may be a promising therapeutic tool to fight against acute thrombosis by enhancing rtPA fibrinolytic efficiency.

Keywords

chimaera - clot - fucoidan - lysine - P-selectin - arterial thrombosis - venous thrombosis

* Radouane Ghebouli and Stephane Loyau contributed equally to this study.


Supplementary Material

 

  • References

  • 1 Finegold JA, Asaria P, Francis DP. Mortality from ischaemic heart disease by country, region, and age: statistics from World Health Organisation and United Nations. Int J Cardiol 2013; 168 (02) 934-945
    CrossrefPubMedGoogle Scholar
  • 2 Go AS, Mozaffarian D, Roger VL. , et al; American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Executive summary: heart disease and stroke statistics--2014 update: a report from the American Heart Association. Circulation 2014; 129 (03) 399-410
    CrossrefPubMedGoogle Scholar
  • 3 National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group. Tissue plasminogen activator for acute ischemic stroke. N Engl J Med 1995; 333 (24) 1581-1587
    CrossrefPubMedGoogle Scholar
  • 4 Chapman KM, Woolfenden AR, Graeb D. , et al. Intravenous tissue plasminogen activator for acute ischemic stroke: A Canadian hospital's experience. Stroke 2000; 31 (12) 2920-2924
    CrossrefPubMedGoogle Scholar
  • 5 Collen D, Lijnen HR. The tissue-type plasminogen activator story. Arterioscler Thromb Vasc Biol 2009; 29 (08) 1151-1155
    CrossrefPubMedGoogle Scholar
  • 6 Docagne F, Parcq J, Lijnen R, Ali C, Vivien D. Understanding the functions of endogenous and exogenous tissue-type plasminogen activator during stroke. Stroke 2015; 46 (01) 314-320
    CrossrefPubMedGoogle Scholar
  • 7 Hacke W, Kaste M, Bluhmki E. , et al; ECASS Investigators. Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med 2008; 359 (13) 1317-1329
    CrossrefPubMedGoogle Scholar
  • 8 Sussman ES, Connolly Jr ES. Hemorrhagic transformation: a review of the rate of hemorrhage in the major clinical trials of acute ischemic stroke. Front Neurol 2013; 4: 69
    CrossrefPubMedGoogle Scholar
  • 9 Cesarman-Maus G, Hajjar KA. Molecular mechanisms of fibrinolysis. Br J Haematol 2005; 129 (03) 307-321
    CrossrefPubMedGoogle Scholar
  • 10 Medcalf RL. Fibrinolysis, inflammation, and regulation of the plasminogen activating system. J Thromb Haemost 2007; 5 (Suppl. 01) 132-142
    CrossrefPubMedGoogle Scholar
  • 11 Mutch NJ, Thomas L, Moore NR, Lisiak KM, Booth NA. TAFIa, PAI-1 and alpha-antiplasmin: complementary roles in regulating lysis of thrombi and plasma clots. J Thromb Haemost 2007; 5 (04) 812-817
    CrossrefPubMedGoogle Scholar
  • 12 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
    CrossrefPubMedGoogle Scholar
  • 13 Silva MM, Thelwell C, Williams SC, Longstaff C. Regulation of fibrinolysis by C-terminal lysines operates through plasminogen and plasmin but not tissue-type plasminogen activator. J Thromb Haemost 2012; 10 (11) 2354-2360
    CrossrefPubMedGoogle Scholar
  • 14 Kaneko M, Sakata Y, Matsuda M, Mimuro J. Interactions between the finger and kringle-2 domains of tissue-type plasminogen activator and plasminogen activator inhibitor-1. J Biochem 1992; 111 (02) 244-248
    CrossrefPubMedGoogle Scholar
  • 15 Foxall C, Watson SR, Dowbenko D. , et al. The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide. J Cell Biol 1992; 117 (04) 895-902
    CrossrefPubMedGoogle Scholar
  • 16 Polley MJ, Phillips ML, Wayner E. , et al. CD62 and endothelial cell-leukocyte adhesion molecule 1 (ELAM-1) recognize the same carbohydrate ligand, sialyl-Lewis x. Proc Natl Acad Sci U S A 1991; 88 (14) 6224-6228
    CrossrefPubMedGoogle Scholar
  • 17 Bachelet L, Bertholon I, Lavigne D. , et al. Affinity of low molecular weight fucoidan for P-selectin triggers its binding to activated human platelets. Biochim Biophys Acta 2009; 1790 (02) 141-146
    CrossrefPubMedGoogle Scholar
  • 18 Rouzet F, Bachelet-Violette L, Alsac JM. , et al. Radiolabeled fucoidan as a p-selectin targeting agent for in vivo imaging of platelet-rich thrombus and endothelial activation. J Nucl Med 2011; 52 (09) 1433-1440
    CrossrefPubMedGoogle Scholar
  • 19 Bonnard T, Yang G, Petiet A. , et al. Abdominal aortic aneurysms targeted by functionalized polysaccharide microparticles: a new tool for SPECT imaging. Theranostics 2014; 4 (06) 592-603
    CrossrefPubMedGoogle Scholar
  • 20 Suzuki M, Bachelet-Violette L, Rouzet F. , et al. Ultrasmall superparamagnetic iron oxide nanoparticles coated with fucoidan for molecular MRI of intraluminal thrombus. Nanomedicine (Lond) 2015; 10 (01) 73-87
    CrossrefPubMedGoogle Scholar
  • 21 Dische Z. New color reactions for determination of sugars in polysaccharides. Methods Biochem Anal 1955; 2: 313-358
    PubMedGoogle Scholar
  • 22 Roth M. Fluorescence reaction for amino acids. Anal Chem 1971; 43 (07) 880-882
    CrossrefPubMedGoogle Scholar
  • 23 Schoergenhofer C, Buchtele N, Schwameis M, Bartko J, Jilma B, Jilma-Stohlawetz P. The use of frozen plasma samples in thromboelastometry. Clin Exp Med 2017
    PubMedGoogle Scholar
  • 24 Dekker SE, Viersen VA, Duvekot A. , et al. Lysis onset time as diagnostic rotational thromboelastometry parameter for fast detection of hyperfibrinolysis. Anesthesiology 2014; 121 (01) 89-97
    CrossrefPubMedGoogle Scholar
  • 25 Boulaftali Y, Ho-Tin-Noe B, Pena A. , et al. Platelet protease nexin-1, a serpin that strongly influences fibrinolysis and thrombolysis. Circulation 2011; 123 (12) 1326-1334
    CrossrefPubMedGoogle Scholar
  • 26 Li W, McIntyre TM, Silverstein RL. Ferric chloride-induced murine carotid arterial injury: a model of redox pathology. Redox Biol 2013; 1: 50-55
    CrossrefPubMedGoogle Scholar
  • 27 von Brühl ML, Stark K, Steinhart A. , et al. Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo. J Exp Med 2012; 209 (04) 819-835
    CrossrefPubMedGoogle Scholar
  • 28 Clauss A. [Rapid physiological coagulation method in determination of fibrinogen]. Acta Haematol 1957; 17 (04) 237-246
    CrossrefPubMedGoogle Scholar
  • 29 Sun X, Berthiller J, Trouillas P, Derex L, Diallo L, Hanss M. Early fibrinogen degradation coagulopathy: a predictive factor of parenchymal hematomas in cerebral rt-PA thrombolysis. J Neurol Sci 2015; 351 (1-2): 109-114
    CrossrefPubMedGoogle Scholar
  • 30 Trouillas P, Derex L, Philippeau F. , et al. Early fibrinogen degradation coagulopathy is predictive of parenchymal hematomas in cerebral rt-PA thrombolysis: a study of 157 cases. Stroke 2004; 35 (06) 1323-1328
    CrossrefPubMedGoogle Scholar
  • 31 El Amki M, Lerouet D, Coqueran B. , et al. Experimental modeling of recombinant tissue plasminogen activator effects after ischemic stroke. Exp Neurol 2012; 238 (02) 138-144
    CrossrefPubMedGoogle Scholar
  • 32 Francis CW, Kornberg A. Fibrinogen- and fibrin-degradation products during fibrinolytic therapy. Ann N Y Acad Sci 1992; 667: 310-323
    CrossrefPubMedGoogle Scholar
  • 33 Loscalzo J, Vaughan DE. Tissue plasminogen activator promotes platelet disaggregation in plasma. J Clin Invest 1987; 79 (06) 1749-1755
    CrossrefPubMedGoogle Scholar
  • 34 Terres W, Umnus S, Mathey DG, Bleifeld W. Effects of streptokinase, urokinase, and recombinant tissue plasminogen activator on platelet aggregability and stability of platelet aggregates. Cardiovasc Res 1990; 24 (06) 471-477
    CrossrefPubMedGoogle Scholar
  • 35 Desilles JP, Loyau S, Syvannarath V. , et al. Alteplase reduces downstream microvascular thrombosis and improves the benefit of large artery recanalization in stroke. Stroke 2015; 46 (11) 3241-3248
    CrossrefPubMedGoogle Scholar
  • 36 Runge MS, Bode C, Matsueda GR, Haber E. Antibody-enhanced thrombolysis: capture of tissue plasminogen activator by a bispecific antibody and direct targeting by an antifibrin-tissue plasminogen activator conjugate in vivo. Trans Assoc Am Physicians 1987; 100: 250-255
    PubMedGoogle Scholar
  • 37 Ma YH, Wu SY, Wu T, Chang YJ, Hua MY, Chen JP. Magnetically targeted thrombolysis with recombinant tissue plasminogen activator bound to polyacrylic acid-coated nanoparticles. Biomaterials 2009; 30 (19) 3343-3351
    CrossrefPubMedGoogle Scholar
  • 38 McCarthy JR, Sazonova IY, Erdem SS. , et al. Multifunctional nanoagent for thrombus-targeted fibrinolytic therapy. Nanomedicine (Lond) 2012; 7 (07) 1017-1028
    CrossrefPubMedGoogle Scholar
  • 39 Absar S, Gupta N, Nahar K, Ahsan F. Engineering of plasminogen activators for targeting to thrombus and heightening thrombolytic efficacy. J Thromb Haemost 2015; 13 (09) 1545-1556
    CrossrefPubMedGoogle Scholar
  • 40 Zhu H, Fan X, Yu Z. , et al. Annexin A2 combined with low-dose tPA improves thrombolytic therapy in a rat model of focal embolic stroke. J Cereb Blood Flow Metab 2010; 30 (06) 1137-1146
    CrossrefPubMedGoogle Scholar
  • 41 Zhang B, Jiang T, She X. , et al. Fibrin degradation by rtPA enhances the delivery of nanotherapeutics to A549 tumors in nude mice. Biomaterials 2016; 96: 63-71
    CrossrefPubMedGoogle Scholar