Genetic variation in S-nitrosoglutathione reductase (GSNOR) and childhood asthma

doi:10.1016/j.jaci.2007.04.022

Journal of Allergy and Clinical Immunology

Volume 120, Issue 2, August 2007, Pages 322-328

https://doi.org/10.1016/j.jaci.2007.04.022 Get rights and content

Background

S-nitrosothiols are potent endogenous bronchodilators depleted in asthmatic airway lining fluid. S-nitrosoglutathione reductase (GSNOR; also known as alcohol dehydrogenase 5 or formaldehyde dehydrogenase) catalyzes the metabolism of S-nitrosoglutathione (GSNO) and controls intracellular levels of S-nitrosothiols. GSNOR knockout mice have increased lung S-nitrosothiol levels and are therefore protected from airway hyperresponsiveness after methacholine or allergen challenge.

Objective

We sought to investigate whether genetic variation in GSNOR is associated with childhood asthma and atopy.

Methods

We genotyped 5 tagging and 2 additional single nucleotide polymorphisms (SNPs) in GSNOR in 532 nuclear families consisting of asthmatic children aged 4 to 17 years and both parents in Mexico City. Atopy was determined by means of skin prick testing.

Results

Carrying 1 or 2 copies of the minor allele of SNP rs1154404 was associated with decreased risk of asthma (relative risk [RR], 0.77; 95% CI, 0.61-0.97; P = .028 for 1 copy and RR, 0.66; 95% CI, 0.44-0.99; P = .046 for 2 copies). Homozygosity for the minor allele of SNP rs28730619 was associated with increased risk of asthma (RR, 1.60; 95% CI, 1.13-2.26; P = .0077). Haplotype analyses supported the single SNP findings. GSNOR SNPs were not associated with the degree of atopy.

Conclusion

This is the first study of genetic polymorphisms in GSNOR and asthma. These data suggest that genetic variation in GSNOR might play a role in asthma susceptibility.

Clinical implications

The association of GSNOR polymorphisms with asthma suggests a potential therapeutic target.

Section snippets

Study design and subject enrollment

We used the case-parent triad design.12, 13 The study population included 532 case-parent triads with adequate DNA samples for genotyping of at least 1 single nucleotide polymorphism (SNP) for the GSNOR genes. The cases were children aged 4 to 17 years with asthma diagnosed by a pediatric allergist at the allergy referral clinic of a large public pediatric hospital in central Mexico City (Hospital Infantil de México, Federico Gómez). Children and parents provided blood samples as sources of

Results

Characteristics of the asthmatic children with genotyping data are presented in Table I. The mean age of patients was 9.0 years (range, 4-17 years). Most had mild (71.7%) as opposed to moderate or severe (28.3%) asthma. Nearly all patients (98.1%) had used medication for asthma in the past 12 months. Wheezing in the past 12 months was reported by 90.3% of the patients, and chronic dry cough was reported by 65.0%. For 73.5% of the patients, asthma symptoms had interfered with daily activities or

Discussion

Based on accumulating experimental evidence,2, 10, 11 we examined GSNOR as a potential asthma candidate gene. Carrying 1 or 2 copies of the minor A allele of the rs1154404 SNP decreased the risk of childhood asthma. This finding translates to an increased risk of asthma among homozygotes for the major T allele of this SNP. For the rs28730619 SNP, homozygotes for the minor G allele also had an increased risk of asthma development. Results of haplotype analysis agreed with the single SNP

References (40)

S.A. Kharitonov et al.
Increased nitric oxide in exhaled air of asthmatic patients
Lancet
(1994)
Q. Hamid et al.
Induction of nitric oxide synthase in asthma
Lancet
(1993)
J.S. Stamler et al.
Nitrosylation. the prototypic redox-based signaling mechanism
Cell
(2001)
B. Gaston et al.
Bronchodilator S-nitrosothiol deficiency in asthmatic respiratory failure
Lancet
(1998)
E.M. Henderson et al.
SNOR and wheeze: the asthma enzyme?
Trends Mol Med
(2005)
C.R. Weinberg et al.
A log-linear approach to case-parent-triad data: assessing effects of disease genes that act either directly or through maternal effects and that may be subject to parental imprinting
Am J Hum Genet
(1998)
M. Stephens et al.
A new statistical method for haplotype reconstruction from population data
Am J Hum Genet
(2001)
C.S. Carlson et al.
Selecting a maximally informative set of single-nucleotide polymorphisms for association analyses using linkage disequilibrium
Am J Hum Genet
(2004)
H. Li et al.
Genetic polymorphisms in arginase I and II and childhood asthma and atopy
J Allergy Clin Immunol
(2006)
J.R. O'Connell et al.
PedCheck: a program for identification of genotype incompatibilities in linkage analysis
Am J Hum Genet
(1998)

D.M. Umbach et al.

The use of case-parent triads to study joint effects of genotype and exposure

Am J Hum Genet

(2000)

H.J. Edenberg

Regulation of the mammalian alcohol dehydrogenase genes

Prog Nucleic Acid Res Mol Biol

(2000)

M.W. Hur et al.

Cloning and characterization of the ADH5 gene encoding human alcohol dehydrogenase 5, formaldehyde dehydrogenase

Gene

(1992)

T.H. Leung et al.

One nucleotide in a kappaB site can determine cofactor specificity for NF-kappaB dimers

Cell

(2004)

R.M. Kacmarek et al.

Inhaled nitric oxide. A bronchodilator in mild asthmatics with methacholine-induced bronchospasm

Am J Respir Crit Care Med

(1996)

L.G. Que et al.

Protection from experimental asthma by an endogenous bronchodilator

Science

(2005)

G. Folkerts et al.

Nitric oxide in asthma therapy

Curr Pharm Des

(2006)

B. Gaston et al.

Endogenous nitrogen oxides and bronchodilator S-nitrosothiols in human airways

Proc Natl Acad Sci U S A

(1993)

L. Liu et al.

A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans

Nature

(2001)

C. Gerard

Biomedicine. Asthmatics breathe easier when it's SNO-ing

Science

(2005)

Cited by (64)

GSNOR deficiency attenuates MPTP-induced neurotoxicity and autophagy by facilitating CDK5 S-nitrosation in a mouse model of Parkinson's disease
2022, Free Radical Biology and Medicine
The S-nitrosoglutathione reductase (GSNOR) is a key denitrosating enzyme that regulates protein S-nitrosation, a process which has been found to be involved in the pathogenesis of Parkinson's disease (PD). However, the physiological function of GSNOR in PD remains unknown. In a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model, we found that GSNOR expression was significantly increased and accompanied by autophagy mediated by MPTP-induced cyclin dependent kinase 5 (CDK5), behavioral dyskinesias and dopaminergic neuron loss. Whereas, knockout of GSNOR, or treatment with the GSNOR inhibitor N6022, alleviated MPTP-induced PD-like pathology and neurotoxicity. Mechanistically, deficiency of GSNOR inhibited MPTP-induced CDK5 kinase activity and CDK5-mediated autophagy by increasing S-nitrosation of CDK5 at Cys83. Our study indicated that GSNOR is a key regulator of CDK5 S-nitrosation and is actively involved in CDK5-mediated autophagy induced by MPTP.
S-nitrosylation is required for β<inf>2</inf>AR desensitization and experimental asthma
2022, Molecular Cell
The β₂-adrenergic receptor (β₂AR), a prototypic G-protein-coupled receptor (GPCR), is a powerful driver of bronchorelaxation, but the effectiveness of β-agonist drugs in asthma is limited by desensitization and tachyphylaxis. We find that during activation, the β₂AR is modified by S-nitrosylation, which is essential for both classic desensitization by PKA as well as desensitization of NO-based signaling that mediates bronchorelaxation. Strikingly, S-nitrosylation alone can drive β₂AR internalization in the absence of traditional agonist. Mutant β₂AR refractory to S-nitrosylation (Cys265Ser) exhibits reduced desensitization and internalization, thereby amplifying NO-based signaling, and mice with Cys265Ser mutation are resistant to bronchoconstriction, inflammation, and the development of asthma. S-nitrosylation is thus a central mechanism in β₂AR signaling that may be operative widely among GPCRs and targeted for therapeutic gain.
The Precision Interventions for Severe and/or Exacerbation-Prone (PrecISE) Asthma Network: An overview of Network organization, procedures, and interventions
2022, Journal of Allergy and Clinical Immunology
Citation Excerpt :
These are done by bronchoscopy117,120,121 or breath condensate.128 Because neither procedure is anticipated in the PrecISE Network for screening all subjects, we will instead perform genotyping for GSNOR/adh5 SNPs associated with asthma123,125,126 and with β2-agonist responsiveness.125,126 To validate this approach, we have queried asthma phenotypes in the SARP population.
Asthma is a heterogeneous disease, with multiple underlying inflammatory pathways and structural airway abnormalities that impact disease persistence and severity. Recent progress has been made in developing targeted asthma therapeutics, especially for subjects with eosinophilic asthma. However, there is an unmet need for new approaches to treat patients with severe and exacerbation-prone asthma, who contribute disproportionately to disease burden. Extensive deep phenotyping has revealed the heterogeneous nature of severe asthma and identified distinct disease subtypes. A current challenge in the field is to translate new and emerging knowledge about different pathobiologic mechanisms in asthma into patient-specific therapies, with the ultimate goal of modifying the natural history of disease. Here, we describe the Precision Interventions for Severe and/or Exacerbation-Prone Asthma (PrecISE) Network, a groundbreaking collaborative effort of asthma researchers and biostatisticians from around the United States. The PrecISE Network was designed to conduct phase II/proof-of-concept clinical trials of precision interventions in the population with severe asthma, and is supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health. Using an innovative adaptive platform trial design, the PrecISE Network will evaluate up to 6 interventions simultaneously in biomarker-defined subgroups of subjects. We review the development and organizational structure of the PrecISE Network, and choice of interventions being studied. We hope that the PrecISE Network will enhance our understanding of asthma subtypes and accelerate the development of therapeutics for severe asthma.
The Emerging Role of GSNOR in Oxidative Stress Regulation
2021, Trends in Plant Science
Citation Excerpt :
Microevolution of GSNOR, resulting in natural variants of regulatory SNPs in the promoter region and the introns of GSNOR within species, was observed in both the animal and plant kingdoms [23,109]. This microevolution of GSNOR has been observed to associate with human disease [109] and the plant Fe toxicity response [23]. Moreover, natural variants of GSNOR have been shown to cause the variation of plant tolerance to Fe-dependent oxidative damage through cis-regulatory element-based transcriptional regulation [23] (Figure 4).
Oxidative stress is a common event in aerobic organisms and a fundamental and unavoidable cost of the aerobic lifestyle. Reactive oxygen and nitrogen species (ROS/RNS) and iron (Fe) are the most common agents that trigger oxidative stress. A conserved enzyme in the S-nitrosoglutathione (GSNO) metabolism, GSNO reductase (GSNOR), modulates a multitude of abiotic and biotic stress responses. In this review, we focus on the emerging role of GSNOR as a master regulator in oxidative stress through its regulation of the interaction of ROS, RNS, and Fe, and highlight recent discoveries in post-translational modifications of GSNOR and functional variations of natural GSNOR variants during oxidative stress. Recent advances in understanding GSNOR regulation show promise for the modulation of oxidative stress in plants.
Chronicles of a reductase: Biochemistry, genetics and physio-pathological role of GSNOR
2017, Free Radical Biology and Medicine
Citation Excerpt :
GSNOR-mediated regulation of β-adrenergic response is also implicated in respiratory efficiency, where increase of lung SNOs, due to GSNOR ablation, stimulates bronchodilation and protects from airway hyper-responsiveness in experimental asthma [15] (Fig. 7). In line with these results, it was observed a genotype/phenotype correlation between GSNOR polymorphisms and childhood asthma susceptibility [101]. Alongside the regulation of adrenergic response, S-nitrosylation has been reported to have protective effects in a variety of cardiovascular diseases, mainly via the induction and/or stabilization of the hypoxia inducible factor 1 (HIF1), a major regulator of angiogenesis during hypoxia.
S-nitrosylation is a major redox posttranslational modification involved in cell signaling. The steady state concentration of S-nitrosylated proteins depends on the balance between the relative ability to generate nitric oxide (NO) via NO synthase and to reduce nitrosothiols by denitrosylases. Numerous works have been published in last decades regarding the role of NO and S-nitrosylation in the regulation of protein structure and function, and in driving cellular activities in vertebrates. Notwithstanding an increasing number of observations indicates that impairment of denitrosylation equally affects cellular homeostasis, there is still no report providing comprehensive knowledge on the impact that denitrosylation has on maintaining correct physiological processes and organ activities.
Among denitrosylases, S-nitrosoglutathione reductase (GSNOR) represents the prototype enzyme to disclose how denitrosylation plays a crucial role in tuning NO-bioactivity and how much it deeply impacts on cell homeostasis and human patho-physiology.
In this review we attempt to illustrate the history of GSNOR discovery and provide the evidence so far reported in support of GSNOR implications in development and human disease.
O-Aminobenzoyl-S-nitrosoglutathione: A fluorogenic, cell permeable, pseudo-substrate for S-nitrosoglutathione reductase
2017, Free Radical Biology and Medicine
S-nitrosoglutathione reductase (GSNOR) is a multifunctional enzyme. It can catalyze NADH-dependent reduction of S-nitrosoglutathione (GSNO); as well as NAD⁺-dependent oxidation of hydroxymethylglutathione (HMGSH; an adduct formed by the spontaneous reaction between formaldehyde and glutathione). While initially recognized as the enzyme that is involved in formaldehyde detoxification, increasing amount of evidence has shown that GSNOR also plays a significant role in nitric oxide mediated signaling through its modulation of protein S-nitrosothiol signaling. In humans, GSNOR/S-nitrosothiols have been implicated in the etiology of several diseases including lung cancer, cystic fibrosis, asthma, pulmonary hypertension, and neuronal dysfunction. Currently, it is not possible to monitor the activity of GSNOR in live cells. In this article, we present a new compound, O-aminobenzoyl-S-nitrosoglutathione (OAbz-GSNO), which acts as a fluorogenic pseudo-substrate for GSNOR with an estimated K_m value of 320 µM. The weak OAbz-GSNO fluorescence increases by approximately 14 fold upon reduction of its S-NO moiety. In live cell imaging studies, OAbz-GSNO is readily taken up by primary pulmonary endothelial cells and localizes to the same perinuclear region as GSNOR. The perinuclear OAbz-GSNO fluorescence increases in a time dependent manner and this increase in fluorescence is abolished by siRNA knockdown of GSNOR or by treatment with GSNOR-specific inhibitors N6022 and C3. Taken together, these data demonstrate that OAbz-GSNO can be used as a tool to monitor the activity of GSNOR in live cells.

View all citing articles on Scopus

Supported by the Division of Intramural Research, National Institute of Environmental Health Sciences (Z01 ES49019), National Institutes of Health, Department of Health and Human Services, and the National Council of Science and Technology (Grant 26206-M), Mexico. Dr Romieu is supported by the National Center for Environmental Health at the Centers for Disease Control.

Disclosure of potential conflict of interest: The authors have declared that they have no conflict of interest.

View full text

Mechanisms of asthma and allergic inflammationGenetic variation in S-nitrosoglutathione reductase (GSNOR) and childhood asthma

Background

Objective

Methods

Results

Conclusion

Clinical implications

Section snippets

Study design and subject enrollment

Results

Discussion

Lancet

Lancet

Cell

Lancet

Trends Mol Med

Am J Hum Genet

Am J Hum Genet

Am J Hum Genet

J Allergy Clin Immunol

Am J Hum Genet

Am J Hum Genet

Prog Nucleic Acid Res Mol Biol

Gene

Cell

Inhaled nitric oxide. A bronchodilator in mild asthmatics with methacholine-induced bronchospasm

Am J Respir Crit Care Med

Protection from experimental asthma by an endogenous bronchodilator

Science

Nitric oxide in asthma therapy

Curr Pharm Des

Endogenous nitrogen oxides and bronchodilator S-nitrosothiols in human airways

Proc Natl Acad Sci U S A

A metabolic enzyme for S-nitrosothiol conserved from bacteria to humans

Nature

Biomedicine. Asthmatics breathe easier when it's SNO-ing

Science

Mechanisms of asthma and allergic inflammation
Genetic variation in S-nitrosoglutathione reductase (GSNOR) and childhood asthma