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- Li, PengPeking University (author)
Transcriptional reactivation of OTX2, RX1 and SIX3 during reprogramming contributes to the generation of RPE cells from human iPSCs
- Article/chapterEnglish2016
Publisher, publication year, extent ...
- Ivyspring International Publisher,2016
- 13 s.
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- LIBRIS-ID:oai:lup.lub.lu.se:4417d02f-dfb7-45cc-b9bd-e80deebf4aba
- https://lup.lub.lu.se/record/4417d02f-dfb7-45cc-b9bd-e80deebf4abaURI
- https://doi.org/10.7150/ijbs.14212DOI
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- Language:English
- Summary in:English
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- Directed differentiation of human induced pluripotent stem cells (iPSCs) into retinal pigmented epithelium (RPE) holds great promise in cell replacement therapy for patients suffering from degenerative eye diseases, including age-related macular degeneration (AMD). In this study, we generated iPSCs from human dermal fibroblasts (HDFs) by electroporation with episomal plasmid vectors encoding OCT4, SOX2, KLF4, L-MYC together with p53 suppression. Intriguingly, cell reprogramming resulted in a metastable transcriptional activation and selective demethylation of neural and retinal specification-associated genes, such as OTX2, RX1 and SIX3. In contrast, RPE progenitor genes were transcriptionally silent in HDFs and descendant iPSCs. Overexpression of OCT4 and SOX2 directly stimulated the expression of OTX2, RX1 and SIX3 in HDFs and iPSCs. Luciferase and chromatin immunoprecipitation (ChIP) assays further identified an OCT4- and two SOX2-binding sites located in the proximal promoter of OTX2. Histone acetylation and methylation on the local promoter also participated in the reactivation of OTX2. The transcriptional conversion of RX1 and SIX3 genes partially attributed to DNA demethylation. Subsequently, iPSCs were induced into the RPE cells displaying the characteristics of polygonal shapes and pigments, and expressing typical RPE cell markers. Taken together, our results establish readily efficient and safe protocols to produce iPSCs and iPSC-derived RPE cells, and underline that the reactivation of anterior neural transcription factor OTX2, eye field transcription factor RX1 and SIX3 in iPSCs is a feature of pluripotency acquisition and predetermines the potential of RPE differentiation.
Subject headings and genre
- MEDICIN OCH HÄLSOVETENSKAP Medicinska och farmaceutiska grundvetenskaper Medicinsk genetik hsv//swe
- MEDICAL AND HEALTH SCIENCES Basic Medicine Medical Genetics hsv//eng
- Epigenetic modification
- Induced pluripotent stem cells
- Reprogramming
- Retinal pigment epithelium
Added entries (persons, corporate bodies, meetings, titles ...)
- Sun, XiaofengHunan University of Traditional Chinese Medicine (author)
- Ma, ZhizhongPeking University (author)
- Liu, YinanPeking University (author)
- Jin, YingPeking University (author)
- Ge, RuiminLund University,Lunds universitet,Neural stamcellsbiologi och terapi,Forskargrupper vid Lunds universitet,Stamcellscentrum (SCC),Avdelningen för stamcellsforskning,Institutionen för laboratoriemedicin,Medicinska fakulteten,Neural stem cell biology and therapy,Lund University Research Groups,Stem Cell Center,Division of stem cell research,Department of Laboratory Medicine,Faculty of Medicine,Skåne University Hospital(Swepub:lu)med-rge (author)
- Hao, LiminCellonis Biotechnologies Co.Ltd (author)
- Ma, YanlingCellonis Biotechnologies Co.Ltd (author)
- Han, ShuoPeking University (author)
- Sun, HaojiePeking University (author)
- Zhang, MingzhiPeking University (author)
- Li, RuizhiPeking University (author)
- Li, TaoZhejiang Normal University (author)
- Shen, LiPeking University (author)
- Peking UniversityHunan University of Traditional Chinese Medicine (creator_code:org_t)
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- In:International Journal of Biological Sciences: Ivyspring International Publisher12:5, s. 505-5171449-2288
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