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Sustained and Widespread Gene Delivery to the Corneal Epithelium via in Situ Transduction of Limbal Epithelial Stem Cells, Using Lentiviral and Adeno-associated Viral Vectors

Basche, M; Kampik, D; Kawasaki, S; Branch, MJ; Robinson, M; Larkin, DF; Smith, AJ; (2018) Sustained and Widespread Gene Delivery to the Corneal Epithelium via in Situ Transduction of Limbal Epithelial Stem Cells, Using Lentiviral and Adeno-associated Viral Vectors. Human Gene Therapy , 29 (10) pp. 1140-1152. 10.1089/hum.2018.115. Green open access

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Sustained and widespread gene delivery to the corneal epithelium via in situ transduction of limbal epithelial stem cells using lentiviral and AAV vectors - COMPILED.pdf - Accepted Version

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Abstract

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.

Type: Article
Title: Sustained and Widespread Gene Delivery to the Corneal Epithelium via in Situ Transduction of Limbal Epithelial Stem Cells, Using Lentiviral and Adeno-associated Viral Vectors
Open access status: An open access version is available from UCL Discovery
DOI: 10.1089/hum.2018.115
Publisher version: http://doi.org/10.1089/hum.2018.115
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology
URI: https://discovery.ucl.ac.uk/id/eprint/10060479
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