title: ADAM17 Inhibitors Attenuate Corneal Epithelial Detachment Induced by Mustard Exposure.
abstract: OBJECTIVE
  Sulfur mustard, nitrogen mustard (NM), and 2-chloroethyl ethyl sulfide all cause corneal injury with epithelial-stromal separation, differing only by degree. Injury can resolve in a few weeks or develop into chronic corneal problems. These vesicants induce microbullae at the epithelial-stromal junction, which is partially caused by cleavage of transmembranous hemidesmosomal collagen XVII, a component anchoring the epithelium to the stroma. ADAM17 is an enzyme involved in wound healing and is able to cleave collagen XVII. The activity of ADAM17 was inhibited in vesicant-exposed corneas by four different hydroxamates, to evaluate their therapeutic potential when applied 2 hours after exposure, thereby allowing ADAM17 to perform its early steps in wound healing.
  
  
  METHODS
  Rabbit corneal organ cultures exposed to NM for 2 hours were washed, then incubated at 37°C for 22 hours, with or without one of the four hydroxamates (dose range, 0.3-100 nmol in 20 μL, applied four times). Corneas were analyzed by light and immunofluorescence microscopy, and ADAM17 activity assays.
  
  
  RESULTS
  Nitrogen mustard-induced corneal injury showed significant activation of ADAM17 levels accompanying epithelial-stromal detachment. Corneas treated with hydroxamates starting 2 hours post exposure showed a dose-dependent ADAM17 activity inhibition up to concentrations of 3 nmol. Of the four hydroxamates, NDH4417 (N-octyl-N-hydroxy-2-[4-hydroxy-3-methoxyphenyl] acetamide) was most effective for inhibiting ADAM17 and retaining epithelial-stromal attachment.
  
  
  CONCLUSIONS
  Mustard exposure leads to corneal epithelial sloughing caused, in part, by the activation of ADAM17 at the epithelial-stromal junction. Select hydroxamate compounds applied 2 hours after NM exposure mitigated epithelial-stromal separation.
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Andrea
authors.initials: A
authors.lastname: DeSantis-Rodrigues
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Yoke-Chen
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Rita A
authors.initials: RA
authors.lastname: Hahn
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Iris P
authors.initials: IP
authors.lastname: Po
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Peihong
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania, United States.
authors.forename: C Jeffrey
authors.initials: CJ
authors.lastname: Lacey
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania, United States.
authors.forename: Abhilash
authors.initials: A
authors.lastname: Pillai
authors.affiliation: Department of Chemistry, Muhlenberg College, Allentown, Pennsylvania, United States.
authors.forename: Sherri
authors.initials: S
authors.lastname: C Young
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania, United States.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Flowers
authors.affiliation: Department of Environmental and Occupational Health, Robert Wood Johnson Medical School, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gallo
authors.affiliation: Department of Environmental and Occupational Health, Robert Wood Johnson Medical School, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Laskin
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Donald R
authors.initials: DR
authors.lastname: Gerecke
authors.affiliation: Department of Biomedical Sciences, Baylor College of Dentistry, Texas A&M Health Science Center, Dallas, Texas, United States.
authors.forename: Kathy K H
authors.initials: KK
authors.lastname: Svoboda
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania, United States.
authors.forename: Ned D
authors.initials: ND
authors.lastname: Heindel
authors.affiliation: Department of Pharmacology and Toxicology Ernest Mario School of Pharmacy, Rutgers University, Piscataway, New Jersey, United States.
authors.forename: Marion K
authors.initials: MK
authors.lastname: Gordon
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072198:ADAM17 Protein; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D003319:Corneal Stroma / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D019573:Epithelium, Corneal / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008466:Mechlorethamine / Q000633:toxicity*; D011817:Rabbits; D041623:Tomography, Optical Coherence; D014409:Tumor Necrosis Factor-alpha
chemical_list: D014409:Tumor Necrosis Factor-alpha; D008466:Mechlorethamine; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human