Postexposure Application of Fas Receptor Small-Interfering RNA to Suppress Sulfur Mustard-Induced Apoptosis in Human Airway Epithelial Cells: Implication for a Therapeutic Approach

Abstract

Sulfur mustard (SM) is a vesicant chemical warfare and terrorism agent. Besides skin and eye injury, respiratory damage has been mainly responsible for morbidity and mortality after SM exposure. Previously, it was shown that suppressing the death receptor (DR) response by the dominant-negative Fas-associated death domain protein prior to SM exposure blocked apoptosis and microvesication in skin. Here, we studied whether antagonizing the Fas receptor (FasR) pathway by small-interfering RNA (siRNA) applied after SM exposure would prevent apoptosis and, thus, airway injury. Normal human bronchial/tracheal epithelial (NHBE) cells were used as an in vitro model with FasR siRNA, FasR agonistic antibody CH11, and FasR antagonistic antibody ZB4 as investigative tools. In NHBE cells, both SM (300 mM) and CH11 (100 ng/ml) induced caspase-3 activation, which was inhibited by FasR siRNA and ZB4, indicating that SM-induced apoptosis was via the Fas response. FasR siRNA inhibited SM-induced caspase-3 activation when added to NHBE cultures up to 8 hours after SM. Results using annexin V/propidium iodidestained cells showed that both apoptosis and necrosis were involved in cell death due to SM; FasR siRNA decreased both apoptotic and necrotic cell populations. Bronchoalveolar lavage fluid (BALF) of rats exposed to SM (1 mg/kg, 50 minutes) revealed a significant (P , 0.05) increase in soluble Fas ligand and active caspase-3 in BALF cells. These findings suggest an intervention of Fas-mediated apoptosis as a postexposure therapeutic strategy with a therapeutic window for SM inhalation injury and possibly other respiratory diseases involving the Fas response.

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Document Details

Document Type
Technical Report
Publication Date
Jan 01, 2013
Accession Number
ADA574486

Entities

People

  • Ashley Appell
  • Betty Benton
  • Brian M. Keyser
  • Chris Carpin
  • Dana R. Anderson
  • Danielle Paradiso
  • Devon K. Andres
  • Eric Nealley
  • Radharaman Ray
  • Wesley W. Holmes
  • William J. Smith

Organizations

  • United States Army Medical Research Institute of Chemical Defense

Tags

DTIC Thesaurus Topics

  • Acute Respiratory Distress Syndrome
  • Biological Factors
  • Cell Physiological Processes
  • Cells
  • Chemical Warfare
  • Chemical Warfare Agents
  • Chemical Weapons
  • Chemistry
  • Culture Techniques
  • Epithelial Cells
  • Lung
  • Lung Diseases
  • Peptides
  • Proteins
  • Rodents
  • Therapy
  • Wounds And Injuries

Fields of Study

  • Biology
  • Medicine

Readers

  • Cellular and Molecular Pathways of Apoptosis.
  • Geochemistry
  • Immunology and Pathology