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Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids
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 gro...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5363513/ https://www.ncbi.nlm.nih.gov/pubmed/28325297 http://dx.doi.org/10.1016/j.omtn.2017.02.002 |
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author | Thumann, Gabriele Harmening, Nina Prat-Souteyrand, Cécile Marie, Corinne Pastor, Marie Sebe, Attila Miskey, Csaba Hurst, Laurence D. Diarra, Sabine Kropp, Martina Walter, Peter Scherman, Daniel Ivics, Zoltán Izsvák, Zsuzsanna Johnen, Sandra |
author_facet | Thumann, Gabriele Harmening, Nina Prat-Souteyrand, Cécile Marie, Corinne Pastor, Marie Sebe, Attila Miskey, Csaba Hurst, Laurence D. Diarra, Sabine Kropp, Martina Walter, Peter Scherman, Daniel Ivics, Zoltán Izsvák, Zsuzsanna Johnen, Sandra |
author_sort | Thumann, Gabriele |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5363513 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-53635132017-03-24 Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids Thumann, Gabriele Harmening, Nina Prat-Souteyrand, Cécile Marie, Corinne Pastor, Marie Sebe, Attila Miskey, Csaba Hurst, Laurence D. Diarra, Sabine Kropp, Martina Walter, Peter Scherman, Daniel Ivics, Zoltán Izsvák, Zsuzsanna Johnen, Sandra Mol Ther Nucleic Acids Original Article 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. American Society of Gene & Cell Therapy 2017-03-17 2017-02-10 /pmc/articles/PMC5363513/ /pubmed/28325297 http://dx.doi.org/10.1016/j.omtn.2017.02.002 Text en © 2017 The Author(s) 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 | Original Article Thumann, Gabriele Harmening, Nina Prat-Souteyrand, Cécile Marie, Corinne Pastor, Marie Sebe, Attila Miskey, Csaba Hurst, Laurence D. Diarra, Sabine Kropp, Martina Walter, Peter Scherman, Daniel Ivics, Zoltán Izsvák, Zsuzsanna Johnen, Sandra Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids |
title | Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids |
title_full | Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids |
title_fullStr | Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids |
title_full_unstemmed | Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids |
title_short | Engineering of PEDF-Expressing Primary Pigment Epithelial Cells by the SB Transposon System Delivered by pFAR4 Plasmids |
title_sort | engineering of pedf-expressing primary pigment epithelial cells by the sb transposon system delivered by pfar4 plasmids |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5363513/ https://www.ncbi.nlm.nih.gov/pubmed/28325297 http://dx.doi.org/10.1016/j.omtn.2017.02.002 |
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