title: In vivo gene transfer into the ocular ciliary muscle mediated by ultrasound and microbubbles.
abstract: This study aimed to assess application of ultrasound (US) combined with microbubbles (MB) to transfect the ciliary muscle of rat eyes. Reporter DNA plasmids encoding for Gaussia luciferase, β-galactosidase or the green fluorescent protein (GFP), alone or mixed with 50% Artison MB, were injected into the ciliary muscle, with or without US exposure (US set at 1 MHz, 2 W/cm(2), 50% duty cycle for 2 min). Luciferase activity was measured in ocular fluids at 7 and 30 days after sonoporation. At 1 week, the US+MB treatment showed a significant increase in luminescence compared with control eyes, injected with plasmid only, with or without MB (×2.6), and, reporter proteins were localized in the ciliary muscle by histochemical analysis. At 1 month, a significant decrease in luciferase activity was observed in all groups. A rise in lens and ciliary muscle temperature was measured during the procedure but did not result in any observable or microscopic damages at 1 and 8 days. The feasibility to transfer gene into the ciliary muscle by US and MB suggests that sonoporation may allow intraocular production of proteins for the treatment of inflammatory, angiogenic and/or degenerative retinal diseases.
authors.affiliation: Inserm U872, Physiopathology of Ocular Diseases: Therapeutic Innovations, Paris, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Kowalczuk
authors.forename: Michèle
authors.initials: M
authors.lastname: Boudinet
authors.forename: Mohamed
authors.initials: M
authors.lastname: El Sanharawi
authors.forename: Elodie
authors.initials: E
authors.lastname: Touchard
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Naud
authors.forename: Amena
authors.initials: A
authors.lastname: Saïed
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Jeanny
authors.forename: Francine
authors.initials: F
authors.lastname: Behar-Cohen
authors.forename: Pascal
authors.initials: P
authors.lastname: Laugier
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002924:Ciliary Body / Q000166:cytology / Q000378:metabolism*; D005128:Eye Diseases / Q000235:genetics / Q000628:therapy; D005240:Feasibility Studies; D005260:Female; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D045423:Microbubbles; D010957:Plasmids; D051381:Rats; D011917:Rats, Inbred Lew; D013010:Sonication; D018709:Statistics, Nonparametric; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D049452:Green Fluorescent Proteins; D008156:Luciferases; D001616:beta-Galactosidase