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1 General aspects (0)   1.1 Epidemiology (2)   1.2 Population genetics (1)   1.3 Pathogenesis (3)   1.4 Quality of life (2)   1.5 Glaucomas as cause of blindness (3)   1.6 Prevention and screening (5) 2 Anatomical structures in glaucoma (0)   2.1 Conjunctiva (0)   2.2 Cornea (19)   2.3 Sclera (0)   2.4 Anterior chamber angle (1)   2.5 Meshwork (0)     2.5.1 Trabecular meshwork (12)     2.5.2 Schlemms canal (1)     2.5.3 Scleral outlet channels (0)   2.6 Aqueous humor dynamics (2)     2.6.1 Production (0)     2.6.2 Outflow (1)       2.6.2.1 Trabecular meshwork (0)       2.6.2.2 Uveoscleral (0)     2.6.3 Compostion (0)   2.7 Episcleral veins and venous pressure (0)   2.8 Iris (0)   2.9 Ciliary body (0)   2.10 Lens (0)   2.11 Vitreous body (0)   2.12 Choroid, peripapillary choroid, peripapillary atrophy (1)   2.13 Retina and retinal nerve fibre layer (8)   2.14 Optic disc (10)   2.15 Optic nerve (2)   2.16 Chiasma and retrochiasmal central nervous system (0)   2.17 Stem cells (0)   2.20 Other (0) 3 Laboratory methods (0)   3.1 Microscopy (1)   3.2 Electron microscopy (3)   3.3 Immunohistochemistry (5)   3.4 Molecular genetics (0)     3.4.1 Linkage studies (10)     3.4.2 Gene studies (6)   3.5 Molecular biology incl. 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Abstract #13697 Published in IGR 8-2

Lentiviral mediated gene delivery to the anterior chamber of rodent eyes

Challa P; Luna C; Liton PB; Chamblin B; Wakefield J; Ramabhadran R; Epstein DL; Gonzalez P
Molecular Vision 2005; 11: 425-430

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PURPOSE: To study the in vivo efficiency of lentiviral vectors in delivering genes to the trabecular meshwork (TM) of rodent eyes. METHODS: Lentiviral vectors were constructed using the elongation factor 1 α (EF-1α) promoter driving expression of the green fluorescent protein (GFP) gene. The viral construct was injected intracamerally through the peripheral cornea into the anterior chamber of live rodent eyes. Several variables were evaluated to determine the optimal conditions for TM cell transduction. These parameters included viral concentration, injection volume, needle rotation, and the modulation of anterior chamber current convections. Changes in intraocular pressures (IOPs) were monitored using a Tonopen XL. Signs of inflammation and corneal neovascularization were evaluated by slit lamp observation. Three weeks after injection, the eyes were enucleated and analyzed for GFP expression and distribution. RESULTS: A single intracameral viral dose between 10⁷ and 10⁸ pfu produced a high and evenly distributed expression of GFP in the TM and corneal endothelial cells. The cornea remained clear and no signs of inflammation were present during the course of the experiment. Moreover, no significant changes in IOPs were observed. CONCLUSIONS: A high transduction efficiency of TM and corneal endothelial cells can be effectively obtained after a single dose of recombinant lentivirus. The EF-1α promoter induces high expression of the reporter gene and is a reliable alternative to the CMV promoter when stable, long term expression is desired.

Dr. P. Challa, Department of Ophthalmology, Duke University, Durham, NC 27710, USA. Chall001@mc.duke.edu

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Classification:

11.9 Gene therapy (Part of: 11 Medical treatment)

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