Lentivirus-mediated gene transfer to the rat, ovine and human cornea

D. G.A. Parker, C. Kaufmann, H. M. Brereton, D. S. Anson, L. Francis-Staite, C. F. Jessup, K. Marshall, C. Tan, R. Koldej, D. J. Coster, Keryn Williams

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Gene therapy of the cornea shows promise for modulating corneal transplant rejection but the most appropriate vector for gene transfer has yet to be determined. We investigated a lentiviral vector (LV) for its ability to transduce corneal endothelium. A lentivector expressing enhanced yellow fluorescent protein (eYFP) under the control of the Simian virus type 40 early promoter (LV-SV40-eYFP) transduced 80-90% of rat, ovine and human corneal endothelial cells as detected by fluorescence microscopy. The kinetics of gene expression varied among species, with ovine corneal endothelium showing a relative delay in detectable reporter gene expression compared with the rat or human corneal endothelium. Vectors containing the myeloproliferative sarcoma virus promoter or the phosphoglycerate kinase promoter were not significantly more effective than LV-SV40-eYFP. The stability of eYFP expression in rat and ovine corneas following ex vivo transduction of the donor cornea was assessed following orthotopic corneal transplantation. Following transduction ex vivo, eYFP expression was maintained in corneal endothelial cells for at least 28 days after corneal transplantation in the sheep and >60 days in the rat. Thus, rat, ovine and human corneal endothelial cells were efficiently transduced by the LV, and gene expression appeared stable over weeks in vivo.

Original languageEnglish
Pages (from-to)760-767
Number of pages8
JournalGene Therapy
Volume14
Issue number9
DOIs
Publication statusPublished - 1 May 2007
Externally publishedYes

Keywords

  • Corneal endothelium
  • Corneal transplantation
  • Lentiviral vector

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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