Scientific publications

Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization

Nov 9, 2019 | Magazine: Molecular Therapy-Methods & Clinical Development

Maria Hernandez, Sergio Recalde, Laura Garcia-Garcia, Jaione Bezunartea, Csaba Miskey, Sandra Johnen, Sabine Diarra, Attila Sebe, Juan Roberto Rodriguez-Madoz, Severine Pouillot, Corinne Marie, Zsuzsanna Izsvák, Daniel Scherman, Martina Kropp, Felipe Prosper, Gabriele Thumann, Zoltán Ivics, Alfredo Garcia-Layana, Patricia Fernandez-Robredo


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.

CITATION  Mol Ther Methods Clin Dev. 2019 Nov 9;15:403-417. doi: 10.1016/j.omtm.2019.10.013. eCollection 2019 Dec 13.

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Dr. Juan Roberto Rodríguez Madoz