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Laser Capture Microdissection of Highly Pure Trabecular Meshwork from Mouse Eyes for Gene Expression Analysis
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High Glucose-induced transcriptomic changes in human trabecular meshwork cells.

Shivendra Singh1, Srimathi Raghavan1, Niketa A Patel2

  • 1Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA.

Molecular Biology Reports
|April 25, 2025
PubMed
Summary

High glucose levels in diabetes mellitus induce oxidative stress and fibrosis in human trabecular meshwork cells, contributing to glaucoma pathogenesis. These findings suggest targeting these pathways may prevent diabetic glaucoma.

Keywords:
ApoptosisAutophagyFibrosisGlaucomaHyperglycemiaOxidative stressTrabecular meshwork

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Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Diabetology

Background:

  • Glaucoma is a leading cause of irreversible blindness, often linked to elevated intraocular pressure (IOP) from trabecular meshwork (TM) dysfunction.
  • Diabetes mellitus (DM) is a risk factor for glaucoma, but the underlying molecular mechanisms of hyperglycemia's effect on TM remain unclear.

Purpose of the Study:

  • To investigate the impact of high glucose on gene expression in human TM (HTM) cells.
  • To identify molecular pathways contributing to TM dysfunction and glaucoma pathogenesis in diabetic conditions.

Main Methods:

  • Primary HTM cells cultured under normoglycemic and hyperglycemic conditions.
  • mRNA sequencing (mRNA-seq) to identify differentially expressed genes.
  • Quantitative PCR (qPCR) and STRING network analysis for validation and interaction prediction.

Main Results:

  • High glucose significantly altered gene expression in HTM cells, with 25 differentially expressed genes identified.
  • Upregulated genes indicated increased oxidative stress (TXNIP), apoptosis, immune response (CCL7, CHI3L1), and fibrosis (SNAI1, FGF7, KRT19).
  • Downregulation of autophagy genes (HSPA6, LAMP3) suggested impaired protein quality control, and increased ECM proteins indicated TM fibrosis.

Conclusions:

  • Hyperglycemia-induced oxidative stress, apoptosis, inflammation, and fibrosis in TM cells can impair aqueous humor outflow and elevate IOP.
  • Metabolic changes in diabetes contribute to TM dysfunction, increasing glaucoma risk.
  • Targeting oxidative stress and fibrosis pathways presents potential therapeutic strategies for diabetic glaucoma.