A novel ELISA-based diagnosis of acquired von Willebrand disease with increased VWF proteolysis.

Abstract

Von Willebrand disease-type 2A (VWD-2A) and acquired von Willebrand syndrome (AVWS) due to aortic stenosis (AS) or left ventricular assist device (LVAD) are associated with an increased proteolysis of von Willebrand factor (VWF). Analysis of VWF multimeric profile is the most sensitive way to assess such increased VWF-proteolysis. However, several technical aspects hamper a large diffusion among routine diagnosis laboratories. This makes early diagnosis and early appropriate care of increased proteolysis challenging. In this context of unmet medical need, we developed a new ELISA aiming a quick, easy and reliable assessment of VWF-proteolysis. This ELISA was assessed successively in a LVAD-model, healthy subjects (n=39), acquired TTP-patients (n=4), VWD-patients (including VWD-2A(IIA), n=22; VWD-2B, n=26; VWD-2A(IIE), n=21; and VWD-1C, n=8) and in AVWS-patients (AS, n=9; LVAD, n=9; and MGUS, n=8). A standard of VWF-proteolysis was specifically developed. Extent of VWF-proteolysis was expressed as relative percentage and as VWF proteolysis/VWF:Ag ratio. A speed-dependent increase in VWF-proteolysis was assessed in the LVAD model whereas no proteolysis was observed in TTP-patients. In VWD-patients, VWF-proteolysis was significantly increased in VWD-2A(IIA) and VWD-2B and significantly decreased in VWD-2A(IIE) versus controls (p< 0.0001). In AVWS-patients, VWF-proteolysis was significantly increased in AS- and LVAD-patients compared to controls (p< 0.0001) and not detectable in MGUS-patients. A significant increase in VWF-proteolysis was detected as soon as three hours after LVAD implantation (p< 0.01). In conclusion, we describe a new ELISA allowing a rapid and accurate diagnosis of VWF-proteolysis validated in three different clinical situations. This assay represents a helpful alternative to electrophoresis-based assay in the diagnosis and management of AVWS with increased VWF-proteolysis.

• Authors: Antoine Rauch, Claudine Caron, Flavien Vincent, Emmanuelle Jeanpierre, Catherine Ternisien, Pierre Boisseau, Christophe Zawadzki, Edith Fressinaud, Annie Borel-Derlon, Sylvie Hermoire, Camille Paris, Cécile Lavenu-Bombled, Agnès Veyradier, Alexandre Ung, André Vincentelli, Eric van Belle, Peter J Lenting, Jenny Goudemand, Sophie Susen
• Journal: Thrombosis and haemostasis (Thromb Haemost) Vol. 115 Issue 5 Pg. 950-9 (05 02 2016) ISSN: 2567-689X [Electronic] Germany
• PMID: 26791163 (Publication Type: Journal Article, Validation Study)

Chemical References

• von Willebrand Factor

Topics

• Amino Acid Substitution
• Aortic Valve Stenosis (complications)
• Case-Control Studies
• Enzyme-Linked Immunosorbent Assay (methods)
• Heart-Assist Devices (adverse effects)
• Humans
• Mutation, Missense
• Protein Multimerization
• Proteolysis
• von Willebrand Disease, Type 2 (blood, diagnosis)
• von Willebrand Diseases (blood, diagnosis, etiology)
• von Willebrand Factor (chemistry, genetics, metabolism)