title: AAV-mediated gene delivery in Dp71-null mouse model with compromised barriers.
abstract: Formation and maintenance of the blood-retinal barrier (BRB) is required for proper vision and breaching of this barrier contributes to the pathology in a wide variety of retinal conditions such as retinal detachment and diabetic retinopathy. Dystrophin Dp71 being a key membrane cytoskeletal protein, expressed mainly in Müller cells, its absence has been related to BRB permeability through delocalization and down-regulation of the AQP4 and Kir4.1 channels. Dp71-null mouse is thus an excellent model to approach the study of retinal pathologies showing blood-retinal barrier permeability. We aimed to investigate the participation of Müller cells in the BRB and in the inner limiting membrane of Dp71-null mice compared with wild-type mice in order to understand how these barriers work in this model of permeable BRB. To this aim, we used an Adeno-associated virus (AAV) variant, ShH10-GFP, engineered to target Müller cells specifically. ShH10 coding GFP was introduced by intravitreal injection and Müller cell transduction was studied in Dp71-null mice in comparison to wild-type animals. We show that Müller cell transduction follows a significantly different pattern in Dp71-null mice indicating changes in viral cell-surface receptors as well as differences in the permeability of the inner limiting membrane in this mouse line. However, the compromised BRB of the Dp71-null mice does not lead to virus leakage into the bloodstream when the virus is injected intravitreally - an important consideration for AAV-mediated retinal gene therapy.
authors.affiliation: Institut de la Vision/INSERM/UPMC, Univ Paris 06/CNRS/CHNO des Quinze-Vingts, Paris, France.
authors.forename: Ophélie
authors.initials: O
authors.lastname: Vacca
authors.forename: Marie
authors.initials: M
authors.lastname: Darche
authors.forename: David V
authors.initials: DV
authors.lastname: Schaffer
authors.forename: John G
authors.initials: JG
authors.lastname: Flannery
authors.forename: José-Alain
authors.initials: JA
authors.lastname: Sahel
authors.forename: Alvaro
authors.initials: A
authors.lastname: Rendon
authors.forename: Deniz
authors.initials: D
authors.lastname: Dalkara
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001813:Blood-Retinal Barrier / Q000503:physiopathology*; D004195:Disease Models, Animal; D016189:Dystrophin / Q000172:deficiency* / Q000235:genetics; D063928:Ependymoglial Cells / Q000378:metabolism*; D005654:Fundus Oculi; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010786:Photoreceptor Cells / Q000473:pathology; D012160:Retina / Q000378:metabolism / Q000473:pathology*; D012164:Retinal Diseases / Q000235:genetics / Q000473:pathology*; D041623:Tomography, Optical Coherence; D014795:Visual Pathways / Q000473:pathology / Q000503:physiopathology
chemical_list: D016189:Dystrophin; C085339:apo-dystrophin 1; D049452:Green Fluorescent Proteins
keywords: AAV; Dystrophin Dp71; Müller cell; blood-retinal barrier; inner limiting membrane