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In vivo gene transfer into murine corneal endothelial and trabecular meshwork cells
D L Budenz1, J Bennett, L Alonso
1Scheie Eye Institute, F. M. Kirby Center for Molecular Ophthalmology, University of Pennsylvania School of Medicine, Philadelphia, USA.
Purpose:
To determine whether a reporter gene can be introduced into adult mammalian corneal endothelial and trabecular meshwork cells in vivo using a recombinant replication-deficient adenovirus.
Methods:
Purified replication-deficient adenovirus containing the cytomegalovirus-promoted Escherichia coli reporter gene, lacZ, was injected into the vitreous cavities or anterior chambers of 30 adult CD-1 mice using the contralateral eyes as controls. LacZ expression was assessed histochemically in enucleated eyes from 2 to 21 days after injection using the beta-Galactosidase (beta-Gal) assay.
Results:
LacZ expression was demonstrated in corneal endothelial and trabecular meshwork cells for as long as 14 days after injection. beta-Gal activity was also observed in lens and iris epithelial cells. There was no toxicity of the adenoviral vector demonstrated histologically, and no nonocular tissues expressed lacZ as measured by beta-Gal assay.
Conclusions:
A functional gene can be transferred in vivo into adult mammalian corneal endothelial and trabecular meshwork cells using a replication-defective adenoviral vector. Gene expression is relatively short-lived compared to that demonstrated previously in other ocular tissues (photoreceptors and retinal pigment epithelium). Adenoviral vectors may be a viable means for short-term delivery of therapeutic genes in vivo to cells in the anterior segment of the eye.