Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

A comprehensive functional and clinical analysis of ABCC2 and its impact on treatment response to carbamazepine

The Pharmacogenomics Journal volume 14, pages 481–487 (2014)Cite this article

Subjects

Abstract

At the blood–brain barrier, overexpression of the drug efflux transporter ABCC2 (also known as MRP2) has been proposed as a mechanism for impaired carbamazepine (CBZ) treatment response in epilepsy. However, investigation of the impact of ABCC2 polymorphisms on CBZ treatment efficacy has produced conflicting and inconclusive results. A series of in vitro cell efflux and plasma membrane vesicle uptake assays were undertaken to investigate whether CBZ was an ABCC2 substrate. In addition, the effect of three common ABCC2 polymorphisms, −24C>T, c.1249G>A and c.3972C>T, on the efficacy of CBZ in epilepsy (assessed using the clinical end points time to first seizure and time to 12-month remission from the SANAD (Standard and New Antiepileptic Drugs) trial) was determined. CBZ was found not to be a substrate for human ABCC2 in vitro. Clinically, no significant association was observed for the ABCC2 genetic variants and CBZ treatment outcomes. This comprehensive analysis does not support a role for ABCC2 in CBZ treatment efficacy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. WHO (2009). Epilepsy fact sheet no. 999; http://www.who.int/mediacentre/factsheets/fs999/en/index.html.

  2. NICE (2012). The epilepsies: the diagnosis and management of the epilepsies in adults and children in primary and secondary care (NICE clinical guideline 137); http://www.nice.org.uk/.

  3. Kwan P, Arzimanoglou A, Berg AT, Brodie MJ, Allen HW, Mathern G et al. Definition of drug resistant epilepsy: consensus proposal by the ad hoc Task Force of the ILAE Commission on Therapeutic Strategies. Epilepsia 2010; 51: 1069–1077.

    Article  CAS  PubMed  Google Scholar 

  4. Aronica E, Gorter JA, Ramkema M, Redeker S, Ozbas-Gerceker F, van Vliet EA et al. Expression and cellular distribution of multidrug resistance-related proteins in the hippocampus of patients with mesial temporal lobe epilepsy. Epilepsia 2004; 45: 441–451.

    Article  CAS  PubMed  Google Scholar 

  5. Dombrowski SM, Desai SY, Marroni M, Cucullo L, Goodrich K, Bingaman W et al. Overexpression of multiple drug resistance genes in endothelial cells from patients with refractory epilepsy. Epilepsia 2001; 42: 1501–1506.

    Article  CAS  PubMed  Google Scholar 

  6. Kubota H, Ishihara H, Langmann T, Schmitz G, Stieger B, Wieser HG et al. Distribution and functional activity of P-glycoprotein and multidrug resistance-associated proteins in human brain microvascular endothelial cells in hippocampal sclerosis. Epilepsy Res 2006; 68: 213–228.

    Article  CAS  PubMed  Google Scholar 

  7. Sisodiya SM, Lin WR, Harding BN, Squier MV, Thom M . Drug resistance in epilepsy: expression of drug resistance proteins in common causes of refractory epilepsy. Brain 2002; 125 (Pt 1): 22–31.

    Article  CAS  PubMed  Google Scholar 

  8. Tishler DM, Weinberg KI, Hinton DR, Barbaro N, Annett GM, Raffel C . MDR1 gene expression in brain of patients with medically intractable epilepsy. Epilepsia 1995; 36: 1–6.

    Article  CAS  PubMed  Google Scholar 

  9. Baltes S, Gastens AM, Fedrowitz M, Potschka H, Kaever V, Loscher W . Differences in the transport of the antiepileptic drugs phenytoin, levetiracetam and carbamazepine by human and mouse P-glycoprotein. Neuropharmacology 2007; 52: 333–346.

    Article  CAS  PubMed  Google Scholar 

  10. Cerveny L, Pavek P, Malakova J, Staud F, Fendrich Z . Lack of interactions between breast cancer resistance protein (bcrp/abcg2) and selected antiepileptic agents. Epilepsia 2006; 47: 461–468.

    Article  PubMed  Google Scholar 

  11. Crowe A, Teoh YK . Limited P-glycoprotein mediated efflux for anti-epileptic drugs. J Drug Target 2006; 14: 291–300.

    Article  CAS  PubMed  Google Scholar 

  12. Dickens D, Yusof SR, Abbott NJ, Weksler B, Romero IA, Couraud PO et al. A multi-system approach assessing the interaction of anticonvulsants with P-gp. PloS One 2013; 8: e64854.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Luna-Tortos C, Fedrowitz M, Loscher W . Several major antiepileptic drugs are substrates for human P-glycoprotein. Neuropharmacology 2008; 55: 1364–1375.

    Article  CAS  PubMed  Google Scholar 

  14. Luna-Tortos C, Fedrowitz M, Loscher W . Evaluation of transport of common antiepileptic drugs by human multidrug resistance-associated proteins (MRP1, 2 and 5) that are overexpressed in pharmacoresistant epilepsy. Neuropharmacology 2010; 58: 1019–1032.

    Article  CAS  PubMed  Google Scholar 

  15. Owen A, Pirmohamed M, Tettey JN, Morgan P, Chadwick D, Park BK . Carbamazepine is not a substrate for P-glycoprotein. Br J Clin Pharmacol 2001; 51: 345–349.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ufer M, von SC, Muhle H, Haenisch S, Remmler C, Majed A et al. Impact of ABCC2 genotype on antiepileptic drug response in Caucasian patients with childhood epilepsy. Pharmacogenet Genomics 2011; 21: 624–630.

    Article  CAS  PubMed  Google Scholar 

  17. Kim WJ, Lee JH, Yi J, Cho YJ, Heo K, Lee SH et al. A nonsynonymous variation in MRP2/ABCC2 is associated with neurological adverse drug reactions of carbamazepine in patients with epilepsy. Pharmacogenet Genomics 2010; 20: 249–256.

    CAS  PubMed  Google Scholar 

  18. Grover S, Gourie-Devi M, Bala K, Sharma S, Kukreti R . Genetic association analysis of transporters identifies ABCC2 loci for seizure control in women with epilepsy on first-line antiepileptic drugs. Pharmacogenet Genomics 2012; 22: 447–465.

    Article  CAS  PubMed  Google Scholar 

  19. Qu J, Zhou BT, Yin JY, Xu XJ, Zhao YC, Lei GH et al. ABCC2 Polymorphisms and Haplotype are Associated with Drug Resistance in Chinese Epileptic Patients. CNS Neurosci Ther 2012; 18: 647–651.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ufer M, Mosyagin I, Muhle H, Jacobsen T, Haenisch S, Hasler R et al. Non-response to antiepileptic pharmacotherapy is associated with the ABCC2 -24C>T polymorphism in young and adult patients with epilepsy. Pharmacogenet Genomics 2009; 19: 353–362.

    Article  CAS  PubMed  Google Scholar 

  21. Grover S, Kukreti R . A systematic review and meta-analysis of the role of ABCC2 variants on drug response in patients with epilepsy. Epilepsia 2013; 54: 936–945.

    Article  CAS  PubMed  Google Scholar 

  22. Hilger E, Reinthaler EM, Stogmann E, Hotzy C, Pataraia E, Baumgartner C et al. Lack of association between ABCC2 gene variants and treatment response in epilepsy. Pharmacogenomics 2012; 13: 185–190.

    Article  CAS  PubMed  Google Scholar 

  23. Kim DW, Lee SK, Chu K, Jang IJ, Yu KS, Cho JY et al. Lack of association between ABCB1, ABCG2, and ABCC2 genetic polymorphisms and multidrug resistance in partial epilepsy. Epilepsy Res 2009; 84: 86–90.

    Article  CAS  PubMed  Google Scholar 

  24. Kwan P, Wong V, Ng PW, Lui CH, Sin NC, Wong KS et al. Gene-wide tagging study of the association between ABCC2, ABCC5 and ABCG2 genetic polymorphisms and multidrug resistance in epilepsy. Pharmacogenomics 2011; 12: 319–325.

    Article  CAS  PubMed  Google Scholar 

  25. Seo T, Ishitsu T, Oniki K, Abe T, Shuto T, Nakagawa K . ABCC2 haplotype is not associated with drug-resistant epilepsy. J Pharm Pharmacol 2008; 60: 631–635.

    Article  CAS  PubMed  Google Scholar 

  26. Sporis D, Bozina N, Basic S, Lovric M, Babic T, Susak I et al. Lack of association between polymorphism in ABCC2 gene and response to antiepileptic drug treatment in Croatian patients with epilepsy. Collegium Antropologicum 2013; 37: 41–45.

    CAS  PubMed  Google Scholar 

  27. Marson AG, Al-Kharusi AM, Alwaidh M, Appleton R, Baker GA, Chadwick DW et al. The SANAD study of effectiveness of carbamazepine, gabapentin, lamotrigine, oxcarbazepine, or topiramate for treatment of partial epilepsy: an unblinded randomised controlled trial. Lancet 2007; 369: 1000–1015.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Brough R, Papanastasiou AM, Porter AC . Stringent and reproducible tetracycline-regulated transgene expression by site-specific insertion at chromosomal loci with pre-characterised induction characteristics. BMC Mol Biol 2007; 8: 30.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Arlanov R, Porter A, Strand D, Brough R, Karpova D, Kerb R et al. Functional characterization of protein variants of the human multidrug transporter ABCC2 by a novel targeted expression system in fibrosarcoma cells. Hum Mutation 2012; 33: 750–762.

    Article  CAS  Google Scholar 

  30. Evers R, Kool M, van Deemter L, Janssen H, Calafat J, Oomen LC et al. Drug export activity of the human canalicular multispecific organic anion transporter in polarized kidney MDCK cells expressing cMOAT (MRP2) cDNA. J Clin Invest 1998; 101: 1310–1319.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Pratt S, Chen V, Perry WI 3rd, Starling JJ, Dantzig AH . Kinetic validation of the use of carboxydichlorofluorescein as a drug surrogate for MRP5-mediated transport. Eur J Pharm Sci 2006; 27: 524–532.

    Article  CAS  PubMed  Google Scholar 

  32. Roelofsen H, Vos TA, Schippers IJ, Kuipers F, Koning H, Moshage H et al. Increased levels of the multidrug resistance protein in lateral membranes of proliferating hepatocyte-derived cells. Gastroenterology 1997; 112: 511–521.

    Article  CAS  PubMed  Google Scholar 

  33. Keppler D, Jedlitschky G, Leier I . Transport function and substrate specificity of multidrug resistance protein. Methods Enzymol 1998; 292: 607–616.

    Article  CAS  PubMed  Google Scholar 

  34. Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD et al. Measurement of protein using bicinchoninic acid. Anal Biochem 1985; 150: 76–85.

    Article  CAS  PubMed  Google Scholar 

  35. Leier I, Jedlitschky G, Buchholz U, Keppler D . Characterization of the ATP-dependent leukotriene C4 export carrier in mastocytoma cells. Eur J Biochem 1994; 220: 599–606.

    Article  CAS  PubMed  Google Scholar 

  36. Speed D, Hoggart C, Petrovski S, Tachmazidou I, Coffey A, Jorgensen A et al. A genome-wide association study and biological pathway analysis of epilepsy prognosis in a prospective cohort of newly treated epilepsy. Hum Mol Genet 2013; 23: 247–258.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Kleinbaum DG, Klein M . Survival Analysis [Electronic Book]: A Self-Learning Text. Springer: New York, NY, c2012 3rd edn 2012.

    Book  Google Scholar 

  38. Colombo F, Poirier H, Rioux N, Montecillo MA, Duan J, Ribadeneira MD . A membrane vesicle-based assay to enable prediction of human biliary excretion. Xenobiotica 2013; 43: 915–919.

    Article  CAS  PubMed  Google Scholar 

  39. Heredi-Szabo K, Kis E, Molnar E, Gyorfi A, Krajcsi P . Characterization of 5(6)-carboxy-2,'7'-dichlorofluorescein transport by MRP2 and utilization of this substrate as a fluorescent surrogate for LTC4. J Biomol Screen 2008; 13: 295–301.

    Article  CAS  PubMed  Google Scholar 

  40. Kidron H, Wissel G, Manevski N, Hakli M, Ketola RA, Finel M et al. Impact of probe compound in MRP2 vesicular transport assays. Eur J Pharm Sci 2012; 46: 100–105.

    Article  CAS  PubMed  Google Scholar 

  41. Lechner C, Reichel V, Moenning U, Reichel A, Fricker G . Development of a fluorescence-based assay for drug interactions with human multidrug resistance related protein (MRP2; ABCC2) in MDCKII-MRP2 membrane vesicles. Eur J Pharm Biopharm 2010; 75: 284–290.

    Article  CAS  PubMed  Google Scholar 

  42. Lehmann T, Kohler C, Weidauer E, Taege C, Foth H . Expression of MRP1 and related transporters in human lung cells in culture. Toxicology 2001; 167: 59–72.

    Article  CAS  PubMed  Google Scholar 

  43. Chen ZS, Hopper-Borge E, Belinsky MG, Shchaveleva I, Kotova E, Kruh GD . Characterization of the transport properties of human multidrug resistance protein 7 (MRP7, ABCC10). Mol Pharmacol 2003; 63: 351–358.

    Article  CAS  PubMed  Google Scholar 

  44. Rius M, Hummel-Eisenbeiss J, Keppler D . ATP-dependent transport of leukotrienes B4 and C4 by the multidrug resistance protein ABCC4 (MRP4). J Pharmacol Exp Ther 2008; 324: 86–94.

    Article  CAS  PubMed  Google Scholar 

  45. Bodo A, Bakos E, Szeri F, Varadi A, Sarkadi B . Differential modulation of the human liver conjugate transporters MRP2 and MRP3 by bile acids and organic anions. J Biol Chem 2003; 278: 23529–23537.

    Article  CAS  PubMed  Google Scholar 

  46. Zelcer N, Huisman MT, Reid G, Wielinga P, Breedveld P, Kuil A et al. Evidence for two interacting ligand binding sites in human multidrug resistance protein 2 (ATP binding cassette C2). J Biol Chem 2003; 278: 23538–23544.

    Article  CAS  PubMed  Google Scholar 

  47. Pedersen JM, Matsson P, Bergstrom CA, Norinder U, Hoogstraate J, Artursson P . Prediction and identification of drug interactions with the human ATP-binding cassette transporter multidrug-resistance associated protein 2 (MRP2; ABCC2). J Med Chem 2008; 51: 3275–3287.

    Article  CAS  PubMed  Google Scholar 

  48. Jemnitz K, Heredi-Szabo K, Janossy J, Ioja E, Vereczkey L, Krajcsi P . ABCC2/Abcc2: a multispecific transporter with dominant excretory functions. Drug Metabol Rev 2010; 42: 402–436.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thankfully acknowledge excellent technical assistance from Sabine Rekersbrink. SR is an EU Marie-Curie Early Stage Researcher within the ‘Fighting Drug Failure’ network and would like to thank the European Commission for funding (grant agreement number 238132). MRJ received a proportion of funding from the Department of Health´s NIHR Biomedical Research Centres funding scheme. MP is a NIHR Senior Investigator and would also like to thank the NHS Chair of Pharmacogenetics programme, MRC Centre for Drug Safety Science and Wolfson Foundation for their support. TL, RA and MS were supported by the Robert Bosch Stiftung, Stuttgart, Germany, RA and TL were supported by the German Research Foundation (DFG, grant number LA 2406/2-1) and MS was supported by the Federal Ministry for Education and Research (BMBF, Germany; grant number 03 IS 2061C) and the IZEPHA project 18-0-0 (University of Tuebingen, Germany).

Author information

Authors and Affiliations

  1. Department of Molecular and Clinical Pharmacology, The Wolfson Centre for Personalised Medicine, Institute of Translational Medicine, University of Liverpool, Liverpool, UK

    S Rädisch, D Dickens, A G Marson & M Pirmohamed

  2. Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, Germany

    T Lang, R Arlanov & M Schwab

  3. Department of Biostatistics and Department of Clinical Infection, Microbiology and Immunology, Institute of Translational Medicine, University of Liverpool, Liverpool, UK

    L Bonnett

  4. Department of Medicine, Division of Brain Science, Imperial College London, London, UK

    M R Johnson

  5. Department of Clinical Pharmacology, Institute of Experimental and Clinical Pharmacology and Toxicology, University Hospital Tübingen, Tübingen, Germany

    M Schwab

Authors
  1. S Rädisch
    View author publications

    Search author on:PubMed Google Scholar

  2. D Dickens
    View author publications

    Search author on:PubMed Google Scholar

  3. T Lang
    View author publications

    Search author on:PubMed Google Scholar

  4. L Bonnett
    View author publications

    Search author on:PubMed Google Scholar

  5. R Arlanov
    View author publications

    Search author on:PubMed Google Scholar

  6. M R Johnson
    View author publications

    Search author on:PubMed Google Scholar

  7. M Schwab
    View author publications

    Search author on:PubMed Google Scholar

  8. A G Marson
    View author publications

    Search author on:PubMed Google Scholar

  9. M Pirmohamed
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to M Pirmohamed.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the The Pharmacogenomics Journal website

Supplementary information

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rädisch, S., Dickens, D., Lang, T. et al. A comprehensive functional and clinical analysis of ABCC2 and its impact on treatment response to carbamazepine. Pharmacogenomics J 14, 481–487 (2014). https://doi.org/10.1038/tpj.2014.5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/tpj.2014.5

Keywords

This article is cited by

Search

Advanced search

Quick links