Abstract
Exposure to cigarettes and other nicotine-based products results in persistent inflammation in the lung. In recent years, electronic cigarettes (e-cigs) have become extremely popular among adults and youth alike. E-cigarette vapor-induced oxidative stress promotes protein breakdown, DNA damage and cell death, culminating in a variety of respiratory diseases. The proteasome, a multi-catalytic protease, superintends protein degradation within the cell. When cells are stimulated with inflammatory cytokines such as IFN-γ and TNF-α, the constitutive catalytic proteasome subunits are replaced by the inducible subunits-low-molecular mass polypeptide (LMP)2 (β1i), multi-catalytic endopeptidase complex-like (MECL)1 (β2i), and LMP7 (β5i), which are required for the production of certain MHC class I-restricted T-cell epitopes. In this study, we used human alveolar epithelial cells (A549) and exposed them to filtered air or (1%) tobacco-flavored (TF) electronic cigarette vapor condensate (ECVC) ± nicotine (6 mg/ml) (TF-ECVC ± N) for 24 h. We observed an increase in the levels of IFN-γ, TNF-α, and inducible proteasome subunits (LMP7/PSMB8, LMP2/PSMB9, MECL1/PSMB10), and a reduced expression of constitutive proteasome subunits (β1/PSMB6 and β2/PSMB7) in challenged A549 cells. Interestingly, knockdown of the inducible proteasome subunit LMP7 reversed ECVC-induced expression of NADPH oxidase and immunoproteasome subunits in A549 cells. In addition, pre-exposure to an LMP7 inhibitor (ONX-0914) abrogated the mRNA expression of several NOX subunits and rescued the excessive production/release of inflammatory cytokines/chemokines (IL-6, IL-8, CCL2, and CCL5) in ECVC-challenged cells. Our findings suggest an important role of LMP7 in regulating the expression of inflammatory mediators during ECVC exposure. Overall, our results provide evidence for proteasome-dependent ROS-mediated inflammation in ECVC-challenged cells.
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The data that support the findings of this study are available on request from the first and corresponding author. The data are not publicly available due to privacy or ethical restrictions.
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This work was supported by Young Clinical Scientist Award from the Flight Attendant Medical Research Institute (FAMRI- 123253_YCSA_Faculty) and by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number 5P20 GM103424-18 and 3 P20 GM103424-15S1 to SB.
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RB and ST performed the experiments and analyzed the data. RB prepared the figures. ST helped RB in drafting the manuscript. SB provided funding support, laboratory space and directed the project. All authors read, discussed, and approved the final manuscript.
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Begum, R., Thota, S. & Batra, S. Interplay between proteasome function and inflammatory responses in e-cig vapor condensate-challenged lung epithelial cells. Arch Toxicol 97, 2193–2208 (2023). https://doi.org/10.1007/s00204-023-03504-5
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DOI: https://doi.org/10.1007/s00204-023-03504-5