Volume 15 Supplement 2
Evaluation of spirometry values in relation to beta-2-adrenergic receptor gene polymorphism
© I. Holzapfel Publishers 2010
Published: 4 November 2010
The vagus nerve plays a special role in the control of respiratory system activity which represents the parasympathetic part of the autonomic nervous system. A small bronchial innervation by the sympathetic system also is observed, and there is a significant expression of adrenergic receptors, in particular β2 receptors, in the airways. The development of genetics and molecular biology allows for a detailed study which can clarify the essential elements in the pathogenesis of many types of lung disease, as well as the physiological phenomena - bronchial smooth muscle tone and their contractile mechanism.
Materials and methods
The study involved 148 healthy male volunteers aged 20-26. In all subjects, gene polymorphism at nucleotide position 46 and 79 of β2-adrenergic receptor (β2-ADR) was assessed. According to the gene polymorphism data, we divided the whole examined population of males into 6 groups for further studies. Moreover, in all the subjects, we performed spirometry testing to verify their pulmonary functions.
The basic values of spirometry tests in all subjects were in the range of normal values. The frequency of different genotypes in the gene polymorphism of the β2-adrenergic receptor at nucleotide positions 46 and 79 were typical for the Caucasian population. Analysis of the output values of spirometry, conducted in the particular groups based on their genotype, showed significant inter-group differences in the selected spirometry tests.
Our results may be useful in explaining the differences in the measured values of spirometric indices in healthy subjects in relation to the polymorphism of β2-ADR, and may also contribute to the verification of standards for spirometric indices for this selected group of young males in the Polish population.
The most common polymorphism of the β2-ADR gene concerns codons 16 and 27. The polymorphism at codon 16 is associated with the potential replacement of adenine with guanine (g. 46 A→ G) in nucleotide 46 of the gene (g. 46), which results in the replacement of arginine with glycine at position 16 of the receptor polypeptide chain. A more common polymorphism at codon 27 of β2-ADR, caused by the replacement of cytosine with guanine at position g. 79 (g.79 C→ G), results in the replacement of glutamine with glutamate acid at position 27 of the receptor chain. Polymorphism at codon 16 β2-ADR is significant for signal transduction by this receptor. The objective of the present study was to evaluate the frequency of β2-ADR gene polymorphism at codon g. 46 and g. 79 and its effect on the values of spirometric indices in healthy men.
Materials and methods
Polymorphism of the β2-ADR encoding gene was evaluated at nucleotide positions 46 and 79. Based on the observed changes subjects were divided into 6 groups: g. 46AA - homozygote, presence of adenine and adenine (at receptor 16Arg/16Arg); g. 46GG - homozygote, presence of guanine and guanine (at receptor 16Gly/16Gly); g. 46AG - heterozygote, presence of adenine and guanine (at receptor 16Arg/16Gly); g. 79CC - homozygote, presence of cytosine and cytosine (at receptor 27Gln/27Gln); g. 79GG - homozygote, presence of guanine and guanine (at receptor 27Glu/27Glu); g. 79GC - heterozygote, presence of guanine and cytosine (at receptor 27Glu/27Gln). Spirometric testing was conducted with Lungtest 1000 apparatus from MES (Cracow, Poland). The obtained values of spirometry indices are presented with relation to the age, sex and body height of examined subjects. The spirometer was calibrated, and current data were entered (temperature, humidity and ambient pressure conditions) before measurements. Testing was carried out according to the recommendations of the American Thoracic Society (ATS) and the European Respiratory Society (ERS) with volumes reported at BTPS parameters (body temperature, pressure saturated) [3, 9, 35]. Vital capacity (VC) and flow/volume loop were measured. The highest measured values of spirometric indices were selected for further analysis.
The Chi2 test was used to compare the frequency of genotypes and allele of the gene encoding the β2-adrenergic receptor. The significance of the differences between the values of measured indices was evaluated in consideration of their correlation and characteristics of sets. Normality of distribution was analyzed with the Shapiro-Wilk test. A non-parametric Mann-Whitney U test was used for distributions other than normal, and a parametric t-test for normally distributed data. Significance of correlations between variables was evaluated with Spearman's rank coefficient of correlation.
The distribution of genotypes in the studied group of 148 subjects conformed to the Hardy-Weinberg equilibrium. It was also found that both polymorphisms (SNP) g. 46 and g.v79 are closely correlated: D' = 0.900655; P < 0.0001. Three genotypes were found within the nucleotide 46 and three within the nucleotide 79. The distribution of their frequency was as follows: g. 46AA n = 20 (13.5%); g. 46GG n = 65 (43.9%); g. 46AG n = 63 (42.6%); g. 79CC n = 38 (25.7%); g. 79GC n = 31 (21%); and g. 79GG n = 79 (53.3%).
FVCex and FEV1 values expressed in % of predicted in the individual groups g.46 and g.79.
g. 46/g. 79
Significance of differences (P) for the values of spirometric indices in individual groups g.46.
Significance of differences between groups within the polymorphism of nucleotide 46
AA and GG
AA and AG
GG and AG
Significance of differences (P) for the values of spirometric indices in individual groups g.79.
Significance of differences between groups within the polymorphism of nucleotide 79
CC and GG
CC and GC
GC and GG
The presence of β2-ADR genotypes, with attempts to link them with clinical symptoms, has been analyzed in patients with chronic obstructive pulmonary disease, asthma, atopy, hypertension, obesity, dyslipidaemia and diabetes type 2 [5, 12, 19, 21, 31, 36]. Small et al  reported that subjects with g. 79GG, particularly women, usually have body mass higher by 20 kg and have by 50% of fatty cells more than subjects with the genotype g. 79CC. Moreover, special importance is attributed to the polymorphism of the β2-ADR encoding gene at the position of nucleotides 46 and 79 (g. 46 and g. 79) [6, 7, 28, 31, 38]. It has been claimed that these polymorphic positions (g. 46 A→G and g. 79 G→C) may predispose people to the development of pulmonary and circulatory diseases, as well as to diverse responses to treatment [10, 31, 36]. It should be emphasized, however, that the frequency of the genotypes in studied groups of healthy subjects and patients is comparable [13, 34].
In the present investigation, distribution of the studied genotypes conformed to the Hardy-Weinberg equilibrium. Their close correlation was found (D' = 0.9; P < 0.0001), which conforms to the results obtained by other authors [13, 14, 16, 31, 39, 40]. Subjects were divided into 6 groups: three based on the polymorphism of the β2-ADR encoding gene at nucleotide position 46, and three based on the polymorphism of the β2-ADR encoding gene at nucleotide position 79.
The frequency of genotype g. 46AA was comparable to the results obtained by Taylor et al , Hall et al , Israel et al  Martinez et al , Lima et al , Liggett , and Aynacioglu et al . In studies conducted by Hopes et al  and Azis et al  this genotype had a lower frequency (6.3-9.3%), while in studies by Xie et al  and Kotani et al , it was significantly higher - 26.6 and 27.7%, respectively. Differences in the evaluation of the frequency of genotype g. 46AA are significant. However, it is claimed that this genotype has the lowest frequency within g. 46 . Frequencies of genotype g. 46GG reported by Taylor et al , Lima et al , Hopes et al , Martinez et al , and Azis et al  are comparable to those observed in our study and ranged from 38.3 to 47.2%. A slightly different range (from 29.1 to 36%) concerned data presented by Hall et al , Xie et al , Kotani et al , Israel et al , and Aynacioglu et al . Only Liggett  found genotype g. 46GG in 57.3% of studied subjects. In this study, the percentage of subjects heterozygous at codon 16 - g. 46AG, as in the study by Xie et al , was comparable with that for homozygotes g. 46GG. Usually, the percentage of individuals with g. 46AG is estimated at 38-49%, which corresponds with the results obtained in our present study [12, 14, 19, 26, 29, 34]. Aynacioglu et al , Israel et al , and Azis et al  recorded a significantly higher frequency of the studied genotype (50%), while Liggett , Taylor et al  recorded a significantly lower one (29-37%). Individuals with g. 79CC, as in our study, in studies by Hall et al , Lima et al , Hopes et al , and Liggett  accounted for 25.7%. Individuals with g. 79GG accounted for 21%. According to Hall et al , Lima et al , Hopes et al , Xie et al , and Israel et al , their frequency was between 15.4 and 24%. In populations studied by Aynacioglu et al  and Martinez et al , genotype g. 79GG was significantly less frequent and accounted for, respectively, 10.6% and 13.8%, while in studies by Azis et al  and Liggett  significantly more frequently (37.5 and 28%). Subjects heterozygous at codon 27 accounted for over a half of the studied population (53.3%), which conforms with results obtained by Hall et al , Lima et al , Hopes et al , Israel et al , and Reihsaus et al  - range from 49.2 to 59%. In contrast to that, in studies presented by Liggett , Martinez et al , Xie et al , Aynacioglu et al , Azis et al , and Kotani et al , the percentage of individuals with g. 79GC was lower (13.6 - 45%). Data obtained in our present study show, similarly to the cited references, that g. 79GG is the genotype g. 79 of the lowest frequency. Differences in the frequency of the analyzed β2-ADR genotypes may result from the fact that the studies covered representatives of different ethnic groups, including Caucasian, Hispanic and Latino American, African-American, and Asian . In this study, similar to the available literature data, no individuals were observed with coexisting g. 46AA and g. 79GG .
In the present study, spirometric measurements demonstrated the functional efficiency of the respiratory system in all subjects. Values of the analyzed indices did not differ from standard values established with respect to the age and body height of subjects. The mean value of FVCex was 108% and FEV1 was 102% of predicted; the mean value of FEV1 %FVCex index was 82%. In all six groups, the output values of basic indices were within the normal range; FVCex 105-111% of predicted, FEV1 98 -105% of predicted value. Currently in Poland, there are no well documented studies available which cover a group of healthy subjects of a wide age range. Therefore, it is recommended to use a set of predicted values according to different authors. However, the most popular predicted values recommended by ERS (unchanged for over 20 years) are not fully representative for the Polish , Greek , Finish  Croatian , German , or Spain  populations.
A comparative analysis of the values of spirometric indices was carried out separately for the genotype-groups. The groups g. 46AA and g. 46GG, as well as g. 46AA and g. 46AG significantly differed for FEV0.5, FEV1, and FEV2 values: with the lowest indices recorded in the group g. 46AA and the highest in the group g. 46GG. Peak expiratory flow (PEF) was different only for the groups g. 46AA and g. 46GG, and had the highest values in subjects from the group g. 46GG. From among indices describing the maximum expiratory flow only MEF75 was different for the groups g. 46AA and g. 46GG, as well as g. 46AA and g. 46AG: the lowest values were measured in subjects from the group g. 46AA, and the highest from the group g.46GG. Similar to the above results, Aex and FEF25/75 values also varied.
The groups g. 79CC and g. 79GG demonstrated significant differences in FVCex values. FEV0.5 and FEV1 were significantly higher in the group g. 79GC when compared with the group g. 79CC. The FEV2 index was different for individuals with g. 79CC and g. 79GG, similar to MEF75, which was also varied for the genotypes g. 79CC and g. 79GC. The highest values of the analyzed indices were measured in the group g. 79GG, intermediate - g. 79GC and the lowest for g. 79CC. The present data conform to the results obtained by Joos , who found that individuals with g. 46GG achieve the highest FEV1 and FVCex expressed as the percent of the predicted value. Israel et al  also recorded the highest FEV1 values in the group g. 46GG. In contrast, Martinez et al , in the analysis of polymorphism of g. 79, found the highest FEV1 values in the group g. 79GG. Taylor et al , in the analysis of the polymorphism of g. 46 in asthmatic patients, recorded the highest FEV1 values in the group g. 46AA, slightly lower in g. 46AG, and the lowest in g. 46GG. Additionally, Martinez et al  recorded the highest FEV1 values in children with g. 46AA, intermediate in g. 46GG, and the lowest in g. 46AG. When analysing the g. 79 polymorphism in the present study we found that, similar to observations by Israel et al , heterozygotes achieve intermediate values of spirometric indices, while the highest values are achieved by homozygotes g. 79GG and the lowest by g. 79CC.
In conclusion, we believe the present study may be useful in explaining the differences in normal values of spirometric indices in healthy subjects in relation to the polymorphism of the β2-ADR gene. Considering the fact that the measured spirometric indices exceeded standard values, further spirometric studies on healthy subjects may contribute to the establishment of new predicted values for relevant age groups in the Polish population.
Conflicts of interest
The authors declare that they have no competing interests.
- Abraham G, Kottke C, Dhein S, Ungemach FR: Pharmacological and biochemical characterization of the beta-adrenergic signal transduction pathway in different segments of the respiratory tract. Biochem Pharmacol 2003,66(6):1067–81. 10.1016/S0006-2952(03)00460-XView ArticlePubMedGoogle Scholar
- Alexandraki S, Koutsilieris M, Siafakas N, Katsardis C: Spirometric reference values in Greek children and adolescents. Vivo 2010,24(2):195–200.Google Scholar
- American Thoracic Society: Standardization of Spirometry 1994 Update. Am J Respir Crit Care Med 1995,152(3):1107–36.View ArticleGoogle Scholar
- Aynacioglu AS, Cascorbi I, Güngör K, Ozkur M, Bekir N, Roots I, Brockm J: Population frequency, mutation linkage and analytical methodology for the Arg16Gly, Gln27Glu and Thr164Ile polymorphisms in the β 2 -adrenergic receptor among Turks. Br J Clin Pharmacol 1999,48(5):761–4.PubMed CentralView ArticlePubMedGoogle Scholar
- Aziz I, Hall IP, McFarlane LC, Lipworth BJ: β 2 -adrenoceptor regulation and bronchodilator sensitivity after regular treatment with formoterol in subjects with stable asthma. J Allergy Clin Immunol 1998,101(3):337–41. 10.1016/S0091-6749(98)70245-3View ArticlePubMedGoogle Scholar
- Bleecker ER, Postma DS, Lawrance RM, Meyers DA, Ambrose HJ, Goldman M: Effect of ADRB2 polymorphisms on response to long acting β 2 -agonist therapy: a pharmacogenetic analysis of two randomized studies. Lancet 2007,370(9605):2118–25. 10.1016/S0140-6736(07)61906-0View ArticlePubMedGoogle Scholar
- Carroll CL, Stoltz P, Schramm CM, Zucker AR: Beta2adrenergic receptor polymorphisms affect response to treatment in children with severe asthma exacerbations. Chest 2009,135(5):1186–92. 10.1378/chest.08-2041View ArticlePubMedGoogle Scholar
- Dixon RAP, Kobilka BK, Strader DJ, Benovic JL, Dohlman HG, Frielle T, Bolanowski MA, Bennett CD, Rands E, Diehl RE, Mumford RA, Slater EE, Sigal IS, Caron MG, Lefkowitz RJ, Strader CDl: Cloning of the gene and cDNA for mammalian β 2 -adrenergic receptor and homology with rhodopsin. Nature 1986, 321: 75–9. 10.1038/321075a0View ArticlePubMedGoogle Scholar
- Enright PL, Beck KC, Sherrill DL: Repeatability of spirometry in 18,000 adult patients. Am J Respir Crit Care Med 2004,169(2):235–8.View ArticlePubMedGoogle Scholar
- Green AS, Turki J, Hall IP, Ligget SB: Implications of genetic variability of human β 2 -adrenergic receptor structure. Pulm Pharmacol 1995,8(1):1–10. 10.1006/pulp.1995.1001View ArticlePubMedGoogle Scholar
- Gonzalez Barcala FJ, Gonzalez Barcala FJ, Cadarso Suarez C, Valdes Cuadrado L, Leis R, Cabanas R, Tojo R: Lung function reference values in children and adolescents aged 6 to years in Galicia. Arch Bronconeumol 2008,44(6):295–302.View ArticlePubMedGoogle Scholar
- Hall IP, Wheatley A, Wilding P, Liggett SB: Association of Glu 27 β 2 -adrenoceptor polymorphism with lower airway reactivity in asthmatic subjects. Lancet 1995,345(8959):1213–4. 10.1016/S0140-6736(95)91994-5View ArticlePubMedGoogle Scholar
- Hancox RJ, Sears MR, Taylor DR: Polymorphism of the β 2 -adrenoceptor and the 3esponse to long-term β 2 -agonist therapy in asthma. Eur Respir J 1998,11(3):589–93.PubMedGoogle Scholar
- Hopes E, McDougall C, Christie G, Wheatley A, Hall IP, Helms PJ: Association of glutamine 27 polymorphism of β 2 -adrenoceptor with reported childhood asthma: population based study. BMJ 1998,316(7132):664. 10.1136/bmj.316.7132.664PubMed CentralView ArticlePubMedGoogle Scholar
- Israel E, Drazen JM, Liggett SB, Boushey HA, Cherniack RM, Chinchilli VM, Cooper DM, Fahy JV, Fish JE, Ford JG, Kraft M, Kunselman S, Lazarus SC, Lemanske RF Jr, Martin RJ, McLean DE, Peters SP, Silverman EK, Sorkness CA, Szefler SJ, Weiss ST, Yandava CN: Effect of polymorphism of the β 2 -adrenergic receptor on response to regular use of albuterol in asthma. Int Arch Allergy Immunol 2001,124(1–3):183–6. 10.1159/000053705View ArticlePubMedGoogle Scholar
- Joos L, Weir TD, Connett JE, Anthonisen NR, Woods R, Pare PD, Sandford AJ: Polymorphisms in the β 2 -adrenergic receptor and bronchodilator response, bronchial hyperresponsiveness, and rate of decline in lung function in smokers. Thorax 2003,58(8):703–7. 10.1136/thorax.58.8.703PubMed CentralView ArticlePubMedGoogle Scholar
- Kainu A, Lindqvist A, Sarna S, Sovijärvi A: Spirometric and anthropometric determinants of forced expiratory time in a general population. Clin Physiol Funct Imaging 2008,28(1):38–42.View ArticlePubMedGoogle Scholar
- Koch B, Schäper C, Ittermann T, Völzke H, Felix SB, Ewert R, Gläser S: Reference values for lung function testing in adults - results from the study of health in Pomerania. Dtsch Med Wochenschr 2009,134(46):2327–32. 10.1055/s-0029-1242688View ArticlePubMedGoogle Scholar
- Kotani Y, Nishimura Y, Maeda H, Yokoyama M: β 2 adrenergic receptor polymorphisms affect airway responsiveness to salbutamol in asthmatics. J Asthma 1999,36(7):583–90. 10.3109/02770909909087295View ArticlePubMedGoogle Scholar
- Kulminski AM, Culminskaya I, Ukraintseva SV, Arbeev KG, Land KC, Yashin AI: β 2 -adrenergic receptor gene polymorphisms as systemic determinants of healthy aging in an evolutionary context. Mech Ageing Dev 2010,131(5):338–45. 10.1016/j.mad.2010.04.001PubMed CentralView ArticlePubMedGoogle Scholar
- Leineweber K, Büscher R, Bruck H, Brodde OE: β-adrenoceptor polymorphisms Naunyn Schmiedebergs. Arch Pharmacol 2004,369(1):1–22. 10.1007/s00210-003-0824-2View ArticleGoogle Scholar
- Liebhart J, Malolepszy J, Dor A: Role of autonomic nervous system (adrenergic, cholinergic, and non-adrenergic non-cholinergic) in the regulation of the functional state of airways. Postepy Hig Med Dosw 1995,49(3):395–407. (Article in Polish)PubMedGoogle Scholar
- Liggett SB: Pharmacogenetics of β 1 -and β 2 -adrenergic receptors. Pharmacology 2000,61(3):167–73. 10.1159/000028397View ArticlePubMedGoogle Scholar
- Liggett SB: Pharmacogenetics of relevant targets in asthma. Clin Exp Allergy 1998,28(Suppl 1):77–9.View ArticlePubMedGoogle Scholar
- Liggett SB: Polymorphisms of the β 2 -adrenergic receptor and asthma. Am J Respir Crit Care Med 1997, 56: 156–62.View ArticleGoogle Scholar
- Lima JJ, Thomason DB, Mohamed MH, Eberle LV, Self TH, Johnson JA: Impact of genetic polymorphisms of the β 2 -adrenergic receptor on albuterol bronchdilator pharmacodynamics. Clin Pharmacol Ther 1999,65(5):519–25. 10.1016/S0009-9236(99)70071-8View ArticlePubMedGoogle Scholar
- Lubinski W, Golczewski T: Physiologically interpretable prediction equations for spirometric indices. J Appl Physiol 2010. January 21, 2010. doi:10.1152/japplphysiol.01211.2009Google Scholar
- Martin AC, Zhang G, Rueter K, Khoo SK, Bizzintino J, Hayden CM, Geelhoed GC, Goldblatt J, Laing IA, Le Souëf PN: β 2 -adrenoceptor polymorphisms predict response to β 2 -agonists in children with acute asthma. J Asthma 2008,45(5):383–8. 10.1080/02770900801971792View ArticlePubMedGoogle Scholar
- Martinez FD, Graves PE, Baldini M: Association between genetic polymorphisms of the β 2 -adrenoceptor and response to albuterol in children with and without a history of wheezing. J Clin Invest 1997,100(12):3184–8. 10.1172/JCI119874PubMed CentralView ArticlePubMedGoogle Scholar
- McGraw DW, Liggett SB: Coding block and 5' leader cistron polymorphisms of the β 2 -adrenergic receptor. Clin Exp Allergy 1999,29(Suppl 4):43–5.PubMedGoogle Scholar
- Memon MA, Sandila MP, Ahmed ST: Spirometic reference values in healthy, non smoking Urban Pakistani population. J Pak Med Assoc 2007,57(4):193–5.PubMedGoogle Scholar
- Ortega VE, Hawkins GA, Peters SP, Bleecker ER: Pharmacogenetics of the β 2 -adrenergic receptor gene. Immunol Allergy Clin North Am 2007,27(4):665–84. 10.1016/j.iac.2007.09.007PubMed CentralView ArticlePubMedGoogle Scholar
- Ramsay CE, Hayden CM, Tiller KJ, Burton PR, Goldblatt J, Lesouef PN: Polymorphisms in the β 2 -ADR gene are associated with decreased airway responsiveness. Clin Exp Allergy 1999,29(9):1195–203. 10.1046/j.1365-2222.1999.00570.xView ArticlePubMedGoogle Scholar
- Reihsaus E, Innis M, MacIntyre N, Liggett SB: Mutations in the gene encoding for the β 2 -adrenergic receptor in normal and asthmatic subjects. Am J Respir Cell Mol Biol 1993,8(3):334–9.View ArticlePubMedGoogle Scholar
- Ruppel GL: Spirometry. Respir Care Clin N Am 1997,3(2):155–81.PubMedGoogle Scholar
- Small KM, McGraw DW, Liggett SB: Pharmacology and physiology of human adrenergic receptor polymorphisms. Annu Rev Pharmacol Toxicol 2003, 43: 381–411. 10.1146/annurev.pharmtox.43.100901.135823View ArticlePubMedGoogle Scholar
- Smolej Narancic N, Pavlovic M, Zuskin E, Milicic J, Skaric-Juric T, Barbalic M, Rudan P: New reference equations for forced spirometry in elderly persons. Respir Med 2009,103(4):621–8. 10.1016/j.rmed.2008.10.013View ArticlePubMedGoogle Scholar
- Taylor DR: β-adrenergic receptor polymorphisms relationship to the beta agonist controversy and clinical implications. Expert Opin Pharmacother 2007,8(18):3195–203. 10.1517/146565220.127.116.1195View ArticlePubMedGoogle Scholar
- Taylor DR, Drazen JM, Herbison GP: Asthma exacerbations during long term β-agonist use: influence of β 2 -adrenoceptor polymorphism. Thorax 2000,55(9):762–7. 10.1136/thorax.55.9.762PubMed CentralView ArticlePubMedGoogle Scholar
- Xie HG, Stein CM, Kim RB, Xiao ZS, He N, Zhou HH, Gainer JV, Brown NJ, Haines JL, Wood AJ: Frequency of functionally important β 2 -adrenoceptor polymorphisms varies markedly among African-American, Caucasian and Chinese individuals. Pharmacogenetics 1999,9(4):511–6.PubMedGoogle Scholar