Integrative Analysis of Myofibroblast Transformation During Corneal Fibrosis

The transparent corneal stroma, which consists of regularly packed collagen fibrils and keratocytes, comprises 90% of the corneal thickness. Injury response in corneal stroma compromises corneal transparency due to overproduction of abnormal extracellular matrix by keratocyte-myofibroblast transformation. Corneal transplantation is considered the optimal solution to restore vision. Whether the keratocyte-myofibroblast transformation can be reversed remains unclear. In this study, we first developed a mouse corneal mechanical injury model. At 24h after the injury, α-SMA expression significantly increased, and SHG signals changed with respect to the collagen orientation. In addition, we isolated primary human keratocytes in advance; thereafter, we can decipher the keratocytes-myofibroblast transformation by adding TGF-β1 in the future. This proposal established three platforms, including a mouse injury model, primary human cell culture, and transgenic mice for targeting keratocytes. We will integrate these systems with single-cell transcriptomics, ATAC-sequencing, and proteomics to comprehensively explore keratocyte-myofibroblast transformation, and eventually, identify therapeutic targets for treating corneal fibrosis.