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Genetic variants in the GATA3 gene are not associated with asthma and atopic diseases in German children

2009; Elsevier BV; Volume: 123; Issue: 5 Linguagem: Inglês

10.1016/j.jaci.2009.02.008

1097-6825

Kathrin Suttner, Martin Depner, Norman Klopp, Thomas Illig, Christian Vogelberg, Jerzy Adamski, Erika von Mutius, Michael Kabesch,

IL-33, ST2, and ILC Pathways

To the Editor: In asthma and atopy a disturbed regulation of T cells can lead to an imbalance between TH1 and TH2 cells and toward an increased TH2 response. The transcription factor GATA3 is necessary and sufficient for the differentiation of naive T cells into TH2 cells and also the maintenance of TH2 cytokine expression (IL-4, IL-5, and IL-13) in already differentiated TH2 cells.1Zheng W. Flavell R.A. The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells.Cell. 1997; 89: 587-596Abstract Full Text Full Text PDF PubMed Scopus (1867) Google Scholar GATA3 is not only involved in T-cell regulation but directly influences asthma symptoms and development. Selectively silencing GATA3, Sel et al2Sel S. Wegmann M. Dicke T. Sel S. Henke W. Yildirim A.O. et al.Effective prevention and therapy of experimental allergic asthma using a GATA-3-specific DNAzyme.J Allergy Clin Immunol. 2008; 121: 910-916Abstract Full Text Full Text PDF PubMed Scopus (86) Google Scholar recently demonstrated the essential involvement of GATA3 in the development of asthma in a murine model of acute experimental allergic asthma. Thus the aim of the present study was to investigate whether polymorphisms in the GATA3 gene exist that might affect TH cell differentiation and influence the pathogenesis of asthma and allergy. By using the HapMap database, GATA3 polymorphisms (minor allele frequency ≥3%) were identified and genotyped by means of matrix-assisted laser desorption/ionization time of flight mass spectrometry in German children from Munich (n = 1159) and Dresden (n = 1940) recruited in the cross-sectional International Study of Asthma and Allergy in Childhood phase II3Weiland S.K. von Mutius E. Hirsch T. Duhme H. Fritzsch C. Werner B. et al.Prevalence of respiratory and atopic disorders among children in the East and West of Germany five years after unification.Eur Respir J. 1999; 14: 862-870Crossref PubMed Scopus (240) Google Scholar and children from Leipzig (n = 1165) phenotyped with a similar protocol.4von Mutius E. Weiland S.K. Fritzsch C. Duhme H. Keil U. Increasing prevalence of hay fever and atopy among children in Leipzig, East Germany.Lancet. 1998; 351: 862-866Abstract Full Text Full Text PDF PubMed Scopus (522) Google Scholar The prevalence of asthma and allergies was assessed by means of self-administered questionnaires and objective measurements, such as lung function tests, skin prick tests, and serum IgE measurements. Further information on primer sequences, technical details, or population characteristics are available from the corresponding author on request. Based on the linkage disequilibrium structure of the GATA3 gene obtained from the HapMap database, 3 single nucleotide polymorphisms (SNPs; rs3781093, rs3824662, and rs11567931) and 6 tagging SNPs (rs1399180, rs3802604, rs10752126, rs3802600, rs406103, and rs1058240) capturing the genetic information of the entire gene locus were genotyped. As indicated by Fig 1, all identified polymorphisms were located in intronic regions of the gene, with the exception of SNP rs1058240, which was located in the 3′ untranslated region of the gene. The associations between the genotyped SNPs and asthma, hay fever, eczema, and atopy were investigated by using pooled data from Dresden, Munich, and Leipzig (total n = 4264, Table I). None of the SNPs showed any significant associations with these phenotypes in an allelic model.5Sasieni P.D. From genotypes to genes: doubling the sample size.Biometrics. 1997; 53: 1253-1261Crossref PubMed Scopus (746) Google Scholar An equivalence test was performed to test for statistical significance of the absence of associations between GATA3 SNPs and these atopic phenotypes (Table I).6Wellek S. Schumann G. Statistical confirmation of negative results of association studies in genetic epidemiology.Am J Med Genet B Neuropsychiatr Genet. 2004; 128: 126-130Crossref Scopus (8) Google Scholar By using these tests, it can be measured how likely it is that an effect falls within a certain predefined odds ratio (OR) range near a null effect or is absent. For our study, this range was defined with an OR from 0.8 to 1.25. For most of the tested GATA3 polymorphisms, the equivalence test rejected the hypothesis of association, even excluding relatively small contributions of GATA3 to disease development defined in the set range. Because the allelic model captures more or less the information of a dominant model, we also investigated GATA3 SNP effects using a recessive model. By using this approach, SNP rs3824662 showed borderline nominal significant associations with eczema (OR, 1.61; 95% CI, 1.07-2.42; P = .0214) and hay fever (OR, 1.69; 95% CI, 1.00-2.84; P = .0474), not withstanding correction for multiple testing. Additionally, haplotype analyses of the 9 SNPs were conducted, but no associations with asthma or atopy could be detected (data not shown).Table ITests for associations and equivalence tests for all GATA3 SNPs with asthma, hay fever, eczema, and atopy in the pooled cross-sectional study population from Dresden, Leipzig, and Munich (total n = 4264)CLs for the OREquivalence limits (using an OR of 0.8-1.25)Equivalence decisionORLower CLUpper CLP value, χ2 testC1(s)C2(s)XAsthmars13991801.180.971.44.0954114123134Nors37810930.970.781.22.80619710498Yesrs38026041.030.871.21.7457233256248Yesrs38246621.030.831.26.8171114123120Yesrs107521261.000.851.18.9881262288275Yesrs38026001.020.841.25.8222120131127Yesrs4061030.970.801.18.7505132144134Yesrs115679311.060.691.63.7812222324Nors10582401.120.921.36.2589125136141NoHay feverrs13991801.020.831.26.8614110119116Yesrs37810931.040.831.30.747594101100Yesrs38026041.020.871.21.7790225247239Yesrs38246621.120.921.38.2637110118124Nors107521261.030.871.21.7240250274266Yesrs38026001.080.881.32.4800116126127Nors4061030.970.801.19.7983126138129Yesrs115679311.290.871.93.2073222328Nors10582401.060.871.29.5772121131131YesEczema∗Eczema is a pooled variable: physician's diagnosis of atopic dermatitis in Munich and Dresden and ever eczema plus ever itchy dermal changes in Leipzig.rs13991800.890.751.04.1354216245211Nors37810931.040.881.22.6857188212204Yesrs38026040.900.801.02.1152450509454Yesrs38246621.080.921.26.3427218247244Yesrs107521260.920.821.05.2078501564512Yesrs38026001.000.861.17.9872230261245Yesrs4061030.930.801.08.3328252287256Yesrs115679311.340.991.82.0545444656Nors10582401.010.871.18.8691239272257YesAtopyrs13991801.000.881.14.9994333381356Yesrs37810931.000.861.15.9455289330308Yesrs38026041.020.921.13.7161691784744Yesrs38246621.040.911.19.5575336385368Yesrs107521261.050.951.16.3742778877844Yesrs38026001.030.911.17.6568357410389Yesrs4061031.010.891.14.8940389447419Yesrs115679311.100.841.45.5007677073Nors10582401.120.991.27.0705366420420YesCL, Confidence limit.∗ Eczema is a pooled variable: physician's diagnosis of atopic dermatitis in Munich and Dresden and ever eczema plus ever itchy dermal changes in Leipzig. Open table in a new tab CL, Confidence limit. Polymorphisms in the GATA3 gene locus were also specifically investigated in an additional sample of 728 asthmatic children from the Multicenter Asthma Genetic in Childhood study and 694 reference children (randomly selected form the Munich and Dresden International Study of Asthma and Allergy in Childhood populations), as previously published in a genome-wide association study (GWA), to further strengthen the analysis.7Moffatt M.F. Kabesch M. Liang L. Dixon A.L. Strachan D. Heath S. et al.Genetic variants regulating ORMDL3 expression contribute to the risk of childhood asthma.Nature. 2007; 448: 470-473Crossref PubMed Scopus (1219) Google Scholar Eight of the 9 GATA3 SNPs investigated in the present study had also been genotyped in the GWA study (rs1399180, rs3781093, rs3802604, rs3824662, rs570613 [in linkage disequilibrium with tagging SNP rs10752126], rs3802600, rs569421 [in linkage disequilibrium with tagging SNP rs406103], and rs1058240). By using information of the allelic model, none of these GATA3 SNPs showed a significant association with asthma in the GWA study, confirming our negative results. Thus we could exclude major contributions of GATA3 SNPs to asthma development in our study population. When we tried to replicate previous associations from Finnish populations8Pykalainen M. Kinos R. Valkonen S. Rydman P. Kilpelainen M. Laitinen L.A. et al.Association analysis of common variants of STAT6, GATA3, and STAT4 to asthma and high serum IgE phenotypes.J Allergy Clin Immunol. 2005; 115: 80-87Abstract Full Text Full Text PDF PubMed Scopus (82) Google Scholar between GATA3 polymorphisms and highly increased total serum IgE levels in our populations, rs1399180 (OR, 1.27; 95% CI, 1.06-1.52; P = .0085) and rs11567931 (OR, 1.55; 95% CI, 1.10-2.19; P = .0114) were found to be associated with total serum IgE levels beyond the 90% percentile (>457 IU/mL), not withstanding correction for multiple testing with the Bonferroni correction. Interestingly, effects on asthma were previously observed in genetic studies of the major counterparts of GATA3 in T-cell regulation: T-box expressed in T cells (TBX21) and H. 20-like homeobox 1 (HLX1). Polymorphisms in these genes significantly modify asthma risk in our studies9Suttner K, Rosenstiel P, Depner M, Pinto LA, Ruether A, Klopp N, et al. T-bet gene variants increase childhood asthma risk in combination with HLX1 variants. J Allergy Clin Immunol. In press 2009.Google Scholar, 10Suttner K. Ruoss I. Rosenstiel P. Depner M. Pinto L.A. Schedel M. et al.HLX1 gene variants influence the development of childhood asthma.J Allergy Clin Immunol. 2009; 123: 82-88Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar and other's studies.11Akahoshi M. Obara K. Hirota T. Matsuda A. Hasegawa K. Takahashi N. et al.Functional promoter polymorphism in the TBX21 gene associated with aspirin-induced asthma.Hum Genet. 2005; 117: 16-26Crossref PubMed Scopus (71) Google Scholar, 12Munthe-Kaas M.C. Carlsen K.H. Haland G. Devulapalli C.S. Gervin K. Egeland T. et al.T cell-specific T-box transcription factor haplotype is associated with allergic asthma in children.J Allergy Clin Immunol. 2008; 121: 51-56Abstract Full Text Full Text PDF PubMed Scopus (34) Google Scholar, 13Raby B.A. Hwang E.S. Van Steen K. Tantisira K. Peng S. Litonjua A. et al.T-bet polymorphisms are associated with asthma and airway hyperresponsiveness.Am J Respir Crit Care Med. 2006; 173: 64-70Crossref PubMed Scopus (69) Google Scholar These results suggest that relatively small genetic variation in TH1 signals affect asthma development, whereas genetic alterations in TH2-related signals show no such consequences. Support for this hypothesis derives from previous animal models, in which it was shown that TBX21 transgenic mice had an asthmatic phenotype very similar to that of mice with a complete loss of TBX21 already when TBX21 expression is reduced in mice heterozygous for TBX21.14Finotto S. Neurath M.F. Glickman J.N. Qin S. Lehr H.A. Green F.H. et al.Development of spontaneous airway changes consistent with human asthma in mice lacking T-bet.Science. 2002; 295: 336-338Crossref PubMed Scopus (532) Google Scholar Furthermore, it was recently proposed by Usui et al15Usui T. Preiss J.C. Kanno Y. Yao Z.J. Bream J.H. O'Shea J.J. et al.T-bet regulates Th1 responses through essential effects on GATA-3 function rather than on IFNG gene acetylation and transcription.J Exp Med. 2006; 203: 755-766Crossref PubMed Scopus (246) Google Scholar that GATA3 directs naive T cells toward TH2 differentiation only until its effects are overruled by TBX21. In that regard it is intriguing that a recent twin study suggested the TH1 response to be predominantly genetically determined, whereas this could not be shown for the initiation of the TH2 response.16Hohler T. Reuss E. Adams P. Bartsch B. Weigmann B. Worns M. et al.A genetic basis for IFN-gamma production and T-bet expression in humans.J Immunol. 2005; 175: 5457-5462PubMed Google Scholar Taken together, it might be speculated that genetic variations in the major TH2-associated transcription factor GATA3 have no major influence on the origin of asthma because an increased TH2 response is initiated by a weak or absent TH1 signal rather than by an augmented TH2 signal. We thank Sonja Zeilinger for her expert technical assistance in genotyping. This work is part of the PhD thesis of Kathrin Suttner.