title: A Comparison of Gene Expression Profiles between Glucocorticoid Responder and Non-Responder Bovine Trabecular Meshwork Cells Using RNA Sequencing.
abstract: The most common ocular side effect of glucocorticoid (GC) therapy is GC-induced ocular hypertension (OHT) and GC-induced glaucoma (GIG). GC-induced OHT occurs in about 40% of the general population, while the other 60% are resistant. This study aims to determine the genes and pathways involved in differential GC responsiveness in the trabecular meshwork (TM). Using paired bovine eyes, one eye was perfusion-cultured with 100nM dexamethasone (DEX), while the fellow eye was used to establish a bovine TM (BTM) cell strain. Based on maximum IOP change in the perfused eye, the BTM cell strain was identified as a DEX-responder or non-responder strain. Three responder and three non-responder BTM cell strains were cultured, treated with 0.1% ethanol or 100nM DEX for 7 days. RNA and proteins were extracted for RNA sequencing (RNAseq), qPCR, and Western immunoblotting (WB), respectively. Data were analyzed using the human and bovine genome databases as well as Tophat2 software. Genes were grouped and compared using Student's t-test. We found that DEX induced fibronectin expression in responder BTM cells but not in non-responder cells using WB. RNAseq showed between 93 and 606 differentially expressed genes in different expression groups between responder and non-responder BTM cells. The data generated by RNAseq were validated using qPCR. Pathway analyses showed 35 pathways associated with differentially expressed genes. These genes and pathways may play important roles in GC-induced OHT and will help us to better understand differential ocular responsiveness to GCs.
authors.affiliation: North Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd. Fort Worth, TX, United States of America.
authors.forename: Jaclyn Y
authors.initials: JY
authors.lastname: Bermudez
authors.affiliation: North Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd. Fort Worth, TX, United States of America.
authors.forename: Hannah C
authors.initials: HC
authors.lastname: Webber
authors.affiliation: Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, IA, United States of America.|Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.|Department of Biomedical Engineering, College of Engineering, University of Iowa, Iowa City, IA, United States of America.
authors.forename: Bartley
authors.initials: B
authors.lastname: Brown
authors.affiliation: North Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd. Fort Worth, TX, United States of America.|Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, IA, United States of America.|Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.|Department of Biomedical Engineering, College of Engineering, University of Iowa, Iowa City, IA, United States of America.
authors.forename: Terry A
authors.initials: TA
authors.lastname: Braun
authors.affiliation: North Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd. Fort Worth, TX, United States of America.
authors.forename: Abbot F
authors.initials: AF
authors.lastname: Clark
authors.affiliation: North Texas Eye Research Institute, University of North Texas Health Science Center, 3500 Camp Bowie Blvd. Fort Worth, TX, United States of America.
authors.forename: Weiming
authors.initials: W
authors.lastname: Mao
mesh_terms: D000818:Animals; D002417:Cattle; D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D005938:Glucocorticoids / Q000494:pharmacology*; D059014:High-Throughput Nucleotide Sequencing; D025941:Protein Interaction Mapping; D017423:Sequence Analysis, RNA; D015398:Signal Transduction; D014129:Trabecular Meshwork / Q000166:cytology* / Q000378:metabolism; D059467:Transcriptome
chemical_list: D005938:Glucocorticoids