Abstract
Sulfur mustard (SM) is a highly toxic war chemical that causes significant morbidity and mortality and lacks any effective therapy. Rats exposed to aerosolized CEES (2-chloroethyl ethyl sulfide; 10% in ethanol), an analog of SM, developed acute respiratory distress syndrome (ARDS), which is characterized by increased inflammation, hypoxemia and impaired gas exchange. We observed elevated levels of extracellular nucleic acids (eNA) in the bronchoalveolar lavage fluid (BALF) of CEES-exposed animals. eNA can induce inflammation, coagulation and barrier dysfunction. Treatment with hexadimethrine bromide (HDMBr; 10 mg/kg), an eNA neutralizing agent, 2 h post-exposure, reduced lung injury, inhibited disruption of alveolar–capillary barrier, improved blood oxygenation (PaO2/FiO2 ratio), thus reversing ARDS symptoms. HDMBr treatment also reduced lung inflammation in the CEES-exposed animals by decreasing IL-6, IL-1A, CXCL-1 and CCL-2 mRNA levels in lung tissues and HMGB1 protein in BALF. Furthermore, HDMBr treatment also reduced levels of lung tissue factor and plasminogen activator inhibitor-1 indicating reduction in clot formation and increased fibrinolysis. Fibrin was reduced in BALF of the HDMBr-treated animals. This was further confirmed by histology that revealed diminished airway fibrin, epithelial sloughing and hyaline membrane in the lungs of HDMBr-treated animals. HDMBr completely rescued the CEES-associated mortality 12 h post-exposure when the survival rate in CEES-only group was just 50%. Experimental eNA treatment of cells caused increased inflammation that was reversed by HDMBr. These results demonstrate a role of eNA in the pathogenesis of CEES/SM-induced injury and that its neutralization can serve as a potential therapeutic approach in treating SM toxicity.
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Acknowledgements
This work is supported by National Institutes of Health Office of the Director (NIH OD), the National Institute of Environmental Health Sciences (NIEHS) grant numbers U01ES025069 (AA) and U01ES028182 (SA) and the National Heart Lung and Blood Institute (NHLBI) grant numbers R01HL114933 (AA). VS and KGS are supported by the CAAP program of the Department of Anesthesiology and Perioperative Medicine of the University of Alabama at Birmingham. The authors would also like to thank Stacy Miller, Tara Hendry-Hofer, Jacqueline Rioux, Rhonda Garlick and Joan Loader from the University of Colorado Denver for help in setup of initial studies. The authors are also grateful to Dr. Carl White and Dr. Raymond Rancourt for their valuable advice.
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AA conceived the idea and designed experiments. NM, MH, IZ, VS, SA and AA carried out the experiments, acquired and analyzed data and wrote the manuscript. KS and JFP analyzed data and wrote the manuscript. DRC acquired and analyzed data.
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A patent application for the use of nucleic acid scavengers in chemical-induced ARDS has been filed (AA). All other authors declare that they have no conflict of interest.
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Mariappan, N., Husain, M., Zafar, I. et al. Extracellular nucleic acid scavenging rescues rats from sulfur mustard analog-induced lung injury and mortality. Arch Toxicol 94, 1321–1334 (2020). https://doi.org/10.1007/s00204-020-02699-1
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DOI: https://doi.org/10.1007/s00204-020-02699-1