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 small interfering RNAs (siRNAs) (selfdeliverable 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. a v integrins play a central role in myofibroblast pathological adhesion, overcontraction, and transforming growth factor b (TGF-b) activation. Previously, we demonstrated that a v 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.

FSTL1-USP10-Notch1 Signaling Axis Protects Against Cardiac Dysfunction Through Inhibition of Myocardial Fibrosis in Diabetic Mice

“…Previous studies established the cross-talk between USP10 and fibrosis in multiple tissues, which are down-regulated in cancers, Parkinson disease (PD), and keloids, and specifically render the heart susceptible to hypertrophy injury ( Anisimov et al, 2019 ; Deng et al, 2019 ; Boumil et al, 2020 ; Zhang et al, 2020 ). These studies provided compelling evidence confirming the role of USP10 in cardiac hypertrophy, and indicated that USP10 inhibition exacerbated pressure overload-induced cardiac dysfunction and facilitated angiotensin II-induced cardiomyocyte injury.…”

Recent Advancements in Molecular Therapeutics for Corneal Scar Treatment

“…It also decreased fibrotic marker (fibronectin and α-SMA) expression and αVβ1,5 integrin expression on the surface of corneal fibroblasts [ 70 ]. Bowmil et al [ 71 ] used a self-deliverable siRNA (siRNA) targeting USP10 conjugated to cys3 and cholesterol to treat wounded rabbit corneas. These siRNAs could penetrate to a depth of 324 µm in the corneal stromal layer within 24 h of transfection and reduce the expression of collagen III and fibrotic markers.…”

Hydroxycamptothecin and Substratum Stiffness Synergistically Regulate Fibrosis of Human Corneal Fibroblasts

“…4 Myofibroblasts are the main contributor to excessive extracellular matrix deposition leading to scar formation and fibrosis. 5 Upon injury, transforming growth factor-β1 (TGF-β1) is released into the corneal stroma 6 and activates keratocytes, the quiescent resident cells in the corneal stroma. Activated keratocytes are differentiated into fibroblasts and further matured into myofibroblasts.…”