Current Projects & Grant Support

The role of oxidative stress in disease and the development of therapeutic approaches to alleviate oxidative stress in lung diseases associated with environmental particulate matter exposures.

My laboratory has made discoveries showing the importance of reactive species generated from desert particulate matter (PM) driving airway hyperresponsiveness seen in deploynment related lung disease (DRLD). A major goal of these studies is to explore the mechanisms by which PM stimulates reactive oxygen and nitrogen species (ROS/RNS) through Toll-like receptor (TLR) activation and their ability to stimulate the activation and release of IL-33. These studies have the potential for defining a role for oxidants in the pathogenesis of PM-mediated airway hyperreactivity.

Berman, R., Downey, G.P., Dakhama, A., *Day, B.J., and *Chu, H.W. Afghanistan particulate matter enhances pro-inflammatory responses in IL-13-exposed human airway epithelium via TLR2-signaling. Toxicol Sci 166:345-353, 2018. (*equal senior authors)
Berman, R., Kopf, K.W., Min, E., Huang, J., Downey, G.P., Alam, R., *Chu, H.W., and *Day, B.J. IL-33/ST2 signaling modulates Afganistan particulate matter-induced airway reactivity in mice. Toxicol Appl Pharmacol 404:115186, 2020. (*equal senior authors)
Berman R, Min E, Huang J, Kopf K, Downey GP, Riemondy K, Smith HA, Rose CS, Seibold MA, Chu HW*, Day BJ* (*equal senior authors). Single Cell RNA Sequencing Reveals a Unique Monocyte Population in Bronchoalveolar Lavage Cells of Mice Challenged with Afghanistan Particulate Matter and Allergen. Toxicol Sci. Online ahead of print. doi: 10.1093/toxsci/kfab065, 2021.

Grant Support:

DOD Grant Contract #W81XWH-16-2-0018
Impact of cigarette smoke on APM-induced airway epithelial injury and exacerbation of asthma and bronchiolitis in deployed military personnel.
PI Greg Downey/ Brian J Day Project Leader
Total Award Period: 9/30/16 – 9/29/21

PR201498 DOD expansion award
Innate Immunity and Deployment-related lung disease
PI Greg Downey/ Brian J Day Project leader
Total Award Period: 9/30/2021 – 08/31/2024

Development of antioxidants as medical countermeasures against chemical weapons

I have been developing manganese containing meso-porphyrins as catalytic antioxidants over the last twenty years. I am an inventor on over a dozen composition of matter and use patents for development of metallporphyrins as therapeutics. I was the first to show that these compounds could protect mammalian cells from oxidant-induced injury and was also the first to use them in animal models. I was the first to report their ability to act as catalase mimics, prevent lipid peroxidation, and scavenge peroxynitrite. I was the first to show that metalloporphyrins were protective against sulfur and nitrogen mustards in both lung and skin rodent animal models. I am currently developing them as medical countermeasures against radiation-induced lung injury and as medical countermeasures against chemical weapons.

O’Neill, H.C., White, C.W., Veress, L.A., Hendry-Hofer, T.B., Loader, J.E., Min, E., Huang, J., Rancourt, R.C., and Day, B.J. Treatment with catalytic metalloporphyrin AEOL 10150 reduces inflammation and oxidative stress due to inhalation of sulfur mustard analog 2-chloroethyl ethyl sulfide (CEES). Free Radic. Biol. Med., 48:1188-1196, 2010. PMCID:PMC2824750.
McGovern, T., Day, B.J., White, C.W., Powell, W.S., and Martin, J.G. AEOL10150: A novel therapeutic for rescue treatment after toxic gas lung injury. Free Radic. Biol. Med. 50:602-608, 2011. PMCID:PMC4026011
McElroy, C.S., Min, E., Huang, J., Loader, J.E., Hendry-Hofer, T.B., Garlick, R.B., Rioux, J.S., Veress, L.A., Smith, R., Osborne, C., Anderson, D.R., Holmes, W.W., Paradiso, D.C., White, C.W., and Day, B.J. Catalytic antioxidant rescue of inhaled sulfur mustard toxicity. Toxicol. Sci. 154:341-353, 2016. PMCID:PMC5139068.
McElroy, C. and Day B.J. Antioxidants as potential medical countermeasures for chemical warfare agents and toxic industrial chemicals. Biochem. Pharmacol. 100:1-11, 2016.
Liang, L.P., Pearson-Smith, J.N., Huang, J., McElroy, P., Day, B.J., and Patel, M. Neuroprotective effects of AEOL10150 in a rat organophosphate model. Toxicol. Sci. 162:611-621, 2018.
Liang, L.P., Pearson-Smith, J.N., Huang, J., Day, B.J., and Patel, M. Neuroprotective effects of a catalytic antioxidant in a rat nerve agent model. Redox Biol. 20, 275-284, 2019.

Grant Support:
UO1 ES029430
Optimization of AEOL10150 treatment of sulfur mustard-induced lung toxidrome in a pig model
P.I. Brian J Day
Total Award Period: 7/1/2018 – 6/30/2024

NIH U01 NS083422-01 (CounterACT)
Evaluation of neuroprotective effects of AEOL 10150
P.I. Manisha Patel/ Co-P.I. Brian J Day
Total Award Period: 8/1/2013 - 7/31/2018

NIH U54 ES015678 (CounterACT)
Novel Antioxidant Therapeutics for Sulfur Mustard Toxicity
Director Carl White/Project Leader Brian J. Day
Total Award Period: 9/1/11 – 8/31/16

Glutathione adaptive response to lung stressors, aging and disease.

I have been fascinated by the unique relationship the lung has with extracellular GSH levels which are 100-fold higher in the lung epithelial lining fluid (ELF) than the plasma. Amazingly, these levels are not static and can change during challenge or disease. For instance, lung ELF GSH levels are lower in COPD subjects experiencing acute exacerbations, in subjects with idiopathic pulmonary fibrosis, acute respiratory distress syndrome, lung transplantation, cystic fibrosis, HIV positive individuals and elderly people. My laboratory was the first to characterize that 50% of the ELF GSH is controlled by protein defective in cystic fibrosis, CFTR. My laboratory has further characterized the lung GSH adaptive responses to cigarette smoke exposure and showed these are impaired with infection and age that results in increased lung oxidative stress and inflammation. My lab has identified that alveolar macrophages set their intracellular GSH levels based on ELF GSH levels. As a consequence of this, macrophage modulate their response to danger signals that in turn modulates inflammatory cytokine responses. Our studies shed light on how the lung controls inflammation. My laboratory has demonstrated investigate this may become dysfunctional and contribute to a host of lung diseases including COPD and has developed novel therapies to correct this redox imbalance.

Gould, N.S., Min, E., Gauthier, S., Chu, H.W., Martin, R., and Day, B.J. Aging adversely affects the cigarette smoke induced glutathione adaptive response in the lung. Am. J. Respir. Crit. Care Med.182:1114-1122, 2010. PMCID:PMC3001254.
Gould, N.S. and Day B.J. Targeting maladaptive glutathione responses in lung disease. Biochem. Pharmacol. 81:187-193, 2011. PMCID:PMC3039114.
Gould, N.S., Min, E., and Day, B.J. Macropinocytosis of extracellular glutathione ameliorates tumor necrosis factor release in activated macrophages. PLoS ONE 6:e25704, 2011. PMCID:PMC3185039.
Gould, N.S., Min, E., Huang, J., Chu, H.W., Good, J., Martin, R.J., and Day, B.J. Glutathione depletion accelerates cigarette smoke induced inflammation and airspace enlargement. Toxicol. Sci. 147:466-474, 2015. PMCID:PMC4707200.

Grant Support:
NIH RO1 HL075523
Glutathione Transport, Oxidative Stress, and Lung Injury
P.I. Brian J. Day
Total Award Period: 12/15/03-11/30/08

NIH RO1 HL084469
Adaptive glutathione responses to cigarette smoke in COPD
P.I. Brian J. Day
Total Award Period: 12/1/07 – 11/31/11

Antioxidant dysregulation and oxidative stress in CF lung disease.

My research in CF began by examining antioxidant imbalance in the CFTR KO mice. Initial work was done looking at changes in lung airway surface fluid glutathione (GSH) levels and lung oxidative stress in CFTR KO mice. These studies revealed 50% decrease in GSH lung lining fluid in the CFTR KO mice corresponding increases in markers of lung oxidative damage. These studies were followed with a discovery that the lung mounts an antioxidant adaptive response during P. aeruginosa lung infection that includes the elevation of airway fluid GSH levels in wild type mice and that this adaptive response was blunted in P. aeruginosa infected CFTR KO mice. Additional novel findings were the reporting of mitochondrial oxidative stress in the lungs of CFTR KO mice where the CFTR KO mice had lower mitochondrial GSH levels and evidence of elevated mitochondrial reactive oxidation species production and markers of oxidative damage. My group was the first to show a role for CFTR in pharmacologically elevating lung GSH levels by oral glutathione administration. More recently my group has reported that CFTR deficient human lung epithelial cells have decreased apical levels of both GSH and SCN and this sensitizes them to hypochlorite-induced injury. We have also shown that the commonly used hypertonic saline treatment in CF increases the airway surface fluid levels of both GSH and SCN in wild type mice and that this is impaired in the CFTR KO mice.

Velsor L, van Heeckeran A, and Day BJ. Antioxidant imbalance in the lungs of the cystic fibrosis transmembrane conductance regulator protein mutant mouse. Am. J. Physiol. 281:L31-L38, 2001. PMID: 11404242.
Day BJ, van Heeckeren AM, Min E, and Velsor L. Role for CFTR in a glutathione response to bronchopulmonary pseudomonas infection. Infect. Immun. 72:2045-2051, 2004. PMCID: PMC375208.
Kariya, C., Leitner, H., Min, E., van Heeckeren, C., van Heeckeren, A., and Day, B.J. A role for CFTR in the elevation of glutathione levels in the lung by oral glutathione administration. Am. J. Physiol. Lung Cell. Mol. Physiol.292:L1590-L1597, 2007. PMCID: PMC3983954.
Gould NS, Gauthier S, Kariya CT, Min E, Huang J, and Day BJ. Hypertonic saline increases lung epithelial lining fluid glutathione and thiocyanate: Two protective CFTR-dependent thiols against oxidative injury. Respir. Res. 11:119, 2010. PMCID: PMC2936384.

Grant Support:
CFF Research Grant
Modulation of lung inflammation by CFTR-dependent thiols
P.I. Brian J Day
Total Award Period: 4/1/12 – 3/31/14

Identified a new function for mammalian thioredoxin reductase in the selective metabolism of HOSCN that promotes lung innate immunity and antioxidant defense.

We have recently identified a novel role of mammalian thioredoxin reductase (H-TrxR) in promoting innate immunity while protecting the host against excessive inflammation and oxidative stress. We discovered that H-TrxR could metabolize the anti-microbial agent HOSCN to SCN and water, but that the bacterial forms of L-TrxR could not and in fact were a target for HOSCN-mediated inactivation. The key feature of H-TrxR that provides this differential effect is the presence of a selenocysteine in the C-terminal tail that is lacking in the bacterial form L-TrxR. TrxR is found in all living creatures and is essential in providing reducing equivalents to ribonucleotide reductase for the synthesis of DNA. We demonstrated that a number of gram positive bacteria including several late disease antibiotic-resistant CF clinical isolates of P. aeruginosa were killed by HOSCN under the same conditions that did not change the viability of human bronchiolar epithelial cells. We also demonstrated that these human bronchiolar epithelial cells lost their resistance to HOSCN if H-TrxR was inhibited. To test the utility of our finding of the selective metabolism of HOSCN by mammalian H-TrxR we utilized several animal models of lung inflammation and infection. We demonstrated that 24 hour delayed treatment with SCN was effective in attenuating both spontaneous lung inflammation in the ENaC Tg mouse and in PA infected wild type and ENaC Tg mice. Furthermore, these studies revealed a marked attenuation of biomarkers of oxidative stress in SCN treated mice. These early studies laid the ground work with pharmacokinetic analysis of nebulized isotonic solution of 0.5% NaSCN along with determining effective concentrations of SCN in the blood, lung and epithelial lining fluid. These studies provide the preclinical basis for considering clinical trials for testing SCN for human efficacy in CF subjects.

Chandler, J.D. and Day, B.J. Thiocyanate: A potentially useful therapeutic agent with host defense and antioxidant properties. Biochem. Pharmacol. 84:1381-1387, 2012. PMCID: PMC3983948.
Chandler JD, Nichols DP, Nick JA, Hondal RJ, Day BJ. Selective metabolism of hypothiocyanous acid by mammalian thioredoxin reductase promotes lung innate immunity and antioxidant defense. J Biol Chem. 288:18421-8, 2013. PMCID: PMC3689984.
Chandler JD, Min E, Huang J, Nichols DP, Day BJ. Nebulized thiocyanate improves lung infection outcomes in mice. Br J Pharmacol. 169:1166-77, 2013. PMCID: PMC3696337.
Chandler JD, Min E, Huang J, McElroy CS, Dickerhof N, Mocatta T, Fletcher AA, Evans CM, Liang L, Patel M, Kettle AJ, Nichols DP*, Day BJ* (shared senior author). Anti-Inflammatory and Anti-Microbial Effects of Thiocyanate in a Cystic Fibrosis Mouse Model. Am J Respir Cell Mol Biol.53:193-205, 2015. PMCID:PMC4566044.

Grant Support:
CFF Research Grant
Anti-inflammatory effects of thiocyanate in treating emerging CF pathogens
P.I. Brian J Day
Total Award Period: 4/1/16 – 3/31/18

Selenocyanate as a novel treatment of cystic fibrosis lung disease

We discovered a novel function of thiocyanate serving as both an antioxidant and a prodrug for an antimicrobial agent hypothiocyanous acid that can be selectively metabolized by the host. We also uncovered CF pathogens that have developed resistant to the endogenous antimicrobial oxidant hypothiocyanous acid. Based on this discovery we are developing an analog of thiocyanate, selenocyanate and its antimicrobial oxidant, hyposelenocyanous acid as a way to overcome antimicrobial resistance to treat CF lung pathogens in a mouse model of lung inflammation and infection.

Grant Support:
CFF Research Grant
Selenocyanate manganese porphyrins as a novel antimicrobial therapy against CF pathogens
P.I. Brian J day
Total Award Period: 4/1/2018 – 3/31/2021

RO1 HL41146-A1
Selenocyanate as a novel treatment of cystic fibrosis lung disease
M.P.I. Brian J Day & Robert Hondal
Total Award Period: 1/1/2019 – 11/31/2023