Identification and Validation of Key Biomarkers in the Proximal Aqueous Humor Outflow Pathway.

Area of Science:

• Ophthalmology and Molecular Biology
• Genomics and Transcriptomics

Background:

• Glaucoma is a primary cause of irreversible blindness, strongly linked to elevated intraocular pressure (IOP).
• Dysfunction of the trabecular meshwork (TM) and Schlemm's canal endothelial (SCE) cells, key components of aqueous humor outflow, significantly contributes to IOP elevation.

Purpose of the Study:

• To identify and validate specific biomarkers for human TM and SCE cells.
• To gain insights into the molecular mechanisms underlying glaucoma pathophysiology.

Main Methods:

• Transcriptome profiling of human TM and SCE cells using Microarray.
• Differential gene expression analysis and cross-referencing with single-cell RNA sequencing (scRNA-Seq) data.
• Validation of candidate biomarkers using quantitative Polymerase Chain Reaction (qPCR).

Main Results:

• Microarray analysis identified 341 differentially expressed genes (DEGs).
• TM cells showed enrichment in metabolic and signaling pathways, while SCE cells were enriched in adhesion, immune, and morphogenesis pathways.
• CTTNBP2 and MGARP confirmed as TM markers; JAM2, PODXL, and IFI27 identified as novel SCE markers.

Conclusions:

• Validated TM and SCE cell biomarkers provide critical insights into glaucoma pathogenesis.
• These biomarkers are foundational for developing targeted therapeutic strategies for glaucoma.
• The study enhances understanding of the aqueous humor outflow pathway in ocular health and disease.