title: Vimentin knockdown decreases corneal opacity.
abstract: OBJECTIVE
  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.
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Subrata K
authors.initials: SK
authors.lastname: Das
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Isha
authors.initials: I
authors.lastname: Gupta
authors.affiliation: St. Vincent's Hospital, College of Medicine, The Catholic University of Korea, Suwon, South Korea.
authors.forename: Yang Kyung
authors.initials: YK
authors.lastname: Cho
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Hironori
authors.initials: H
authors.lastname: Uehara
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Muddana
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Ashlie A
authors.initials: AA
authors.lastname: Bernhisel
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Bonnie
authors.initials: B
authors.lastname: Archer
authors.affiliation: Department of Ophthalmology, Moran Eye Center, University of Utah, Salt Lake City, Utah, United States.
authors.forename: Balamurali K
authors.initials: BK
authors.lastname: Ambati
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003315:Cornea / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D060527:Corneal Keratocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003318:Corneal Opacity / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D004195:Disease Models, Animal; D005131:Eye Injuries / Q000378:metabolism / Q000473:pathology; D005260:Female; D005355:Fibrosis / Q000235:genetics / Q000473:pathology / Q000628:therapy; D005500:Follow-Up Studies; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D012313:RNA / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D014746:Vimentin / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use*; D014945:Wound Healing
chemical_list: D014746:Vimentin; D049452:Green Fluorescent Proteins; D012313:RNA
keywords: corneal fibroblasts; corneal wound healing; keratocytes