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Oxidative stress Leading to Decreased Lung Function
Point of Contact
- Karsta Luettich
- Frazer Lowe
- Monita Sharma
- Hasmik Yepiskoposyan
- Damien Breheny
- Rex FitzGerald
|Handbook Version||OECD status||OECD project|
This AOP was last modified on April 29, 2023 16:03
|Cilia Beat Frequency, Decreased||September 10, 2021 01:38|
|Mucociliary Clearance, Decreased||September 10, 2021 07:19|
|Decrease, Lung function||September 08, 2021 04:54|
|Oxidative Stress||March 21, 2023 15:16|
|Oxidative Stress leads to CBF, Decreased||March 24, 2023 06:04|
|CBF, Decreased leads to MCC, Decreased||March 24, 2023 08:17|
|MCC, Decreased leads to Decreased lung function||March 24, 2023 08:27|
|Acrolein||September 28, 2021 08:23|
|Ozone||September 28, 2021 08:26|
|Cigarette smoke||September 28, 2021 09:07|
|Nitrogen dioxide||September 28, 2021 08:54|
|Diesel engine exhaust||September 28, 2021 08:55|
We propose here an AOP that attempts to delineate how exposure to oxidative insults lead to decreased lung function (Luettich et al., 2021). This AOP evaluates one of the major processes known to be involved in regulating efficient mucociliary clearance (MCC)—ciliary function. MCC is a key aspect of the innate immune defense against airborne pathogens and inhaled chemicals and is governed by the concerted action of its functional components, the cilia and the airway surface liquid (ASL), which is composed of mucus and periciliary layers (Bustamante-Marin and Ostrowski, 2017). Disturbances in any of the processes regulating ciliary function can cause MCC dysfunction. Impaired MCC is linked to airway diseases such as chronic obstructive pulmonary disease (COPD) or asthma, both of which are characterized by decreased lung function and bear a significant risk of increased morbidity and mortality. Given the individual and public health burden of the consequences of lung function impairment, gaining a greater understanding of the underlying mechanisms is extremely important in the risk assessment of respiratory toxicants.
The KE proposed here are moderately to highly essential, and we judge the overall biological plausibility of this AOP as strong. The KER Oxidative stress leading to decreased CBF is supported by multiple studies across different species with ample empirical evidence reflecting both dose-response and time concordance. The KER Decreased CBF leading to decreased MCC lacks this expanse of empirical evidence, or the evidence does not fully support the causality between the KE even though the relationship is logical and plausible. Overall, our quantitative understanding of the AOP is moderate.
AOP Development Strategy
With a surface area of ~100 m2 and ventilated by 10,000 to 20,000 liters of air per day (National Research Council, 1988; Frohlich et al., 2016), the lungs are a major barrier that protect the body from a host of external factors that enter the respiratory system and may cause lung pathologies. Mucociliary clearance (MCC) is a key aspect of the innate immune defense against airborne pathogens and inhaled particles and is governed by the concerted action of its functional components, the cilia and the airway surface liquid (ASL), which comprises mucus and the periciliary layer (Bustamante-Marin and Ostrowski, 2017). In healthy subjects, ≥10 mL airway secretions are continuously produced and transported daily by the mucociliary escalator. Disturbances in any of the processes regulating ASL volume, mucus production, mucus viscoelastic properties, or ciliary function can cause MCC dysfunction and are linked to airway diseases such as chronic obstructive pulmonary disease (COPD) or asthma, both of which bear a significant risk of increased morbidity and mortality. The mechanism by which exposure to inhaled toxicants might lead to mucus hypersecretion and thereby impact pulmonary function has already been mapped in AOP148 on decreased lung function. However, whether an exposure-related decline in lung function is solely related to excessive production of mucus is debatable, particularly in light of the close relationship between mucus, ciliary function, and efficient MCC. To date, no single event has been attributed to MCC impairment, and it is likely that events described in this AOP as well as in AOP148, AOP424, and AOP425 have to culminate to lead to decreased lung function.
Summary of the AOP
Molecular Initiating Events (MIE)
Key Events (KE)
Adverse Outcomes (AO)
Relationships Between Two Key Events (Including MIEs and AOs)
Life Stage Applicability
|All life stages|
|Homo sapiens||Homo sapiens||High||NCBI|
Overall Assessment of the AOP
The experimental evidence to support the biological plausibility of the KERs from MIE to AO is moderate to strong overall for the AOP presented here, while there is a moderate concordance of dose-response relationships. In terms of essentiality, we have rated all of the KEs as either moderate or high.
AOPs such as this one can play a central role in risk assessment strategies for a wide variety of regulatory purposes by providing mechanistic support to an integrated approach to testing and assessment (IATA; (Clippinger et al., 2018)). IATAs are flexible frameworks that can be adapted to best address the regulatory question or purpose at hand. More specifically, this AOP can be applied to the risk assessment of inhaled toxicants, by enabling the development of testing strategies through the assembly of existing information and the generation of new data where they are currently lacking. Targeted approaches to fill data gaps can be developed using new approach methodologies (NAMs) informed by this AOP.
Domain of Applicability
All KE proposed in this AOP occur and are measurable in several species, including frogs, mice, rats, guinea pigs, ferrets, cats, dogs, cows, monkeys, and humans. The majority of the supporting empirical evidence derives from studies in rodent and human systems, and experimental findings in animals appear to be highly translatable to humans.
Data regarding the applicability of KE to all life-stages from birth to adulthood are available for the MIE (Oxidative Stress), KE1908 (Cilia Beat Frequency, Decreased), KE1909 (Mucociliary Clearance, Decreased), and AO (Decreased Lung Function), and indicate that they apply to all life stages. It is also worth noting here that age-dependent decreases in CBF, MCC, and lung function have been demonstrated in several species (e.g., guinea pigs, mice, and humans) and reflect normal physiological aging processes (Bailey et al., 2014; Grubb et al., 2016; Ho et al., 2001; Joki and Saano, 1997; Paul et al., 2013; Sharma and Goodwin, 2006).
Gender-specific data relevant to the AOP network are not as widely available as species-specific data, and to our knowledge, the role of gender has not been systematically evaluated for all KE described here. Informative evidence on gender differences stems from patients with chronic pulmonary diseases, such as cystic fibrosis, asthma, COPD, and bronchiectasis, that are characterized by decreased lung function. Considering the importance of efficient MCC—brought about by the interactions of ciliary function, ASL homeostasis and mucus properties—for normal physiological function, we consider this AOP applicable to both genders.
Essentiality of the Key Events
The definition of essentiality implies that the modulation of upstream KEs impacts the downstream KEs in an expected fashion. If blocked or failing to occur, the KEs in the current AOP will not necessarily stop the progression to subsequent KEs. Due to the complex biology of motile cilia formation and function, ASL homeostasis, mucus properties and their concerted impact on MCC, the KEs and AO may be triggered because of alternative pathways or biological redundancies. However, when exacerbated, the KEs promote the occurrence of downstream events eventually leading to the AO. The causal pathway starting from the exposure to oxidants and leading to decreased lung function involves parallel routes with KEs, each of which is sufficient to cause the downstream KE to occur. Based on the evidence we judge the MIE (Oxidative Stress), KE1908 (Cilia Beat Frequency, Decreased), and KE1909 (Mucociliary Clearance, Decreased) as highly essential.
We judge the overall biological plausibility of this AOP as strong. The KER Oxidative stress leading to decreased CBF is supported by multiple studies across different species with ample empirical evidence reflecting both dose-response and time concordance. The KER Decreased CBF leading to decreased MCC lacks this expanse of empirical evidence, or the evidence does not fully support the causality between the KE even though the relationship is logical and plausible.
Known Modulating Factors
Overall, our quantitative understanding of the AOP network is moderate.
There is robust evidence that provides an insight into the KER Oxidative stress leading to decreased cilia beat frequency and Decreased cilia beat frequency leading to decreased MCC, and the dose response and temporal relationship between the two KE in question are well described and quantified for different stressors across different test systems. In some instances, we are less confident in our quantitative understanding. For example, dose response data as well as data supportive of the KE causality are limited for the KER Decreased MCC leading to decreased lung function.
Considerations for Potential Applications of the AOP (optional)
Given the individual and public health burden of the consequences of lung function impairment, gaining a greater understanding of the underlying mechanisms is extremely important in the risk assessment of respiratory toxicants. An integrated assessment of substances with the potential to be inhaled, either intentionally or unintentionally, could incorporate inhalation exposure and dosimetry modelling to inform an in vitro approach with appropriate exposure techniques and cell systems to assess KEs in this AOP (EPA’s Office of Chemical Safety and Pollution Prevention, 2019). Standardization and robustness testing of assays against explicit performance criteria using suitable reference materials can greatly increase the level of confidence in their use for KE assessment (Petersen et al., 2021). Much of the empirical evidence that supports the KERs in the qualitative AOP described here was obtained from in vitro studies using well-established methodologies for biological endpoint assessment. Being chemical agnostic, this AOP can be applied to a variety of substances that share the AO. For example, impaired MCC and decreased lung function have a long-known relationship with smoking, but little is known about the consequences of long-term use of alternative inhaled nicotine delivery products such as electronic cigarettes and heated tobacco products. This AOP can form the basis of an assessment strategy to evaluate the effects of exposure to aerosol from these products based on the KEs identified here.
Bailey, K.L., Bonasera, S.J., Wilderdyke, M., Hanisch, B.W., Pavlik, J.A., DeVasure, J., et al. (2014). Aging causes a slowing in ciliary beat frequency, mediated by PKCε. Am. J. Physiol. Lung Cell. Mol. Physiol. 306, L584-L589.
Bustamante-Marin, X.M., and Ostrowski, L.E. (2017a). Cilia and Mucociliary Clearance. Cold Spring Harb. Persp. Biol. 9, a028241.
Clippinger, A.J., Allen, D., Behrsing, H., BéruBé, K.A., Bolger, M.B., Casey, W., et al. (2018). Pathway-based predictive approaches for non-animal assessment of acute inhalation toxicity. Toxicol. In Vitro 52, 131-145.
EPA’s Office of Chemical Safety and Pollution Prevention (2019). "FIFRA Scientific Advisory Panel Meeting Minutes and Final Report No. 2019-01 Peer Review on Evaluation of a Proposed Approach to Refine the Inhalation Risk Assessment for Point of Contact Toxicity: A Case Study Using a New Approach Methodology (NAM) December 4 and 6, 2018 FIFRA Scientific Advisory Panel Meeting". U.S. Environmental Protection Agency).
Frohlich, E., Mercuri, A., Wu, S., and Salar-Behzadi, S. (2016). Measurements of Deposition, Lung Surface Area and Lung Fluid for Simulation of Inhaled Compounds. Front. Pharmacol. 7, 181.
Grubb, B.R., Livraghi-Butrico, A., Rogers, T.D., Yin, W., Button, B., and Ostrowski, L.E. (2016). Reduced mucociliary clearance in old mice is associated with a decrease in Muc5b mucin. Am. J. Physiol. Lung Cell. Mol. Physiol. 310, L860-L867.
Ho, J.C., Chan, K.N., Hu, W.H., Lam, W.K., Zheng, L., Tipoe, G.L., et al. (2001). The effect of aging on nasal mucociliary clearance, beat frequency, and ultrastructure of respiratory cilia. Am. J. Respir. Crit. Care Med. 163, 983-988.
Joki, S., and Saano, V. (1997). Influence of ageing on ciliary beat frequency and on ciliary response to leukotriene D4 in guinea-pig tracheal epithelium. Clin. Exp. Pharmacol. Physiol. 24, 166-169.
Luettich, K., Sharma, M., Yepiskoposyan, H., Breheny, D., and Lowe, F. J. (2021). An Adverse Outcome Pathway for Decreased Lung Function Focusing on Mechanisms of Impaired Mucociliary Clearance Following Inhalation Exposure. Frontiers in Toxicology, 55.
National Research Council (1988). Air Pollution, the Automobile, and Public Health. Washington, DC: The National Academies Press.
Paul, P., Johnson, P., Ramaswamy, P., Ramadoss, S., Geetha, B., and Subhashini, A. (2013). The effect of ageing on nasal mucociliary clearance in women: a pilot study. ISRN 2013, 598589.
Petersen, E.J., Sharma, M., Clippinger, A.J., Gordon, J., Katz, A., Laux, P., et al. (2021). Use of Cause-and-Effect Analysis to Optimize the Reliability of In Vitro Inhalation Toxicity Measurements Using an Air–Liquid Interface. Chem. Res. Toxicol. 34, 1370–1385.
Sharma, G., and Goodwin, J. (2006). Effect of aging on respiratory system physiology and immunology. Clin. Interv. Aging 1, 253-260.