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

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Item Type:Article
Title:Preclinical evaluation of a cell-based gene therapy using the Sleeping Beauty transposon system in choroidal neovascularization
Creators Name:Hernandez, M. and Recalde, S. and Garcia-Garcia, L. and Bezunartea, J. and Miskey, C. and Johnen, S. and Diarra, S. and Sebe, A. and Rodriguez-Madoz, J.R. and Pouillot, S. and Marie, C. and Izsvák, Z. and Scherman, D. and Kropp, M. and Prosper, F. and Thumann, G. and Ivics, Z. and Garcia-Layana, A. and Fernandez-Robredo, P.
Abstract: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.
Keywords:Angiogenesis, Iris Pigment Epithelial Cells (IPE), Retinal Pigment Epithelium (RPE), Pigment Epithelium Derived Factor (PEDF), Vascular Endothelial Growth Factor (VEGF), SB100X Transposase, Animals, Rats
Source:Molecular Therapy - Methods and Clinical Development
Publisher:Cell Press
Page Range:403-417
Date:13 December 2019
Official Publication:https://doi.org/10.1016/j.omtm.2019.10.013
PubMed:View item in PubMed

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