Thiopurine S-methyltransferase alleles, TPMT*2, *3B and *3C, and genotype frequencies in an Indian population

Murugesan,Raju; Vahab,Saadi A.; Patra,Satyajit; Rao,Rekha; Rao,Jyothi; Rai,Padmalatha; Gopinath,P. M.; Satyamoorthy,Kapaettu

doi:10.3892/etm_00000021

January-February 2010 Volume 1 Issue 1

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January-February 2010 Volume 1 Issue 1

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Article

Thiopurine S-methyltransferase alleles, TPMT2, 3B and *3C, and genotype frequencies in an Indian population

Authors:
- Raju Murugesan
- Saadi A. Vahab
- Satyajit Patra
- Rekha Rao
- Jyothi Rao
- Padmalatha Rai
- P. M. Gopinath
- Kapaettu Satyamoorthy
View Affiliations / Copyright

Affiliations: Manipal Life Sciences Center, Manipal University, Manipal 576104, India
Pages: 121-127
|
Published online on: January 1, 2010

https://doi.org/10.3892/etm_00000021
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Abstract

Thiopurine S-methyltransferase (TPMT) catalyzes the S-methylation of aromatic and heterocyclic sulfhydryl compounds including thiopurine drugs such as 6-mercapto-purine, 6-thioguanine and azathioprine. TPMT activity exhibits genetic variation and shows tri-modal distribution with 89-94% of individuals possessing high activity, 6-11% intermediate activity and approximately 0.3% low activity. Patients with intermediate or deficient TPMT activity exposed to thiopurine drugs show severe hematopoietic toxicity. Three single nucleotide polymorphisms (SNPs) in TPMT (NM_000367.2:c.238G>C, NM_000367.2:c.460G>A and NM_000367.2:c.719A>G) define the most prevalent mutant alleles associated with loss of catalytic activity reported in several populations. The present study investigated, for the first time, the frequency distribution of these three SNPs of TPMT, their alleles and genotypes in a Southern Indian population. Peripheral blood was obtained from 326 individuals of a Southern Indian population, and genomic DNA was isolated from total peripheral white blood cells. The genotypes at the polymorphic loci were determined by allele-specific polymerase chain reaction, restriction fragment length polymorphism and confirmatory DNA sequencing. The estimated genotype frequency for homozygous TPMT*1/*1 was 97.24%, for heterozygous TPMT*1/*2 and TPMT*1/*3B, 0.61% each, and for heterozygous TPMT*1/*3C, 1.53%. The frequency of heterozygous mutants in the studied Indian population was 2.76%. This study demonstrated significant variations in TPMT gene polymorphisms in an Indian population in relation to other human populations and may help to predict both clinical efficacy and drug toxicity of thiopurine drugs.

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1.	Remy CN: Metabolism of thiopyrimidines and thiopurines. S-Methylation with S-adenosylmethionine transmethylase and catabolism in mammalian tissues. J Biol Chem. 238:1078–1084. 1963.
2.	Tidd DM and Paterson AR: A biochemical mechanism for the delayed cytotoxic reaction of 6-mercaptopurine. Cancer Res. 34:738–746. 1974.
3.	Lennard L, van Loon JA, Lilleyman JS and Weinshilboum RM: Thiopurine pharmacogenetics in leukemia: correlation of erythrocyte thiopurine methyltransferase activity and 6-thioguanine nucleotide concentrations. Clin Pharmacol Ther. 41:18–25. 1987.
4.	Krynetski EY and Evans WE: Drug methylation in cancer therapy: lessons from the TPMT polymorphism. Oncogene. 22:7403–7413. 2003.
5.	Woodson LC, Dunnette JH and Weinshilboum RM: Pharmacogenetics of human thiopurine methyltransferase: kidney-erythrocyte correlation and immunotitration studies. J Pharmacol Exp Ther. 222:174–181. 1982.
6.	Krynetski EY, Tai HL, Yates CR, Fessing MY, Loennechen T, Schuetz JD, Relling MV and Evans WE: Genetic polymorphism of thiopurine S-methyltransferase: clinical importance and molecular mechanisms. Pharmacogenetics. 6:279–290. 1996.
7.	Snow JL and Gibson LE: The role of genetic variation in thiopurine methyltransferase activity and the efficacy and/or side effects of azathioprine therapy in dermatologic patients. Arch Dermatol. 131:193–197. 1995.
8.	Lee D, Szumlanski C, Houtman J, Honchel R, Rojas K, Overhauser J, Wieben ED and Weinshilboum RM: Thiopurine methyltransferase pharmacogenetics. Cloning of human liver cDNA and a processed pseudogene on human chromosome 18q21.1. Drug Metab Dispos. 23:398–405. 1995.
9.	Hon YY, Fessing MY, Pui C-H, Relling MV, Krynetski EY and Evans WE: Polymorphism of the thiopurine S-methyltransferase gene in African-Americans. Hum Mol Genet. 8:371–376. 1999.
10.	Evans WE, Hon YY, Bomgaars L, Coutre S, Holdsworth M, Janco R, Kalwinsky D, Keller F, Khatib Z, Margolin J, Murray J, Quinn J, Ravindranath Y, Ritchey K, Roberts W, Rogers ZR, Schiff D, Steuber C, Tucci F, Kornegay N, Krynetski EY and Relling MV: Preponderance of thiopurine S-methyltransferase deficiency and heterozygosity among patients intolerant to mercaptopurine or azathioprine. J Clin Oncol. 19:2293–2301. 2001.
11.	Lindqvist M, Haglund S, Almer S, Peterson C, Taipalensu J, Hertervig E, Lyrenäs E and Söderkvist P: Identification of two novel sequence variants affecting thiopurine methyltransferase enzyme activity. Pharmacogenetics. 14:261–265. 2004.
12.	Krynetski EY, Schuetz JD, Galpin AJ, Pui C-H, Relling MV and Evans WE: A single point mutation leading to loss of catalytic activity in human thiopurine S-methyltransferase. Proc Natl Acad Sci USA. 92:949–953. 1995.
13.	Tai HL, Krynetski EY, Yates CR, Loennechen T, Fessing MY, Krynetskaia NF and Evans WE: Thiopurine S-methyltransferase deficiency: two nucleotide transitions define the most prevalent mutant allele associated with loss of catalytic activity in Caucasians. Am J Hum Genet. 58:694–702. 1996.
14.	Otterness D, Szumlanski C, Lennard L, Klemetsdal B, Aarbakke J, Park-Hah JO, Iven H, Schmiegelow K, Branum E, O'Brien J and Weinshilboum R: Human thiopurine methyltransferase pharmacogenetics: gene sequence polymorphisms. Clin Pharmacol Ther. 62:60–73. 1997.
15.	Otterness DM, Szumlanski CL, Wood TC and Weinshilboum RM: Human thiopurine methyltransferase pharmacogenetics. Kindred with a terminal exon splice junction mutation that results in loss of activity. J Clin Invest. 101:1036–1044. 1998.
16.	Maniatis T, Fritsch EF and Sambrook J: Analysis and cloning of eukaryotic genomic DNA. Molecular Cloning. 2nd edition. A Laboratory Manual. Cold Spring Harbor; New York: pp. 9.16–9.23. 1989
17.	Lennard L, Lilleyman JS, van Loon J and Weinshilboum RM: Genetic variation in response to 6-mercaptopurine for childhood acute lymphoblastic leukaemia. Lancet. 336:225–229. 1990.
18.	Weinshilboum R: Thiopurine pharmacogenetics: clinical and molecular studies of thiopurine methyltransferase. Drug Metab Dispos. 29:601–605. 2001.
19.	Coulthard SA, Hogarth LA, Little M, Matheson EC, Redfern CPF, Minto L and Hall AG: The effect of thiopurine methyltransferase expression on sensitivity to thiopurine drugs. Mol Pharmacol. 62:102–109. 2002.
20.	Yates CR, Krynetski EY, Loennechen T, Fessing MY, Tai HL, Pui CH, Relling MV and Evans WE: Molecular diagnosis of thiopurine S-methyltransferase deficiency: genetic basis for azathioprine and mercaptopurine intolerance. Ann Intern Med. 126:608–614. 1997.
21.	Black AJ, McLeod HL, Capell HA, Powrie RH, Matowe LK, Pritchard SC, Collie-Duguid ESR and Reid RM: Thiopurine methyltransferase genotype predicts therapy-limiting severe toxicity from azathioprine. Ann Intern Med. 129:716–718. 1998.
22.	Kham SK, Tan PL, Tay AH, Heng CK, Yeoh AE and Quah TC: Thiopurine methyltransferase polymorphisms in a multiracial Asian population and children with acute lymphoblastic leukemia. J Pediatr Hematol Oncol. 24:353–359. 2002.
23.	Ameyaw MM, Collie-Duguid ES, Powrie RH, Ofori-Adjei D and McLeod HL: Thiopurine methyltransferase alleles in British and Ghanaian populations. Hum Mol Genet. 8:367–370. 1999.
24.	Leipold G, Schütz E, Haas JP and Oellerich M: Azathioprine-induced severe pancytopenia due to a homozygous two-point mutation of the thiopurine methyltransferase gene in a patient with juvenile HLA-B27-associated spondylarthritis. Arthritis Rheum. 40:1896–1898. 1997.
25.	Laróvere LE, de Kremer RD, Lambooy LH and de Abreu RA: Genetic polymorphism of thiopurine S-methyltransferase in Argentina. Ann Clin Biochem. 40:388–393. 2003.
26.	Reis M, Santoro A and Suarez-Kurtz G: Thiopurine methyltransferase phenotypes and genotypes in Brazilians. Pharmacogenetics. 13:371–373. 2003.
27.	McLeod HL, Pritchard SC, Githang'a J, Indalo A, Ameyaw MM, Powrie RH, Booth L and Collie-Duguid ES: Ethnic differences in thiopurine methyltransferase pharmacogenetics: evidence for allele specificity in Caucasian and Kenyan individuals. Pharmacogenetics. 9:773–776. 1999.
28.	Collie-Duguid ES, Pritchard SC, Powrie RH, Sludden J, Collier DA, Li T and McLeod HL: The frequency and distribution of thiopurine methyltransferase alleles in Caucasian and Asian populations. Pharmacogenetics. 9:37–42. 1999.
29.	Isaza C, Henao J, López AM and Cacabelos R: Allelic variants of the thiopurine methyltransferase (TPMT) gene in the Colombian population. Methods Find Exp Clin Pharmacol. 25:423–429. 2003.
30.	Hamdy SI, Hiratsuka M, Narahara K, Endo N, El-Enany M, Moursi N, Ahmed MS-E and Mizugaki M: Genotype and allele frequencies of TPMT, NAT2, GST, SULT1A1 and MDR-1 in the Egyptian population. Br J Clin Pharmacol. 55:560–569. 2003.
31.	Ganiere-Monteil C, Medard Y, Lejus C, Bruneau B, Pineau A, Fenneteau O, Bourin M and Jacqz-Aigrain E: Phenotype and genotype for thiopurine methyltransferase activity in the French Caucasian population: impact of age. Eur J Clin Pharmacol. 60:89–96. 2004.
32.	Rossi AM, Bianchi M, Guarnieri C, Barale R and Pacifici GM: Genotype-phenotype correlation for thiopurine S-methyltransferase in healthy Italian subjects. Eur J Clin Pharmacol. 57:51–54. 2001.
33.	Hiratsuka M, Inoue T, Omori F, Agatsuma Y, Kishikawa Y and Mizugaki M: Detection assay of rare variants of the thiopurine methyltransferase gene by PCR-RFLP using a mismatch primer in a Japanese population. Biol Pharm Bull. 23:1090–1093. 2000.
34.	Wei H, Zhou S, Li C, Zhang J, Wu J and Huang M: Phenotyping and genotyping studies of thiopurine S-methyltransferase in Kazaks. Pharm Res. 22:1762–1766. 2005.
35.	Loennechen T, Utsi E, Hartz I, Lysaa R, Kildalsen H and Aarbakke J: Detection of one single mutation predicts thiopurine S-methyltransferase activity in a population of Saami in northern Norway. Clin Pharmacol Ther. 70:183–188. 2001.
36.	Alves S, Prata MJ, Ferreira F and Amorim A: Thiopurine methyltransferase pharmacogenetics: alternative molecular diagnosis and preliminary data from Northern Portugal. Pharmacogenetics. 9:257–261. 1999.
37.	Kurzawski M, Gawronska-Szklarz B and Drozdzik M: Frequency distribution of thiopurine S-methyltransferase alleles in a polish population. Ther Drug Monit. 26:541–545. 2004.
38.	Chang JG, Lee LS, Chen CM, Shih MC, Wu MC, Tsai FJ and Liang DC: Molecular analysis of thiopurine S-methyltransferase alleles in South-east Asian populations. Pharmacogenetics. 12:191–195. 2002.
39.	Haglund S, Lindqvist M, Almer S, Peterson C and Taipalensuu J: Pyrosequencing of TPMT alleles in a general Swedish population and in patients with inflammatory bowel disease. Clin Chem. 50:288–295. 2004.
40.	Srimartpirom S and Tassaneeyakul W, Kukongviriyapan V and Tassaneeyakul W: Thiopurine S-methyltransferase genetic polymorphism in the Thai population. Br J Clin Pharmacol. 58:66–70. 2004.
41.	Vesell ES: Therapeutic lessons from pharmacogenetics. Ann Intern Med. 126:653–655. 1997.