Aim: To investigate genes and pathways involved in differential glucocorticoid (GC) responsiveness in human trabecular meshwork (HTM) cells using RNA sequencing. Methods: Using paired human donor eyes, human organ-cultured anterior segment (HOCAS) was established in one eye to characterize GC responsiveness based on intra ocular pressure (IOP) change and, in the other eye, primary HTM cell culture was established. For RNA sequencing, total RNA extracted from GC-responder (GC-R) and non-responder (GC-NR) cells after dexamethasone (DEX) or ethanol (ETH) treatment for 7d was used. Differentially expressed genes (DEGs) were compared among five groups and validated. Results: In total, 616 and 216 genes were identified as significantly dysregulated in Group #1 and #2 (#1: ETH vs. DEX-treated GC-R; #2: ETH vs. DEX-treated GC-NR), respectively. Around 80 genes were commonly dysregulated in Group #3 (overlapping DEGs between #1 and #2), whereas 536 and 136 genes were uniquely expressed in GC-R (#4) and GC-NR HTM (#5) cells, respectively. Pathway analysis revealed that WNT signaling, drug metabolism cytochrome p450, cell adhesion, TGF-β signaling, and MAPK signaling were associated with GC responsiveness. Conclusion: This is the first study reporting distinct gene signatures and their associated pathways for GC-R and GC-NR HTM cells. WNT and MAPK signaling are potential therapeutic targets for the management of GC-induced glaucoma.
|Early online date||15 May 2022|
|Publication status||Published online - 15 May 2022|
Bibliographical noteFunding Information:
Funding: This study was supported by the Department of Biotechnology (DBT)-Wellcome Trust/India Alliance fellowship ([grant number: IA/I/16/2/502694] awarded to Dr. Senthilkumari Srinivasan).
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
- glucocorticoid-induced ocular hypertension
- human perfusion-cultured anterior segment
- trabecular meshwork cells
- gene expression
- candidate genes
- Intraocular Pressure
- Trabecular Meshwork - metabolism
- Gene Expression Profiling
- Transcriptome - genetics
- Glucocorticoids - metabolism - pharmacology