Vimentin Knockdown Decreases Corneal Opacity

Abstract

PURPOSE. Wound induced corneal fibrosis can lead to permanent visual impairment. Keratocyte activation and differentiation play a key role in fibrosis, and vimentin, a major structural type III intermediate filament, is a required component of this process. The purpose of our study was to develop a nonviral therapeutic strategy for treating corneal fibrosis in which we targeted the knockdown of vimentin. METHODS.To determine the duration of plasmid expression in corneal keratocytes, we injected a naked plasmid expressing green fluorescent protein (GFP; pCMV-GFP) into an unwounded mouse corneal stroma. We then injected pCMV-GFP or plasmids expressing small hairpin RNA in the corneal wound injury model (full-thickness corneal incision) to evaluate opacification.RESULTS. GFP expression peaked between days 1 and 3 and had prominent expression for 15 days. In the corneal wound injury model, we found that the GFP-positive cells demonstrated extensive dendritic-like processes that extended to adjacent cells, whereas the vimentin knockdown model showed significantly reduced corneal opacity.CONCLUSIONS. These findings suggest that a nonviral gene therapeutic approach has potential for treating corneal fibrosis and ultimately reducing scarring.

S100A4 Silencing Facilitates Corneal Wound Healing After Alkali Burns by Promoting Autophagy via Blocking the PI3K/Akt/mTOR Signaling Pathway

“…1 C). After corneal damage, corneal stromal cell fibrosis is the main cause of visual impairment, 17 so the level of α-smooth muscle actin (ɑ-SMA) in the cornea was measured by Western blot assay. The results showed that the protein level of ɑ-SMA in the cornea of rabbits with alkali burn was significantly reduced after S100A4 silencing (all P < 0.05) ( Fig.…”

Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding

“…HNE, 15-deoxy-Δ 12 14 -prostaglandin J 2 (15d-PGJ 2 ), prostaglandin A 1 (PGA 1 ) and their biotinylated derivatives were from Cayman Chemical. Diamide, DBB, Iac-B, TPEN, Z-LLL o MG132, Zinquin acid and Zinquin ethyl ester, anti-actin, anti-tubulin and anti-vimentin antibodies were obtained from Sigma.…”

“…Previously considered as a marker for epithelial–mesenchymal transition, it is now recognized as an active factor in this process, and therefore in cancer cell dissemination 8 9 . Vimentin is essential in wound healing, but it can also promote excessive scarring 10 11 12 . Moreover, it plays a facilitating role in certain viral infections 13 14 and as an autoantigen in the pathogenesis of rheumatic fever 15 and rheumatoid arthritis 16 .…”

Vimentin intermediate filament assembly regulates fibroblast invasion in fibrogenic lung injury

“…A lack of vimentin is associated with defects in wound healing (12). Inhibiting vimentin protects from fibrotic injuries (13,14), whereas overexpression of vimentin promotes increased invasiveness and excessive scarring (10). Our previous studies have demonstrated that modified forms of vimentin target signaling pathways that include transcription of profibrotic genes in pulmonary fibrosis (15,16).…”