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R Krishnan Kutty, William Samuel, Cynthia Jaworski, Todd Duncan, Chandrasekharam N Nagineni, T. Michael Redmond; Altered expression of long non-Coding RNAs is associated with retinal pigment epithelial dysfunction induced by proinflammatory cytokines. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5788.
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© ARVO (1962-2015); The Authors (2016-present)
Proinflammatory cytokines secreted by infiltrating macrophages and lymphocytes may play a role in retinal pigment epithelial (RPE) dysfunction associated with age-related macular degeneration (AMD). We have observed earlier that proinflammatory cytokines IFN-γ, TNF-α and IL-1β induced epithelial-mesenchymal transition (EMT) and decreased RPE characteristics in RPE (ARPE-19) cells. Recent studies have shown that long non-coding RNAs (lncRNAs), non-protein coding transcripts with a minimum length of 200 nucleotides, regulate many pathophysiological processes including EMT. The purpose of the present study was to elucidate the potential role of lncRNAs in modulating the deleterious effect of proinflammatory cytokines on RPE cells.
ARPE-19 cells were cultured until they exhibited epithelial morphology and expressed visual cycle genes RPE65 and RDH5. The differentiated cells were treated with a mixture containing proinflammatory cytokines IFN-γ (100 u/ml), TNF-α (10 ng/ml) and IL-1β (10 ng/ml) for 4 days in a medium containing serum. Expression of protein-coding gene transcripts, lncRNAs, and miRNAs was analyzed by real-time PCR.
Differentiated ARPE-19 cells responded to proinflammatory cytokines by highly increasing the expression of chemokines and cytokines while markedly decreasing the expression of RPE-specific genes (RPE65, RDH5, RDH10, MITF, TYR and MERTK), epithelial marker gene (CDH1) and RPE-characteristic miRNAs (miR-204 and miR-211). ARPE-19 cells were found to express a number of the 84 well-characterized human lncRNAs that we tested. Proinflammatory cytokines increased the expression of BANCR, an lncRNA recently shown to regulate EMT and metastasis, by >100-fold. The treatment also altered the expression of many other lncRNAs: HAR1B, LUCAT1 and ST7-AS1 were highly upregulated while DANCR, HAR1A, HULC, KEASP1 and UCA1 were greatly downregulated.
The EMT-like changes and the loss of RPE characteristics in RPE cells exposed to proinflammatory cytokines IFN-γ, TNF-α and IL-1β are accompanied by the altered expression of BANCR and several other lncRNAs. Thus, lncRNAs due to their ability to regulate numerous cellular functions could potentially contribute to RPE dysfunction originating from aberrant inflammatory response and, therefore, may play a role in AMD pathology.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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