Molecular genetics in the chronic myeloid leukemia diagnostics and therapy

Czech version

Authors: K. Machová Poláková; K. Zemanová; M. Součková; A. Broučková; H. Klamová
Authors‘ workplace: Ústav hematologie a krevní transfuze Praha, ředitel prof. MUDr. Marek Trněný, CSc.
Published in: Vnitř Lék 2012; 58(Suppl 2): 38-45
Category:

Overview

This overview discusses an importance of molecular diagnostics of chronic myeloid leukemia, molecular monitoring of treatment efficacy, residual disease and resistance to therapy and the role of the National reference laboratory ÚHKT in these issues. The qualitative detection based on the multiplex reverse transcriptase PCR confirms the presence of mRNA of the fusion gene BCR-ABL in the examined sample, thus a diagnosis of chronic myeloid leukemia. Characterization of the type of BCR-ABL rearrangement is also important for the subsequent monitoring based on the quantification of BCR-ABL transcripts. The quantitative determination of BCR-ABL transcripts at regular intervals monitors the kinetics of the disease during the treatment at the molecular level. A milestone in the successful management of chronic myeloid leukemia by tyrosine kinase inhibitors is the achievement of the major molecular response, which corresponds to the levels of BCR-ABL transcripts ≤ 0.1%^IS. Thus, a fundamental aim is national and international harmonization of BCR-ABL transcripts quantification among laboratories. Currently, definition and monitoring of the complete molecular remission or deep molecular response rates is currently intensively studied worldwide, because of a higher number of patients achieving complete molecular response under 2nd generation TKI. The most studied and proved mechanism of the resistance to TKI therapy are mutations in the kinase domain of BCR-ABL. Sanger sequencing is the gold standard for the routine detection and characterization of BCR-ABL mutations. At present, mutation studies starting with using of the second-generation sequencing, which is expected to help in understanding of mutation development and clonal evolution under the pressure of TK inhibitors and the potential impact of this extremely sensitive technology for the prognosis.

Key words:
BCR-ABL – chronic myeloid leukemia – CMR – tyrosine kinase inhibitors – quantification – multiplex RT-PCR – mutations – real-time RT-PCR – resistance – Sanger sequencing – next generation sequencing – standardization

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