title: Gene therapy in ophthalmology: validation on cultured retinal cells and explants from postmortem human eyes.
abstract: Gene therapy studies in primates can provide important information regarding vector tropism, specific cellular expression, biodistribution, and safety prior to clinical trials. In this study, we report the assessment of transduction efficiency of recombinant adeno-associated virus (rAAV) vectors using human postmortem retina. Transductions were performed using two in vitro models prepared from human tissue: dissociated cell cultures and retinal explants. These models were used to assess cellular tropism and selectivity of rAAV vectors encoding for fluorescent proteins under the control of different promoters. These promoters were a ubiquitous cytomegalovirus promoter and a cell type-specific promoter targeting expression in ON bipolar cells. The results demonstrate that this in vitro approach can limit the use of living primates for the validation of gene therapy in vision and ophthalmology.
authors.affiliation: INSERM, U968, Paris, 75 012, France.
authors.forename: Mathias
authors.initials: M
authors.lastname: Fradot
authors.forename: Volker
authors.initials: V
authors.lastname: Busskamp
authors.forename: Valérie
authors.initials: V
authors.lastname: Forster
authors.forename: Thérèse
authors.initials: T
authors.lastname: Cronin
authors.forename: Thierry
authors.initials: T
authors.lastname: Léveillard
authors.forename: Jean
authors.initials: J
authors.lastname: Bennett
authors.forename: José-Alain
authors.initials: JA
authors.lastname: Sahel
authors.forename: Botond
authors.initials: B
authors.lastname: Roska
authors.forename: Serge
authors.initials: S
authors.lastname: Picaud
mesh_terms: D002478:Cells, Cultured; D000229:Dependovirus; D005453:Fluorescence; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000627:therapeutic use*; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D009885:Ophthalmology / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012160:Retina / Q000166:cytology*; D012164:Retinal Diseases / Q000235:genetics / Q000628:therapy*; D014161:Transduction, Genetic / Q000379:methods*