Abstract
Purpose :
To examine the localization of distinctive expressed genes and proteins in keratoconic corneas.
Methods :
Corneal stromal cells were cultured from three keratoconic corneas obtained from patients who underwent keratoplasty and three healthy corneas. Proteome and transcriptome analyses were conducted. Expression variation analysis was performed using transcriptome FASTQ data from 44 keratoconic corneas and 26 control corneas obtained from the Gene Expression Omnibus database. Genes consistently showing increased expression (ALDH1A1, SCIN) and decreased expression (PKP2, KRT18, SFRP1, MEST, PTGIS) were subjected to immunofluorescence staining using two keratoconic corneas and three control corneas. Additionally, spatial transcriptome analysis using Visium (10X) was performed on excised keratoconic corneas to confirm the localization of these expressed genes.
Results :
Immunofluorescence staining revealed strong expression of ALDH1A1 and SCIN in the corneal epithelium and superficial stroma, while PKP2, MEST, PTGIS, and SFRP1 showed overall low gene expression, but their expression was observed in the corneal epithelium through immunostaining. KRT18 exhibited relatively strong gene expression in the corneal epithelium.
Conclusions :
Immunofluorescence staining and spatial transcriptome analysis were conducted to investigate the localization of genes that showed consistent expression variations in cultured corneal stromal cells and excised corneal tissues of keratoconic patients. Genes with high expression were predominantly localized in the epithelium and superficial stroma, whereas genes with low expression, such as PKP2, exhibited protein expression in the epithelium. The localization analysis provided new insights into the expression patterns of these genes in keratoconic corneas.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.