USP10 Targeted Self-Deliverable siRNA to Prevent Scarring in the Cornea

• Published in: Molecular therapy. Nucleic acids Vol. 21; pp. 1029 - 1043
• Main Authors: Boumil, Edward F., Castro, Nileyma, Phillips, Andrew T., Chatterton, Jon E., McCauley, Sean M., Wolfson, Alexey D., Shmushkovich, Taisia, Ridilla, Marc, Bernstein, Audrey M.
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 04.09.2020; American Society of Gene & Cell Therapy; Elsevier
• Subjects: cornea; fibrosis; integrin; myofibroblast; ocular; Original; scarring; stroma; wound healing; wound healing; scarring; myofibroblast; cornea; stroma; fibrosis; integrin; ocular
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1016/j.omtn.2020.07.032

Ocular scarring after surgery, trauma, or infection leads to vision loss. The transparent cornea is an excellent model system to test anti-scarring therapies. Cholesterol-conjugated fully modified asymmetric small interfering RNAs (siRNAs) (self-deliverable siRNAs [sdRNAs]) are a novel modality for in vivo gene knockdown, transfecting cells and tissues without any additional formulations. Myofibroblasts are a main contributor to scarring and fibrosis. αv integrins play a central role in myofibroblast pathological adhesion, overcontraction, and transforming growth factor β (TGF-β) activation. Previously, we demonstrated that αv integrins are protected from intracellular degradation after wounding by upregulation of the deubiquitinase (DUB) ubiquitin-specific protease 10 (USP10), leading to integrin cell surface accumulation. In this study, we tested whether knockdown of USP10 with a USP10-targeting sdRNA (termed US09) will reduce scarring after wounding a rabbit cornea in vivo. The wounded corneal stroma was treated once with US09 or non-targeting control (NTC) sdRNA. At 6 weeks US09 treatment resulted in faster wound closure, limited scarring, and suppression of fibrotic markers and immune response. Specifically, fibronectin-extra domain A (EDA), collagen III, and a-smooth muscle actin (p < 0.05), CD45+ cell infiltration (p < 0.01), and apoptosis at 24 (p < 0.01) and 48 h (p < 0.05) were reduced post-wounding. Corneal thickness and cell proliferation were restored to unwounded parameters. Targeting the DUB, USP10 is a novel strategy to reduce scarring. This study indicates that ubiquitin-mediated pathways should be considered in the pathogenesis of fibrotic healing. [Display omitted] Boumil, Castro, et al. demonstrate that treatment of a corneal wound in rabbits with a self-delivery siRNA targeting the deubiquitinase USP10 significantly reduces scarring following severe injury. These findings reveal a novel scarring pathway and indicate that ubiquitin-mediated pathways play a role in the pathogenesis of fibrotic healing.