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.
Highlights
Glucocorticoids (GCs) are anti-inflammatory agents used to treat ocular diseases such as uveitis and macular edema
Prolonged ocular application of GCs may lead to GCinduced ocular hypertension (OHT) and GC-induced glaucoma (GIG), a severe side effect that can lead to permanent visual loss
Three GC responder and non-responder bovine TM (BTM) cell strains were established using the approach described in “Methods.” Only confluent BTM cell cultures were used to mimic in vivo conditions [37,43]
Summary
Glucocorticoids (GCs) are anti-inflammatory agents used to treat ocular diseases such as uveitis and macular edema. Prolonged ocular application of GCs may lead to GCinduced ocular hypertension (OHT) and GC-induced glaucoma (GIG), a severe side effect that can lead to permanent visual loss. GC-OHT can occur with other non-ocular routes of administration such as systemic application of GCs and endogenous elevation of cortisol that can lead to Cushing’s syndrome/disease, the incidence of GC-induced OHT is lower than with topical GC application [1]. GC responders are at greater risk for developing POAG [7,8,9]. These studies further suggest the correlation between POAG and GIG
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