1.Direct conversion of human fibroblasts into retinal pigment epithelium-like cells by defined factors.
Kejing ZHANG ; Guang-Hui LIU ; Fei YI ; Nuria MONTSERRAT ; Tomoaki HISHIDA ; Concepcion Rodriguez ESTEBAN ; Juan Carlos IZPISUA BELMONTE
Protein & Cell 2014;5(1):48-58
The generation of functional retinal pigment epithelium (RPE) is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-related macular degeneration (AMD). Although RPE cells can be produced from either embryonic stem cells or induced pluripotent stem cells, direct cell reprogramming driven by lineage-determining transcription factors provides an immediate route to their generation. By monitoring a human RPE specific Best1::GFP reporter, we report the conversion of human fibroblasts into RPE lineage using defined sets of transcription factors. We found that Best1::GFP positive cells formed colonies and exhibited morphological and molecular features of early stage RPE cells. Moreover, they were able to obtain pigmentation upon activation of Retinoic acid (RA) and Sonic Hedgehog (SHH) signaling pathways. Our study not only established an ideal platform to investigate the transcriptional network regulating the RPE cell fate determination, but also provided an alternative strategy to generate functional RPE cells that complement the use of pluripotent stem cells for disease modeling, drug screening, and cell therapy of retinal degeneration.
Animals
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Bestrophins
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Cell Differentiation
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Cell Line
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Cell Lineage
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Chloride Channels
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genetics
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metabolism
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Embryonic Stem Cells
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cytology
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metabolism
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Eye Proteins
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genetics
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metabolism
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Fibroblasts
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cytology
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metabolism
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Genes, Reporter
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Green Fluorescent Proteins
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genetics
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metabolism
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Humans
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Mice
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Pigmentation
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Retinal Pigment Epithelium
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cytology
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metabolism
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Transcription Factors
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metabolism
2.Mutations in foregut SOX2 cells induce efficient proliferation via CXCR2 pathway.
Tomoaki HISHIDA ; Eric VAZQUEZ-FERRER ; Yuriko HISHIDA-NOZAKI ; Ignacio SANCHO-MARTINEZ ; Yuta TAKAHASHI ; Fumiyuki HATANAKA ; Jun WU ; Alejandro OCAMPO ; Pradeep REDDY ; Min-Zu WU ; Laurie GERKEN ; Reuben J SHAW ; Concepcion RODRIGUEZ ESTEBAN ; Christopher BENNER ; Hiroshi NAKAGAWA ; Pedro GUILLEN GARCIA ; Estrella NUÑEZ DELICADO ; Antoni CASTELLS ; Josep M CAMPISTOL ; Guang-Hui LIU ; Juan Carlos IZPISUA BELMONTE
Protein & Cell 2019;10(7):485-495
Identification of the precise molecular pathways involved in oncogene-induced transformation may help us gain a better understanding of tumor initiation and promotion. Here, we demonstrate that SOX2 foregut epithelial cells are prone to oncogenic transformation upon mutagenic insults, such as Kras and p53 deletion. GFP-based lineage-tracing experiments indicate that SOX2 cells are the cells-of-origin of esophagus and stomach hyperplasia. Our observations indicate distinct roles for oncogenic KRAS mutation and P53 deletion. p53 homozygous deletion is required for the acquisition of an invasive potential, and Kras expression, but not p53 deletion, suffices for tumor formation. Global gene expression analysis reveals secreting factors upregulated in the hyperplasia induced by oncogenic KRAS and highlights a crucial role for the CXCR2 pathway in driving hyperplasia. Collectively, the array of genetic models presented here demonstrate that stratified epithelial cells are susceptible to oncogenic insults, which may lead to a better understanding of tumor initiation and aid in the design of new cancer therapeutics.