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Status |
Public on Jul 06, 2015 |
Title |
Reversal of persistent wound-induced retinal pigmented epithelial-to-mesenchymal transition by the TGFb pathway inhibitor, A-83-01. |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Age-related macular degeneration (AMD) is a leading cause of blindness. Most vision loss occurs following the transition from a disease of deposit formation and inflammation to a disease of neovascular fibrosis and/or cell death. Here, we investigate how repeated wound stimulus leads to seminal changes in gene expression and the onset of a perpetual state of stimulus-independent wound response in retinal pigmented epithelial (RPE) cells, a cell-type central to the etiology of AMD. Using a human fetal RPE cell culture model that considers monolayer disruption and subconfluent culture as a proxy for wound stimulus, we have shown that prolonged wound stimulus leads to terminal acquisition of a mesenchymal phenotype post-confluence and altered expression of more than 40% of the transcriptome (see GEO:GSE62224). In contrast, at subconfluence fewer than 5% of expressed transcripts have 2-fold or greater expression differences after repeated passage. Protein-protein and pathway interaction analysis of the genes with passage-dependent expression levels in subconfluent cultures reveals a 158-node interactome comprised of two interconnected modules with functions pertaining to wound response and cell division. Among the wound response genes are the TGFb pathway activators: TGFB1, TGFB2, INHBA, INHBB, GDF6, CTGF, and THBS1. Significantly, inhibition of TGFBR1/ACVR1B mediated signaling using receptor kinase inhibitors both forestalls and reverses the passage-dependent loss of epithelial potential. In this RNA-Seq based transcriptome analysis we show that the TGFb receptor kinase inhibitor, A-83-01, largely reverses the effects of passage and restores the transcriptome profile of Passage 4 RPE highly similar to that seen in differentiated Passage 0 RPE.
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Overall design |
Examination of mRNA expression in three different primary fetal RPE donor lines in 32 day old passage 0, passage 3, and passage 3 treated with 500 nM A-83-01 culutres
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Contributor(s) |
Radeke MJ, Radeke CM, Shih Y, Coffey P |
Citation(s) |
26150894 |
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Submission date |
Apr 15, 2015 |
Last update date |
May 15, 2019 |
Contact name |
Monte J. Radeke |
E-mail(s) |
radeke@ucsb.edu
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Phone |
805-893-3695
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Organization name |
University of California, Santa Barbara
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Department |
Neuroscience Research Institute
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Lab |
Center for the Study of Macular Degeneration
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Street address |
Neuroscience Research Institiute
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City |
Santa Barbara |
State/province |
CA |
ZIP/Postal code |
93106-5060 |
Country |
USA |
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Platforms (2) |
GPL17301 |
Ion Torrent PGM (Homo sapiens) |
GPL17303 |
Ion Torrent Proton (Homo sapiens) |
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Samples (10)
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This SubSeries is part of SuperSeries: |
GSE67899 |
Delay and restoration of persistent wound-induced retinal pigmented epithelial-to-mesenchymal transition by TGF-beta pathway inhibitors: Implications for age-related macular degeneration |
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Relations |
BioProject |
PRJNA281448 |
SRA |
SRP057295 |