Sripathi, Srinivasa R.Hu, Ming-WenLiu, Melissa M.Wan, JunCheng, JieDuan, YukanMertz, Joseph L.Wahlin, Karl J.Maruotti, JulienBerlinicke, Cynthia A.Qian, JiangZack, Donald J.2022-08-262022-08-262021-04Sripathi SR, Hu MW, Liu MM, et al. Transcriptome Landscape of Epithelial to Mesenchymal Transition of Human Stem Cell-Derived RPE. Invest Ophthalmol Vis Sci. 2021;62(4):1. doi:10.1167/iovs.62.4.1https://hdl.handle.net/1805/29898Purpose: RPE injury often induces epithelial to mesenchymal transition (EMT). Although RPE-EMT has been implicated in a variety of retinal diseases, including proliferative vitroretinopathy, neovascular and atrophic AMD, and diabetic retinopathy, it is not well-understood at the molecular level. To contribute to our understanding of EMT in human RPE, we performed a time-course transcriptomic analysis of human stem cell-derived RPE (hRPE) monolayers induced to undergo EMT using 2 independent, yet complementary, model systems. Methods: EMT of human stem cell-derived RPE monolayers was induced by either enzymatic dissociation or modulation of TGF-β signaling. Transcriptomic analysis of cells at different stages of EMT was performed by RNA-sequencing, and select findings were confirmed by reverse transcription quantitative PCR and immunostaining. An ingenuity pathway analysis (IPA) was performed to identify signaling pathways and regulatory networks associated with EMT. Results: Proteocollagenolytic enzymatic dissociation and cotreatment with TGF-β and TNF-α both induce EMT in human stem cell-derived RPE monolayers, leading to an increased expression of mesenchymal factors and a decreased expression of RPE differentiation-associated factors. Ingenuity pathway analysis identified the upstream regulators of the RPE-EMT regulatory networks and identified master switches and nodes during RPE-EMT. Of particular interest was the identification of widespread dysregulation of axon guidance molecules during RPE-EMT progression. Conclusions: The temporal transcriptome profiles described here provide a comprehensive resource of the dynamic signaling events and the associated biological pathways that underlie RPE-EMT onset. The pathways defined by these studies may help to identify targets for the development of novel therapeutic targets for the treatment of retinal disease.en-USAttribution 4.0 InternationalStem cellsDifferentiationRetinal pigment epitheliumEpithelial–mesenchymal transitionTGF-β/TNF-αTranscriptomicsTranscriptome Landscape of Epithelial to Mesenchymal Transition of Human Stem Cell–Derived RPEArticle