September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Efficacy of Rat Primary Cells transfected with Pigment Epithelium-Derived Factor using the Sleeping Beauty Transposon system in choroidal neovascularization.
Author Affiliations & Notes
  • Maria Hernandez
    Experimental Ophthalmology laboratory, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Laura Garcia-Garcia
    Experimental Ophthalmology laboratory, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Sergio Recalde
    Experimental Ophthalmology laboratory, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Patricia Fernandez
    Experimental Ophthalmology laboratory, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Juan Roberto Rodriguez
    Cell Therapy Area. Division of Cancer, Center of Aplied Medical Research (CIMA), Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Jaione Bezunartea-Bezunartea
    Experimental Ophthalmology laboratory, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Daniel Scherman
    Unité de Technologies Chimiques et Biologiques pour la Santé, INSERM U1022 – CNRS UMR8258, Paris, France
  • Sandra Johnen
    Department of Ophthalmology, University Hospital RWTH Aachen, Aachen, Germany
  • Gabriele Thumann
    Département des neurosciences cliniques, Service d’ophtalmologie, Hôpitaux universitaires de Genève, Genève, Switzerland
  • Alfredo Garcia-Layana
    Experimental Ophthalmology laboratory, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
    Idisna, Navarra Institute for Health Research, Pamplona, Navarra, Spain
  • Footnotes
    Commercial Relationships   Maria Hernandez, None; Laura Garcia-Garcia, None; Sergio Recalde, None; Patricia Fernandez, None; Juan Rodriguez, None; Jaione Bezunartea-Bezunartea, None; Daniel Scherman, None; Sandra Johnen, None; Gabriele Thumann, None; Alfredo Garcia-Layana, Alcon (C), Allergan (C), Bayer (C), Novartis (C), Thea (C)
  • Footnotes
    Support  FP7 HEALTH 2012-305134. LGG received ADA predoctoral grant from University of Navarra
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 271. doi:
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      Maria Hernandez, Laura Garcia-Garcia, Sergio Recalde, Patricia Fernandez, Juan Roberto Rodriguez, Jaione Bezunartea-Bezunartea, Daniel Scherman, Sandra Johnen, Gabriele Thumann, Alfredo Garcia-Layana; Efficacy of Rat Primary Cells transfected with Pigment Epithelium-Derived Factor using the Sleeping Beauty Transposon system in choroidal neovascularization.. Invest. Ophthalmol. Vis. Sci. 2016;57(12):271.

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

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Abstract

Purpose : The objective was to analyze the effect of PEDF release using the non-viral Sleeping Beauty (SB100X) transposon system (pFAR4-ITRs CMV PEDF BGH/pFAR4-CMV SB100x SV40) in rat primary cells in a model of laser-induced choroidal neovascularization (CNV).

Methods : Retinal pigment epithelia cells (RPEs) and iris pigment epithelia cells (IPEs) were transfected with pFAR4-pigment epithelium-derived factor (PEDF) plasmid. Laser-induced CNV was performed and 48h later the transfected cells were injected in the subretinal space of Brown Norway rats. Follow up of the lesions was assessed by fluorescein angiographies (FA) at 7 and 14 days post laser. The animals were sacrified at 5 and 12 days post injection. Flat mounts were labeled with isolectin, retinal cross sections were subjected to immunofluorescence technique with microglial/macrophage markers (isolectin, Iba1, CD68) and visualized under a confocal microscope.

Results : Transplantation of PEDF-transfected cells subretinally showed feasibility of the procedure. A significant reduction of FA leakage was observed in cell injected groups mainly 14 days after laser. Microglial and macrophage cell activation was localized in the angiogenesis area after subretinal injection. Finally, a significant reduction of neovascularization was observed in the CNV area labelled with isolectin marker.

Conclusions : The non-viral Sleeping Beauty (SB100X) transposon system for PEDF secretion was effective in reducing the area of leakage and CNV volume and area. This is an important step for the establishment of a safe, specific and effective gene therapeutic treatment for angiogenic retinal diseases such as exudative age-related macular degeneration.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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