Corneal Keratocytes, Fibroblasts, and Myofibroblasts Exhibit Distinct Transcriptional Profiles In Vitro

Area of Science:

• Ophthalmology
• Cell Biology
• Molecular Biology

Background:

• Corneal stromal injury triggers keratocyte activation into fibroblasts and myofibroblasts.
• Persistent myofibroblast phenotype causes corneal fibrosis and scarring, leading causes of blindness.
• Understanding these cellular transitions is key to developing treatments for corneal scarring.

Purpose of the Study:

• To establish comprehensive transcriptional profiles of cultured corneal keratocytes, fibroblasts, and myofibroblasts.
• To identify key genes and signaling pathways involved in corneal cell phenotypic changes.
• To uncover novel molecular markers for each cell type.

Main Methods:

• Cultured rabbit corneal keratocytes in serum-free media, serum-containing media, or TGF-β1-supplemented media to induce specific phenotypes.
• Performed bulk RNA sequencing and bioinformatic analyses to identify differentially expressed genes (DEGs) and enriched pathways.

Main Results:

• Distinct gene expression patterns confirmed keratocyte, fibroblast, and myofibroblast phenotypes.
• Identified novel DEGs and enriched pathways related to proliferation, apoptosis, extracellular matrix (ECM) synthesis, and cell mechanics.
• Highlighted differences in cytokine signaling and cell-ECM interactions among the cell types.

Conclusions:

• Demonstrated significant transcriptional differences between corneal keratocyte, fibroblast, and myofibroblast phenotypes.
• Identified genes and pathways involved in mechanotransduction and ECM biology relevant to keratocyte differentiation.
• Revealed novel molecular markers and potential therapeutic targets for modulating corneal wound healing.