Alcohol abuse has long been known to impair lung host defenses increasing the risk in heavy drinkers for bronchitis, pneumonia and acute respiratory distress syndrome. Our laboratory has focused on alcohol's impact on mucociliary clearance in the conducting airways of the lung, which provides the first line of defense of the lung against inhaled infectious agents, particles and debris. We have established two key observations that drive this proposal: 1) brief modest alcohol intake stimulates airway ciliary motility;and 2) sustained heavy alcohol intake impairs mucociliary clearance increasing the risk for airway injury and pneumonia. We have termed this impairment """"""""Alcohol-Induced Ciliary Dysfunction"""""""" (AICD). Our mechanistic studies have demonstrated that both the brief alcohol stimulation and sustained AICD involve ciliated cell mechanisms dependent on the regulation of nitric oxide (NO), cilia-associated cyclases and cyclic nucleotide-dependent kinases (PKG &PKA). Recently, we published that these alcohol-triggered ciliary regulation pathways are distinctly localized to the basal body of each airway cilium in what we have called the alcohol-responsive """"""""ciliary metabolon"""""""". This pathway is exquisitely sensitive to very low concentrations of alcohol (1-10 mM, which is less than half of the legal intoxication limit) and is fully functionl in isolated airway cilia organelles. This leads us to hypothesize that: Alcohol causes reversible airway ciliary dysfunction due to modified nitric oxide signaling and altered regulation of key cila proteins, including the dynein motors and the mechanisms that regulate dyneins. In this proposal, we outline experiments designed to answer four new questions: 1) How does brief alcohol exposure stimulate NO production in airway cilia? We explore this question in Aim #1 by examining the role alcohol plays in activating nitric oxide synthase (eNOS) by enhancing the chaperone function of eNOS by heat shock protein 90 (HSP90). 2) Why does sustained alcohol exposure deplete ciliary NO levels? We address this question in Aim #2 by defining how eNOS becomes uncoupled by sustained alcohol exposure due to L- arginine depletion and the generation of reactive oxygen species (ROS). We think increased ciliary ROS leads to depletion of the eNOS cofactor tetrahydrobiopterin (BH4) resulting in cilia desensitization. 3) How do cilia regulatory enzymes interact to modify motility? In Aim #3 we will determine how sustained alcohol exposure activates protein phosphatase 1 (PP1), which leads to dephosphorylation of HSP90 and other key cilia activation proteins such as PKA. 4) What downstream cilia motor molecules are modified by alcohol exposure? Using the model genetic system of the alcohol-sensitive motile organism, Chlamydomonas, Aim #4 will focus on how sustained alcohol modifies ciliary outer dynein arms, which are the motor proteins that make cilia beat. The studies we propose will greatly extend our knowledge of how to prevent and treat AICD and will expand our insight into how alcohol alters cilia molecules critical for airway cilia function. 1

Public Health Relevance

Cilia, the wave-producing finger-like projections of the bronchial tubes that clear mucus and inhaled particles from the lungs, are normally activated by stress events to clear mucus faster. Catching a cold, choking on food or breathing in dirty air cause cilia to produce nitric oxide, which makes them beat faster to clear out the lungs. After heavy alcohol drinking, lung cilia become unresponsive to stress, because they can no longer make nitric oxide, which leads to mucus congestion and infections such as bronchitis &pneumonia. Our research is focused on how alcohol shuts off nitric oxide production by lung cilia with an emphasis on discovering ways to reverse and/or prevent alcohol-related cilia damage to avoid lung infections. 1

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
2R01AA008769-20A1
Application #
8372198
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Jung, Kathy
Project Start
1991-03-01
Project End
2017-05-31
Budget Start
2012-08-20
Budget End
2013-05-31
Support Year
20
Fiscal Year
2012
Total Cost
$482,689
Indirect Cost
$134,825
Name
University of Nebraska Medical Center
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
168559177
City
Omaha
State
NE
Country
United States
Zip Code
68198
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Yang, Fan; Scarbrough, Chasity; Sisson, Joseph H et al. (2018) PKA, PP1, and DC1 phosphorylation mediate alcohol-induced ciliary dysfunction in Chlamydomonas reinhardtii. Alcohol 75:31-38
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Price, Michael E; Pavlik, Jacqueline A; Liu, Miao et al. (2017) Alcohol drives S-nitrosylation and redox activation of protein phosphatase 1, causing bovine airway cilia dysfunction. Am J Physiol Lung Cell Mol Physiol 312:L432-L439
Warren, Kristi J; Simet, Samantha M; Pavlik, Jacqueline A et al. (2016) RSV-specific anti-viral immunity is disrupted by chronic ethanol consumption. Alcohol 55:35-42
Gerald, Carresse L; Romberger, Debra J; DeVasure, Jane M et al. (2016) Alcohol Decreases Organic Dust-Stimulated Airway Epithelial TNF-Alpha Through a Nitric Oxide and Protein Kinase-Mediated Inhibition of TACE. Alcohol Clin Exp Res 40:273-83
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