title: Ocular cell transfection with the human basic fibroblast growth factor gene delays photoreceptor cell degeneration in RCS rats.
abstract: Based on the K8/JTS-1-mediated transfection technique, we developed an in vivo protocol for an efficient transfer of plasmid DNA to ocular cells. As determined with condensed plasmids containing reporter genes for either beta-galactosidase (pcDNA-lacZ) or enhanced green fluorescent protein (pREP-EGFP), the immortalized human retinal epithelial cells RPE D407 and human embryonic kidney 293 cells can be transfected with typical efficiencies of 11 and 19%, respectively. Unlike 293 cells, RPE D407 cells had a reduced viability on transfection with both plasmids. In vivo, subretinal injections of DNA-K8/JTS-1 complexes revealed reporter gene expression in choroidal and RPE cells of normal pink-eyed Royal College of Surgeons (RCS) rats. The validity of this transfection technique in terms of retinal cell survival in RCS rats was then examined by using pREP-hFGF2 plasmid, which encodes the human basic fibroblast growth factor isoforms (hFGF2). Subretinal injection of pREP-hFGF2-K8/JTS-1 complexes into 3-week-old dystrophic RCS rat eyes reveals a delayed photoreceptor cell degeneration 60 days postinjection. In this case, the average analyzed field points with delayed cell dystrophy represent 14 to 17% of the retinal surface as compared with 2.6 and 4% in pREP5beta and vehicle-injected eyes, respectively. Peptide-mediated in oculo transfection thus appears to be a promising technique for the treatment of retinal cell and photoreceptor degenerations.
authors.affiliation: Centre de Recherche Thérapeutique en Ophtalmologie, Equipe d'Accueil no. 2502 du Ministère de la Recherche et de l'Enseignement Supérieur, Université René Descartes Paris V, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Neuner-Jehle
authors.forename: L V
authors.initials: LV
authors.lastname: Berghe
authors.forename: S
authors.initials: S
authors.lastname: Bonnel
authors.forename: Y
authors.initials: Y
authors.lastname: Uteza
authors.forename: F
authors.initials: F
authors.lastname: Benmeziane
authors.forename: J S
authors.initials: JS
authors.lastname: Rouillot
authors.forename: D
authors.initials: D
authors.lastname: Marchant
authors.forename: A
authors.initials: A
authors.lastname: Kobetz
authors.forename: J L
authors.initials: JL
authors.lastname: Dufier
authors.forename: M
authors.initials: M
authors.lastname: Menasche
authors.forename: M
authors.initials: M
authors.lastname: Abitbol
mesh_terms: D000818:Animals; D001483:Base Sequence; D002470:Cell Survival / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008845:Microinjections; D008969:Molecular Sequence Data; D020419:Photoreceptor Cells, Vertebrate / Q000166:cytology / Q000473:pathology* / Q000502:physiology; D010857:Pigment Epithelium of Eye / Q000166:cytology; D010957:Plasmids / Q000235:genetics; D051381:Rats; D012162:Retinal Degeneration / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase