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Finny Monickaraj, Prakasha Kempaiah, Paul McGuire, Arup Das; Differential Transcriptome Profile of human retinal endothelial cells in response to Inflammation and Glycation. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3563.
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© ARVO (1962-2015); The Authors (2016-present)
Advanced glycation end products (AGEs), which accumulate under hyperglycemic conditions and retinal inflammation by chemokine and cytokine production and leukostasis play an important role in the pathogenesis of diabetic retinopathy. The purpose of this present study was to examine the influence of inflammation and glycation in the transcritptome profile and molecular signaling pathways of human retinal endothelial cells.
Human retinal endothelial cells (HRECs) were treated with macrophage-conditioned media (MCM) overnight, or 200μg of advanced glycation end-products (AGE) for 96 hrs. Total RNA was extracted from the samples, followed by quality check and library preparation using Illumina TruSeq RNA Sample Prep Kit and sequenced by Illumina HiSeq 2500. Differential expression was determined using DESeq, edgeR from the curated reads of the RNA-Seq data. Further characterization of functional annotation of differentially expressed genes in MCM and AGE-treated cells were performed using MetaCoreTM (Genego.Inc.) software suite.
Differential expression analysis revealed that 4804 genes were significantly down-regulated and 4730 genes were up-regulated in MCM treated cells, and 2978 and 3586 genes were significantly down and up-regulated respectively in the AGE-treated cells relative to untreated cells. Hierarchically clustered heatmap analysis of transcripts between MCM and AGE-treated cells revealed a significantly differential expression pattern in MCM treated cells compared to than AGE-treated cells. Further characterization of the differentially expressed genes in MCM and AGE-treated cells using the MetaCoreTM database to assess signaling pathway enrichment, revealed 82 signaling pathways in MCM and 13 in AGE-treated cells specifically related to cytokine/chemokine signaling and cell-cell adhesion pathways.
Overall the transcriptomics profiling gave us insight on the effect of MCM or AGE on the retinal endothelial cells in culture. The profiling also revealed the exacerbated impact of inflammation on the expression pattern in the endothelial cells in comparison with the glycation product.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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