ADAMTS13 exerts a thrombolytic effect in microcirculation

Summary

Recombinant tissue plasminogen activator (r-tPA) is the drug of choice for thrombolysis, but it is associated with a significant risk of bleeding and is not always successful. By cleaving von Willebrand factor (VWF), the metalloprotease ADAMTS13 (a disintegrin-like and metalloprotease with thrombospondin type I repeats-13) down-regulates thrombus formation in injured vessels. We investigated whether recombinant ADAMTS13 (r-ADAMTS13) induces thrombolysis in vivo in mice. Thrombosis was produced by ferric chloride-induced (FeCl₃) injury in the venules of a dorsal skinfold chamber. Phosphate-buffered saline (PBS, vehicle), r-tPA or r-ADAMTS13, supplemented with hirudin (to stop ongoing thrombin generation), was directly applied onto the occluded vessel, and thrombus dissolution was evaluated by intravital microscopy. The incidence of blood flow restoration significantly increased 30 minutes (min) after r-ADAMTS13 vs. PBS treatment (60% vs. 0%, p<0.05) and 60 min after r-tPA treatment (75% vs. 17%, p<0.05). Both r-tPA and r-ADAMTS13 significantly reduced thrombus size 60 min after their superfusion (53.2% and 62.3% of the initial thrombus size, p<0.05 and p<0.01, respectively). Bleeding occurred in all r-tPA-treated chambers, while it was absent in mice treated with r-ADAMTS13 or PBS. We observed that, similar to r-tPA, r-ADAMTS13 can dissolve occlusive thrombi induced by FeCl₃ injury in venules. In contrast to r-tPA, the in vivo thrombolytic effect of ADAMTS13 was not associated with any signs of haemorrhage. ADAMTS13 could represent a new therapeutic option for thrombolysis.

Keywords

ADAMS/ADAMTS 13 - thrombolysis / thrombolytic agents - plasminogen activators - intravital microscopy

^* M.C. and G.M.T. contributed equally to this study.

• References

• 1 Marder VJ. Foundations of thrombolytic therapy. In: Hemostasis and Thrombosis: Basic Principles and Clinical Practice. 5th ed Lippincott: Williams & Wilkins; 2006: 1739-1752.
• 2 Collen D, Lijnen HR. Thrombolytic agents. Thromb Haemost 2005; 93: 627-630.
• 3 Fibrinolytic Therapy Trialists' (FTT) Collaborative Group. Indications for fibrinolytic therapy in suspected acute myocardial infarction: collaborative overview of early mortality and major morbidity results from all randomised trials of more than 1000 patients. Lancet 1994; 343: 311-322.
• 4 Lees KR, Bluhmki E, von Kummer R. et al. Time to treatment with intravenous alteplase and outcome in stroke: an updated pooled analysis of ECASS, ATLANTIS, NINDS, and EPITHET trials. Lancet 2010; 375: 1695-1703.
• 5 Wechsler LR. Intravenous thrombolytic therapy for acute ischemic stroke. N Engl J Med 2011; 364: 2138-2146.
• 6 Marder VJ. Historical perspective and future direction of thrombolysis research: the rediscovery of plasmin. J Thromb Haemost 2011; 09 (Suppl. 01) 364-373.
• 7 Brill A, Fuchs TA, Chauhan AK. et al. von Willebrand factor-mediated platelet adhesion is critical for deep vein thrombosis in mouse models. Blood 2011; 117: 1400-1407.
• 8 De Meyer SF, Schwarz T, Deckmyn H. et al. Binding of von Willebrand factor to collagen and glycoprotein Ibalpha, but not to glycoprotein IIb/IIIa, contributes to ischemic stroke in mice--brief report. Arterioscler Thromb Vasc Biol 2010; 30: 1949-1951.
• 9 Ruggeri ZM. The role of von Willebrand factor in thrombus formation. Thromb Res 2007; 120 (Suppl. 01) S5-9.
• 10 Ni H, Denis CV, Subbarao S. et al. Persistence of platelet thrombus formation in arterioles of mice lacking both von Willebrand factor and fibrinogen. J Clin Invest 2000; 106: 385-392.
• 11 Plaimauer B, Zimmermann K, Volkel D. et al. Cloning, expression, and functional characterization of the von Willebrand factor-cleaving protease (ADAMTS13). Blood 2002; 100: 3626-3632.
• 12 Chauhan AK, Motto DG, Lamb CB. et al. Systemic antithrombotic effects of ADAMTS13. J Exp Med 2006; 203: 767-776.
• 13 Zhao BQ, Chauhan AK, Canault M. et al. von Willebrand factor-cleaving protease ADAMTS13 reduces ischemic brain injury in experimental stroke. Blood 2009; 114: 3329-3334.
• 14 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: 1326-1334.
• 15 Lehr HA, Leunig M, Menger MD. et al. Dorsal skinfold chamber technique for intravital microscopy in nude mice. Am J Pathol 1993; 143: 1055-1062.
• 16 Boulaftali Y, Lamrani L, Rouzaud MC. et al. The mouse dorsal skinfold chamber as a model for the study of thrombolysis by intravital microscopy. Thromb Haemost 2012; 107: 962-971.
• 17 Thomas GM, Panicot-Dubois L, Lacroix R. et al. Cancer cell-derived microparticles bearing P-selectin glycoprotein ligand 1 accelerate thrombus formation in vivo. J Exp Med 2009; 206: 1913-1927.
• 18 Dubois C, Atkinson B, Furie BC. et al. Real-time in vivo imaging of platelets during thrombus formation. In: Platelets. Elsevier/Academic Press; 2006: 611-628.
• 19 Yamamoto J, Kawano M, Hashimoto M. et al. Adjuvant effect of antibodies against von Willebrand Factor, fibrinogen, and fibronectin on staphylokinase-induced thrombolysis as measured using mural thrombi formed in rat mesenteric venules. Thromb Res 2000; 97: 327-333.
• 20 Gurevitz O, Goldfarb A, Hod H. et al. Recombinant von Willebrand factor fragment AR545C inhibits platelet aggregation and enhances thrombolysis with rtPA in a rabbit thrombosis model. Arterioscler Thromb Vasc Biol 1998; 18: 200-207.
• 21 Mazighi M, Serfaty JM, Labreuche J. et al. Comparison of intravenous alteplase with a combined intravenous-endovascular approach in patients with stroke and confirmed arterial occlusion (RECANALISE study): a prospective cohort study. Lancet Neurol 2009; 08: 802-809.
• 22 Ouriel K. Current status of thrombolysis for peripheral arterial occlusive disease. Ann Vasc Surg 2002; 16: 797-804.
• 23 Alesh I, Kayali F, Stein PD. Catheter-directed thrombolysis (intrathrombus injection) in treatment of deep venous thrombosis: a systematic review. Catheter Cardiovasc Interv 2007; 70: 143-148.
• 24 Kim HS, Preece SR, Black JH. et al. Safety of catheter-directed thrombolysis for deep venous thrombosis in cancer patients. J Vasc Surg 2008; 47: 388-394.
• 25 Mewissen MW, Seabrook GR, Meissner MH. et al. Catheter-directed thrombolysis for lower extremity deep venous thrombosis: report of a national multicenter registry. Radiology 1999; 211: 39-49.
• 26 Kuo WT, van den Bosch MA, Hofmann LV. et al. Catheter-directed embolectomy, fragmentation, and thrombolysis for the treatment of massive pulmonary embolism after failure of systemic thrombolysis. Chest 2008; 134: 250-254.
• 27 Kuo WT, Gould MK, Louie JD. et al. Catheter-directed therapy for the treatment of massive pulmonary embolism: systematic review and meta-analysis of modern techniques. J Vasc Interv Radiol 2009; 20: 1431-1440.
• 28 Korninger C, Collen D. Studies on the specific fibrinolytic effect of human extrinsic (tissue-type) plasminogen activator in human blood and in various animal species in vitro. Thromb Haemost 1981; 46: 561-565.
• 29 Lijnen HR, van Hoef B, Beelen V. et al. Characterization of the murine plasma fibrinolytic system. Eur J Biochem 1994; 224: 863-871.
• 30 Zhu Y, Carmeliet P, Fay WP. Plasminogen activator inhibitor-1 is a major determinant of arterial thrombolysis resistance. Circulation 1999; 99: 3050-3055.
• 31 Kilic E, Hermann DM, Hossmann KA. Recombinant tissue-plasminogen activator-induced thrombolysis after cerebral thromboembolism in mice. Acta Neuropathol 2000; 99: 219-222.
• 32 Orset C, Macrez R, Young AR. et al. Mouse model of in situ thromboembolic stroke and reperfusion. Stroke 2007; 38: 2771-2778.
• 33 Lapergue B, Moreno JA, Dang BQ. et al. Protective effect of high-density lipoprotein-based therapy in a model of embolic stroke. Stroke 2010; 41: 1536-1542.
• 34 Grines CL, Serruys P, O'Neill WW. Fibrinolytic therapy: is it a treatment of the past?. Circulation 2003; 107: 2538-2542.
• 35 Wahlgren N, Ahmed N, Davalos A. et al. Thrombolysis with alteplase 3-4.5 h after acute ischaemic stroke (SITS-ISTR): an observational study. Lancet 2008; 372: 1303-1309.
• 36 Westerhout CM, Bonnefoy E, Welsh RC. et al. The influence of time from symptom onset and reperfusion strategy on 1-year survival in ST-elevation myocardial infarction: a pooled analysis of an early fibrinolytic strategy versus primary percutaneous coronary intervention from CAPTIM and WEST. Am Heart J 2011; 161: 283-290.
• 37 Crawley JT, de Groot R, Xiang Y. et al. Unraveling the scissile bond: how ADAMTS13 recognizes and cleaves von Willebrand factor. Blood 2011; 118: 3212-3221.
• 38 Jin R, Yang G, Li G. Molecular insights and therapeutic targets for blood-brain barrier disruption in ischemic stroke: critical role of matrix metalloproteinases and tissue-type plasminogen activator. Neurobiol Dis 2010; 38: 376-385.