title: A nonionic amphiphile agent promotes gene delivery in vivo to skeletal and cardiac muscles.
abstract: Direct injection of naked DNA into skeletal or cardiac muscle induces detectable gene expression. Although this provides a practical system for transgene expression, the reported efficacy is too low to confer a therapeutic benefit. By following a rational strategy based on the supramolecular structures adopted by active complexes, we have discovered a novel nonionic amphiphile synthetic agent [poly(ethyleneoxide)(13)-poly(propyleneoxide)(30)-poly(ethyleneoxide)(13) block copolymer; PE6400] that enables gene expression in up to 35% of muscle fibers from mouse tibial cranial muscle. PE6400 abolishes the ceiling effect on transgene expression of increasing amounts of naked DNA and permits long-term expression of the beta-galactosidase reporter gene in immunologically tolerant transgenic rats. This improvement in gene expression over naked DNA was observed irrespective of the reporter gene, ranging from 0.7 to 3.4 kb, and of the animal model used. In skeletal muscle, the PE6400 formulation led to a level of transfection efficiency similar to that obtained by electrotransfer. PE6400 also promotes high transgene expression in cardiac muscle. In contrast, PE6400-DNA formulations were inefficient in vitro in established cell lines and in isolated cardiomyocytes. When microinjected into the cell cytoplasm, PE6400 promotes DNA trafficking into the nucleus and induces gene expression. PE6400 provides a simple gene delivery system for skeletal and myocardial gene transfer. We propose that the PE6400 formulation could serve for the treatment of diseases primarily affecting muscle or for the expression of therapeutic proteins for local or systemic benefit.
authors.affiliation: Inserm U533, Faculté de Médecine, 44000 Nantes, France. bruno.pitard@nantes.inserm.fr
authors.forename: Bruno
authors.initials: B
authors.lastname: Pitard
authors.forename: Hélène
authors.initials: H
authors.lastname: Pollard
authors.forename: Onnik
authors.initials: O
authors.lastname: Agbulut
authors.forename: Olivier
authors.initials: O
authors.lastname: Lambert
authors.forename: Jean-Thomas
authors.initials: JT
authors.lastname: Vilquin
authors.forename: Yan
authors.initials: Y
authors.lastname: Cherel
authors.forename: Jérôme
authors.initials: J
authors.lastname: Abadie
authors.forename: Jane-Lise
authors.initials: JL
authors.lastname: Samuel
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Rigaud
authors.forename: Severine
authors.initials: S
authors.lastname: Menoret
authors.forename: Ignacio
authors.initials: I
authors.lastname: Anegon
authors.forename: Denis
authors.initials: D
authors.lastname: Escande
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D019556:COS Cells / Q000378:metabolism; D002522:Chlorocebus aethiops; D020285:Cryoelectron Microscopy; D004274:DNA, Recombinant / Q000008:administration & dosage* / Q000235:genetics; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D016325:HIV Long Terminal Repeat; D006321:Heart; D007267:Injections; D007273:Injections, Intramuscular; D007763:Lac Operon; D008081:Liposomes; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008845:Microinjections; D018482:Muscle, Skeletal / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D004274:DNA, Recombinant; D008081:Liposomes; D011993:Recombinant Fusion Proteins; C472329:poly(ethyleneoxide)(13)-poly(propyleneoxide)(30)-poly(ethyleneoxide)(13) block copolymer; D011092:Polyethylene Glycols; D001616:beta-Galactosidase