Tobacco and alcohol are the two most used and abused drugs in the world. Together, they are responsible for millions of deaths each year worldwide. Despite the enormous amount of research carried out to understand how these agents affect the lung, new information leading to significant advances at the clinic has been disappointing. Because of the high prevalence of tobacco and alcohol exposure in veterans, the Roman lab is exploring the mechanisms by which these agents affect the lung. Our research revealed that both alcohol and nicotine, an alkaloid present in tobacco, promote alterations in lung matrix composition that we have termed 'transitional' remodeling, a process characterized by subtle alterations in the relatively composition of the lung extracellular matrix. Although the exact mechanisms linking these events to lung disease remain incompletely understood, we believe that transitional remodeling renders the host susceptible to lung disease, and this belief is supported by published research performed both in vitro and in vivo. Considering the above, we set out to identify the cellular/molecular mechanisms responsible for these events. Using cultured primary lung fibroblasts, we found that nicotine induces cellular proliferation and matrix expression through the activation of specific 7 nicotinic acetylcholine receptors (7 nAChRs). The importance of these receptors was confirmed in vivo showing that prenatal exposure to nicotine affects lung development and induces airway thickening and hyperreactivity in wildtype animals, but not in 7 nAChR knockout mice. Using the same culture model, we discovered that alcohol acts on lung fibroblasts via another set of nAChRs, 4 nAChRs. In rodent lungs, the expression of these receptors is increased by chronic alcohol exposure which, in turn, predisposes to acute lung injury. This work suggests that the cholinergic system plays an important role in lung homeostasis and response to injury, and points to nAChRs as potential targets for intervention. More recently, we found that nAChRs are activated through both ligand-dependent and -independent pathways. Specifically, while nicotine acts on 7 nAChRs in a traditional ligand-dependent fashion, alcohol works, at least in part, through induction of a particular form of oxidant stress (oxidation of the cysteine/cystine redox potential or Eh Cys/CySS) which, together with cysteine residues strategically located on the subunits of certain nAChR pentamers, activates nAChR signaling via ligand-independent mechanisms of action. Importantly, we have generated preliminary data to suggest that susceptibility to injury in aging might activate nAChRs through similar pathways thereby promoting transitional remodeling and increased susceptibility to lung injury. Considering the above, we propose that aging (as observed for nicotine and ethanol exposure) is associated with lung transitional remodeling via distinct nAChRs. nAChRs can be activated via ligand-dependent and -independent pathways, the latter triggered by oxidant stress induced by oxidation of the Eh Cys/CySS. Specifically, oxidant stress affects cysteines strategically located in the ? subunits of nAChRs which, in turn, triggers signal transduction and promotes lung transitional remodeling and increase susceptibility to disrepair after lung injury. To test this hypothesis, we propose experiments designed to:
Aim 1. Investigate the role of specific subunit cysteines in nAChR activation in lung cells and in vivo.
Aim 2. Determine the role of oxidant stress, especially, oxidation of the Eh Cys/CySS in nAChR activation both in vitro and in vivo.
Aim 3. Examine the role of nAChR activation in lung disease related to aging and how this is affected by tobacco and alcohol exposure.

Public Health Relevance

Aging and tobacco and alcohol abuse are important problems worldwide and many disorders observed in veterans are associated with these states. This work will study how aging affects lung cells in an attempt to identify potential targets for intervention, and determine how tobacco and alcohol exposure influence these events. Specifically, the research will focus on a special kind of cell receptors termed nicotinic acetylcholine receptors (nAChRs). Studies will be performed in cultured cells as well as in animal models to study the role of these receptors in aging and in tobacco- and alcohol-related lung disease.

Agency
National Institute of Health (NIH)
Institute
Veterans Affairs (VA)
Type
Non-HHS Research Projects (I01)
Project #
5I01BX000216-08
Application #
9501671
Study Section
Respiration (PULM)
Project Start
2009-04-01
Project End
2018-09-30
Budget Start
2017-10-01
Budget End
2018-09-30
Support Year
8
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Louisville VA Medical Medical Center
Department
Type
DUNS #
086765245
City
Louisville
State
KY
Country
United States
Zip Code
40206
Zheng, Yuxuan; Ritzenthaler, Jeffrey D; Burke, Tom J et al. (2018) Age-dependent oxidation of extracellular cysteine/cystine redox state (Eh(Cys/CySS)) in mouse lung fibroblasts is mediated by a decline in Slc7a11 expression. Free Radic Biol Med 118:13-22
Zelko, Igor N; Zhu, Jianxin; Roman, Jesse (2018) Role of SOD3 in silica-related lung fibrosis and pulmonary vascular remodeling. Respir Res 19:221
Watson, Walter H; Ritzenthaler, Jeffrey D; Roman, Jesse (2016) Lung extracellular matrix and redox regulation. Redox Biol 8:305-15
Watson, Walter H; Burke, Tom J; Zelko, Igor N et al. (2016) Differential Regulation of the Extracellular Cysteine/Cystine Redox State (EhCySS) by Lung Fibroblasts from Young and Old Mice. Oxid Med Cell Longev 2016:1561305
Zelko, Igor N; Zhu, Jianxin; Ritzenthaler, Jeffrey D et al. (2016) Pulmonary hypertension and vascular remodeling in mice exposed to crystalline silica. Respir Res 17:160