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In vivo adenovirus-mediated gene transfer into ocular tissues
B Mashhour1, D Couton, M Perricaudet
1CIF 90-03 Institut National de la Santé et de la Recherche Médicale, Institut Cochin de Génétique Moléculaire (ICGM), Paris, France.
Abstract:
Replication-deficient adenoviruses have been successfully used to transfer foreign DNA into a variety of cells including post-mitotic cells, in vivo. In the eyes, most of the cells are quiescent or slowly dividing. They constitute the obligatory targets of gene transfer for a number of ocular diseases that have been elucidated at the molecular level and are potential targets for gene therapy. We have therefore analysed the ability of an adenovirus vector to transfer in vivo the Escherichia coli lacZ gene into ocular cells of mice. Injection of up to 3 x 10(7) p.f.u. into the vitreous body, the anterior chamber or the peribulbar space, did not result in any detectable cytopathic effect and was associated with endocytosis of viral particles in corneal endothelial, photoreceptor, bipolar, ganglionic and oculomotor muscle cells, depending on the administration route. At the viral titer used (3 x 10(7) p.f.u.), the expression was detected for at least 50 days. These results open new prospects for the treatment of some retinal hereditary disorders and acquired corneal or retinal alterations due to inflammatory disease.
Insights
Adenovirus vectors efficiently transfer genes into mouse ocular cells, including quiescent cells, without causing harm. This gene transfer method shows sustained expression, offering promise for treating inherited retinal diseases and corneal or retinal damage.
Area of Science:
- Ophthalmology
- Molecular Biology
- Gene Therapy
Background:
- Replication-deficient adenoviruses are effective for in vivo gene transfer.
- Ocular cells, often quiescent, are key targets for gene therapy of various eye diseases.
Purpose of the Study:
- To evaluate adenovirus vector-mediated gene transfer into mouse ocular cells.
- To assess the safety and duration of gene expression in ocular tissues following adenovirus delivery.
Main Methods:
- Adenovirus vectors carrying the Escherichia coli lacZ gene were injected into mouse eyes (vitreous, anterior chamber, peribulbar space).
- Viral titers up to 3 x 10(7) plaque-forming units (p.f.u.) were administered.
- Cellular uptake via endocytosis and gene expression were analyzed in various ocular cell types.
Main Results:
- No cytopathic effects were observed at the tested viral titers.
- Adenovirus particles were endocytosed by corneal endothelial, photoreceptor, bipolar, ganglionic, and oculomotor muscle cells.
- Sustained lacZ gene expression was detected for at least 50 days post-injection.
Conclusions:
- Adenovirus vectors can safely deliver genes to diverse ocular cells in vivo.
- This approach holds potential for treating hereditary retinal disorders and inflammatory corneal/retinal conditions.
- Gene transfer into quiescent ocular cells is feasible, expanding therapeutic options for eye diseases.