5′ Flanking region polymorphism of the gene encoding leukotriene C4 synthase does not correlate with the aspirin-intolerant asthma phenotype in the United States

doi:10.1067/mai.2000.107603

Journal of Allergy and Clinical Immunology

Volume 106, Issue 1, Part 1, July 2000, Pages 72-76

https://doi.org/10.1067/mai.2000.107603 Get rights and content

Abstract

Background: Approximately 10% of patients with asthma have a distinct clinical entity in which their symptoms are exacerbated by aspirin and most other nonsteroidal anti-inflammatory agents. These individuals typically have significant basal overproduction of cysteinyl leukotrienes, and within their biosynthetic pathway, the terminal enzyme, leukotriene C₄ synthase (LTC₄S), is significantly overexpressed. A single nucleotide polymorphism consisting of an adenine (A) to cytosine (C) transversion –444 nucleotides upstream of the ATG translation start site in the LTC ₄S gene has been associated with a relative risk of 3.89 for the aspirin-intolerant phenotype in Polish patients. Objective: These studies were undertaken to further investigate the functional effect of this allele in LTC ₄S gene expression and subsequently to determine whether an association between the presence of this polymorphism and aspirin-intolerant asthma existed within patients of the United States. Methods: Functionality of the C-444 allele was assessed by using promoter-reporter constructs and transient transfection assays in the THP-1 monocytic cell line. Genotyping was performed on 137 unaffected control subjects, 33 patients with aspirin-tolerant asthma, and 61 patients with aspirin-intolerant asthma from the United States. Results: Promoter-reporter constructs containing the C-444 allele revealed no significant upregulatory or downregulatory effects in the transcription of the LTC ₄S gene. The LTC ₄S genotype distribution was consistent with the Hardy-Weinberg equilibrium in patients with aspirin-tolerant asthma and unaffected control subjects but not in patients with aspirin-intolerant asthma; however, the distributions were not significantly different among the phenotype groups. Conclusions: Our data demonstrate that the C-444 allele in the LTC4S gene is not statistically different among patients with the aspirin-intolerant asthmatic phenotype, patients with the aspirin-tolerant asthmatic phenotype, and unaffected control subjects in the United States. This finding, along with the lack of functionality of this polymorphism, suggest that it is not related to a specific asthma phenotype and may represent a population-stratified polymorphism within patients of eastern European descent. (J Allergy Clin Immunol 2000;106:72-6.)

Section snippets

Assessment of the functional effect of the C allele in the –444 position of the 5′ flanking region of the LTC₄S gene

To assess the relative functional contribution of the C allele, a 550-bp DNA fragment of the proximal 5′ flanking region of the LTC ₄S gene was produced by means of restriction enzyme digest of a P1 plasmid containing the human LTC ₄S gene. This DNA fragment encompasses the 5′ flanking region of the gene extending 3′ to the ATG translation start site and was used as a template to further create a site-directed mutagenic fragment that incorporated the polymorphic cytosine transversion at

Functional effect of the C allele in the –444 position of the 5′ flanking region of the LTC₄S gene

The C allele at the –444 position within the context of the 550-bp construct of the 5′ flanking region of the LTC ₄S gene in THP-1 cells provided luciferase activity that was equal to the activity of the wild-type construct (n = 7, Fig 2).

. Effect of the C-444 single-nucleotide polymorphism in LTC ₄S gene expression. Luciferase activity of THP-1 cells after transient transfection with a promoterless and enhancerless luciferase control is shown (as fold increase) in comparison with constructs

DISCUSSION

Typically, individuals with AIA have significantly increased levels of cysteinyl LTs in their urine, and immunohistochemical analysis has revealed that the level of LTC₄S overexpression in the aspirin-intolerant asthmatic phenotype correlates with overproduction of cysteinyl LTs and bronchial hyperreactivity to lysine aspirin. Furthermore, of the proteins in LT and prostanoid biosynthetic pathways, only LTC₄S is significantly overexpressed, which suggests that a defect in the regulation of this

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Citation Excerpt :
A number of SNPs were reported to be associated with AERD from this pathway from several genes, including 5-lipoxygenase (ALOX5), COX2 (COX2), cysteinyl leukotriene receptors (CYSLTR1/CYSLTR2), leukotriene C4 synthase (LTC4S), prostaglandin E2 receptor (PTGER/EP2), thromboxane A2 receptor (TBXA2R), and thromboxane A synthase (TBXAS1; Table I).86 However, several other studies have not confirmed some of these associations in different populations, particularly for LTC4S.45-49 In subsequent years, SNPs involving numerous other genes involved in innate immunity (eg, TAP2 and TLR3), dysfunction of epithelial cells (eg, SPINK5), biochemical signaling pathways in inflammatory cells (eg, ACE and PPARG), TH2 pathways (eg, IL13 and IL41), and aspirin metabolism (eg, NAT2 and CYP2C19) have also been associated with AERD and have been reviewed elsewhere.86
Adverse drug reactions (ADRs) are a relatively common cause of morbidity and mortality. Many factors can contribute to ADRs, including genetics. The degree to which genetics contributes to ADRs is not entirely clear and varies by drug, as well as the type of ADR. Pharmacogenetics and, more recently, pharmacogenomics have been applied to the field of ADRs for both predictable ADRs and hypersensitivity drug reactions. Evaluations for glucose-6-phosphate dehydrogenase and thiopurine S-methyltransferase are commonplace clinical tests to reduce hematologic problems associated with drugs, such as dapsone and azathioprine, respectively. Numerous pharmacogenetic associations have been discovered for immediate hypersensitivity reactions to β-lactams, aspirin, and nonsteroidal anti-inflammatory drugs; however, the clinical utility of testing for these genetic associations has not been established. In contrast, pharmacogenetic testing for HLA-B*1502 before carbamazepine in patients of certain Asian ethnicities and testing for HLA-B*5701 before abacavir treatment are recommended. This review will focus on pharmacogenetics and pharmacogenomics and their role in reducing ADRs, especially those caused by drug hypersensitivity reactions.
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Aspirin-exacerbated respiratory disease (AERD) is a complex clinical syndrome characterised by severe asthmatic attack upon treatment with aspirin and/or non-steroidal anti-inflammatory drugs (NSAIDs). Genetic predisposition has been considered as a crucial determinant and candidate genes have concentrated especially on cysteinyl leukotrienes (LTs)-related genes as the inhibitory action of aspirin and NSAIDs on cyclooxygenase activity may cause overproduction of cysteinyl LTs. However, conflicting results have been reported, in parallel with replication studies in different ethnic groups. Thus, future areas of investigations need to focus on comprehensive approaches towards the discovery of other genetic biomarkers. Unfortunately, few papers have been reported about gene polymorphisms in Japanese patients with AERD. Here, we described on our recent genetic investigations on B2ADR, IL-13, IL-17A, CYP2C19, TBXA2R, CRTH2 and HSP70. This review indicates potential genetic biomarkers contributing to the early diagnosis of AERD, which may include CYP2C19 and HSP70 gene polymorphisms, and future validation studies in independent population are required to provide reassurance about our findings.
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Citation Excerpt :
There was no association between A-444C and asthma, atopy, or total IgE concentrations. An increased prevalence of a genetic polymorphism in the LTC4S promoter region has been identified in Polish patients with aspirin-induced asthma [60c], although no polymorphisms in the flanking region of the LTC4S gene were discovered [61c,62c]. Two single nucleotide polymorphisms in the LTC4S promoter region, –1702G>A and –444A>C, were not associated with aspirin hypersensitivity in a study in 110 Korean patients with aspirin-induced asthma, 125 aspirin-tolerant patients with asthma, and 125 controls [63c].
The Side Effects of Drugs Annuals form a series of volumes in which the adverse effects of drugs and adverse reactions to them are surveyed. The series supplements the contents of Meyler’s Side Effects of Drugs: the International Encyclopedia of Adverse Drug Reactions and Interactions. This review of the 2011 publications on adverse reactions to anti-inflammatory and antipyretic analgesics and drugs used in gout covers metamizole, paracetamol (acetaminophen), non-steroidal anti-inflammatory drugs (the anthranilic acid derivative mefenamic acid, the arylalkanoic acid derivatives bufexamac, diclofenac, ibuprofen, and ketorolac, the COX-2 selective inhibitors celecoxib, etoricoxib, rofecoxib, the oxicam meloxicam, and the pyrazolone derivative phenylbutazone), acetylsalicylic acid (aspirin), benzydamine (benzindamine), nefopam, and drugs used in the treatment of gout (allopurinol, colchicine, febuxostat, and rasburicase). Special features include discussion of the supposed link between paracetamol and allergic disorders and a special review on genetic polymorphisms associated with adverse reactions to salicylates, including studies on ADAM33, adenosine receptors, chemokine CC motif receptors, complement, CYP isoenzymes, EMID2, FANCC, G protein signalling, growth factors, HLA, indoleamine-pyrrole 2,3 dioxygenase, interleukins, kinesin, leukotrienes, MS4A2, N-acetyl transferase-2, N-methyl D-aspartate receptors, prostaglandin receptors, purinergic receptors, the renin–angiotensin system, thromboxanes, transporters, and tumor necrosis factor-α.

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^☆: Supported by National Institutes of Health grant K08-HL03208-05 and, for the GCRC at Scripps Clinic, National Institutes of Health grant #M01RR00633.

^☆☆: Reprint requests: John F. Penrose, MD, Brigham and Women’s Hospital, Smith Building, Room 626C, 1 Jimmy Fund Way, Boston, MA 02115.

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Asthma, rhinitis, other respiratory diseases5′ Flanking region polymorphism of the gene encoding leukotriene C4 synthase does not correlate with the aspirin-intolerant asthma phenotype in the United States☆,☆☆

Abstract

Section snippets

Assessment of the functional effect of the C allele in the –444 position of the 5′ flanking region of the LTC4S gene

Functional effect of the C allele in the –444 position of the 5′ flanking region of the LTC4S gene

DISCUSSION

Cell

Arch Biochem Biophys

J Biol Chem

Biochem Biophys Res Commun

J Allergy Clin Immunol

Mechanism of aspirin-induced asthma

Allergy

Urinary excretion of leukotriene E4 and 11-dehydro-thromboxane B2 in response to bronchial provocations with allergen, aspirin, leukotriene D4 and histamine in asthmatics

Am Rev Respir Dis

Urinary leukotriene E4 concentration increase after aspirin challenge in aspirin-sensitive asthmatic subjects

Am Rev Respir Dis

Increased excretion of leukotriene E4 during aspirin-induced asthma

J Lab Clin Med

The leukotriene-receptor antagonist MK-0679 blocks airway obstruction induced by inhaled lysine-aspirin in aspirin-sensitive asthmatics

Eur Respir J

The pivotal role of 5-lipoxygenase products in the reaction of aspirin-sensitive asthmatics to aspirin

Am Rev Respir Dis

Effect of the leukotriene receptor antagonist MK-0679 on baseline pulmonary function in aspirin sensitive asthmatic subjects

Thorax

Asthma, rhinitis, other respiratory diseases
5′ Flanking region polymorphism of the gene encoding leukotriene C₄ synthase does not correlate with the aspirin-intolerant asthma phenotype in the United States☆,☆☆

Assessment of the functional effect of the C allele in the –444 position of the 5′ flanking region of the LTC₄S gene

Functional effect of the C allele in the –444 position of the 5′ flanking region of the LTC₄S gene

Urinary excretion of leukotriene E₄ and 11-dehydro-thromboxane B₂ in response to bronchial provocations with allergen, aspirin, leukotriene D₄ and histamine in asthmatics

Urinary leukotriene E₄ concentration increase after aspirin challenge in aspirin-sensitive asthmatic subjects

Increased excretion of leukotriene E₄ during aspirin-induced asthma