Preclinical evaluation of a cell-based gene therapy using the sleeping beauty transposon system in choroidal neovascularization
Keywords: 
Angiogenesis
Iris pigment epithelial cells (IPE)
Retinal pigment epithelium (RPE)
Pigment epithelium derived factor (PEDF)
Vascular endothelial growth factor (VEGF)
SB100X transposase
Issue Date: 
2019
Publisher: 
Elsevier BV
ISSN: 
2329-0501
Note: 
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Citation: 
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
Abstract
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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