Preclinical evaluation of a cell-based gene therapy using the Sleeping Beauty transposon system in choroidal neovascularization


  • M. Hernandez
  • S. Recalde
  • L. Garcia-Garcia
  • J. Bezunartea
  • C. Miskey
  • S. Johnen
  • S. Diarra
  • A. Sebe
  • J.R. Rodriguez-Madoz
  • S. Pouillot
  • C. Marie
  • Z. Izsvák
  • D. Scherman
  • M. Kropp
  • F. Prosper
  • G. Thumann
  • Z. Ivics
  • A. Garcia-Layana
  • P. Fernandez-Robredo


  • Molecular Therapy - Methods and Clinical Development


  • Mol Ther Methods Clin Dev 15: 403-417


  • Age-related macular degeneration (AMD) is a progressiveretinal disorder characterized by imbalanced pro- andantiangiogenic signals. The aim of this study was to evaluatethe effect ofex vivocell-based gene therapy with stable expres-sion of human pigment epithelium-derived factor (PEDF)release using the non-viralSleeping Beauty(SB100X) trans-poson system delivered by miniplasmids free of antibiotic resis-tance markers (pFAR4). Retinal pigment epithelial (RPE) cellsand iris pigment epithelial (IPE) cells were co-transfected withpFAR4-inverted terminal repeats (ITRs) CMV-PEDF-BGHand pFAR4-CMV-SB100X-SV40 plasmids. Laser-inducedchoroidal neovascularization (CNV) was performed in rats,and transfected primary cells (transfected RPE [tRPE] andtransfected IPE [tIPE] cells) were injected into the subretinalspace. The leakage and CNV areas, vascular endothelial growthfactor (VEGF), PEDF protein expression, metalloproteinases 2and 9 (MMP-2/9), and microglial/macrophage markers weremeasured. Injection with tRPE/IPE cells significantly reducedthe leakage area at 7 and 14 days and the CNV area at 7 days.There was a significant increase in PEDF and the PEDF/VEGF ratio with tRPE cells and a reduction in the MMP-2activity. Our data demonstrated thatex vivonon-viral genetherapy reduces CNV and could be an effective and safe thera-peutic option for angiogenic retinal diseases.