Stable and efficient intraocular gene transfer using pseudotyped EIAV lentiviral vectors.
BACKGROUND
We have developed minimal non-primate lentiviral vectors based on the equine infectious anaemia virus (EIAV). We evaluated the in vivo expression profiles of these vectors delivered regionally to ocular tissues to define their potential utility in ocular gene therapy.


METHODS
EIAV vectors pseudotyped with VSV-G or rabies-G envelope proteins were delivered subretinally, intravitreally or into the anterior chambers (intracameral administration) in mice. Reporter gene (eGFP) expression was analysed using in vivo retinal imaging or histological examination of eyes and brains at intervals between 3 days and 16 months. We investigated the effects of vector titre, pseudotype, genome configuration, site of intraocular administration, intentional retinal trauma and the degree of retinal maturation on the spatial and temporal expression profiles of these vectors.


RESULTS
Subretinal vector delivery resulted in efficient and stable transduction of retinal pigment epithelial (RPE) cells and variable transduction of photoreceptors up to 16 months post-injection. Retinal trauma facilitated the local transduction of neurosensory retinal cells. Intracameral administration of VSV-G- but not rabies-G-pseudotyped vectors produced stable eGFP expression in corneal endothelial cells and trabecular meshwork.


CONCLUSIONS
The cellular tropism and expression kinetics of optimised EIAV vectors after intraocular administration make them attractive vehicles for delivering therapeutic genes in the management of inherited and acquired retinal and anterior segment disorders.
Balaggan
K S
KS
Division of Molecular Therapy, Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
Binley
K
K
Esapa
M
M
Iqball
S
S
Askham
Z
Z
Kan
O
O
Tschernutter
M
M
Bainbridge
J W B
JW
Naylor
S
S
Ali
R R
RR
D000818:Animals; D005260:Female; D020869:Gene Expression Profiling; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D004860:Infectious Anemia Virus, Equine / Q000235:genetics*; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D010786:Photoreceptor Cells; D010857:Pigment Epithelium of Eye / Q000502:physiology; D012160:Retina / Q000293:injuries; D012164:Retinal Diseases / Q000235:genetics / Q000628:therapy*; D014161:Transduction, Genetic; D018523:Tropism
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins