Skip to main content

Advertisement

Log in

Refinement of the prediction of N-acetyltransferase 2 (NAT2) phenotypes with respect to enzyme activity and urinary bladder cancer risk

  • Toxicogenomics
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

Polymorphisms of N-acetyltransferase 2 (NAT2) are well known to modify urinary bladder cancer risk as well as efficacy and toxicity of pharmaceuticals via reduction in the enzyme’s acetylation capacity. Nevertheless, the discussion about optimal NAT2 phenotype prediction, particularly differentiation between different degrees of slow acetylation, is still controversial. Therefore, we investigated the impact of single nucleotide polymorphisms and their haplotypes on slow acetylation in vivo and on bladder cancer risk. For this purpose, we used a study cohort of 1,712 bladder cancer cases and 2,020 controls genotyped for NAT2 by RFLP-PCR and for the tagSNP rs1495741 by TaqMan® assay. A subgroup of 344 individuals was phenotyped by the caffeine test in vivo. We identified an ‘ultra-slow’ acetylator phenotype based on combined *6A/*6A, *6A/*7B and *7B/*7B genotypes containing the homozygous minor alleles of C282T (rs1041983, *6A, *7B) and G590A (rs1799930, *6A). ‘Ultra-slow’ acetylators have significantly about 32 and 46 % lower activities of caffeine metabolism compared with other slow acetylators and with the *5B/*5B genotypes, respectively (P < 0.01, both). The ‘ultra-slow’ genotype showed an association with bladder cancer risk in the univariate analysis (OR = 1.31, P = 0.012) and a trend adjusted for age, gender and smoking habits (OR = 1.22, P = 0.082). In contrast, slow acetylators in general were not associated with bladder cancer risk, neither in the univariate (OR = 1.02, P = 0.78) nor in the adjusted (OR = 0.98, P = 0.77) analysis. In conclusion, this study suggests that NAT2 phenotype prediction should be refined by consideration of an ‘ultra-slow’ acetylation genotype.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Agúndez JA (2003) NAT2 genotyping: equilibrium between accuracy and feasibility in routine analyses. J Appl Res 3:1–6

    Google Scholar 

  • Agúndez JA (2008) Polymorphisms of human N-acetyltransferases and cancer risk. Curr Drug Metab 9:520–531

    Article  PubMed  Google Scholar 

  • Agúndez JA, Golka K, Martínez C, Selinski S, Blaszkewicz M, García-Martín E (2008) Unraveling ambiguous NAT2 genotyping data. Clin Chem 54:1390–1394

    Article  PubMed  Google Scholar 

  • An HR, Wu XQ, Wang ZY, Zhang JX, Liang Y (2012) NAT2 and CYP2E1 polymorphisms associated with antituberculosis drug-induced hepatotoxicity in Chinese patients. Clin Exp Pharmacol Physiol 39:535–543

    Article  PubMed  CAS  Google Scholar 

  • Arylamine N-Acetyltransferase Nomenclature Committee. Update July 22, 2011. http://www.louisville.edu/medschool/pharmacology/NAT.html. Accessed Feb 7, 2013

  • Blaszkewicz M (2004) N-acetyltransferase 2 (phenotyping: caffeine test). In: Angerer J, Müller M, Weiss T et al. (eds) Analyses of hazardous substances in biological materials, vol 9. Special issue: Markers of susceptibility. Wiley-VCH, Weinheim, pp 165–182

  • Blaszkewicz M, Dannappel D, Thier R, Lewalter J (2004) N-acetyltransferase 2 (genotyping). In: Angerer J, Müller M, Weiss T et al. (eds) Analyses of hazardous substances in biological materials, vol 9. Special issue: markers of susceptibility. Wiley-VCH, Weinheim, pp 135–163

  • Bolt HM (2013a) Human bladder cancer risk calculation based on genome-wide analysis of genetic variants. Arch Toxicol 87:397–399

    Article  PubMed  CAS  Google Scholar 

  • Bolt HM (2013b) Relevance of genetic disposition versus environmental exposure for cancer risk: an old controversy revisited with novel methods. EXCLI J 12:79–80

    Google Scholar 

  • Bolt HM, Selinski S, Dannappel D, Blaszkewicz M, Golka K (2005) Re-investigation of the concordance of human NAT2 phenotypes and genotypes. Arch Toxicol 79:196–200

    Article  PubMed  CAS  Google Scholar 

  • Cai Y, Yi J, Zhou C, Shen X (2012) Pharmacogenetic study of drug-metabolising enzyme polymorphisms on the risk of anti-tuberculosis drug-induced liver injury: a meta-analysis. PLoS ONE 7:e47769

    Article  PubMed  CAS  Google Scholar 

  • Cascorbi I, Drakoulis N, Brockmöller J, Maurer A, Sperling K, Roots I (1995) Arylamine N-acetyltransferase (NAT2) mutations and their allelic linkage in unrelated Caucasian individuals: correlation with phenotypic activity. Am J Hum Genet 57:581–592

    Article  PubMed  CAS  Google Scholar 

  • Deitz AC, Rothman N, Rebbeck TR, Hayes RB, Chow WH, Zheng W et al (2004) Impact of misclassification in genotype-exposure interaction studies: example of N-acetyltransferase 2 (NAT2), smoking, and bladder cancer. Cancer Epidemiol Biomarkers Prev 13:1543–1546

    PubMed  CAS  Google Scholar 

  • DerSimonian R, Laird N (1986) Meta-analysis in clinical trials. Control Clin Trials 7:177–188

    Article  PubMed  CAS  Google Scholar 

  • García-Closas M, Malats N, Silverman D, Dosemeci M, Kogevinas M, Hein DW et al (2005) NAT2 slow acetylation, GSTM1 null genotype, and risk of bladder cancer: results from the Spanish Bladder Cancer Study and meta-analyses. Lancet 366:649–659

    Article  PubMed  Google Scholar 

  • García-Closas M, Hein DW, Silverman D, Malats N, Yeager M, Jacobs K et al (2011) A single nucleotide polymorphism tags variation in the arylamine N-acetyltransferase 2 phenotype in populations of European background. Pharmacogenet Genomics 21:231–236

    PubMed  Google Scholar 

  • Golka K, Prior V, Blaszkewicz M, Cascorbi I, Schöps W, Kierfeld G et al (1996) Occupational history and genetic N-acetyltransferase polymorphism in urothelial cancer patients of Leverkusen, Germany. Scand J Work Environ Health 22:332–338

    Article  PubMed  CAS  Google Scholar 

  • Golka K, Prior V, Blaszkewicz M, Bolt HM (2002) The enhanced bladder cancer susceptibility of NAT2 slow acetylators towards aromatic amines: a review considering ethnic differences. Toxicol Lett 128:229–241

    Article  PubMed  CAS  Google Scholar 

  • Golka K, Blaszkewicz M, Samimi M, Bolt HM, Selinski S (2008) Reconstruction of N-acetyltransferase 2 haplotypes using PHASE. Arch Toxicol 82:265–270

    Article  PubMed  CAS  Google Scholar 

  • Golka K, Hermes M, Selinski S, Blaszkewicz M, Bolt HM, Roth G et al (2009) Susceptibility to urinary bladder cancer: relevance of rs9642880[T], GSTM1 0/0 and occupational exposure. Pharmacogenet Genomics 19:903–906

    Article  PubMed  CAS  Google Scholar 

  • Grant DM, Hughes NC, Janezic SA, Goodfellow GH, Chen HJ, Gaedigk A et al (1997) Human acetyltransferase polymorphisms. Mutat Res 376:61–70

    Article  PubMed  CAS  Google Scholar 

  • Gross M, Kruisselbrink T, Anderson K, Lang N, McGovern P, Delongchamp R et al (1999) Distribution and concordance of N-acetyltransferase genotype and phenotype in an American population. Cancer Epidemiol Biomarkers Prev 8:683–692

    PubMed  CAS  Google Scholar 

  • He YJ, Shapero MH, McLeod HL (2012) Novel tagging SNP rs1495741 and 2-SNPs (rs1041983 and rs1801280) yield a high prediction of the NAT2 genotype in HapMap samples. Pharmacogenet Genomics 22:322–324

    Article  PubMed  CAS  Google Scholar 

  • Hein DW (2002) Molecular genetics and function of NAT1 and NAT2: role in aromatic amine metabolism and carcinogenesis. Mutat Res 506–507:65–77

    Article  PubMed  Google Scholar 

  • Hein DW (2006) N-acetyltransferase 2 genetic polymorphism: effects of carcinogen and haplotype on urinary bladder cancer risk. Oncogene 25:1649–1658

    Article  PubMed  CAS  Google Scholar 

  • Hein DW (2009) N-acetyltransferase SNPs: emerging concepts serve as a paradigm for understanding complexities of personalized medicine. Expert Opin Drug Metab Toxicol 5:353–366

    Article  PubMed  CAS  Google Scholar 

  • Hein DW, Doll MA (2012a) A four-SNP NAT2 genotyping panel recommended to infer human acetylator phenotype. Pharmacogenomics 13:855

    Article  CAS  Google Scholar 

  • Hein DW, Doll MA (2012b) Accuracy of various human NAT2 SNP genotyping panels to infer rapid, intermediate and slow acetylator phenotypes. Pharmacogenomics 13:31–41

    Article  PubMed  CAS  Google Scholar 

  • Hein DW, Boukouvala S, Grant DM, Minchin RF, Sim E (2008) Changes in consensus arylamine N-acetyltransferase gene nomenclature. Pharmacogenet Genomics 18:367–368

    Article  PubMed  CAS  Google Scholar 

  • Hengstler JG, Arand M, Herrero ME, Oesch F (1998) Polymorphisms of N-acetyltransferases, glutathione S-transferases, microsomal epoxide hydrolase and sulfotransferases: influence on cancer susceptibility. Recent Results Cancer Res 154:47–85

    Article  PubMed  CAS  Google Scholar 

  • Huang YS, Chern HD, Su WJ, Wu JC, Lai SL, Yang SY et al (2002) Polymorphism of the N-acetyltransferase 2 gene as a susceptibility risk factor for antituberculosis drug-induced hepatitis. Hepatology 35:883–889

    Article  PubMed  CAS  Google Scholar 

  • Hung RJ, Boffetta P, Brennan P, Malaveille C, Hautefeuille A, Donato F et al (2004) GST, NAT, SULT1A1, CYP1B1 genetic polymorphisms, interactions with environmental exposures and bladder cancer risk in a high-risk population. Int J Cancer 110:598–604

    Article  PubMed  CAS  Google Scholar 

  • Lee SW, Chung LS, Huang HH, Chuang TY, Liou YH, Wu LS (2010) NAT2 and CYP2E1 polymorphisms and susceptibility to first-line anti-tuberculosis drug-induced hepatitis. Int J Tuberc Lung Dis 14:622–626

    PubMed  Google Scholar 

  • Lehmann ML, Selinski S, Blaszkewicz M, Orlich M, Ovsiannikov D, Moormann O et al (2010) Rs710521[A] on chromosome 3q28 close to TP63 is associated with increased urinary bladder cancer risk. Arch Toxicol 84:967–978

    Article  PubMed  CAS  Google Scholar 

  • Leiro-Fernandez V, Valverde D, Vázquez-Gallardo R, Botana-Rial M, Constenla L, Agúndez JA et al (2011) N-acetyltransferase 2 polymorphisms and risk of anti-tuberculosis drug-induced hepatotoxicity in Caucasians. Int J Tuberc Lung Dis 15:1403–1408

    Article  PubMed  CAS  Google Scholar 

  • Li N, Stephens M (2003) Modelling linkage disequilibrium, and identifying recombination hotspots using SNP data. Genetics 165:2213–2233

    PubMed  CAS  Google Scholar 

  • Moore LE, Baris DR, Figueroa JD, Garcia-Closas M, Karagas MR, Schwenn MR et al (2011) GSTM1 null and NAT2 slow acetylation genotypes, smoking intensity and bladder cancer risk: results from the New England bladder cancer study and NAT2 meta-analysis. Carcinogenesis 32:182–189

    Article  PubMed  CAS  Google Scholar 

  • Possuelo LG, Castelan JA, de Brito TC, Ribeiro AW, Cafrune PI, Picon PD et al (2008) Association of slow N-acetyltransferase 2 profile and anti-TB drug-induced hepatotoxicity in patients from Southern Brazil. Eur J Clin Pharmacol 64:673–681

    Article  PubMed  CAS  Google Scholar 

  • R Development Core Team (2008) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna

    Google Scholar 

  • Rothman N, Garcia-Closas M, Chatterjee N, Malats N, Wu X, Figueroa JD et al (2010) A multi-stage genome-wide association study of bladder cancer identifies multiple susceptibility loci. Nat Genet 42:978–984

    Article  PubMed  CAS  Google Scholar 

  • Ruiz JD, Martínez C, Anderson K, Gross M, Lang NP, García-Martín E et al (2012) The differential effect of NAT2 variant alleles permits refinement in phenotype inference and identifies a very slow acetylation genotype. PLoS ONE 7:e44629

    Article  PubMed  CAS  Google Scholar 

  • Sabbagh A, Darlu P, Crouau-Roy B, Poloni ES (2011) Arylamine N-acetyltransferase 2 (NAT2) genetic diversity and traditional subsistence: a worldwide population survey. PLoS ONE 6:e18507

    Article  PubMed  CAS  Google Scholar 

  • Sanderson S, Salanti G, Higgins J (2007) Joint effects of the N-acetyltransferase 1 and 2 (NAT1 and NAT2) genes and smoking on bladder carcinogenesis: a literature-based systematic HuGE review and evidence synthesis. Am J Epidemiol 166:741–751

    Article  PubMed  Google Scholar 

  • Santos NP, Callegari-Jacques SM, Santos AK, Silva CA, Vallinoto AC, Fernandes DC et al (2013) N-acetyltransferase 2 and cytochrome P450 2E1 genes and isoniazid-induced hepatotoxicity in Brazilian patients. Int J Tuberc Lung Dis 17:499–504

    Article  PubMed  CAS  Google Scholar 

  • SAS/STAT® software, version 9.2. Copyright © 2002–2008, SAS Institute Inc., Cary

  • Schwender H, Selinski S, Blaszkewicz M, Marchan R, Ickstadt K, Golka K et al (2012) Distinct SNP combinations confer susceptibility to urinary bladder cancer in smokers and non-smokers. PLoS ONE 7:e51880

    Article  PubMed  CAS  Google Scholar 

  • Selinski S (2012) Genetic variants confer susceptibility to urinary bladder cancer: an updated list of confirmed polymorphisms. EXCLI J 11:743–747

    Google Scholar 

  • Selinski S, Blaszkewicz M, Lehmann ML, Ovsiannikov D, Moormann O, Guballa C et al (2011) Genotyping NAT2 with only two SNPs (rs1041983 and rs1801280) outperforms the tagging SNP rs1495741 and is equivalent to the conventional 7-SNP NAT2 genotype. Pharmacogenet Genomics 21:673–678

    Article  PubMed  CAS  Google Scholar 

  • Selinski S, Lehmann ML, Blaszkewicz M, Ovsiannikov D, Moorman O, Guballa C et al (2012) Rs11892031[A] on chromosome 2q37 in an intronic region of the UGT1A locus is associated with urinary bladder cancer risk. Arch Toxicol 86:1369–1378

    Article  PubMed  CAS  Google Scholar 

  • Selinski S, Blaszkewicz M, Agúndez JA, Martínez C, García-Martín E, Hengstler JG et al (2013a) Clarifying haplotype ambiguity of NAT2 in multi-national cohorts. Front Biosci (Schol Ed) 5:672–684

    Article  Google Scholar 

  • Selinski S, Blaszkewicz M, Ickstadt K, Hengstler JG, Golka K (2013b) Improvements in algorithms for phenotype inference: The NAT2 example. Curr Drug Metab (in press)

  • Stephens M, Donnelly P (2003) A comparison of Bayesian methods for haplotype reconstruction. Am J Hum Genet 73:1162–1169

    Article  PubMed  CAS  Google Scholar 

  • Stephens M, Smith NJ, Donnelly P (2001) A new statistical method for haplotype reconstruction from population data. Am J Hum Genet 68:978–989

    Article  PubMed  CAS  Google Scholar 

  • Suarez-Kurtz G, Sortica VA, Vargens DD, Bruxel EM, Petzl-Erler ML, Tsuneto LT et al (2012a) Impact of population diversity on the prediction of 7-SNP NAT2 phenotypes using the tagSNP rs1495741 or paired SNPs. Pharmacogenet Genomics 22:305–309

    Article  PubMed  CAS  Google Scholar 

  • Suarez-Kurtz G, Vargens DD, Sortica VA, Hutz MH (2012b) Accuracy of NAT2 SNP genotyping panels to infer acetylator phenotypes in African, Asian, Amerindian and admixed populations. Pharmacogenomics 13:851–854

    Article  PubMed  CAS  Google Scholar 

  • Teixeira RL, Morato RG, Cabello PH, Muniz LM, Moreira Ada S, Kritski AL et al (2011) Genetic polymorphisms of NAT2, CYP2E1 and GST enzymes and the occurrence of antituberculosis drug-induced hepatitis in Brazilian TB patients. Mem Inst Oswaldo Cruz 106:716–724

    PubMed  CAS  Google Scholar 

  • Vineis P, Marinelli D, Autrup H, Brockmöller J, Cascorbi I, Daly AK et al (2001) Current smoking, occupation, N-acetyltransferase-2 and bladder cancer: a pooled analysis of genotype-based studies. Cancer Epidemiol Biomarkers Prev 10:1249–1252

    PubMed  CAS  Google Scholar 

  • Walraven JM, Zang Y, Trent JO, Hein DW (2008) Structure/function evaluations of single nucleotide polymorphisms in human N-acetyltransferase 2. Curr Drug Metab 9:471–486

    Article  PubMed  CAS  Google Scholar 

  • Zang Y, Doll MA, Zhao S, States JC, Hein DW (2007) Functional characterization of single-nucleotide polymorphisms and haplotypes of human N-acetyltransferase 2. Carcinogenesis 28:1665–1671

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank Ms. Doris Dannappel, Ms. Kirsten Liesenhoff-Henze, Ms. Marion Page, Ms. Claudia Schulte-Dahmann and Mr. Roland Wycislok for excellent technical support.

Conflict of interest

None.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Silvia Selinski.

Additional information

Jan G. Hengstler and Klaus Golka have shared senior authorship.

Electronic supplementary material

Below is the link to the electronic supplementary material.

204_2013_1157_MOESM1_ESM.docx

Supplementary Materials and Methods.doc Detailed description of the study groups and the genotyping and phenotyping methods. Supplementary material 1 (DOCX 46 kb)

204_2013_1157_MOESM2_ESM.doc

Supplementary Tables S1-S12 and Figure S1.doc Study group characteristics and a more detailed analysis on individual study group level. Supplementary material 2 (DOC 1855 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Selinski, S., Blaszkewicz, M., Ickstadt, K. et al. Refinement of the prediction of N-acetyltransferase 2 (NAT2) phenotypes with respect to enzyme activity and urinary bladder cancer risk. Arch Toxicol 87, 2129–2139 (2013). https://doi.org/10.1007/s00204-013-1157-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-013-1157-7

Keywords

Navigation