Abstract
Bronchiolitis obliterans (BO) is a devastating lung disease seen commonly after lung transplant, following severe respiratory tract infection or chemical inhalation exposure. Diacetyl (DA; 2,3-butanedione) is a highly reactive alpha-diketone known to cause BO when inhaled, however, the mechanisms of how inhalation exposure leads to BO development remains poorly understood. In the current work, we combined two clinically relevant models for studying the pathogenesis of DA-induced BO: (1) an in vivo rat model of repetitive DA vapor exposures with recovery and (2) an in vitro model of primary human airway epithelial cells exposed to pure DA vapors. Rats exposed to 5 consecutive days 200 parts-per-million DA 6 h per day had worsening survival, persistent hypoxemia, poor weight gain, and histologic evidence of BO 14 days after DA exposure cessation. At the end of exposure, increased expression of the ubiquitin stress protein ubiquitin-C accumulated within DA-exposed rat lung homogenates and localized primarily to the airway epithelium, the primary site of BO development. Lung proteasome activity increased concurrently with ubiquitin-C expression after DA exposure, supportive of significant proteasome stress. In primary human airway cultures, global proteomics identified 519 significantly modified proteins in DA-exposed samples relative to controls with common pathways of the ubiquitin proteasome system, endosomal reticulum transport, and response to unfolded protein pathways being upregulated and cell–cell adhesion and oxidation–reduction pathways being downregulated. Collectively, these two models suggest that diacetyl inhalation exposure causes abundant protein damage and subsequent ubiquitin proteasome stress prior to the development of chemical-induced BO pathology.
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All proteomics data are uploaded to ftp://massive.ucsd.edu/MSV000086874/, and will be made publically available at the time of publication.
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Acknowledgements
The authors thank the University of Rochester Medical Center’s Inhalation Exposure Facility, specifically Director Alison Elder, PhD, David Chalupa, MS, and Robert Gelein, MS, for their continued support with diacetyl inhalation exposures. Authors also thank Jon Oldach and Anna Maione with MatTek, Corporation, for support on the project as well as Meghan O’Neil (URMC) for her staining of human samples for Ubq-C. Proteomics experiments were performed in the Environmental Molecular Sciences Laboratory (EMSL), a national scientific user facility sponsored by the Department of Energy (DOE) and located at Pacific Northwest National Laboratory, which is operated by Battelle Memorial Institute for the DOE under Contract DE-AC05-76RL0 1830.
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Grants from the National Institute of Environmental Health Sciences P30 ES001247 (TJM, JNF, MDM) and L40 ES030909-01 (MDM), National Heart Lung and Blood Institute 5R01HL139335 (WJQ, CA, MDM), National Center for Advancing Translational Sciences 2KL2TR001999 (MDM), and the University of Rochester Medical Center’s David H. Smith Fund (MDM).
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Matthew D. McGraw and Jacob N. Finkelstein conceptualized and designed the study; So-Young Kim, Emma House, Heather M. Olson, David Chalupa, and Matthew D. McGraw acquired data for the work; Juan Wang, So-Young Kim, Thomas J Mariani, Wei-Jun Qian, Charles Ansong, Jacob N. Finkelstein and Matthew D. McGraw analyzed and interpreted the findings; Juan Wang and Matthew D. McGraw drafted the manuscript; all authors revised the manuscript for intellectual content, gave final approval for publication and agreed to accountability for all aspects of the work.
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Wang, J., Kim, SY., House, E. et al. Repetitive diacetyl vapor exposure promotes ubiquitin proteasome stress and precedes bronchiolitis obliterans pathology. Arch Toxicol 95, 2469–2483 (2021). https://doi.org/10.1007/s00204-021-03076-2
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DOI: https://doi.org/10.1007/s00204-021-03076-2