Silencing Ubiquitin-Specific Peptidase 10 Alleviates Persistent Corneal Epithelial Defect in Mice

Abstract

Persistent corneal epithelial defect is a condition in which the corneal epithelium fails to heal properly after injury, resulting in vision loss. Previous studies demonstrate that the deubiquitinase, ubiquitin-specific peptidase 10 (USP10), is a key regulator of wound healing and scarring in the cornea. Here, a highly modified in vivo self-delivering siRNA (sdRNA) targeting USP10 in a mouse model of persistent corneal epithelial defect was used. In this model, the corneal epithelium is scraped from the basement membrane and, although the wound initially closes, the epithelium re-opens at day 28, thereby modeling a persistent corneal defect. These studies revealed that one dose of USP10-targeting sdRNA after wounding prevented re-opening of the wound at day 28. Optical coherence tomography images, hematoxylin and eosin staining, and immunohistochemistry demonstrated improved corneal morphology and normal epithelial stratification of the regrown epithelium, with re-establishment of hemidesmosomes and reduced scarring and inflammation (CD45⁺ cells). Correspondingly, RNA sequencing of the regenerated corneal epithelium revealed that USP10 knockdown altered gene expression and pathways controlling tissue repair. Specifically, cell proliferation pathways were enhanced, whereas extracellular matrix, integrin, and immune cell signatures that lead to scar formation were reduced. The results demonstrate that a one-time application of in vivo sdRNA targeting USP10 is a novel method to prevent epithelial wound re-opening and subsequent scarring and inflammation and may be useful to promote regenerative healing in other tissues.