USP10 Silencing Prevents Scarring and Inflammation in a Modified Rabbit Model of Glaucoma Filtration Surgery.

PURPOSE: Trabeculectomy remains the gold standard for lowering intraocular pressure (IOP) in glaucoma, but postoperative scarring and inflammation can cause bleb failure. Because USP10 knockdown is anti-inflammatory and anti-fibrotic in the cornea, we tested whether USP10 silencing reduces bleb scarring and inflammation in a rabbit glaucoma filtration surgery model.

METHODS: Primary rabbit Tenon's fibroblasts (RTFs) were transfected with fully modified in vivo self-delivering siRNA targeting USP10 (sdRNA, US09) or non-targeting control sdRNA (NTC) to assess myofibroblast conversion and fibronectin expression. Female New Zealand White rabbits underwent modified glaucoma filtering surgery with tube insertion and subconjunctival US09, NTC, or mitomycin C (MMC). Bleb health and vascularity were scored using a modified Moorfields system, IOP was measured with TonoVet Plus, and excised blebs were analyzed for tissue width, apoptosis (TUNEL), α-SMA, and CD45+ cell infiltration. Inflammatory cytokines were profiled from aqueous humor by ELISA.

RESULTS: US09 transfection in RTFs reduced USP10 expression and suppressed myofibroblast activation, lowering α-SMA and fibronectin expression and disrupting fibrillar organization. In vivo, a single US09 dose reduced postoperative apoptosis versus NTC and MMC. Although both US09 and MMC inhibited myofibroblast differentiation and early immune infiltration, only US09 preserved bleb architecture, sustained immune suppression, and avoided MMC-associated ischemic vascularity. US09 treatment was also associated with lower mean levels of several aqueous humor cytokines.

CONCLUSIONS: Perioperative USP10-targeting sdRNA reduced myofibroblast development and sustained an anti-inflammatory, tissue-preserving phenotype after rabbit glaucoma filtration surgery, with advantages over MMC in tissue-preservation and inflammatory endpoints. USP10 silencing may represent a promising anti-scarring strategy to improve long-term trabeculectomy outcomes.