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Leukaemias with BCR/ABL fusion gene.
Hovorková, Lenka ; Zuna, Jan (advisor) ; Zemanová, Karla (referee)
Philadelphia (Ph) chromosome, as a result of reciprocal translocation, is in majority of cases connected to two types of leukaemia - chronic myelogenous (CML) and acute lymphoblastic (ALL). The translocation occurs within large intronic sequences of BCR and ABL genes. The breakpoints are specific for individual patient and may be used as a target for monitoring of leukemic burden (MRD, minimal residual disease) during the treatment. In general, MRD is an important prognostic factor, which influences the treatment intensity. Two standardized methods are currently used for its monitoring. The first one is based on the detection of clonal specific Immunoglobulin and/or T-cell receptor genes rearrangements (and thus cannot be used for CML cases) at the DNA level, the second one utilizes detection of the BCR/ABL fusion gene at the mRNA level. Our aim was to optimize and standardize the process to find individual patient breakpoints on Ph chromosome and to use it for MRD quantification. We found the breakpoint in 80 % cases. The MRD data from 15 patients obtained by our method were compared to the levels obtained by standard methods (Ig/TCR and BCR/ABL transcript quantification). In all but 1 patient we found significant discrepancies, raising the questions about leukemic origin and the most accurate method for...
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Leukaemias with BCR/ABL fusion gene.
Hovorková, Lenka ; Zuna, Jan (advisor) ; Zemanová, Karla (referee)
Philadelphia (Ph) chromosome, as a result of reciprocal translocation, is in majority of cases connected to two types of leukaemia - chronic myelogenous (CML) and acute lymphoblastic (ALL). The translocation occurs within large intronic sequences of BCR and ABL genes. The breakpoints are specific for individual patient and may be used as a target for monitoring of leukemic burden (MRD, minimal residual disease) during the treatment. In general, MRD is an important prognostic factor, which influences the treatment intensity. Two standardized methods are currently used for its monitoring. The first one is based on the detection of clonal specific Immunoglobulin and/or T-cell receptor genes rearrangements (and thus cannot be used for CML cases) at the DNA level, the second one utilizes detection of the BCR/ABL fusion gene at the mRNA level. Our aim was to optimize and standardize the process to find individual patient breakpoints on Ph chromosome and to use it for MRD quantification. We found the breakpoint in 80 % cases. The MRD data from 15 patients obtained by our method were compared to the levels obtained by standard methods (Ig/TCR and BCR/ABL transcript quantification). In all but 1 patient we found significant discrepancies, raising the questions about leukemic origin and the most accurate method for...
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Molecular evaluation of novel BCR/ABL kinase domain variants in patients with chronic myeloid leukemia
Dvořáková, Lucie ; Peková, Soňa (advisor) ; Kozák, Tomáš (referee)
1 Abstract BCR/ABL is a constitutively active tyrosine kinase that has been shown to be at the heart of the development of chronic myeloid leukemia (CML) and about 30% of acute lymphoblastic leukemia (ALL). With the recent advent of tyrosine kinase inhibitors (TKIs), exemplified by Imatinib, Nilotinib, Dasatinib and Bosutinib, patients with Ph+ CML or ALL are candidates for the therapy with these agents. From the available TKIs, Imatinib is considered as front-line therapy for CML patients in chronic phase, while for Ph+ ALL patients, 2nd generation TKIs (nilotinib, dasatinib, bosutinib) might be considered as more effective therapeutic option. Since the treatment with TKIs is a long-term affair, a substantial proportion of patients acquire some sort of mutation in kinase domain of BCR- ABL, which could be a reason of treatment failure. To date, over ninety BCR/ABL kinase domain mutations have been identified, affecting over 50 amino acids. Recurrent BCR/ABL kinase domain mutations have already been in vitro tested to approximate for their in vivo behavior. Our goal is to invent in vitro technique that would allow testing TKI sensitivity of novel BCR/ABL kinase domain mutations, identified at very low MRD levels. The technique makes use of site-directed mutagenesis to create the novel BCR/ABL kinase domain...
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