A Comparison of Gene Expression Profiles between Glucocorticoid Responder and Non-Responder Bovine Trabecular Meshwork Cells Using RNA Sequencing

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dc.creatorBermudez, Jaclyn Y.
dc.creatorWebber, Hannah C.
dc.creatorBrown, Bartley
dc.creatorBraun, Terry A.
dc.creatorClark, Abbot F.
dc.creatorMao, Weiming
dc.creator.orcid0000-0003-3594-6560 (Clark, Abbot F.)
dc.date.accessioned2022-09-13T16:34:16Z
dc.date.available2022-09-13T16:34:16Z
dc.date.issued2017-01-09
dc.description.abstractThe 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.
dc.description.sponsorshipThis work was funded by the National Eye Institute R21EY023048 (W.M.), https://nei.nih.gov/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was also funded by a National Institutes of Health training grant in the Neurobiology of Aging T32 AG20494 (J.Y.B & H.C.W.), https://www.nih.gov/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
dc.identifier.citationBermudez, J. Y., Webber, H. C., Brown, B., Braun, T. A., Clark, A. F., & Mao, W. (2017). A Comparison of Gene Expression Profiles between Glucocorticoid Responder and Non-Responder Bovine Trabecular Meshwork Cells Using RNA Sequencing. PloS one, 12(1), e0169671. https://doi.org/10.1371/journal.pone.0169671
dc.identifier.issn1932-6203
dc.identifier.issue1
dc.identifier.urihttps://hdl.handle.net/20.500.12503/31723
dc.identifier.volume12
dc.publisherPLOS
dc.relation.urihttps://doi.org/10.1371/journal.pone.0169671
dc.rights.holder© 2017 Bermudez et al.
dc.rights.licenseAttribution 4.0 International (CC BY 4.0)
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourcePLoS One
dc.subject.meshAnimals
dc.subject.meshCattle
dc.subject.meshComputational Biology / methods
dc.subject.meshGene Expression Profiling
dc.subject.meshGene Expression Regulation / drug effects
dc.subject.meshGlucocorticoids / pharmacology
dc.subject.meshHigh-Throughput Nucleotide Sequencing
dc.subject.meshProtein Interaction Mapping
dc.subject.meshSequence Analysis, RNA
dc.subject.meshSignal Transduction
dc.subject.meshTrabecular Meshwork / cytology
dc.subject.meshTrabecular Meshwork / metabolism
dc.subject.meshTranscriptome
dc.titleA Comparison of Gene Expression Profiles between Glucocorticoid Responder and Non-Responder Bovine Trabecular Meshwork Cells Using RNA Sequencing
dc.typeArticle
dc.type.materialtext

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