April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Non-Viral Gene Therapy and Retinal Pigment Epithelium Tissue Engineering for the Treatment of Age-Related Macular Degeneration
Author Affiliations & Notes
  • A. Subrizi
    Centre for Drug Research,
    Division of Biopharmaceutics and Pharmacokinetics, Faculty of Pharmacy,
    University of Helsinki, Helsinki, Finland
  • S. Van Vlierberghe
    Polymer Chemistry and Biomaterials Research Group, Ghent University, Ghent, Belgium
  • G. Subra
    Equipe Aminoacides Peptides et Proteines, University of Montpellier, Montpellier, France
  • M. Amblard
    Equipe Aminoacides Peptides et Proteines, University of Montpellier, Montpellier, France
  • J. Martinez
    Equipe Aminoacides Peptides et Proteines, University of Montpellier, Montpellier, France
  • E. Schacht
    Polymer Chemistry and Biomaterials Research Group, Ghent University, Ghent, Belgium
  • P. Dubruel
    Polymer Chemistry and Biomaterials Research Group, Ghent University, Ghent, Belgium
  • M. Yliperttula
    Division of Biopharmaceutics and Pharmacokinetics, Faculty of Pharmacy,
    University of Helsinki, Helsinki, Finland
  • A. Urtti
    Centre for Drug Research,
    University of Helsinki, Helsinki, Finland
  • Footnotes
    Commercial Relationships  A. Subrizi, None; S. Van Vlierberghe, None; G. Subra, None; M. Amblard, None; J. Martinez, None; E. Schacht, None; P. Dubruel, None; M. Yliperttula, None; A. Urtti, None.
  • Footnotes
    Support  EU FP6 PolExGene (contract number 019114)
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 4496. doi:https://doi.org/
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      A. Subrizi, S. Van Vlierberghe, G. Subra, M. Amblard, J. Martinez, E. Schacht, P. Dubruel, M. Yliperttula, A. Urtti; Non-Viral Gene Therapy and Retinal Pigment Epithelium Tissue Engineering for the Treatment of Age-Related Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2010;51(13):4496. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Our approach to recovering visual function in AMD is the development of an ex vivo gene therapy in combination with retinal pigment epithelium (RPE) tissue engineering. We have studied the effect of substrate on RPE monolayer morphogenesis and electrophysiology. We have evaluated duration and direction of transgene secretion in RPE monolayers grown on selected scaffolds.

Methods: : The scaffolds were prepared from aminopropylmethacrylamide-modified gelatine and decorated with several bioactive molecules: heparan sulphate, chondroitin sulphate, hyaluronic acid, fibronectin, laminin and the epitope RGD. Screening was carried out with Cell-IQ live cell imaging and analysis platform. Bioelectric measurements of RPE monolayers were performed with glass barrel micro reference Ag/AgCl electrodes and a voltage-current clamp. RPE cells grown on selected scaffolds were clamped inside vertical Ussing chambers and transfected with non-viral vectors by means of conventional and reverse transfection methods. A self-replicating plasmid and a plasmid with RPE tissue-specific promoter were used to prolong the expression of the transgene and to improve the transfection efficacy.

Results: : According to growth curves, morphology and viability of RPE cells, three scaffolds were selected for further studies: hyaluronic acid, heparan sulphate and laminin. The barrier properties of RPE monolayers and their viability were monitored by determining their bioelectric properties. Transgene expression in RPE monolayers lasted for several weeks. At the same time transepithelial electrical potential difference and transepithelial resistance were measured.

Conclusions: : The reconstruction of a functional RPE and nanoparticulate-enhanced gene therapy providing efficient gene replacement may be a promising therapeutic option for the treatment of AMD.

Keywords: retinal pigment epithelium • gene transfer/gene therapy • regeneration 
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