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Influence of dexamethasone-induced matrices on the TM transcriptome.

Keerti Soundappan1, Jingwen Cai1, Hongfang Yu1

  • 1Department of Cellular Biology and Anatomy, Augusta University, Augusta, GA, United States.

Experimental Eye Research
|September 5, 2024
PubMed
Summary

Pathologic extracellular matrix (ECM) from steroid-treated human trabecular meshwork (hTM) cells induces glaucoma-like gene expression changes in healthy hTM cells. This study validates an in vitro model for investigating glaucoma pathogenesis and therapeutic targets.

Keywords:
DexamethasoneExtracellular matrixRNA-SeqTrabecular meshwork cellsddPCR

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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.