title: Inactivation of the miR-183/96/182 Cluster Decreases the Severity of Pseudomonas aeruginosa-Induced Keratitis.
abstract: OBJECTIVE
  The microRNA-183/96/182 cluster (miR-183/96/182) plays important roles in sensory organs. Because the cornea is replete with sensory innervation, we hypothesized that miR-183/96/182 modulates the corneal response to bacterial infection through regulation of neuroimmune interactions.
  
  
  METHODS
  Eight-week-old miR-183/96/182 knockout (ko) mice and their wild-type littermates (wt) were used. The central cornea of anesthetized mice was scarred and infected with Pseudomonas aeruginosa (PA), strain 19660. Corneal disease was graded at 1, 3, and 5 days postinfection (dpi). Corneal RNA was harvested for quantitative RT-PCR. Polymorphonuclear neutrophils (PMN) were enumerated by myeloperoxidase assays; the number of viable bacteria was determined by plate counts, and ELISA assays were performed to determine cytokine protein levels. A macrophage (Mϕ) cell line and elicited peritoneal PMN were used for in vitro functional assays.
  
  
  RESULTS
  MicroRNA-183/96/182 is expressed in the cornea, and in Mϕ and PMN of both mice and humans. Inactivation of miR-183/96/182 resulted in decreased corneal nerve density compared with wt mice. Overexpression of miR-183/96/182 in Mϕ decreased, whereas knockdown or inactivation of miR-183/96/182 in Mϕ and PMN increased their capacity for phagocytosis and intracellular killing of PA. In PA-infected corneas, ko mice showed decreased proinflammatory neuropeptides such as substance P and chemoattractant molecules, MIP-2, MCP1, and ICAM1; decreased number of PMN at 1 and 5 dpi; increased viable bacterial load at 1 dpi, but decreased at 5 dpi; and markedly decreased corneal disease.
  
  
  CONCLUSIONS
  MicroRNA-183/96/182 modulates the corneal response to bacterial infection through its regulation of corneal innervation and innate immunity.
authors.affiliation: Department of Ophthalmology, Kresge Eye Institute, Wayne State University, School of Medicine, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Chithra K
authors.initials: CK
authors.lastname: Muraleedharan
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Sharon A
authors.initials: SA
authors.lastname: McClellan
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Ronald P
authors.initials: RP
authors.lastname: Barrett
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Cui
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Ophthalmology, Kresge Eye Institute, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Daniel
authors.initials: D
authors.lastname: Montenegro
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Thomas
authors.initials: T
authors.lastname: Carion
authors.affiliation: Department of Ophthalmology, Kresge Eye Institute, Wayne State University, School of Medicine, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Berger
authors.affiliation: Department of Ophthalmology, Kresge Eye Institute, Wayne State University, School of Medicine, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Linda D
authors.initials: LD
authors.lastname: Hazlett
authors.affiliation: Department of Ophthalmology, Kresge Eye Institute, Wayne State University, School of Medicine, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan, United States.
authors.forename: Shunbin
authors.initials: S
authors.lastname: Xu
mesh_terms: D000818:Animals; D002460:Cell Line; D015169:Colony Count, Microbial; D003315:Cornea / Q000294:innervation / Q000378:metabolism / Q000382:microbiology; D003320:Corneal Ulcer / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015818:Eye Infections, Bacterial / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D005260:Female; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D007113:Immunity, Innate; D008264:Macrophages / Q000276:immunology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics*; D009504:Neutrophils / Q000276:immunology; D011552:Pseudomonas Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D011550:Pseudomonas aeruginosa / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D012668:Trigeminal Ganglion / Q000276:immunology
chemical_list: D016207:Cytokines; D035683:MicroRNAs; C540524:Mirn182 microRNA, mouse; C543802:Mirn183 microRNA, mouse; C541554:Mirn96 microRNA, mouse