Abstract
Purpose :
We previously reported an RNA-Seq dataset of corneal endothelium derived from healthy subjects (Tokuda Y, et al. Sci Data, 2020). The purpose of this present study was to elucidate the genetic alternations and potential signaling pathways related to the pathophysiology of Fuchs endothelial corneal dystrophy (FECD) by utilizing an RNA-Seq of corneal endothelium obtained from the patients with FECD.
Methods :
Corneal endothelial cells were isolated from patients with FECD (n=10) and healthy controls (n=7). RNA-Seq libraries were generated using the SMARTer® Stranded Total RNA-Seq Kit v2, and then sequenced by using a paired-end 100-bp read protocol. Genes were considered as differentially expressed genes (DEGs) with adjusted P-value (< 0.05) and Log2 Fold Change (≥ 1.0 or ≤ −1.0). Variation or similarity of gene expression was then confirmed by principal component analysis (PCA). For enrichment analysis, gene ontology (GO), Reactome, and KEGG pathways were investigated by use of R software.
Results :
Among a total of 24,636 genes detected by RNA-Seq, 2,366 genes were identified as DEGs in FECD (1,092 upregulated and 1,274 downregulated genes). PCA revealed the presence of two visual groups composed of control and FECD, respectively. For downstream analysis, 1,706 DEGs (696 upregulated and 1,010 downregulated genes) were extracted by confirming gene annotations. GO analysis indicated enrichment of extracellular structure organization, extracellular matrix (ECM) organization, response to oxidative stress, and apoptotic signaling pathway. Consistently, Reactome pathway analysis indicated the dysregulation of ECM related pathways. KEGG pathway analysis demonstrated the involvement of the phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) signaling pathways.
Conclusions :
The current study presented additional evidence that ECM dysregulation and oxidative stress are involved in the pathophysiology of FECD, and that the PI3K/Akt and MAPK signaling pathways might be interesting targets for elucidating the pathophysiology as well as drug discovery, though future in vitro and in vivo FECD-model "wet experiments" will be necessary.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.