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European Journal of Drug Metabolism and Pharmacokinetics
Published in: European Journal of Drug Metabolism and Pharmacokinetics 4/2016

01-08-2016 | Original Paper

Genetic Determinants of Methotrexate Toxicity in Tunisian Patients with Rheumatoid Arthritis: A Study of Polymorphisms Involved in the MTX Metabolic Pathway

Authors: Souhir Chaabane, Sameh Marzouk, Rim Akrout, Mariem Ben Hamad, Yosser Achour, Ahmed Rebai, Leila Keskes, Hela Fourati, Zouhir Bahloul, Abdellatif Maalej

Published in: European Journal of Drug Metabolism and Pharmacokinetics | Issue 4/2016

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Abstract

Background and Objective

Methotrexate (MTX) is a disease-modifying anti-rheumatic drug used in the treatment of rheumatoid arthritis (RA). It is the first line drug in the treatment of this disease. However, MTX-related adverse drug reactions (ADRs) are seen in 40 % of the patients. The aim of this study was to determine the impact of six genetic polymorphisms located in five genes encoding proteins involved in the MTX metabolic pathway in Tunisian RA patients and evaluate its association with MTX toxicity.

Methods

Genotyping of 5,10 methylenetetrahydrofolate reductase (MTHFR C677T and A1298C), dihydrofolate reductase (DHFR 19–base pair deletion allele), thymidylate synthase (TYMS 2R/3R), methionine synthase (MTR A2756G) and methionine synthase reductase (MTRR A66G) was performed using PCR and PCR–RFLP method in 141 RA patients treated with MTX. Demographic and clinical characteristics were obtained and ADRs were recorded. Association analyses with regard to MTX toxicity were performed using the χ 2 test, the toxicogenetic risk index (TRI) and the Mann–Whitney U-test.

Results

The analysis highlighted a significant association of the T/T genotype of MTHFR C677T polymorphism with increased MTX toxicity. However, the MTHFR A1298C, DHFR 19-base pair deletion allele, MTR A2756G and MTRR A66G polymorphisms were not associated with increased MTX toxicity. The TYMS 2R/3R polymorphism had a protective effect against MTX toxicity.

Conclusion

The results demonstrated that the C677T polymorphism in the MTHFR gene is associated with MTX toxicity in Tunisian RA patients. In contrast, the TYMS 2R/3R polymorphism is associated with a protective effect against overall MTX toxicity.
Literature
1.
2.
Smolen JS, Landewe´ R, Breedveld FC, Buch M, Burmester G, Dougados M, et al. EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease- modifying antirheumatic drugs: 2013 update. Ann Rheum Dis. 2014;73:492–509.CrossRefPubMed
3.
Rath T, Rubbert A. Drug combinations with methotrexate to treat rheumatoid arthritis. Clin Exp Rheumatol. 2010;28(5 Suppl 61):S52–7.PubMed
4.
Johnsen AK, Weinblatt ME. Methotrexate. In: Hochberg MC, Silman AJ, Smolen JS, editors. Rheumatology. 5th ed. Philadelphia: Mosby-Elsevier; 2011. p. 509–18.CrossRef
5.
Peters GJ, Hooijberg JH, Kaspers GJ, Jansen G. Folates and antifolates in the treatment of cancer; role of folic acid supplementation on efficacy of folate and non-folate drugs. Food Sci Technol. 2005;16(6–7):289–97.CrossRef
6.
7.
Inoue K, Yuasa H. Molecular basis for pharmacokinetics and pharmacodynamics of methotrexate in rheumatoid arthritis therapy. Drug Metab Pharmacokinet. 2014;29:12–9.CrossRefPubMed
8.
Lima A, Sousa H, Monteiro J, Azevedo R, Medeiros R, Seabra V. Genetic polymorphisms in lowdose methotrexate transporters: current relevance as methotrexate therapeutic outcome biomarkers. Pharmacogenomics. 2014;15(12):1611–35. doi:10.​2217/​pgs.​14.​116.CrossRefPubMed
9.
10.
van Ede AE, Laan RF, Blom HJ, De Abreu RA, van de Putte LB. Methotrexate in rheumatoid arthritis: an update with focus on mechanisms involved in toxicity. Semin Arthritis Rheum. 1998;27(5):277–92.CrossRefPubMed
11.
Lima A, Seabra V, Martins S, Coelho A, Araujo A, Medeiros R. Thymidylate synthase polymorphisms are associated to therapeutic outcome of advanced non-small cell lung cancer patients treated with platinum-based chemotherapy. Mol Biol Rep. 2014;41:3349–57.CrossRefPubMed
12.
13.
Frosst P, Blom HJ, Milos R, Goyette P, Sheppard CA, Matthews RG, et al. A candidate genetic risk factor for vascular disease: a commonmutation in methylenetetrahydrofolate reductase. NatGenet. 1995;10(1):111–3.
14.
van der Put NM, Gabreëls F, Stevens EM, Smeitink JA, Trijbels FJ, Eskes TK, et al. A second common mutation in the methylenetetrahydrofolate reductase gene: an additional risk factor for neural-tube defects? Am J Hum Genet. 1998;62(5):1044–51.CrossRefPubMedPubMedCentral
15.
Johnson WG, Stenroos ES, Spychala JR, Chatkupt S, Ming SX, Buyske S. New 19 bp deletion polymorphism in intron-1 of dihydrofolate reductase (DHFR): a risk factor for spina bifida acting in mothers during pregnancy? Am J Med Genet. 2004;124(4):339–45.CrossRef
16.
Horie N, Aiba H, Oguro K, Hojo H, Takishi K. Functional analysis and DNA polymorphism of the tandemly repeated sequences in the 5′-terminal regulatory region of the human gene for thymidylate synthase. Cell Struct Funct. 1995;20(3):191–7.CrossRefPubMed
17.
Paolo DI, Chu E. The role of thymidylate synthase as a molecular biomarker. Clin Cancer Res. 2004;10(2):411–2.CrossRef
18.
Leclerc D, Campeau E, Goyette P, Adjalla CE, Christensen B, Ross M, et al. Human methionine synthase: cDNA cloning and identification of mutations in patients of the cblG complementation group of folate/cobalamin disorders. Hum Mol Genet. 1996;5(12):1867–74.CrossRefPubMed
19.
Wilson A, Platt R, Wu Q, Leclerc D, Christensen B, Yang H, et al. A common variant in methionine synthase reductase combined with low cobalamin (vitamin B12) increases risk for spina bifida. Mol Genet Metab. 1999;67(4):317–23.CrossRefPubMed
20.
Gaughan DJ, Kluijtmans LA, Barbaux S, McMaster D, Young IS, Yarnell JW, et al. The methionine synthase reductase (MTRR) A66G polymorphism is a novel genetic determinant of plasma homocysteine concentrations. Atherosclerosis. 2001;157(2):451–6.CrossRefPubMed
21.
Ranganathan P, Culverhouse Marsh S, Mody A, Scott-Horton TJ, Brasington R, et al. Methotrexate gene polymorphisms and their effects on MTX toxicity in Caucasian and African American patients with rheumatoid arthritis. J Rheumatol. 2008;5(4):559–69.
22.
Brinker RR, Ranganathan P. Methotrexate pharmacogenetics in rheumatoid arthritis. Clin Exp Rheumatol. 2010;28(5 Suppl 61):S33–9.PubMed
23.
Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988;31(3):315–24.CrossRefPubMed
24.
Gustincich S, Manfioletti G, Del Sal G, Schneider C, Carninci P. A fast method for high quality genomic-DNA extraction from whole human blood. Biotechniques. 1991;11(3):300–2.
25.
Hanson NQ, Aras O, Yang F, Tsai MY. C677T and A1298C polymorphisms of the methylenetetrahydrofolate reductase gene: incidence and effect of combined genotypes on plasma fasting and post-methionine load homocysteine in vascular disease. Clin Chem. 2001;47(4):661–6.PubMed
26.
Matsuo K, Suzuki R, Hamajima N, Ogura M, Kagami Y, Taji H, et al. Association between polymorphisms of folate- and methionine-metabolizing enzymes and susceptibility to malignant lymphoma. Blood. 2001;97(10):3205–9.CrossRefPubMed
27.
Jacques PF, Bostom AG, Selhub J, Rich S, Ellison RC, Eckfeldt JH, et al. Effects of polymorphisms of methionine synthase and methionine synthase reductase on total plasma homocysteine in the NHLBI Family Heart Study. Atherosclerosis. 2003;166(1):49–55.CrossRefPubMed
28.
Hayashi H, Horino M, Tazoe Morishita M, Tsuboi S, Matsuyama T, Kosuge K, et al. Dihydrofolate reductase gene intronic 19-bp deletion polymorphisms in a Japanese population. Drug Metab Pharmacokinet. 2010;25(5):516–8.CrossRefPubMed
29.
Iacopetta B, Grieu F, Joseph D, Elsaleh H. A polymorphism in the enhancer region of the thymidylate synthase promoter influences the survival of colorectal cancer patients treated with 5- fluorouracil. Br J Cancer. 2001;85(6):827–30.CrossRefPubMedPubMedCentral
30.
Lima A, Bernardes M, Azevedo R, Monteiro J, Sousa H, Medeiros R, et al. SLC19A1, SLC46A1 and SLCO1B1 polymorphisms as predictors of methotrexate related toxicity in Portugueserheumatoid arthritis patients. Toxicol Sci. 2014;142(1):196–209. doi:10.​1093/​toxsci/​kfu162.CrossRefPubMed
31.
De Bakker PI, Yelensky R, Pe’er I, Gabriel SB, Daly MJ, Altshuler D. Efficiency and power in genetic association studies. Nat Genet. 2005;37(11):1217–23.CrossRefPubMed
32.
Fujimaki Chihiro, Hayashi Hideki, Tsuboi Seiji, Matsuyama Taiji, Kosuge Kazuhiro, Yamada Hiroshi, et al. Plasma total homocysteine level and methylenetetrahydrofolate reductase 677C>T genetic polymorphism in Japanese patients with rheumatoid arthritis. Biomarkers. 2009;14(1):49–54.CrossRefPubMed
33.
van Ede AE, Laan RF, Blom HJ, Huizinga TW, Haagsma CJ, Giesendorf BA, et al. The C677T mutation in the methylenetetrahydrofolate reductase gene: a genetic risk factor for methotrexate-related elevation of liver enzymes in rheumatoid arthritis patients. Arthritis Rheum. 2001;44(11):2525–30.CrossRefPubMed
34.
Ulrich CM, Yasui Y, Storb R, Schubert MM, Wagner JL, Bigler J, et al. Pharmacogenetics of methotrexate: toxicity among marrow transplantation patients varies with the methylenetetrahydrofolate reductase C677T polymorphism. Blood. 2001;98(1):231–4.CrossRefPubMed
35.
36.
37.
Kumagai K, Hiyama K, Oyama T, Maeda H, Kohno N. Polymorphisms in the thymidylate synthase and methylenetetrahydrofolate reductase genes and sensitivity to the low-dose methotrexate therapy in patients with rheumatoid arthritis. Int J Mol Med. 2003;11(5):593–600.PubMed
38.
Urano W, Taniguchi A, Yamanaka H, Tanaka E, Nakajima H, Matsuda Y, et al. Polymorphisms in the methylenetetrahydrofolate reductase gene were associated with both the efficacy and the toxicity of methotrexate used for the treatment of rheumatoid arthritis, as evidenced by single locus and haplotype analyses. Pharmacogenetics. 2002;12(3):183–90.CrossRefPubMed
39.
Aggarwal P, Naik S, Mishra KP, Aggarwal A, Misra R. Correlation between methotrexate efficacy and toxicity with C677T polymorphism of the methylenetetrahydrofolate gene in rheumatoid arthritis patients on folate supplementation. Indian J Med Res. 2006;124(5):521–6.PubMed
40.
Kurzawski M, Pawlik A, Safranow K, Herczynska M, Drozdzik M. 677C>T and 1298A>CMTHFR polymorphisms affect methotrexate treatment outcome in rheumatoid arthritis. Pharmacogenomics. 2007;8(11):1551–9.CrossRefPubMed
41.
Kim SK, Jun JB, El-Sohemy A, Bae SC. Cost-effectiveness analysis of MTHFR polymorphism screening by polymerase chain reaction in Korean patients with rheumatoid arthritis receiving methotrexate. J Rheumatol. 2006;33(7):1266–74.PubMed
42.
Wessels JA, de Vries-Bouwstra JK, Heijmans BT, Slagboom PE, Goekoop-Ruiterman YP, Allaart CF, et al. Efficacy and toxicity of methotrexate in early rheumatoid arthritis are associated with single-nucleotide polymorphisms in genes coding for folate pathway enzymes. Arthritis Rheum. 2006;54(4):1087–95.CrossRefPubMed
43.
Berkun Y, Levartovsky D, Rubinow A, Orbach H, Aamar S, Grenader T, et al. Methotrexate related adverse effects in patients with rheumatoid arthritis are associated with the A1298C polymorphism of the MTHFR gene. Ann Rheum Dis. 2004;63(10):1227–31.CrossRefPubMedPubMedCentral
44.
Taniguchi A, Urano W, Tanaka E, Furihata S, Kamitsuji S, Inoue E, et al. Validation of the associations between single nucleotide polymorphisms or haplotypes and responses to disease-modifying antirheumatic drugs in patients with rheumatoid arthritis: a proposal for prospective pharmacogenomic study in clinical practice. Pharmacogenet Genomics. 2007;17(6):383–90.CrossRefPubMed
45.
46.
Marsh S, Ameyaw MM, Githang’a J, Indalo A, Ofori-Adjei D, Mcleod HL. Novel thymidylate synthase enhancer region alleles in African populations. Hum Mutat. 2000;16(6):528.CrossRefPubMed
47.
Luo HR, Lü XM, Yao YG, Horie N, Takeishi K, Jorde LB, et al. Length polymorphism of thymidylate synthase regulatory region in Chinese populations and evolution of the novel alleles. Biochem Genet. 2002;40(1–2):41–51.CrossRefPubMed
48.
Weisman MH, Furst DE, Park GS, Kremer JM, Smith KM, Wallace DJ, et al. Risk genotypes in folate-dependent enzymes and their association with methotrexate-related side effects in rheumatoid arthritis. Arthritis Rheum. 2006;54(2):607–12.CrossRefPubMed
49.
Bohanec Grabar P, Logar D, Lestan B, Dolzan V. Genetic determinants of methotrexate toxicity in rheumatoid arthritis patients: a study of polymorphisms affecting methotrexate transport and folate metabolism. Eur J Clin Pharmacol. 2008;64(11):1057–68. doi:10.​1007/​s00228-008-0521-7.CrossRefPubMed
50.
Dervieux T, Greenstein N, Kremer J. Pharmacogenomic and metabolic biomarkers in the folate pathway and their association with methotrexate effects during dosage escalation in rheumatoid arthritis. Arthritis Rheum. 2006;54(10):3095–103.CrossRefPubMed
51.
Hughes LB, Beasley TM, Patel H, Tiwari HK, Morgan SL, Baggott JE, et al. Racial or ethnic differences in allele frequencies of single-nucleotide polymorphisms in the methylenetetrahydrofolate reductase gene and their influence on response to methotrexate in rheumatoid Arthritis. Ann Rheum Dis. 2006;65(9):1213–8.CrossRefPubMedPubMedCentral
52.
53.
Uchida K, Hayashi K, Kawakami K, Schneider S, Yochim JM, Kuramochi H, et al. Loss of heterozygosity at the thymidylate synthase (TS) locus on chromosome 18 affects tumor response and survival in individuals heterozygous for a 28-bp polymorphism in the TS gene. Clin Cancer Res. 2004;10(2):433–9.CrossRefPubMed
54.
Gellekink H, Blom HJ, van der LindenI J, den Heijer M. Molecular genetic analysis of the human dihydrofolate reductase gene: relation with plasma total homocysteine, serum and red blood cell folate levels. Eur J Hum Genet. 2007;15(1):103–9.CrossRefPubMed
55.
Johnson WG, Stenroos ES, Spychala JR, Chatkupt S, Ming SX, Buyske S. New 19 bp deletion polymorphism in intron-1 of dihydrofolate reductase (DHFR): a risk factor for spina bifida acting in mothers during pregnancy? Am J Med Genet. 2004;124A(4):339–45.CrossRefPubMed
56.
Salliot C, van der Heijde D. Long-term safety of methotrexate monotherapy in patients with rheumatoid arthritis: a systematic literature research. Ann Rheum Dis. 2009;68:1100–4.CrossRefPubMed
Metadata
Title
Genetic Determinants of Methotrexate Toxicity in Tunisian Patients with Rheumatoid Arthritis: A Study of Polymorphisms Involved in the MTX Metabolic Pathway
Authors
Souhir Chaabane
Sameh Marzouk
Rim Akrout
Mariem Ben Hamad
Yosser Achour
Ahmed Rebai
Leila Keskes
Hela Fourati
Zouhir Bahloul
Abdellatif Maalej
Publication date
01-08-2016
Publisher
Springer International Publishing
Published in
European Journal of Drug Metabolism and Pharmacokinetics / Issue 4/2016
Print ISSN: 0378-7966
Electronic ISSN: 2107-0180
DOI
https://doi.org/10.1007/s13318-015-0288-z

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