The possibility of gene therapy for the treatment of choroidal neovascularization.
Ophthalmology
; 107(7): 1364-73, 2000 Jul.
Article
em En
| MEDLINE
| ID: mdl-10889114
ABSTRACT
PURPOSE:
Choroidal neovascularization (CNV) is responsible for most cases of severe visual loss in age-related macular degeneration. Recently, the possibility of gene therapy has been proposed for the treatment of CNV. The purpose of this study was to examine the feasibility of ex vivo and in situ gene therapy approaches for CNV.DESIGN:
Experimental study.METHODS:
Human retinal pigment epithelial (RPE) cells were transduced with a retroviral vector coding for beta-galactosidase. Transduced cells were grown on type II collagen sheets and transplanted under the retina of 20 rabbits. Animals were observed for 3 to 56 days, and transplanted cells were examined histologically and with X-gal staining. Bovine choroidal endothelial cells (CEC) were transduced with retroviral vectors coding for tissue inhibitor of metalloproteinase-2 (TIMP-2) or control vector. Production of TIMP-2 by transduced cells was determined by immunohistochemical analysis and enzyme-linked immunosorbent assay. Effect of transduction on in vitro proliferation, migration, and tube formation was examined in response to vascular endothelial growth factor (VEGF). Four CNV lesions were induced in one cynomolgus monkey by laser photocoagulation. Two days later, retroviral vector coding for TIMP-2 or control vector was injected into the subretinal space overlying the CNV lesions. The monkey was observed for 12 weeks using fluorescein angiography.RESULTS:
Transplantation of transduced RPE cells was technically achieved in 10 of 20 animals. In these animals, RPE cells at the site of transplantation formed a monolayer and expressed beta-galactosidase for 14 days. beta-Galactosidase-positive cells were not identified at 56 days. Choroidal endothelial cells transduced with TIMP-2 secrete TIMP-2 into the media and show decreased migration and tube formation in vitro. In the in vivo monkey model, the control CNV lesions (n = 2) showed prominent leakage, whereas the experimental lesions (n = 2) showed minimal hyperfluorescence.CONCLUSIONS:
Retrovirally transduced RPE cells survive in the subretinal space for at least 14 days and continue to express the gene product coded for by the vector. Choroidal endothelial cells retrovirally transduced for TIMP-2 produce TIMP-2 in vitro and show decreased angiogenic responses in vitro in response to VEGF. A preliminary study attempting in situ delivery of TIMP-2 vector to CNV lesions in a monkey eye supports the feasibility of this approach and encourages further study.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Epitélio Pigmentado Ocular
/
Retina
/
Endotélio Vascular
/
Terapia Genética
/
Beta-Galactosidase
/
Inibidor Tecidual de Metaloproteinase-2
/
Neovascularização de Coroide
Tipo de estudo:
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Ophthalmology
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
Estados Unidos