Scientific article
Open access

Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids

Published inMolecular therapy. Nucleic acids, vol. 6, p. 302-314
Publication date2017

Neovascular age-related macular degeneration (nvAMD) is characterized by choroidal blood vessels growing into the subretinal space, leading to retinal pigment epithelial (RPE) cell degeneration and vision loss. Vessel growth results from an imbalance of pro-angiogenic (e.g., vascular endothelial growth factor [VEGF]) and anti-angiogenic factors (e.g., pigment epithelium-derived factor [PEDF]). Current treatment using intravitreal injections of anti-VEGF antibodies improves vision in about 30% of patients but may be accompanied by side effects and non-compliance. To avoid the difficulties posed by frequent intravitreal injections, we have proposed the transplantation of pigment epithelial cells modified to overexpress human PEDF. Stable transgene integration and expression is ensured by the hyperactive Sleeping Beauty transposon system delivered by pFAR4 miniplasmids, which have a backbone free of antibiotic resistance markers. We demonstrated efficient expression of the PEDF gene and an optimized PEDF cDNA sequence in as few as 5 × 10(3) primary cells. At 3 weeks post-transfection, PEDF secretion was significantly elevated and long-term follow-up indicated a more stable secretion by cells transfected with the optimized PEDF transgene. Analysis of transgene insertion sites in human RPE cells showed an almost random genomic distribution. The results represent an important contribution toward a clinical trial aiming at a non-viral gene therapy of nvAMD.

Citation (ISO format)
THUMANN, Gabriele et al. Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids. In: Molecular therapy. Nucleic acids, 2017, vol. 6, p. 302–314. doi: 10.1016/j.omtn.2017.02.002
Main files (1)
Article (Published version)
ISSN of the journal2162-2531

Technical informations

Creation04/21/2017 10:14:00 AM
First validation04/21/2017 10:14:00 AM
Update time03/15/2023 1:42:43 AM
Status update03/15/2023 1:42:42 AM
Last indexation02/12/2024 12:44:32 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack