Page 287 - Haematologica April 2020

Coagulation & its Disorders
Correction of bleeding in experimental severe hemophilia A by systemic delivery of factor VIII-encoding mRNA
Jules Russick,1 Sandrine Delignat,1 Peter Milanov,2 Olivier Christophe,3 Gábor Boros,4 Cécile V. Denis,3 Peter J. Lenting,3 Srinivas V. Kaveri1 and Sébastien Lacroix-Demazes1
1Centre de Recherche des Cordeliers, INSERM, Sorbonne Université, Université de Paris, Paris, France; 2DRK-Blutspendedienst, Institut für Transfusionsmedizin und Immunhämatologie, Frankfurt am Main, Germany; 3HITh, UMR_S1176, INSERM, Université Paris-Saclay, Le Kremlin-Bicêtre, France and 4BioNTech RNA Pharmaceuticals, Mainz, Germany
ABSTRACT
The treatment or prevention of bleeding in patients with hemophilia A relies on replacement therapy with different factor VIII (FVIII)-con- taining products or on the use of by-passing agents, i.e., activated prothrombin complex concentrates or recombinant activated factor VII. Emerging approaches include the use of bispecific anti-factor IXa/factor X antibodies, anti-tissue factor pathway inhibitor antibodies, interfering RNA to antithrombin, and activated protein C-specific serpins or gene therapy. The latter strategies are, however, hampered by the short clinical experi- ence and potential adverse effects including the absence of tight temporal and spatial control of coagulation and the risk of uncontrolled insertional mutagenesis. Systemic delivery of mRNA allows endogenous production of the corresponding encoded protein. Thus, injection of erythropoietin- encoding mRNA in a lipid nanoparticle formulation resulted in increased erythropoiesis in mice and macaques. Here, we demonstrate that a single injection of in vitro transcribed B domain-deleted FVIII-encoding mRNA to FVIII-deficient mice enables endogenous production of pro-coagulant FVIII. Circulating FVIII:C levels above 5% of normal levels were maintained for up to 72 h, with an estimated half-life of FVIII production of 17.9 h, and cor- rected the bleeding phenotype in a tail clipping assay. The endogenously produced FVIII did however exhibit low specific activity and induced a potent neutralizing IgG response upon repeated administration of the mRNA. Our results suggest that the administration of mRNA is a plausible strategy for the endogenous production of proteins characterized by poor translational efficacy. The use of alternative mRNA delivery systems and improved FVIII-encoding mRNA should foster the production of functional molecules and reduce their immunogenicity.
Introduction
Hemophilia A is a rare X-linked hemorrhagic disorder that results from insuffi- cient plasma levels of pro-coagulant factor VIII (FVIII).1 Replacement therapy using exogenous FVIII is to date the most efficient strategy to treat or prevent bleeds. It is however extremely expensive because of the elevated production costs, the short half-life of therapeutic FVIII and the need for life-long treatment. Several alternative strategies to correct bleeding include the use of FVIII by-passing agents, such as activated prothrombin complex concentrates, recombinant factor VIIa or mono- clonal FVIII-mimicking bispecific antibodies,2 the injection of anti-tissue factor pathway inhibitor,3 of interfering RNA to antithrombin (AT)4 or of activated protein C-specific serpins,5 and gene therapy.6 Each of these promising therapies does, however, have intrinsic challenges that may limit broad application.
In vivo production of proteins following the administration of mRNA was demon- strated in the early 1990s in the case of luciferase and β-galactosidase,7 leading to the first clinical trial with mRNA a decade later.8 Concomitantly, both double- and sin-
Ferrata Storti Foundation
Haematologica 2020 Volume 105(4):1129-1137
Correspondence:
SÉBASTIEN LACROIX-DESMAZES
sebastien.lacroix-desmazes@crc.jussieu.fr
Received: October 30, 2018. Accepted: July 5, 2019. Pre-published: July 9, 2019.
doi:10.3324/haematol.2018.210583
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/105/4/1129
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