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Thromb Haemost 2019; 119(12): 1981-1993
DOI: 10.1055/s-0039-1697662
Coagulation and Fibrinolysis
Georg Thieme Verlag KG Stuttgart · New York

A Thrombin-Activatable Factor X Variant Corrects Hemostasis in a Mouse Model for Hemophilia A

Vincent Muczynski
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Sebastien Verhenne
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Caterina Casari 
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Ghislaine Chérel
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Laurence Panicot-Dubois
2   Faculty of Pharmacy, INSERM UMR-S1076, Aix-Marseille University, Marseille, France
,
Paul Gueguen
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Marc Trossaert
3   Centre Régional de Traitement des Hémophiles, CHU de Nantes, Nantes, France
,
Christophe Dubois
2   Faculty of Pharmacy, INSERM UMR-S1076, Aix-Marseille University, Marseille, France
,
Peter J. Lenting
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Cécile V. Denis‡   *
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
,
Olivier D. Christophe‡   *
1   Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, Le Kremlin-Bicêtre, France
› Author Affiliations
Further Information

Publication History

15 February 2019

15 August 2019

Publication Date:
22 October 2019 (online)

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Abstract

Engineered recombinant factor X (FX) variants represent a promising strategy to bypass the tenase complex and restore hemostasis in hemophilia patients. Previously, a thrombin-activatable FX variant with fibrinopeptide-A replacing the activation peptide (FX-delAP/FpA) has been described in this regard. Here we show that FX-delAP/FpA is characterized by a sixfold shorter circulatory half-life compared with wild-type FX, limiting its therapeutical applicability. We therefore designed a variant in which the FpA sequence is inserted C-terminal to the FX activation peptide (FX/FpA). FX/FpA displayed a similar survival to wt-FX in clearance experiments and could be converted into FX by thrombin and other activating agents. In in vitro assays, FX/FpA efficiently restored thrombin generation in hemophilia A and hemophilia B plasmas, even in the presence of inhibitory antibodies. Expression following hydrodynamic gene transfer of FX/FpA restored thrombus formation in FVIII-deficient mice in a laser-induced injury model as well as hemostasis in a tail-clip bleeding model. Hemostasis after tail transection in FVIII-deficient mice was also corrected at 5 and 90 minutes after injection of purified FX/FpA. Our data indicate that FX/FpA represents a potential tenase-bypassing agent for the treatment of hemophilia patients with or without inhibitors.

Keywords

hemophilia - factor X - factor VIII

Authors' Contributions

V.M., S.V., C.C., O.D.C., G.C., L.P-.D., P.G., C.D., P.J.L., and C.V.D. performed experiments and analyzed data. M.T. provided essential reagents for this study. C.V.D., O.D.C., and P.J.L. wrote the manuscript. All authors contributed to the editing of the final manuscript.


* C.V.D. and O.D.C. contributed equally to this study.


Supplementary Material

 

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