Long-read single-cell RNA sequencing analysis of key genes and isoforms during corneal wound healing in cynomolgus monkeys

The repair of corneal damage is essential for maintaining clear vision. Upon corneal epithelial injury, cells at the corneal limbus initiate complex processes such as migration, extracellular matrix remodeling, and proliferation. However, the transcriptional heterogeneity of limbal cell populations during these stages remains understudied. In this study, we used high-throughput long-read single-cell RNA sequencing to analyze five major cell types in the corneal limbus of cynomolgus monkeys at three time points: before injury, and one and three days post-injury. These cell types include terminally differentiated corneal epithelial cells (TDCE), basal cells (BC), transit-amplifying cells (TAC), limbal stem cells (LSC), and conjunctival cells (CC). We identified key regulatory genes and RNA isoforms involved in cell migration, proliferation, and differentiation, including IGF2 , FN1 , LAMC2 , ITGB1 , ITGAV , and keratins ( KRT3 , KRT12 , and KRT6A ). Our findings reveal the critical roles of LSC and BC in corneal repair and provide new insights into the transcriptional landscape during epithelial healing.