1. Improved molecular platform for the gene therapy of rare diseases by liver protein secretion
- Author
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Mickael Quiviger, Aristeidis Giannakopoulos, Sebastien Verhenne, Karen Vanhoorelbeke, Daniel Scherman, Aglaia Athanassiadou, Zsuzsanna Izsvák, Simon F. De Meyer, Corinne Marie, Eleana F. Stavrou, ORANGE, Colette, Unité de Technologies Chimiques et Biologiques pour la Santé (UTCBS - UM 4 (UMR 8258 / U1022)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), University of Patras, School of Medicine, Laboratory for Thrombosis Research [Kortrijk, Belgium], KU Leuven Campus Kulak Kortrijk [Belgium], Max Delbrück Center for Molecular Medicine [Berlin] (MDC), Helmholtz-Gemeinschaft = Helmholtz Association, and Institut de Chimie du CNRS (INC)-Université de Paris (UP)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
0301 basic medicine ,[SDV.BIO]Life Sciences [q-bio]/Biotechnology ,VON-WILLEBRAND-FACTOR ,Genetic enhancement ,Transgene ,Transposases ,VECTOR ,Bioinformatics ,03 medical and health sciences ,DELIVERY ,0302 clinical medicine ,Protein replacement therapy ,Rare Diseases ,Von Willebrand factor ,S/MAR ELEMENT ,PLASMID DNA ,von Willebrand Factor ,Genetics ,Medicine ,Humans ,EPISOMAL TRANSGENE EXPRESSION ,HUMAN ALPHA-1-ANTITRYPSIN ,Gene ,Genetics (clinical) ,Transposase ,Reporter gene ,SLEEPING-BEAUTY ,biology ,business.industry ,General Medicine ,Genetic Therapy ,Sleeping Beauty transposon system ,3. Good health ,[SDV.BIO] Life Sciences [q-bio]/Biotechnology ,030104 developmental biology ,Liver ,HEMOPHILIA-B ,030220 oncology & carcinogenesis ,biology.protein ,DNA Transposable Elements ,EXPRESSION IN-VIVO ,business - Abstract
International audience; Many rare monogenic diseases are treated by protein replacement therapy, in which the missing protein is repetitively administered to the patient. However, in several cases, the missing protein is required at a high and sustained level, which renders protein therapy far from being adequate. As an alternative, a gene therapy treatment ensuring a sustained effectiveness would be particularly valuable. Liver is an optimal organ for the secretion and systemic distribution of a therapeutic transgene product. Cutting edge non-viral gene therapy tools were tested in order to produce a high and sustained level of therapeutic protein secretion by the liver using the hydrodynamic delivery technique. The use of S/MAR matrix attachment region provided a slight, however not statistically significant, increase in the expression of a reporter gene in the liver. We have selected the von Willebrand Factor (vWF) gene as a particularly challenging large gene (8.4 kb) for liver delivery and expression, and also because a high vWF blood concentration is required for disease correction. By using the optimized miniplasmid pFAR free of antibiotic resistance gene together with the Sleeping Beauty transposon and the hyperactive SB100X transposase, we have obtained a sustainable level of vWFblood secretion by the liver, at 65% of physiological level. Our results point to the general use of this plasmid platform using the liver as a protein factory to treat numerous rare disorders by gene therapy.
- Published
- 2018