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Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization
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 u...
Autores principales: | , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6909167/ https://www.ncbi.nlm.nih.gov/pubmed/31890733 http://dx.doi.org/10.1016/j.omtm.2019.10.013 |
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author | Hernandez, Maria Recalde, Sergio Garcia-Garcia, Laura Bezunartea, Jaione Miskey, Csaba Johnen, Sandra Diarra, Sabine Sebe, Attila Rodriguez-Madoz, Juan Roberto Pouillot, Severine Marie, Corinne Izsvák, Zsuzsanna Scherman, Daniel Kropp, Martina Prosper, Felipe Thumann, Gabriele Ivics, Zoltán Garcia-Layana, Alfredo Fernandez-Robredo, Patricia |
author_facet | Hernandez, Maria Recalde, Sergio Garcia-Garcia, Laura Bezunartea, Jaione Miskey, Csaba Johnen, Sandra Diarra, Sabine Sebe, Attila Rodriguez-Madoz, Juan Roberto Pouillot, Severine Marie, Corinne Izsvák, Zsuzsanna Scherman, Daniel Kropp, Martina Prosper, Felipe Thumann, Gabriele Ivics, Zoltán Garcia-Layana, Alfredo Fernandez-Robredo, Patricia |
author_sort | Hernandez, Maria |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6909167 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-69091672019-12-30 Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization Hernandez, Maria Recalde, Sergio Garcia-Garcia, Laura Bezunartea, Jaione Miskey, Csaba Johnen, Sandra Diarra, Sabine Sebe, Attila Rodriguez-Madoz, Juan Roberto Pouillot, Severine Marie, Corinne Izsvák, Zsuzsanna Scherman, Daniel Kropp, Martina Prosper, Felipe Thumann, Gabriele Ivics, Zoltán Garcia-Layana, Alfredo Fernandez-Robredo, Patricia Mol Ther Methods Clin Dev Article 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. American Society of Gene & Cell Therapy 2019-11-09 /pmc/articles/PMC6909167/ /pubmed/31890733 http://dx.doi.org/10.1016/j.omtm.2019.10.013 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Hernandez, Maria Recalde, Sergio Garcia-Garcia, Laura Bezunartea, Jaione Miskey, Csaba Johnen, Sandra Diarra, Sabine Sebe, Attila Rodriguez-Madoz, Juan Roberto Pouillot, Severine Marie, Corinne Izsvák, Zsuzsanna Scherman, Daniel Kropp, Martina Prosper, Felipe Thumann, Gabriele Ivics, Zoltán Garcia-Layana, Alfredo Fernandez-Robredo, Patricia Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization |
title | Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization |
title_full | Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization |
title_fullStr | Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization |
title_full_unstemmed | Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization |
title_short | Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization |
title_sort | preclinical evaluation of a cell-based gene therapy using the sleeping beauty transposon system in choroidal neovascularization |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6909167/ https://www.ncbi.nlm.nih.gov/pubmed/31890733 http://dx.doi.org/10.1016/j.omtm.2019.10.013 |
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