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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, Niketa A Patel2, Avinash Soundararajan1

  • 1Indiana University School of Medicine.

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Summary

High glucose levels in diabetes mellitus damage human trabecular meshwork cells, increasing oxidative stress and fibrosis, which elevates intraocular pressure and glaucoma risk.

Keywords:
Glaucomaapoptosisautophagyfibrosishyperglycemiaoxidative stresstrabecular meshwork

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Area of Science:

  • Ophthalmology
  • Endocrinology
  • Molecular Biology

Background:

  • Glaucoma is a leading cause of irreversible blindness, linked to elevated intraocular pressure (IOP) from trabecular meshwork (TM) dysfunction.
  • Diabetes mellitus (DM) is a risk factor for glaucoma, but the molecular link between hyperglycemia and TM dysfunction is 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 were cultured under normoglycemic and hyperglycemic conditions.
  • mRNA sequencing (mRNA-seq) and quantitative PCR (qPCR) were used to identify differentially expressed genes.
  • STRING network analysis predicted protein interactions.

Main Results:

  • High glucose significantly altered gene expression in HTM cells, with 25 differentially expressed genes identified.
  • Upregulated genes were associated with oxidative stress (e.g., TXNIP), apoptosis, immune response (e.g., CCL7, CHI3L1), and fibrosis (e.g., SNAI1, FGF7, KRT19).
  • Downregulated autophagy genes (e.g., HSPA6, LAMP3) suggest impaired protein quality control.

Conclusions:

  • Hyperglycemia induces oxidative stress, apoptosis, inflammation, and fibrosis in HTM cells, contributing to TM dysfunction.
  • These molecular changes impair aqueous humor outflow, elevate IOP, and increase glaucoma risk in diabetic patients.
  • Targeting oxidative stress and fibrosis pathways may offer therapeutic strategies for diabetic glaucoma.