title: Identification and validation of vesicant therapeutic targets using a high-throughput siRNA screening approach.
abstract: Sulfur mustard [SM, bis-(2-chloroethyl) sulfide] is a highly reactive bifunctional alkylating agent that has been used as a vesicating agent in warfare scenarios to induce severe lung, skin, and eye injury. SM cutaneous lesions are characterized by both vesication and severe inflammation, but the molecular mechanisms that lead to these signs and symptoms are not well understood. There is a pressing need for effective therapeutics to treat this injury. An understanding of the molecular mechanisms of injury and identification of potential therapeutic targets is necessary for rational therapeutic development. We have applied a high-throughput small interfering RNA (siRNA) screening approach to the problem of SM cutaneous injury in an effort to meet these needs. Our siRNA screening efforts have initially focused on SM-induced inflammation in cutaneous injury since chronic inflammation after exposure appears to play a role in progressive clinical pathology, and intervention may improve clinical outcome. Also, targets that mitigate cellular injury should reduce the inflammatory response. Historical microarray data on this injury were mined for targets and pathways implicated in inflammation, and a siRNA library of 2,017 targets was assembled for screening. Primary screening and library deconvolution were performed using human HaCaT keratinocytes and focused on cell death and inflammatory markers as end points. Using this in vitro approach, we have identified and validated novel targets for the potential treatment of SM-induced cutaneous injury.
authors.affiliation: Cell and Molecular Biology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD, 21010-5400, USA. albert.l.ruff2.civ@mail.mil.
authors.forename: Albert L
authors.initials: AL
authors.lastname: Ruff
authors.affiliation: Cell and Molecular Biology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD, 21010-5400, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Beach
authors.affiliation: Cell and Molecular Biology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD, 21010-5400, USA.
authors.forename: John
authors.initials: J
authors.lastname: Lehman
authors.affiliation: Cell and Molecular Biology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD, 21010-5400, USA.
authors.forename: Cristin
authors.initials: C
authors.lastname: Rothwell
authors.affiliation: Cell and Molecular Biology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD, 21010-5400, USA.
authors.forename: James F
authors.initials: JF
authors.lastname: Dillman
mesh_terms: D002460:Cell Line; D002619:Chemical Warfare Agents / Q000633:toxicity; D003877:Dermatitis, Contact / Q000209:etiology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007509:Irritants / Q000633:toxicity*; D015603:Keratinocytes / Q000187:drug effects; D058990:Molecular Targeted Therapy / Q000379:methods*; D009151:Mustard Gas / Q000633:toxicity*; D034741:RNA, Small Interfering; D012867:Skin / Q000187:drug effects
chemical_list: D002619:Chemical Warfare Agents; D007509:Irritants; D034741:RNA, Small Interfering; D009151:Mustard Gas
keywords: Cutaneous injury; High-throughput screening; In vitro models; Inflammation; Sulfur mustard; siRNA