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DOI: 10.1055/s-0039-1697662
A Thrombin-Activatable Factor X Variant Corrects Hemostasis in a Mouse Model for Hemophilia A
Publication History
15 February 2019
15 August 2019
Publication Date:
22 October 2019 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/201912/lookinside/thumbnails/10-1055-s-0039-1697662_190095-1.jpg)
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.
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.
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