USP10 targeted self-deliverable siRNA to prevent scarring in the cornea

Abstract 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 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. Alpha-v integrins play a central role in myofibroblast pathological adhesion, over-contraction, and TGFβ activation. Previously we demonstrated that αv integrins are protected from intracellular degradation after wounding by upregulation of the deubiquitinase USP10, leading to integrin cell surface accumulation. Here, we tested if 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 sdRNA (NTC). At six weeks US09 treatment resulted in faster wound closure, limited scarring, and suppression of fibrotic markers and immune response. Specifically, Fibronectin-EDA, Collagen III, and α-smooth muscle actin (p