title: AAV-mediated RLBP1 gene therapy improves the rate of dark adaptation in Rlbp1 knockout mice.
abstract: Recessive mutations in RLBP1 cause a form of retinitis pigmentosa in which the retina, before its degeneration leads to blindness, abnormally slowly recovers sensitivity after exposure to light. To develop a potential gene therapy for this condition, we tested multiple recombinant adeno-associated vectors (rAAVs) composed of different promoters, capsid serotypes, and genome conformations. We generated rAAVs in which sequences from the promoters of the human RLBP1, RPE65, or BEST1 genes drove the expression of a reporter gene (green fluorescent protein). A promoter derived from the RLBP1 gene mediated expression in the retinal pigment epithelium and Müller cells (the intended target cell types) at qualitatively higher levels than in other retinal cell types in wild-type mice and monkeys. With this promoter upstream of the coding sequence of the human RLBP1 gene, we compared the potencies of vectors with an AAV2 versus an AAV8 capsid in transducing mouse retinas, and we compared vectors with a self-complementary versus a single-stranded genome. The optimal vector (scAAV8-pRLBP1-hRLBP1) had serotype 8 capsid and a self-complementary genome. Subretinal injection of scAAV8-pRLBP1-hRLBP1 in Rlbp1 nullizygous mice improved the rate of dark adaptation based on scotopic (rod-plus-cone) and photopic (cone) electroretinograms (ERGs). The effect was still present after 1 year.
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Vivian W
authors.initials: VW
authors.lastname: Choi
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Chad E
authors.initials: CE
authors.lastname: Bigelow
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Terri L
authors.initials: TL
authors.lastname: McGee
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Akshata N
authors.initials: AN
authors.lastname: Gujar
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Shawn M
authors.initials: SM
authors.lastname: Hanks
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Joanna
authors.initials: J
authors.lastname: Vrouvlianis
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Maker
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Barrett
authors.initials: B
authors.lastname: Leehy
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Yiqin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Jorge
authors.initials: J
authors.lastname: Aranda
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: George
authors.initials: G
authors.lastname: Bounoutas
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: John T
authors.initials: JT
authors.lastname: Demirs
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Junzheng
authors.initials: J
authors.lastname: Yang
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Fort Worth, Texas, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Ornberg
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Fort Worth, Texas, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Preclinical Safety, Alcon , Fort Worth, Texas, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Martin
authors.affiliation: Preclinical Safety, Alcon , Fort Worth, Texas, USA.
authors.forename: Kelly R
authors.initials: KR
authors.lastname: Stout
authors.affiliation: Preclinical Safety, Novartis Institutes for BioMedical Research , East Hanover, New Jersey, USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Argentieri
authors.affiliation: Preclinical Safety, Novartis Institutes for BioMedical Research , East Hanover, New Jersey, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Grosenstein
authors.affiliation: Preclinical Safety, Novartis Institutes for BioMedical Research , East Hanover, New Jersey, USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Diaz
authors.affiliation: Preclinical Safety, Novartis Institutes for BioMedical Research , East Hanover, New Jersey, USA.
authors.forename: Oliver
authors.initials: O
authors.lastname: Turner
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Jaffee
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Seshidhar R
authors.initials: SR
authors.lastname: Police
authors.affiliation: Ophthalmology Disease Area, Novartis Institutes for BioMedical Research , Cambridge, Massachusetts, USA.
authors.forename: Thaddeus P
authors.initials: TP
authors.lastname: Dryja