Non-invasive observation of repeated adenoviral GFP gene delivery to the anterior segment of the monkey eye in vivo.
BACKGROUND
Glaucoma is a group of chronic eye diseases often associated with an elevated intraocular pressure (IOP). If not controlled, the condition leads to blindness. The eye tissue responsible for maintaining aqueous humor resistance and thus normal IOP is the trabecular meshwork (TM). Adenoviral vectors are capable of transducing the TM in several rodent species. Because of the relevance of the non-human primate model in the study of glaucoma, gene transfer to the eyes of cynomolgus monkeys was investigated.


METHODS
Four cynomolgus monkeys were injected with AdenoGFP into the anterior chamber: two monkeys received 10(9) pfu and the other two 10(7) pfu. One monkey received four consecutive injections into the same eye (10(7) pfu in each injection) over a 7-month period. In vivo gene transfer (fluorescence) and IOP were evaluated by standard clinical ophthalmic instruments (slit lamp biomicroscopy, gonioscopy and tonometry). Histopathology and cellular distribution were assessed postmortem.


RESULTS
The first injection of the lower viral dose resulted in marked TM-preferred gene transfer visible non-invasively by in vivo gonioscopy. The expression of the transgene lasted for 3-4 weeks with little or no signs of clinical inflammation. Gene transfer was achieved on three sequential occasions (3-4 weeks each) but failed and induced substantial, albeit reversible, corneal abnormalities on the fourth occasion.


CONCLUSIONS
Gene transfer to the TM and cornea can be monitored non-invasively in non-human primates, allowing correlation of gene transfer with physiological parameters. Because of ocular immune privilege, repeated anterior chamber administrations of adenoviral vectors expressing appropriate genes may have therapeutic potential for glaucoma.
Borrás
T
T
Department of Ophthalmology, Duke University Medical Center, Durham, NC 27710, USA. Borra001@mc.duke.edu
Gabelt
B T
BT
Klintworth
G K
GK
Peterson
J C
JC
Kaufman
P L
PL
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006068:Gonioscopy; D049452:Green Fluorescent Proteins; D007429:Intraocular Pressure; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008252:Macaca fascicularis; D008297:Male; D009924:Organ Culture Techniques; D010781:Photography; D016133:Polymerase Chain Reaction; D014129:Trabecular Meshwork / Q000378:metabolism*; D019076:Transgenes
D017931:DNA Primers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins