title: Vitrectomy Before Intravitreal Injection of AAV2/2 Vector Promotes Efficient Transduction of Retinal Ganglion Cells in Dogs and Nonhuman Primates.
abstract: Recombinant adeno-associated virus (AAV) has emerged as a promising vector for retinal gene delivery to restore visual function in certain forms of inherited retinal dystrophies. Several studies in rodent models have shown that intravitreal injection of the AAV2/2 vector is the optimal route for efficient retinal ganglion cell (RGC) transduction. However, translation of these findings to larger species, including humans, is complicated by anatomical differences in the eye, a key difference being the comparatively smaller volume of the vitreous chamber in rodents. Here, we address the role of the vitreous body as a potential barrier to AAV2/2 diffusion and transduction in the RGCs of dogs and macaques, two of the most relevant preclinical models. We intravitreally administered the AAV2/2 vector carrying the CMV-eGFP reporter cassette in dog and macaque eyes, either directly into the vitreous chamber or after complete vitrectomy, a surgical procedure that removes the vitreous body. Our findings suggest that the vitreous body appears to trap the injected vector, thus impairing the diffusion and transduction of AAV2/2 to inner retinal neurons. We show that vitrectomy before intravitreal vector injection is an effective means of overcoming this physical barrier, improving the transduction of RGCs in dog and macaque retinas. These findings support the use of vitrectomy in clinical trials of intravitreal gene transfer techniques targeting inner retinal neurons.
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Kizito-Tshitoko
authors.initials: KT
authors.lastname: Tshilenge
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Baptiste
authors.initials: B
authors.lastname: Ameline
authors.affiliation: 2 CHU de Nantes, Service d'Ophtalmologie, Nantes, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Weber
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Mendes-Madeira
authors.affiliation: 3 Cellular and Tissular Imaging Core Facility of Nantes University, SFR Santé Francois Bonamy INSERM UMS016/CNRS UMS3556, Nantes, France .
authors.forename: Steven
authors.initials: S
authors.lastname: Nedellec
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Marine
authors.initials: M
authors.lastname: Biget
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Provost
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Lyse
authors.initials: L
authors.lastname: Libeau
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Blouin
authors.affiliation: 4 Emergency and Critical Care Unit, ONIRIS, Nantes-Atlantic College of Veterinary Medicine Food Science and Engineering, Nantes, France .
authors.forename: Jack-Yves
authors.initials: JY
authors.lastname: Deschamps
authors.affiliation: 2 CHU de Nantes, Service d'Ophtalmologie, Nantes, France.
authors.forename: Guylène
authors.initials: G
authors.lastname: Le Meur
authors.affiliation: 5 UMR 703 PAnTher INRA/ONIRIS, Nantes-Atlantic College of Veterinary Medicine Food Science and Engineering, Nantes, France .
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: Colle
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.|6 Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida , Gainesville, Florida.
authors.forename: Philippe
authors.initials: P
authors.lastname: Moullier
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Pichard
authors.affiliation: 1 Atlantic Gene Therapies, INSERM UMR 1089, Université de Nantes, CHU de Nantes, Nantes, France.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Rolling
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics; D004285:Dogs; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use*; D049452:Green Fluorescent Proteins; D006801:Humans; D058449:Intravitreal Injections; D008251:Macaca; D012160:Retina / Q000473:pathology / Q000637:transplantation; D012165:Retinal Ganglion Cells; D014161:Transduction, Genetic; D014821:Vitrectomy
chemical_list: D049452:Green Fluorescent Proteins