SpringerLink
Log in

Thiopurine S-methyltransferase polymorphisms: efficient screening method for patients considering taking thiopurine drugs

  • Pharmacogenetics
  • Published:
European Journal of Clinical Pharmacology Aims and scope Submit manuscript

Abstract

Objective

More than 11% of the Caucasian population are heterozygous or homozygous carriers of thiopurine S-methyltransferase (TPMT) mutants and are at risk for toxic side effects when treated with thiopurine drugs. Therefore, screening for TPMT polymorphisms in a patient prior to prescribing these agents is recommended. The goal of this study was to determine a cut-off concentration of the TPMT activity assay beyond which genoty** of the TPMT gene should be performed.

Methods

The TPMT activity of 240 unrelated Caucasian subjects was measured using high-performance liquid chromatography. Genoty** for the most frequent allelic variants, TPMT*2, *3A, *3B, *3C and *7 was performed by LightCycler technology and sequencing.

Results

The inter-individual TPMT activity showed a range from 23 nmol MTG/g*Hb*h−1 to 97 nmol MTG/g*Hb*h−1 with a median of 56 nmol MTG/g*Hb*h−1. Using a cut-off concentration of 45.5 nmol MTG/g*Hb*h−1, a test sensitivity of 100% and a specificity of 89% were reached for heterozygous carriers of a TPMT mutation. We identified 1 carrier of TPMT*2, 14 carriers of TPMT*3A and 3 carriers of TPMT*3C, resulting in a TPMT heterozygosity prevalence of 7.5%.

Conclusions

This study defines the cut-off value for the TPMT phenoty** assay at 45.5 nmol/g*Hb*h−1, beyond which additional genoty** elucidates the individual risk for drug therapy. Using this cut-off concentration, the number of genoty** assays could be reduced by about 60%.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Woodson LC, Weinshilboum RM (1983) Human kidney thiopurine methyltransferase. Purification and biochemical properties. Biochem Pharmacol 32:819–826

    Article  CAS  PubMed  Google Scholar 

  2. Weinshilboum RM, Sladek SL (1980) Mercaptopurine pharmacogenetics: monogenic inheritance of erythrocyte thiopurine methyltransferase activity. Am J Hum Genet 32:651–662

    CAS  PubMed  Google Scholar 

  3. Tai HL, Krynetski EY, Yates CR, Loennechen T, Fessing MY, Krynetskaia NF, Evans WE (1996) 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

    Google Scholar 

  4. Yates CR, Krynetski EY, Loennechen T, Fessing MY, Tai HL, Pui CH, Relling MV, Evans WE (1997) Molecular diagnosis of thiopurine S-methyltransferase deficiency: genetic basis for azathioprine and mercaptopurine intolerance. Ann Intern Med 126:608–614

    CAS  PubMed  Google Scholar 

  5. Szumlanski C, Otterness D, Her C, Lee D, Brandriff B, Kelsell D, Spurr N, Lennard L, Wieben E, Weinshilboum R (1996) Thiopurine methyltransferase pharmacogenetics: human gene cloning and characterization of a common polymorphism. DNA Cell Biol 15:17–30

    CAS  PubMed  Google Scholar 

  6. Otterness D, Szumlanski C, Lennard L, Klemetsdal B, Aarbakke J, Park-Hah JO, Iven H, Schmiegelow K, Branum E, O’Brien J, Weinshilboum R (1997) Human thiopurine methyltransferase pharmacogenetics: gene sequence polymorphisms. Clin Pharmacol Ther 62:60–73

    Google Scholar 

  7. Krynetski EY, Schuetz JD, Galpin AJ, Pui CH, Relling MV, Evans WE (1995) A single point mutation leading to loss of catalytic activity in human thiopurine S-methyltransferase. Proc Natl Acad Sci U S A 92:949–953

    CAS  PubMed  Google Scholar 

  8. Evans WE, Horner M, Chu YQ, Kalwinsky D, Roberts WM (1991) Altered mercaptopurine metabolism, toxic effects, and dosage requirement in a thiopurine methyltransferase-deficient child with acute lymphocytic leukemia. J Pediatr 119:985–989

    CAS  PubMed  Google Scholar 

  9. Lennard L, Lilleyman JS, Van Loon J, Weinshilboum RM (1990) Genetic variation in response to 6-mercaptopurine for childhood acute lymphoblastic leukaemia. Lancet 336:225–229

    CAS  PubMed  Google Scholar 

  10. Lennard L (1998) Clinical implications of thiopurine methyltransferase—optimization of drug dosage and potential drug interactions. Ther Drug Monit 20:527–531

    CAS  PubMed  Google Scholar 

  11. Colombel JF, Ferrari N, Debuysere H, Marteau P, Gendre JP, Bonaz B, Soule JC, Modigliani R, Touze Y, Catala P, Libersa C, Broly F (2000) Genotypic analysis of thiopurine S-methyltransferase in patients with Crohn’s disease and severe myelosuppression during azathioprine therapy. Gastroenterology 118:1025–1030

    CAS  PubMed  Google Scholar 

  12. Gearry RB, Barclay ML, Burt MJ, Collett JA, Chapman BA, Roberts RL, Kennedy MA (2003) Thiopurine S-methyltransferase (TPMT) genotype does not predict adverse drug reactions to thiopurine drugs in patients with inflammatory bowel disease. Aliment Pharmacol Ther 18:395–400

    Article  CAS  PubMed  Google Scholar 

  13. Schwab M, Schaffeler E, Marx C, Fischer C, Lang T, Behrens C, Gregor M, Eichelbaum M, Zanger UM, Kaskas BA (2002) Azathioprine therapy and adverse drug reactions in patients with inflammatory bowel disease: impact of thiopurine S-methyltransferase polymorphism. Pharmacogenetics 12:429–436

    Article  PubMed  Google Scholar 

  14. Krynetski EY, Fessing MY, Yates CR, Sun D, Schuetz JD, Evans WE (1997) Promoter and intronic sequences of the human thiopurine S-methyltransferase (TPMT) gene isolated from a human PAC1 genomic library. Pharm Res 14:1672–1678

    Article  CAS  PubMed  Google Scholar 

  15. Spire-Vayron de la Moureyre C, Debuysere H, Fazio F, Sergent E, Bernard C, Sabbagh N, Marez D, Lo Guidice JM, D’Halluin J C, Broly F (1999) Characterization of a variable number tandem repeat region in the thiopurine S-methyltransferase gene promoter. Pharmacogenetics 9:189–198

    PubMed  Google Scholar 

  16. Spire-Vayron de la Moureyre C, Debuysere H, Sabbagh N, Marez D, Vinner E, Chevalier ED, Lo Guidice JM, Broly F (1998) Detection of known and new mutations in the thiopurine S-methyltransferase gene by single-strand conformation polymorphism analysis. Hum Mutat 12:177–185

    Article  PubMed  Google Scholar 

  17. Schaeffeler E, Lang T, Zanger UM, Eichelbaum M, Schwab M (2001) High-throughput genoty** of thiopurine S-methyltransferase by denaturing HPLC. Clin Chem 47:548–555

    CAS  PubMed  Google Scholar 

  18. Schutz E, von Ahsen N, Oellerich M (2000) Genoty** of eight thiopurine methyltransferase mutations: three-color multiplexing, “two-color/shared” anchor, and fluorescence-quenching hybridization probe assays based on thermodynamic nearest-neighbor probe design. Clin Chem 46:1728–1737

    CAS  PubMed  Google Scholar 

  19. Weinshilboum RM, Raymond FA, Pazmino PA (1978) Human erythrocyte thiopurine methyltransferase: radiochemical microassay and biochemical properties. Clin Chim Acta 85:323–333

    CAS  PubMed  Google Scholar 

  20. Ganiere-Monteil C, Pineau A, Kergueris MF, Azoulay C, Bourin M (1999) Thiopurine methyl transferase activity: new extraction conditions for high-performance liquid chromatographic assay. J Chromatogr B Biomed Sci Appl 727:235–239

    Article  CAS  PubMed  Google Scholar 

  21. Jacqz-Aigrain E, Bessa E, Medard Y, Mircheva Y, Vilmer E (1994) Thiopurine methyltransferase activity in a French population: HPLC assay conditions and effects of drugs and inhibitors. Br J Clin Pharmacol 38:1–8

    PubMed  Google Scholar 

  22. Kroplin T, Weyer N, Gutsche S, Iven H (1998) Thiopurine S-methyltransferase activity in human erythrocytes: a new HPLC method using 6-thioguanine as substrate. Eur J Clin Pharmacol 54:265–271

    CAS  PubMed  Google Scholar 

  23. Weinshilboum R (1987) Pharmacogenetics of methyl conjugation and thiopurine drug toxicity. Bioessays 7:78–82

    CAS  PubMed  Google Scholar 

  24. Coulthard SA, Hogarth LA, Little M, Matheson EC, Redfern CP, Minto L, Hall AG (2002) The effect of thiopurine methyltransferase expression on sensitivity to thiopurine drugs. Mol Pharmacol 62:102–109

    Article  CAS  PubMed  Google Scholar 

  25. Spire-Vayron de la Moureyre C, Debuysere H, Mastain B, Vinner E, Marez D, Lo Guidice JM, Chevalier D, Brique S, Motte K, Colombel JF, Turck D, Noel C, Flipo RM, Pol A, Lhermitte M, Lafitte JJ, Libersa C, Broly F (1998) Genotypic and phenotypic analysis of the polymorphic thiopurine S-methyltransferase gene (TPMT) in a European population. Br J Pharmacol 125:879–887

    PubMed  Google Scholar 

  26. McLeod HL, Siva C (2002) The thiopurine S-methyltransferase gene locus—implications for clinical pharmacogenomics. Pharmacogenomics 3:89–98

    CAS  PubMed  Google Scholar 

  27. 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, Relling MV (2001) Preponderance of thiopurine S-methyltransferase deficiency and heterozygosity among patients intolerant to mercaptopurine or azathioprine. J Clin Oncol 19:2293–2301

    CAS  PubMed  Google Scholar 

  28. McLeod HL, Krynetski EY, Wilimas JA, Evans WE (1995) Higher activity of polymorphic thiopurine S-methyltransferase in erythrocytes from neonates compared to adults. Pharmacogenetics 5:281–286

    CAS  PubMed  Google Scholar 

  29. Lysaa RA, Giverhaug T, Wold HL, Aarbakke J (1996) Inhibition of human thiopurine methyltransferase by furosemide, bendroflumethiazide and trichlormethiazide. Eur J Clin Pharmacol 49:393–396

    Google Scholar 

Download references

Acknowledgement

We are grateful for the generous donation of genomic DNA with TPMT*2, *3B, *3C and *7 alleles by Dr. med. N. von Ahsen, Uniklinikum Göttingen, Department of Clinical Chemistry, Göttingen, Germany.

Author information

Authors and Affiliations

  1. Institute for Clinical Chemistry, University Hospital Zurich, Rämistrasse 100, 8091, Zurich, Switzerland

    Barbara Wusk, A. von Eckardstein & K. M. Rentsch

  2. Division of Gastroenterology and Hepatology, University Hospital Zurich, 8091, Zurich, Switzerland

    G. A. Kullak-Ublick, C. Rammert & M. Fried

Authors
  1. Barbara Wusk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. A. Kullak-Ublick
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Rammert
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. von Eckardstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Fried
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K. M. Rentsch
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K. M. Rentsch.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wusk, B., Kullak-Ublick, G.A., Rammert, C. et al. Thiopurine S-methyltransferase polymorphisms: efficient screening method for patients considering taking thiopurine drugs. Eur J Clin Pharmacol 60, 5–10 (2004). https://doi.org/10.1007/s00228-004-0728-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00228-004-0728-1

Keywords

Search

Navigation