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

Thromb Haemost. 2014 Nov;112(5):1014-23. doi: 10.1160/TH14-02-0148. Epub 2014 Jul 17.

Abstract

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 blood-perfusion system, mAb508 was able to reduce degradation of high-molecular-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.

Keywords: ADAMTS13; acquired von Willebrand syndrome; antibody therapy; circulatory assist devices; von Willebrand factor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / metabolism*
  • ADAMTS13 Protein
  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / pharmacology*
  • Antigen-Antibody Reactions
  • Dose-Response Relationship, Immunologic
  • Epitopes / immunology
  • Heart-Assist Devices / adverse effects*
  • Hemorheology
  • Hemorrhage / blood
  • Hemorrhage / etiology
  • Hemorrhage / prevention & control*
  • Hemostasis
  • Humans
  • Mice
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / immunology
  • Protein Structure, Tertiary
  • Proteolysis
  • Recombinant Proteins / metabolism
  • von Willebrand Factor / immunology
  • von Willebrand Factor / metabolism*

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • Peptide Fragments
  • Recombinant Proteins
  • von Willebrand Factor
  • ADAM Proteins
  • ADAMTS13 Protein
  • ADAMTS13 protein, human