Antibody-based prevention of von Willebrand factor degradation mediated by circulatory assist devices

 

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Summary

Haemorrhagic episodes in patients carrying circulatory assist devices represent a severe life-threatening clinical complication. These bleeding episodes may originate from a reduced functionality of von Willebrand factor (VWF), a multimeric protein pertinent to the formation of a haemostatic plug. It has been reported that the reduced functionality is due to increased proteolytic degradation by the enzyme ADAMTS13, a phenomenon that is facilitated by device-induced increases in shear stress to which VWF is exposed. Here, we have tested a series of VWF-derived protein fragments and monoclonal murine anti-VWF antibodies for their capacity to reduce shear stress-dependent degradation of VWF. Via direct binding experiments, we identified an anti-VWF antibody that partially blocked VWF-ADAMTS13 interactions (46 ± 14%). Epitope mapping experiments revealed that the antibody, designated mAb508, is directed against the distal portion of the VWF D4-domain (residues 2134–2301) and recognises a synthetic peptide encompassing residues 2158–2169. Consistent with its partial inhibition of VWF-ADAMTS13 interactions in binding assays, mAb508 reduced ADAMTS13-mediated VWF degradation in a vortex-based degradation assay by 48 ± 10%. In a HeartMateII-based whole bloodperfusion system, mAb508 was able to reduce degradation of highmolecular- weight (HMW)-VWF-multimers dose-dependently, with a maximal inhibition (83 ± 8%) being reached at concentrations of 10 μg/ml or higher. In conclusion, we report that partial inhibition of VWF-ADAMTS13 interactions using an anti-VWF antibody can prevent excessive degradation of HMW-VWF multimers. This strategy may be used for the development of therapeutic options to treat bleeding episodes due to shear stress-dependent VWF degradation, for instance in patients carrying circulatory assist devices.

• References

• 1 Zhou YF. et al. Sequence and structure relationships within von Willebrand factor. Blood 2012; 120: 449-458.
  CrossrefPubMedGoogle Scholar
• 2 Sadler JE. von Willebrand factor assembly and secretion. J Thromb Haemost 2009; 7 (Suppl. 01) 24-27.
  CrossrefPubMedGoogle Scholar
• 3 Valentijn KM. et al. Functional architecture of Weibel-Palade bodies. Blood 2011; 117: 5033-5043.
  CrossrefPubMedGoogle Scholar
• 4 Groot E. et al. The active conformation of von Willebrand factor in patients with thrombotic thrombocytopenic purpura in remission. J Thromb Haemost 2009; 7: 962-969.
  CrossrefPubMedGoogle Scholar
• 5 De Ceunynck K. et al. Unwinding the von Willebrand factor strings puzzle. Blood 2013; 121: 270-277.
  CrossrefPubMedGoogle Scholar
• 6 Crawley JT. et al. Unraveling the scissile bond: how ADAMTS13 recognizes and cleaves von Willebrand factor. Blood 2011; 118: 3212-3221.
  CrossrefPubMedGoogle Scholar
• 7 Gao W. et al. Exosite interactions contribute to tension-induced cleavage of von Willebrand factor by the antithrombotic ADAMTS13 metalloprotease. Proc Natl Acad Sci USA 2006; 103: 19099-19104.
  CrossrefPubMedGoogle Scholar
• 8 Zanardelli S. et al. A novel binding site for ADAMTS13 constitutively exposed on the surface of globular VWF. Blood 2009; 112: 2819-2828.
  PubMedGoogle Scholar
• 9 Feys HB. et al. Multi-step binding of ADAMTS-13 to von Willebrand factor. J Thromb Haemost 2009; 7: 2088-2095.
  CrossrefPubMedGoogle Scholar
• 10 Xiang Y. et al. Mechanism of von Willebrand factor scissile bond cleavage by a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13). Proc Natl Acad Sci USA 2011; 108: 11602-11607.
  CrossrefPubMedGoogle Scholar
• 11 Tsai HM. von Willebrand factor, shear stress, and ADAMTS13 in haemostasis and thrombosis. ASAIO J 2012; 58: 163-169.
  PubMedGoogle Scholar
• 12 Dong JF. et al. ADAMTS-13 rapidly cleaves newly secreted ultralarge von Wille-brand factor multimers on the endothelial surface under flowing conditions. Blood 2002; 100: 4033-4039.
  CrossrefPubMedGoogle Scholar
• 13 De Ceunynck K. et al. Local elongation of endothelial cell-anchored von Wille-brand factor strings precedes ADAMTS13 protein-mediated proteolysis. J Biol Chem 2011; 286: 36361-36367.
  CrossrefPubMedGoogle Scholar
• 14 Shim K. et al. Platelet-VWF complexes are preferred substrates of ADAMTS13 under fluid shear stress. Blood 2008; 111: 651-657.
  CrossrefPubMedGoogle Scholar
• 15 Levy GG. et al. Mutations in a member of the ADAMTS gene family cause thrombotic thrombocytopenic purpura. Nature 2001; 413: 488-494.
  CrossrefPubMedGoogle Scholar
• 16 Sadler JE. et al. Update on the pathophysiology and classification of von Wille-brand disease: a report of the Subcommittee on von Willebrand Factor. J Thromb Haemost 2006; 4: 2103-2114.
  CrossrefPubMedGoogle Scholar
• 17 Warkentin TE. et al. Aortic stenosis and bleeding gastrointestinal angiodysplasia: is acquired von Willebrand‘s disease the link?. Lancet 1992; 340: 35-37.
  CrossrefPubMedGoogle Scholar
• 18 Vincentelli A. et al. Acquired von Willebrand syndrome in aortic stenosis. N Engl J Med 2003; 349: 343-349.
  CrossrefPubMedGoogle Scholar
• 19 Geisen U. et al. Non-surgical bleeding in patients with ventricular assist devices could be explained by acquired von Willebrand disease. Eur J Cardiothorac Surg 2008; 33: 679-684.
  CrossrefPubMedGoogle Scholar
• 20 Uriel N. et al. Acquired von Willebrand syndrome after continuous-flow mechanical device support contributes to a high prevalence of bleeding during long-term support and at the time of transplantation. J Am Coll Cardiol 2010; 56: 1207-1213.
  CrossrefPubMedGoogle Scholar
• 21 Crow S. et al. Gastrointestinal bleeding rates in recipients of nonpulsatile and pulsatile left ventricular assist devices. J Thorac Cardiovasc Surg 2009; 137: 208-215.
  CrossrefPubMedGoogle Scholar
• 22 Pareti FI. et al. Proteolysis of von Willebrand factor and shear stress-induced platelet aggregation in patients with aortic valve stenosis. Circulation 2000; 102: 1290-1295.
  CrossrefPubMedGoogle Scholar
• 23 Meyer AL. et al. Acquired von Willebrand syndrome in patients with an axial flow left ventricular assist device. Circ Heart Fail 2010; 3: 675-681.
  CrossrefPubMedGoogle Scholar
• 24 Thompson JL. et al. Risk of recurrent gastrointestinal bleeding after aortic valve replacement in patients with Heyde syndrome. J Thorac Cardiovasc Surg 2012; 144: 112-116.
  CrossrefPubMedGoogle Scholar
• 25 Lenting PJ. et al. An experimental model to study the in vivo survival of von Willebrand factor. Basic aspects and application to the R1205H mutation. J Biol Chem 2004; 279: 12102-12109.
  CrossrefPubMedGoogle Scholar
• 26 Pegon JN. et al. Factor VIII and von Willebrand factor are ligands for the carbohydrate-receptor Siglec-5. Haematologica 2012; 97: 1855-1863.
  CrossrefPubMedGoogle Scholar
• 27 Meyer D. et al. Hybridoma antibodies to human von Willebrand factor. I. Char-acterisation of seven clones. Br J Haematol 1984; 57: 597-608.
  CrossrefPubMedGoogle Scholar
• 28 Meyer D. et al. Hybridoma antibodies to human von Willebrand factor. II. Relative role of intramolecular loci in mediation of platelet adhesion to the subendothelium. Br J Haematol 1984; 57: 609-620.
  CrossrefPubMedGoogle Scholar
• 29 Rayes J. et al. Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS-13. J Thromb Hae-most 2007; 5: 321-328.
  CrossrefPubMedGoogle Scholar
• 30 Han Y. et al. A shear-based assay for assessing plasma ADAMTS13 activity and inhibitors in patients with thrombotic thrombocytopenic purpura. Transfusion 2011; 51: 1580-1591.
  CrossrefPubMedGoogle Scholar
• 31 Frazier OH. et al. First clinical use of the redesigned HeartMate II left ventricular assist system in the United States: a case report. Tex Heart Inst J 2004; 31: 157-159.
  PubMedGoogle Scholar
• 32 Layet S. et al. Evidence that a secondary binding and protecting site for factor VIII on von Willebrand factor is highly unlikely. Biochem J 1992; 282: 129-137.
  CrossrefPubMedGoogle Scholar
• 33 Eckman PM, John R. Bleeding and thrombosis in patients with continuous-flow ventricular assist devices. Circulation 2012; 125: 3038-3047.
  CrossrefPubMedGoogle Scholar
• 34 Lancellotti S. et al. Proteolytic Processing of Von Willebrand Factor by Adamts13 and Leukocyte Proteases. Mediterr J Hematol Infect Dis 2013; 5: e2013058.
  CrossrefPubMedGoogle Scholar
• 35 Wu JJ. et al. Characterisation of a core binding site for ADAMTS-13 in the A2 domain of von Willebrand factor. Proc Natl Acad Sci USA 2006; 103: 18470-18474.
  CrossrefPubMedGoogle Scholar
• 36 Zhang J. et al. A conformation-sensitive monoclonal antibody against the A2 domain of von Willebrand factor reduces its proteolysis by ADAMTS13. PLoS One 2011; 6: e22157.
  CrossrefPubMedGoogle Scholar
• 37 Klaus C. et al. Epitope mapping of ADAMTS13 autoantibodies in acquired thrombotic thrombocytopenic purpura. Blood 2004; 103: 4514-4519.
  CrossrefPubMedGoogle Scholar
• 38 Luken BM. et al. The spacer domain of ADAMTS13 contains a major binding site for antibodies in patients with thrombotic thrombocytopenic purpura. Thromb Haemost 2005; 93: 267-274.
  Article in Thieme ConnectPubMedGoogle Scholar
• 39 Blackshear JL. et al. Hypertrophic obstructive cardiomyopathy, bleeding history, and acquired von Willebrand syndrome: response to septal myectomy. Mayo Clin Proc 2011; 86: 219-224.
  CrossrefPubMedGoogle Scholar
• 40 Favaloro EJ. et al. Different bleeding risk in type 2A and 2M von Willebrand disease: a 2-year prospective study in 107 patients: a rebuttal. J Thromb Haemost 2012; 10: 1455-1458.
  CrossrefPubMedGoogle Scholar
• 41 Rolf N. et al. Essential thrombocythaemia in a teenage girl resulting in acquired von Willebrand syndrome with joint haemorrhage and menorrhagia. Thromb Haemost 2010; 103: 1272-1274.
  Article in Thieme ConnectPubMedGoogle Scholar
• 42 Heilmann C. et al. Acquired von Willebrand syndrome in patients with extra-corporeal life support (ECLS). Intensive Care Med 2012; 38: 62-68.
  CrossrefPubMedGoogle Scholar