Influence of dexamethasone-induced matrices on the TM transcriptome

Area of Science:

• Ophthalmology
• Cell Biology
• Genomics

Background:

• Pathologic interactions between extracellular matrix (ECM) and human trabecular meshwork (hTM) cells contribute to ocular hypertension.
• An in vitro model is crucial for studying these cell-matrix interactions and their impact on outflow homeostasis.

Purpose of the Study:

• To determine if pathogenic ECM from dexamethasone (DEX)-treated hTM cultures induces glaucoma-like transcriptional changes in healthy hTM cells.
• To validate an in vitro model for studying steroid-induced glaucoma.

Main Methods:

• Primary hTM cells were cultured and treated with DEX or ethanol.
• Decellularized ECM from treated cells was used to culture new hTM cells.
• RNA sequencing (RNA-Seq) and Droplet Digital PCR (ddPCR) were performed to analyze gene expression changes.

Main Results:

• DEX-induced ECM significantly altered the transcriptional profile of healthy hTM cells, with 267 up- and 135 down-regulated genes identified (p<0.05, fold change ≥2.0).
• Affected pathways include inflammation, integrin signaling, matrix remodeling, and angiogenesis.
• Seven glaucoma-associated genes were validated using ddPCR.

Conclusions:

• The study supports the use of pathogenic ECM from DEX-treated hTM cells as a valid in vitro model for steroid-induced glaucoma.
• Matrix changes significantly influence hTM cell expression profiles, offering insights into glaucomatous mechanisms.
• Further studies with larger sample sizes are needed to confirm the model's utility.