Antithrombotic potential of a single-domain antibody enhancing the activated protein C-cofactor activity of protein S

Josepha C Sedzro, Frédéric Adam, Claire Auditeau, Elsa Bianchini, Allan De Carvalho, Ivan Peyron, Sadyo Daramé, Sophie Gandrille, Stella Thomassen, Tilman M Hackeng, Olivier D Christophe, Peter J Lenting, Cécile V Denis, Delphine Borgel, François Saller*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

11 Downloads (Pure)

Abstract

Background: Protein S (PS) is a natural anticoagulant acting as a cofactor for activated protein C (APC) in the proteolytic inactivation of activated factors V (FVa) and VIII (FVIIIa), but also for tissue factor pathway inhibitor α (TFPIα) in the inhibition of activated factor X (FXa). Objective: For therapeutic purposes, we aimed at generating single-domain antibodies (sdAbs) that could specifically modulate the APC-cofactor activity of PS in vivo. Methods: A llama-derived immune library of sdAbs was generated and screened on recombinant human PS by phage display. PS binders were tested in a global activated partial thromboplastin time (APTT)-based APC-cofactor activity assay. Results: A PS-specific sdAb (PS003) was found to enhance the APC-cofactor activity of PS in our APTT-based assay, and this enhancing effect was greater for a bivalent form of PS003 (PS003biv). Further characterization of PS003biv demonstrated that PS003biv also enhanced the APC-cofactor activity of PS in a tissue factor (TF)-induced thrombin generation assay and stimulated APC in the inactivation of FVa, but not FVIIIa, in plasma-based assays. Furthermore, PS003biv was directed against the sex hormone-binding globulin (SHBG)-like domain but did not inhibit the binding of PS to C4b-binding protein (C4BP) and did not interfere with the TFPIα-cofactor activity of PS. In mice, PS003biv exerted an antithrombotic effect in a FeCl 3-induced thrombosis model, while not affecting physiological hemostasis in a tail-clip bleeding model. Discussion: Altogether, these results showed that pharmacological enhancement of the APC-cofactor activity of PS through an original anti-PS sdAb might constitute a promising and safe antithrombotic strategy.

Original languageEnglish
Pages (from-to)1653-1664
Number of pages12
JournalJournal of Thrombosis and Haemostasis
Volume20
Issue number7
Early online date21 Apr 2022
DOIs
Publication statusPublished - Jul 2022

Keywords

  • BINDING
  • COMPLEX
  • DEFICIENCY
  • FACTOR-VA
  • FACTOR-XA
  • INACTIVATION
  • INHIBITION
  • MEDIATED CLEAVAGES
  • PROTHROMBINASE
  • TISSUE FACTOR PATHWAY
  • blood coagulation
  • protein C
  • protein S
  • single-domain antibodies
  • thrombosis

Fingerprint

Dive into the research topics of 'Antithrombotic potential of a single-domain antibody enhancing the activated protein C-cofactor activity of protein S'. Together they form a unique fingerprint.

Cite this