Abstract
Cellular drug resistance remains a major concern in cancer therapy and usually results from increased expression of ABC drug transporters. Imatinib mesylate (IM), a competitive inhibitor of BCR/ABL1 tyrosine kinase activity, is the current standard therapy for chronic myeloid leukaemia (CML) which is caused by the BCR/ABL1 gene fusion encoding a constitutively active tyrosine kinase. However, up to 33 % of CML patients do not respond to therapy either initially or due to acquired resistance. Usually, IM resistance is due to the presence of BCR-ABL1 mutations but in many cases resistance is far from being completely understood or from being satisfactorily addressed from a therapeutic standpoint. Although second- and third-generation TKIs (e.g., dasatinib (DA), nilotinib, and bosutinib) were developed to override this phenomenon, resistance remains an unsolved problem. Above all, as more patients are treated with TKIs, more cases of resistance are expected and the discovery of biomarkers of resistance acquires a crucial clinical significance.
We established a valuable in vitro experimental system that mimics the acquired resistance in the absence of mutations. It was developed by the continuous exposure of K562, a human CML-derived cell line expressing BCR-ABL gene, to increasing concentrations of IM and DA (over 36 and 24 weeks, respectively) allowing us to obtain several cell lines with different resistance levels, and therefore to evaluate drug transporters’ role in the dynamic cellular responses allied with resistance evolution. The development of such cell models is fundamental to understand the role of drug transporters in resistance since the majority of previous studies were performed on cell lines engineered to over-express a single transporter.
Drug transporters were overexpressed in the majority of resistant cell lines and cell lines from all levels of resistance had increased expression of more than one drug transporter. However, the transporters that attain higher mRNA overexpression (e.g., ABCB1 and ABCG2) did not substantiate a linear relation with the level of resistance. Also, variation in expression of these genes occurs over time of exposure to the same concentration of IM while maintaining resistance, suggesting that resistance mechanisms could vary dynamically in patients as disease progresses. Indeed, we observed that while responding patients demonstrated stable transporters’ expression signatures in consecutive samples, in IM-resistant patients they vary significantly over time, advising caution when comparing single-point samples from responsive and resistant patients.
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References
Rodrigues AS, Dinis J, Gromicho M et al (2012) Genomics and cancer drug resistance. Curr Pharm Biotechnol 13:651–673
Lage H (2008) An overview of cancer multidrug resistance: a still unsolved problem. Cell Mol Life Sci 65:3145–3167
Gillet J-P, Efferth T, Remacle J (2007) Chemotherapy-induced resistance by ATP-binding cassette transporter genes. Biochim Biophys Acta 1775:237–262
Scheffer GL, Kool M, Heijn M et al (2000) Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies. Cancer Res 60:5269–5277
Bhamidipati PK, Kantarjian H, Cortes J et al (2013) Management of imatinib-resistant patients with chronic myeloid leukemia. Ther Adv Hematol 4:103–117
Jabbour E, Cortes J, Kantarjian H (2011) Long-term outcomes in the second-line treatment of chronic myeloid leukemia: a review of tyrosine kinase inhibitors. Cancer 117:897–906
Hochhaus A, O’Brien SG, Guilhot F et al (2009) Six-year follow-up of patients receiving imatinib for the first-line treatment of chronic myeloid leukemia. Leukemia 23:1054–1061
de Lavallade H, Apperley JF, Khorashad JS et al (2008) Imatinib for newly diagnosed patients with chronic myeloid leukemia: incidence of sustained responses in an intention-to-treat analysis. J Clin Oncol 26:3358–3363
Marin D, Milojkovic D, Olavarria E et al (2008) European LeukemiaNet criteria for failure or suboptimal response reliably identify patients with CML in early chronic phase treated with imatinib whose eventual outcome is poor. Blood 112:4437–4444
Soverini S, Colarossi S, Gnani A et al (2006) Contribution of ABL kinase domain mutations to imatinib resistance in different subsets of Philadelphia-positive patients: by the GIMEMA Working Party on Chronic Myeloid Leukemia. Clin Cancer Res 12:7374–7379
Larson RA, Druker BJ, Guilhot F et al (2008) Imatinib pharmacokinetics and its correlation with response and safety in chronic-phase chronic myeloid leukemia: a subanalysis of the IRIS study. Blood 111:4022–4028
Picard S, Titier K, Etienne G et al (2007) Trough imatinib plasma levels are associated with both cytogenetic and molecular responses to standard-dose imatinib in chronic myeloid leukemia. Blood 109:3496–3499
White DL, Saunders VA, Dang P et al (2007) Most CML patients who have a suboptimal response to imatinib have low OCT-1 activity: higher doses of imatinib may overcome the negative impact of low OCT-1 activity. Blood 110:4064–4072
Clark RE, Davies A, Pirmohamed M et al (2008) Pharmacologic markers and predictors of responses to imatinib therapy in patients with chronic myeloid leukemia. Leuk Lymphoma 49:639–642
Brendel C, Scharenberg C, Dohse M et al (2007) Imatinib mesylate and nilotinib (AMN107) exhibit high-affinity interaction with ABCG2 on primitive hematopoietic stem cells. Leukemia 21:1267–1275
Burger H, van Tol H, Boersma AWM et al (2004) Imatinib mesylate (STI571) is a substrate for the breast cancer resistance protein (BCRP)/ABCG2 drug pump. Blood 104:2940–2942
Houghton PJ (2004) Imatinib mesylate is a potent inhibitor of the ABCG2 (BCRP) transporter and reverses resistance to topotecan and SN-38 in vitro. Cancer Res 64:2333–2337
Nakanishi T, Shiozawa K, Hassel BA et al (2006) Complex interaction of BCRP/ABCG2 and imatinib in BCR-ABL-expressing cells: BCRP-mediated resistance to imatinib is attenuated by imatinib-induced reduction of BCRP expression. Blood 108:678–684
Gromicho M, Dinis J, Magalhães M et al (2011) Development of imatinib and dasatinib resistance: dynamics of expression of drug transporters ABCB1, ABCC1, ABCG2, MVP, and SLC22A1. Leuk Lymphoma 52:1980–1990
Dinis J, Silva V, Gromicho M et al (2012) DNA damage response in imatinib resistant chronic myeloid leukemia K562 cells. Leuk Lymphoma 53:2004–2014
Gromicho M, Magalhães M, Torres F et al (2013) Instability of mRNA expression signatures of drug transporters in chronic myeloid leukemia patients resistant to imatinib. Oncol Rep 29:741–750
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408
Branford S, Hughes T (2006) Detection of BCR-ABL mutations and resistance to imatinib mesylate. Methods Mol Med 125:93–106
Acknowledgments
This work was supported by grant PEst-OE/SAU/UI0009/2014 from Fundação de Ciência e Tecnologia (FCT).
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Gromicho, M., Rueff, J., Rodrigues, A.S. (2016). Dynamics of Expression of Drug Transporters: Methods for Appraisal. In: Rueff, J., Rodrigues, A. (eds) Cancer Drug Resistance. Methods in Molecular Biology, vol 1395. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3347-1_6
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DOI: https://doi.org/10.1007/978-1-4939-3347-1_6
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