| Autor: |
Basche M; 1 Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom; London, United Kingdom ., Kampik D; 1 Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom; London, United Kingdom ., Kawasaki S; 2 Department of Ophthalmology, Kyoto Prefectural University of Medicine , Kyoto, Japan ., Branch MJ; 1 Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom; London, United Kingdom ., Robinson M; 1 Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom; London, United Kingdom ., Larkin DF; 3 Moorfields Eye Hospital, and London, United Kingdom ., Smith AJ; 1 Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom; London, United Kingdom ., Ali RR; 1 Department of Genetics, UCL Institute of Ophthalmology, London, United Kingdom; London, United Kingdom .; 4 NIHR Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom . |
| Abstrakt: |
Corneal epithelial dystrophies are typically characterized by symptoms such as pain, light sensitivity, and corneal opacification leading to impaired vision. The development of gene therapy for such conditions has been hindered by an inability to achieve sustained and extensive gene transfer, as the epithelium is highly replicative and has evolved to exclude foreign material. We undertook a comprehensive study in mice aiming to overcome these impediments. Direct injection of lentiviral vector within the stem cell niche resulted in centripetal streaks of epithelial transgene expression sustained for >1 year, indicating limbal epithelial stem cell transduction in situ. The extent of transgene expression varied markedly but at maximum covered 26% of the corneal surface. After intrastromal injection, adeno-associated viral (AAV) vectors were found to penetrate Bowman's membrane and mediate widespread, but transient (12-16 days), epithelial transgene expression. This was sufficient, when applied within a Cre/lox system, to result in recombined epithelium covering up to approximately 80% of the corneal surface. Lastly, systemic delivery of AAV2/9 in neonatal mice resulted in extensive corneal transduction, despite the relative avascularity of the tissue. These findings provide the foundations of a gene therapy toolkit for the corneal epithelium, which might be applied to correction of inherited epithelial dystrophies. |
