Establishing the Genotyping Method for NAT2 Polymorphism in Vietnamese Tuberculoma Patients

The metabolism of Isoniazid, one of the first-line antituberculosis drugs for TB treatment and prophylaxis, depends on the acetyltransferase 2 acetylation (NAT2) phenotype. Different phenotypes of NAT2 will lead to differences in drug concentration and the risk of uncontrolled side effects, such as hepatitis, peripheral neuropathy, gastrointestinal disorders (nausea, vomiting, and stomach pain). These risks are related to the presence of mutant NAT2 alleles such as NAT2*5 (c.341T> C), *6 (c.590G> A) and *7 (c.857G> A), that reduce the N- acetyltransferase activity. Therefore, the genotyping method for NAT2 polymorphism using RFLP and Sanger sequencing was established. The method was successfully applied to determine the polymorphism of 84 TB patients. This study provides a better tool for analyzing NAT2 gene to assist clinicians in treating isoniazid. Keywords Enzyme NAT2, isoniazid, single nucleotide polymorphism, RFLP, Sanger sequencing. References [1] U.A. Boelsterli, K.K. Lee, Mechanisms of isoniazid-induced idiosyncratic liver injury: emerging role of mitochondrial stress, J. Gastroenterol. Hepatol. 29 (2014) 678–687.[2] A. Zabost, S. Brzezinska, M. Kozinska, M. Blachnio, J. Jagodzinski, Z. Zwolska, E. Augustynowicz-Kopec, Correlation of N-acetyltransferase 2 genotype with isoniazid acetylation in Polish tuberculosis patients, Biomed Res Int. 2013 (2013) 1-5.[3] M. Kinzig-Schippers, D. Tomalik-Scharte, A. Jetter, B. Scheidel, V. Jakob, M. Rodamer, I. Cascorbi, O. Doroshyenko, F. Sorgel, U. Fuhr, Should we use N-acetyltransferase type 2 genotyping to personalize isoniazid doses? Antimicrob Agents Chemother. 49 (2005) 1733-8[4] K. Walker, G. Ginsberg, D. Hattis, D.O. Johns, K.Z. Guyton, B. Sonawane, Genetic polymorphism in N-Acetyltransferase (NAT): Population distribution of NAT1 and NAT2 activity, J Toxicol Environ Health B Crit Rev. 12 (2009) 440-472. [5] G. Ramachandran, S. Swaminathan, Role of pharmacogenomics in the treatment of tuberculosis: a review, Pharmgenomics Pers Med. 5 (2012) 89-98.[6] J. Azuma, M. Ohno, R. Kubota, S. Yokota, T. Nagai, K. Tsuyuguchi, Y. Okuda, T. Takashima, S. Kamimura, Y. Fujio, I. Kawase, Pharmacogenetics-based tuberculosis therapy research group, NAT2 genotype guided regimen reduces isoniazid-induced liver injury and early treatment failure in the 6-month four-drug standard treatment of tuberculosis: a randomized controlled trial for pharmacogenetics-based therapy, Eur J Clin Pharmacol. 69 (2013) 1091-1101.[7] P.S. Adole, P.S. Kharbanda, S. Sharma, N-acetyltransferase 2 (NAT2) gene polymorphism as a predisposing factor for phenytoin intoxication in tuberculous meningitis or tuberculoma patients having seizures - A pilot study, Indian J Med Res. 143 (2016) 581-590.[8] WHO Scientific Group on Pharmacogenetics and World Health Organization, Pharmacogenetics: report of a WHO scientific group,World Health Organization Technical Report Series. (1973)[9] T.D. Da Silva, A.V. Felipe, J.M. De Lima, C.T. Oshima, N.M. Forones, N-Acetyltransferase 2 genetic polymorphisms and risk of colorectal cancer, World J Gastroenterol. 17 (2011) 760-765. [10] E.Y. Lau, J.S. Felton, F.C. Lightstone, Insights into the o-acetylation reaction of hydroxylated heterocyclic amines by human arylamine N-acetyltransferases: a computational study, Chem Res Toxicol. 19 (2006) 182-1190.[11] Ensembl - EBI, http://asia.ensembl.org/Homo_sapiens/Variation/Population?db=core;r=8:18399844-18400844;v=rs1801280;vdb=variation;vf=1243314,2019 (Ensembl release 96 - April 2019).[12] I.B. Kuznetsov, M. McDuffie, R. Moslehi, A web server for inferring the human N-acetyltransferase-2 (NAT2) enzymatic phenotype from NAT2 genotype, Bioinformatics. 25 (2009) 1185-1186. [13] P. Wang, K. Pradhan, X.B. Zhong, X. Ma, Isoniazid metabolism and hepatotoxicity, Acta Pharm Sin B. 6 (2016) 384-392.[14] M. Ohno, I. Yamaguchi, I. Yamamoto, T. Fukuda, S. Yokota, Slow N-acetyltransferase 2 genotype affects the incidence of isoniazid and rifampicin-induced hepatotoxicity, Int J Tuberc Lung Dis. 4 (2000) 256-261. [15] G.M. Lower, T. Nilsson, C.E. Nelson, H. Wolf, T.E. Gamsky, G.T. Bryan, N-acetyltransferase phenotype and risk in urinary bladder cancer: approaches in molecular epidemiology. Preliminary results in Sweden and Denmark, Int J Epidemiol. 36 (2007) 11-18.