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

Article Article scientifique Article Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization eng 2019 author Hernandez Maria author Recalde Sergio author Garcia-Garcia Laura author Bezunartea Jaione author Miskey Csaba author Johnen Sandra author Diarra Sabine 796918 author Sebe Attila author Rodriguez-Madoz Juan Roberto author Pouillot Severine author Marie Corinne author Izsvák Zsuzsanna author Scherman Daniel author Kropp Martina 0000-0001-6861-5851 793095 author Prosper Felipe author Thumann Gabriele 0000-0003-3682-9569 785873 author Ivics Zoltán author Garcia-Layana Alfredo author Fernandez-Robredo Patricia 403-417 10.1016/j.omtm.2019.10.013 31890733 2329-0501 https://linkinghub.elsevier.com/retrieve/pii/S2329050119301214 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. Epithelium-derived factor Retinal-pigment epithelium Endothelial growth-factor Macular degeneration Retroviral integration Tissue inhibitor Site selection RCS rats Iris PEDF Angiogenesis Iris pigment epithelial cells (IPE) Retinal pigment epithelium (RPE) Pigment epithelium derived factor (PEDF) Vascular endothelial growth factor (VEGF) SB100X transposase Dewey 616.8 61 Département des neurosciences cliniques 226 false 582 Ophtalmologie expérimentale 925 Molecular Therapy - Methods & Clinical Development 15 f606304b-c190-47e2-b0fc-e3a722aa4a56 Article (Published version) Public 1-s2.0-S2329050119301214-mmc2.pdf 3732653 application/pdf CC BY-NC-ND openAccess Faculté de médecine / Section de médecine clinique / Département des neurosciences cliniques

Title	Published in	Access level	OA Policy	Year	Views	Downloads

Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization	Molecular Therapy - Methods & Clinical Development			2019	576	458

Archive ouverte UNIGE

Diarra, Sabine