Lentiviral mediated RPE65 gene transfer in healthy hiPSCs-derived retinal pigment epithelial cells markedly increased RPE65 mRNA, but modestly protein level
| العنوان: | Lentiviral mediated RPE65 gene transfer in healthy hiPSCs-derived retinal pigment epithelial cells markedly increased RPE65 mRNA, but modestly protein level |
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| المؤلفون: | Florian Udry, Sarah Decembrini, Corinne Kostic, David M. Gamm, Nicole Déglon, Yvan Arsenijevic |
| المصدر: | Scientific Reports, Vol 10, Iss 1, Pp 1-15 (2020) Scientific Reports Scientific reports, vol. 10, no. 1, pp. 8890 |
| بيانات النشر: | Nature Publishing Group, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | cis-trans-Isomerases, Induced Pluripotent Stem Cells, Cell Culture Techniques, Stem-cell differentiation, lcsh:Medicine, Retinal Pigment Epithelium, Article, Viral vector, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, medicine, Humans, RNA, Messenger, Induced pluripotent stem cell, lcsh:Science, Cells, Cultured, 030304 developmental biology, 0303 health sciences, Messenger RNA, Retina, Multidisciplinary, Retinal pigment epithelium, Chemistry, Lentivirus, lcsh:R, Gene Transfer Techniques, Cell Differentiation, Retinal, eye diseases, Up-Regulation, 3. Good health, Cell biology, medicine.anatomical_structure, RPE65, Cell culture, lcsh:Q, sense organs, Visual system, 030217 neurology & neurosurgery |
| الوصف: | The retinal pigment epithelium (RPE) is a monolayer of cobblestone-like epithelial cells that accomplishes critical functions for the retina. Several protocols have been published to differentiate pluripotent stem cells into RPE cells suitable for disease modelling and therapy development. In our study, the RPE identity of human induced pluripotent stem cell (hiPSC)-derived RPE (iRPE) was extensively characterized, and then used to test a lentiviral-mediated RPE65 gene augmentation therapy. A dose study of the lentiviral vector revealed a dose-dependent effect of the vector on RPE65 mRNA levels. A marked increase of the RPE65 mRNA was also observed in the iRPE (100-fold) as well as in an experimental set with RPE derived from another hiPSC source and from foetal human RPE. Although iRPE displayed features close to bona fide RPE, no or a modest increase of the RPE65 protein level was observed depending on the protein detection method. Similar results were observed with the two other cell lines. The mechanism of RPE65 protein regulation remains to be elucidated, but the current work suggests that high vector expression will not produce an excess of the normal RPE65 protein level. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 2045-2322 |
| DOI: | 10.1038/s41598-020-65657-y |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8ba25eb3c94647c51209f06672c7bd69 http://link.springer.com/article/10.1038/s41598-020-65657-y |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....8ba25eb3c94647c51209f06672c7bd69 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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| DOI: | 10.1038/s41598-020-65657-y |