Premium
Comparison of viral vectors for gene transfer to corneal endothelial cells
Author(s) -
FUCHSLUGER T
Publication year - 2012
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2012.2637.x
Subject(s) - transduction (biophysics) , viral vector , genetic enhancement , green fluorescent protein , microbiology and biotechnology , flow cytometry , biology , gene transfer , corneal endothelium , cornea , gene delivery , cell culture , vectors in gene therapy , gene , virology , genetics , recombinant dna , biochemistry , neuroscience
Purpose Thanks to its anatomical location at the posterior surface of the cornea and its monolayer structure, the corneal endothelium is an ideal target for gene therapy approaches. Lentiviral vectors have been shown by our group to be suitable vectors for the transfer of genes into corneal endothelial cells (EC). Aiming for an alternative to these HIV‐based vectors, it was the goal of this study to determine the suitability of non‐pathogenic adeno‐associated viral vectors (AAV) for gene transfer to EC. Methods Comparison of protein expression after EC transduction using a lentiviral vector or AAV 2/2 with GFP in murine EC (Balb/C) and in human EC (cell line and primary cells)by flow cytometry. Results Following transduction of EC using lentiviral vector, kinetics of the protein expression are considerably different compared to gene transfer using AAV. In contrast to AAV with protein expression showing a plateau after two to three weeks, lentiviral transfer results in a very rapid of reporter protein. Moreover, we detected significant differences in transduction rates between human and murine EC lines as well as betweeen human EC lines and human corneas (plateau at 70% versus 50% GFP‐positive cells with AAV, versus 90‐95% with lentivirus). Conclusion DNA transfer using AAV vectors seems to be an appropriate alternative to lentiviral vectors for gene transfer to EC. Relating to the cultivation of human donor corneas in eye banks over weeks, translation of AAV from bench to bedside, e.g. to reduce apoptosis in corneas, seems to be a promising approach for future gene transfer into donor corneas.