Abstract
Glucocorticoids (GCs) are known to regulate several physiological processes and are the mainstay in the management of inflammatory eye diseases. The long-term use of GC causes raised intraocular pressure (IOP) or ocular hypertension (OHT) in about 30–50% of the susceptible individuals depending on the route of administration, and can lead to steroid-induced secondary glaucoma. The present study aims to understand the role of microRNAs (miRNAs) in differential glucocorticoid (GC) responsiveness in human trabecular meshwork (HTM) cells using small RNA sequencing. The human organ-cultured anterior segment (HOCAS) model was used to identify whether donor eyes were from GC-responders (GC-R; n = 4) or GC-non-responders (GC-NR; n = 4) following treatment with either 100 nM dexamethasone (DEX) or ethanol (ETH) for 7 days. The total RNA was extracted from cultured HTM cells with known GC responsiveness, and the differentially expressed miRNAs (DEMIRs) were compared among the following five groups: Group #1: ETH vs. DEX-treated GC-R; #2: ETH vs. DEX-treated GC-NR; #3: overlapping DEGs between Group #1 and #2; #4: Unique DEMIRs of GC-R; #5: Unique DEMIRs of GC-NR; and validated by RT-qPCR. There were 13 and 21 DEMIRs identified in Group #1 and Group #2, respectively. Seven miRNAs were common miRNAs dysregulated in both GC-R and GC-NR (Group #3). This analysis allowed the identification of DEMIRs that were unique to GC-R (6 miRNAs) and GC-NR (14 miRNAs) HTM cells, respectively. Ingenuity Pathway Analysis identified enriched pathways and biological processes associated with differential GC responsiveness in HTM cells. This is the first study to reveal a unique miRNA signature between GC-R and GC-NR HTM cells, which raises the possibility of developing new molecular targets for the management of steroid-OHT/glaucoma.
Original language | English |
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Pages (from-to) | 1-18 |
Number of pages | 18 |
Journal | Genes |
Volume | 14 |
Issue number | 11 |
Early online date | 27 Oct 2023 |
DOIs | |
Publication status | Published online - 27 Oct 2023 |
Bibliographical note
Publisher Copyright:© 2023 by the authors.
Data Access Statement
Data Access: The raw miRNA sequencing data of HTM cells from each human donor eye used in the present study have been deposited publicly in NCBI-SRA under the BioProject PRJNA982055 (https://www.ncbi.nlm.nih.gov/sra/PRJNA982055, accessed on 9 June 2023). Code Availability: The bioinformatics In-house pipeline used for mRNA sequencing data analysis in the present study have been submitted to GitHub in shell script (https://github.com/SenthilKumariLab/mRNA-seq-Analysis-Pipeline.git, accessed on 14 May 2021).Keywords
- microRNAs
- pathway analysis
- ocular hypertension
- intra-ocular pressure
- miRNA
- glaucoma
- small RNA-Seq
- trabecular meshwork
- glucocorticoids