Direct Comparison of Administration Routes for AAV8-mediated Ocular Gene Therapy

Abstract Purpose: We recently demonstrated that direct subretinal (SR) injection of adeno-associated virus (AAV) type 8 (AAV8) into photoreceptor cells and retinal pigment epithelium (RPE) is a highly efficient model of gene delivery. The current study compared transduction efficiency and expression...

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Bibliographic Details
Published in:Current eye research 2013-05, Vol.38 (5), p.569-577
Main Authors: Igarashi, Tsutomu, Miyake, Koichi, Asakawa, Nagisa, Miyake, Noriko, Shimada, Takashi, Takahashi, Hiroshi
Format: Article
Language:English
Subjects:
AAV vector
Animals
damage
Dependovirus - genetics
Electroretinography
eye disease
Eye Diseases - therapy
Gene therapy
Genetic Therapy - adverse effects
Genetic Therapy - methods
Genetic Vectors - genetics
Green Fluorescent Proteins - genetics
Humans
intravitreal
Intravitreal Injections - adverse effects
Intravitreal Injections - methods
IVIS
Luciferases - genetics
Male
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Photoreceptor Cells, Vertebrate
Random Allocation
subretinal
Subretinal Fluid
Transduction, Genetic - methods
type 8
Vitreous Body
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Summary:Abstract Purpose: We recently demonstrated that direct subretinal (SR) injection of adeno-associated virus (AAV) type 8 (AAV8) into photoreceptor cells and retinal pigment epithelium (RPE) is a highly efficient model of gene delivery. The current study compared transduction efficiency and expression patterns associated with various routes of vector administration. Method: The efficacy of intravitreal (VT), SR and subconjunctival (SC) injections for delivery of AAV8-derived vectors, i.e. those expressing luciferase (Luc) and enhanced green fluorescent protein (GFP) - AAV8/Luc and AAV8/GFP, respectively - were compared in an animal (mouse) model (n = 8 mice/group). Transduction efficiency and expression patterns were examined at post-injection weeks 1 and 2, and months 1, 3, 6 and 12 via in vivo imaging. Results: One year after AAV injection, AAV8/Luc-treated mice exhibited stable and sustained high expression of vector in the VT and SR groups, but not in the SC group (VT:SR:SC = 3,218:2,923:115; 1 × 105 photons/s). Histological analysis showed that GFP expression was observed in the inner retina of VT group mice, and in photoreceptor cells and RPE of SR group mice, whereas no GFP expression was noted in the SC group. Electroretinography (ERG) revealed adverse effects following SR delivery. Conclusions: Results suggest that both SR and VT injections of AAV8 vectors are useful routes for administering ocular gene therapy, and stress the importance of selecting an appropriate administration route, i.e. one that targets specific cells, for treating ocular disorders.
ISSN:0271-3683
1460-2202
DOI:10.3109/02713683.2013.779720