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Do SNPs in folate pharmacokinetic pathway alter levels of intracellular methotrexate polyglutamates and affect response? A prospective study in Indian patients

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Abstract

This study investigated the impact of seven polymorphisms in genes of folate transport and (de)glutamation pathway on methotrexate polyglutamate levels and response in patients with rheumatoid arthritis. This prospective study included patients with rheumatoid arthritis. They were treated with methotrexate (up to 25 mg per week) for 24 weeks and categorized by EULAR response criteria into responders (good and moderate) and non-responders. Using real-time Taqman discrimination assay, SNPs were genotyped—rs1045642 (ABCB1 3435C>T), rs1128503 (ABCB1 1236C>T), rs10106 (FPGS 1994A>G), rs1544105 (FPGS G>A), rs11545078 (GGH 452C>T), rs3758149 (GGH -401C>T), and rs1051266 (RFC1 80G>A). RBC methotrexate polyglutamate1–5(MTX-glu1–5) levels were determined at 4, 8, 16, and 24 weeks using by reverse phase HPLC using C-18 column followed by post column photo-oxidation. This study included 117 patients with rheumatoid arthritis (M:F = 14:103). The mean dose of methotrexate at 24 weeks was 22.0 ± 4.0 mg, with data on DAS28(3) at 24 weeks available in 96 patients—61 responders and 35 non-responders. Minor allele of GGH 452C>T had an association with non-response (odds ratio 2.9, 95% CI 1.4–5.6) and assuming the dominance of C, the recessive genetic model found GGH 452C>T CC genotype (odds ratio 9.5, 95% CI 1.2 to 76.0) was significantly associated with response. However, there was no difference in MTX-glu1–5 levels among the various genotypes of this SNP (p = 0.9). Other SNPs were neither associated with response nor with alteration in methotrexate polyglutamate levels. On logistic regression, GGH 452C>T CC genotype and DAS28(3) at baseline were independent predictors of response. GGH 452C>T CC genotype was associated with response to methotrexate. None of the SNPs affected MTX-glu1–5levels.

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Acknowledgements

Methotrexate tablets were received as an educational grant from Zydus Activa (Zydus Cadilla Inc., Ahmedabad, India). We acknowledge the efforts of Mrs. Nidhi Gupta and Mr. Mohinder Kumar for sample collection. We also acknowledge the all technical staff of central sophisticated cell of PGIMER for the use of instruments.

Funding

This work was funded by a grant from the Asia Pacific League of Associations for Rheumatology (APLAR) (gene polymorphisms) and from the Department of Biotechnology, Government of India [Grant # BT/PR4608/MED/30/800/2012] (methotrexate polyglutamate levels). Mr. Amit Sandhu was supported by ICMR through its Junior Research Fellowship scheme. Mr. Shabeer Ahmad was supported through a Junior Research Fellowship from the Department of Science and Technology (DST), Government of India.

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Authors and Affiliations

  1. Department of Internal Medicine (Rheumatology Unit), Postgraduate Institute of Medical Education and Research, Chandigarh, 160012, India

    Amit Sandhu, Shabeer Ahmad & Varun Dhir

  2. Department of Biochemistry, Panjab University, Chandigarh, 160014, India

    Amit Sandhu & Archana Bhatnagar

  3. Department of Biochemistry, Government Medical College and Hospital Sector 32, Chandigarh, India

    Jasbinder Kaur

  4. Department of Experimental Medicine and Biotechnology, Postgraduate Institute of Medical Education and Research, Chandigarh, 160012, India

    Veena Dhawan

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  1. Amit Sandhu
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  2. Shabeer Ahmad
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  3. Jasbinder Kaur
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  4. Archana Bhatnagar
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  5. Veena Dhawan
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  6. Varun Dhir
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Contributions

Planning study: VDhir, AS; collecting data: AS, VDhir; laboratory work: AS, VDhir, VDhaw, JK, AB; analysis and writing: AS, VDhir, VDhaw, JK, AB.

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Correspondence to Varun Dhir.

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Ethics approval and consent to participate

This study was approved by the institutional ethics committee of Post Graduate Institute of Medical Education and Research. All patients gave written informed consent.

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Sandhu, A., Ahmad, S., Kaur, J. et al. Do SNPs in folate pharmacokinetic pathway alter levels of intracellular methotrexate polyglutamates and affect response? A prospective study in Indian patients. Clin Rheumatol 37, 3221–3228 (2018). https://doi.org/10.1007/s10067-018-4206-z

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  • DOI: https://doi.org/10.1007/s10067-018-4206-z

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