Preclinical evaluation of a cell-based gene therapy using the sleeping beauty transposon system in choroidal neovascularization
Iris pigment epithelial cells (IPE)
Retinal pigment epithelium (RPE)
Pigment epithelium derived factor (PEDF)
Vascular endothelial growth factor (VEGF)
SB100X transposase
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Elsevier BV
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Hernandez, M. (María); Recalde, S. (Sergio); Garcia-Garcia, L. (Laura); et al. "Preclinical evaluation of a cell-based gene therapy using the sleeping beauty transposon system in choroidal neovascularization". Molecular Therapy - Methods & Clinical Development. 15, 2019, 403 - 417
Age-related macular degeneration (AMD) is a progressive retinal disorder characterized by imbalanced pro- and antiangiogenic signals. The aim of this study was to evaluate the effect of ex vivo cell-based gene therapy with stable expression of human pigment epithelium-derived factor (PEDF) release using the non-viral Sleeping Beauty (SB100X) transposon system delivered by miniplasmids free of antibiotic resistance markers (pFAR4). Retinal pigment epithelial (RPE) cells and iris pigment epithelial (IPE) cells were co-transfected with pFAR4-inverted terminal repeats (ITRs) CMV-PEDF-BGH and pFAR4-CMV-SB100X-SV40 plasmids. Laser-induced choroidal neovascularization (CNV) was performed in rats, and transfected primary cells (transfected RPE [tRPE] and transfected IPE [tIPE] cells) were injected into the subretinal space. The leakage and CNV areas, vascular endothelial growth factor (VEGF), PEDF protein expression, metalloproteinases 2 and 9 (MMP-2/9), and microglial/macrophage markers were measured. Injection with tRPE/IPE cells significantly reduced the 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-2 activity. Our data demonstrated that ex vivo non-viral gene therapy reduces CNV and could be an effective and safe therapeutic option for angiogenic retinal diseases.

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