The absorption enhancer sodium deoxycholate promotes high gene transfer in skeletal muscles.
Gene delivery to skeletal muscle is a promising strategy for the treatment of muscle disorders and for the systemic secretion of therapeutic proteins. In addition, muscle is an attractive target tissue because it is easily accessible. However, very few synthetic vectors proved capable of surpassing naked DNA mediated muscle gene transfer. In fact, only neutral copolymers, in particular poloxamers, demonstrated capacities to increase transgene expression in skeletal muscles. Here, we studied in vitro and in vivo behaviour of different bile salts. We report that sodium deoxycholate (DOC) and derivatives thereof increase after intramuscular injection by more than 100-fold the levels of the reporter gene luciferase compared to naked DNA. Using a LacZ expression cassette, we found that more than 20% of the muscle fibers expressed the reporter gene. Prolonged expression of a secreted reporter gene derived from a natural murine alkaline phosphatase enzyme could be documented. Altogether, our results demonstrate that bile salts belong to the most efficient chemicals identified so far for skeletal muscle gene transfer. Importantly, since these compounds are naturally found in the body, there is no risk of immune response against them and in addition several bile salts are already used in human medicine. Bile salt mediated muscle gene transfer may thus have broad applications in gene therapy.
Genethon, BP60, 91002 Evry cedex, France.
Christian
C
Leborgne
Genethon, BP60, 91002 Evry cedex, France.
Debborah
D
Alimi-Guez
Unité de Technologies Chimiques et Biologiques pour la Santé, CNRS UMR8258 - Inserm U1022 - Université Paris Descartes, Chimie ParisTech, 75006 Paris, France.
Nelly
N
El Shafey
Genethon, BP60, 91002 Evry cedex, France.
Laetitia
L
van Wittenberghe
Unité de Technologies Chimiques et Biologiques pour la Santé, CNRS UMR8258 - Inserm U1022 - Université Paris Descartes, Chimie ParisTech, 75006 Paris, France.
Pascal
P
Bigey
Genethon, BP60, 91002 Evry cedex, France; Unité de Technologies Chimiques et Biologiques pour la Santé, CNRS UMR8258 - Inserm U1022 - Université Paris Descartes, Chimie ParisTech, 75006 Paris, France.
Daniel
D
Scherman
Genethon, BP60, 91002 Evry cedex, France; Unité de Technologies Chimiques et Biologiques pour la Santé, CNRS UMR8258 - Inserm U1022 - Université Paris Descartes, Chimie ParisTech, 75006 Paris, France; Laboratoire de Conception et Application de Molécules Bioactives UMR7199 CNRS - Université de Strasbourg, LabEx Medalis, Faculté de Pharmacie, 67401 Illkirch, France. Electronic address: kichler@unistra.fr.
Antoine
A
Kichler
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage*; D003840:Deoxycholic Acid / Q000008:administration & dosage*; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D007273:Injections, Intramuscular; D007763:Lac Operon; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D049409:Luciferases, Firefly / Q000235:genetics; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000378:metabolism*
D003840:Deoxycholic Acid; D004247:DNA; D008156:Luciferases; D049409:Luciferases, Firefly; D000469:Alkaline Phosphatase
Bile salts; Gene transfer; Non-viral vectors; Skeletal muscle; Sodium deoxycholate