- MeSH
- Apoptosis MeSH
- Fusion Proteins, bcr-abl * MeSH
- Drug Resistance, Neoplasm genetics immunology drug effects MeSH
- Leukemia, Myelogenous, Chronic, BCR-ABL Positive * drug therapy MeSH
- Hematopoietic Stem Cells MeSH
- Precision Medicine MeSH
- Protein Kinase Inhibitors * therapeutic use MeSH
- Humans MeSH
- Reverse Transcriptase Polymerase Chain Reaction methods utilization MeSH
- Cell Proliferation MeSH
- Antineoplastic Agents pharmacokinetics therapeutic use MeSH
- Proto-Oncogene Proteins c-abl MeSH
- Proto-Oncogene Proteins c-bcr MeSH
- Protein-Tyrosine Kinases MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Aim. This is a case report of a 51 year old male with marked splenomegaly, basophilia, severe thrombocytopenia, anemia and high SFKL phosphorylation downstream of Bcr-Abl, investigated for association of the e6a2 BCR-ABL fusion gene and marked basophilia. The treatment strategy implications in patients with Philadelphia positive CML are described. Methods. RT-PCR and sequencing were carried out on the peripheral blood leukocytes to detect the type of BCRABL transcript. The BCR-ABL mutational status was assessed using sequencing of the RT-PCR products. The in vitro test of sensitivity to TKIs was based on detecting inhibited phosphorylation of the Crkl and Phospho-Src family kinases (SFK, Tyr416) using immunodetection. Results. The cytogenetics revealed 90% of Ph+ (Philadelphia) cells in the bone marrow aspirate with no additional clonal chromosomal abnormalities at diagnosis. This correlated with an accelerated phase of the CML. Sequencing analysis of reverse transcribed and PCR amplified BCR-ABL transcript revealed a rare e6a2 fusion, with no evidence for Bcr-Abl kinase domain mutation. Western blot analysis showed high phosphorylation (activation) of Crkl and the Src family of kinases (P-SFK). In vitro test of sensitivity of the patients’ leukemic cells to imatinib demonstrated sensitivity of Bcr-Abl tyrosine kinase to imatinib, as assessed by a decrease in phosphorylated Crkl and the disappearance of P-SFK, suggesting that P-Src reflects only the Bcr-Abl–dependent Src activity. The initial treatment strategy was reduced imatinib and search for an unrelated hematopoietic stem cell donor (according to the ELN recommendations). The patient was allografted with peripheral stem cells from an HLA- identical male donor but on day +70 graft failure occurred. He was allografted again with the peripheral stem cells from an HLA-identical female donor, engrafted on day +15 and showed 100% donor chimerism with no evidence of the e6a2 BCR-ABL fusion transcript on day +30. Conclusion. The clinical disease course in patients with the rare e6a2 BCR-ABL transcript variant is aggressive. This may be the result of increased kinase activity due to partial loss of the guanine exchange factor/dbl-like domain which mediates the interaction with several Ras-like G-proteins involved in cell proliferation, signal transduction, and cytoskeletal organization. For the above reasons, these patients should receive stem cell transplant immediately after a short course of treatment with imatinib/ dual Src/Abl kinase inhibitor or they should be registered in clinical trials with experimental agents.
- MeSH
- Basophils MeSH
- Leukemia, Myelogenous, Chronic, BCR-ABL Positive genetics blood therapy MeSH
- Philadelphia Chromosome MeSH
- Middle Aged MeSH
- Humans MeSH
- Oncogene Fusion MeSH
- Leukocyte Count MeSH
- Stem Cell Transplantation MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
- Research Support, Non-U.S. Gov't MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
Přeruš. str. : il., tab. ; 31 cm
Stanovení in vitro senzitivity leukemických buněk u nemocných s chronickou myeloidní leukemií léčenými inhibitory tyrosinové kinázy a její využití při rozhodování o léčbě. Klony leukemických buněk in vitro rezistentní k imatinibu, získané od nemocných sPh+ CML, jsou vyšetřovány technologií CGH mikroarray na skryté genetické změny s cílem rozšířit poznání mechanismů odpovědných za vznik rezistence k imatinibu.; Assessment of in vitro sensitivity of leukemic cells from patients with chronic myeloid leukemia treated with tyrosine kinase inhibitors and its implications for the therapy management. Clones of in vitro imatinib-resistant cells, obtained from patientswith Ph+ CML, are assessed for hidden genetic changes using CGH microarray with the aim to increase our knowledge of the mechanisms responsible for imatinib resistance development.
- MeSH
- Fusion Proteins, bcr-abl antagonists & inhibitors therapeutic use MeSH
- Leukemia, Myelogenous, Chronic, BCR-ABL Positive therapy MeSH
- Cytogenetic Analysis MeSH
- In Situ Hybridization, Fluorescence MeSH
- Drug Resistance MeSH
- Biomarkers, Tumor MeSH
- Polymerase Chain Reaction MeSH
- Sensitivity and Specificity MeSH
- Protein-Tyrosine Kinases antagonists & inhibitors therapeutic use MeSH
- Conspectus
- Biochemie. Molekulární biologie. Biofyzika
- NML Fields
- hematologie a transfuzní lékařství
- onkologie
- biologie
- NML Publication type
- závěrečné zprávy o řešení grantu IGA MZ ČR
Embryonic stem cells (ESCs) proliferate rapidly and have a unique cell-cycle structure with a very short G1 phase. Previous reports suggested that the rapid G1 phase progression of ESCs might be underpinned by high and precocious Cdk2 activity and that Cdk2 activity might be crucial for both cell-cycle regulation and cell-fate decisions in human ESCs. However, the actual role of Cdk2 in cell-cycle progression of mouse ESCs (mESCs) has not been elucidated. In this study, we investigated the effects of down-regulation of Cdk2 activity by olomoucine II in 2 mESC lines. Olomoucine II treatment significantly increased the G1 phase cell numbers, decreased the S phase cell numbers, and inhibited DNA replication in mESCs. In nocodazole-synchronized mESCs, we show that specific down-regulation of Cdk2 activity prolongs G1 phase progression. In addition, down-regulation of Cdk2 activity in mESCs established a somatic cell-like cell cycle and induced expression of differentiation markers. Our results suggest that high Cdk2 activity is essential for rapid G1 phase progression and establishment of ESC-specific cell-cycle structure in mESCs and support the hypothesis of a link between cell-cycle regulation and pluripotency maintenance in ESCs. This study reveals olomoucine II to be an effective tool for manipulation of the cell cycle and pluripotency in ESCs and very likely also for the manipulation of other stem cell types, including cancer stem cells.
- MeSH
- Cell Line MeSH
- Cell Cycle drug effects MeSH
- HT29 Cells MeSH
- Time Factors MeSH
- Cyclin-Dependent Kinase 2 antagonists & inhibitors genetics metabolism MeSH
- Cyclin-Dependent Kinase 9 antagonists & inhibitors genetics metabolism MeSH
- Embryonic Stem Cells cytology drug effects metabolism MeSH
- G1 Phase drug effects MeSH
- Mice, Inbred Strains MeSH
- Inhibitory Concentration 50 MeSH
- Humans MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- CDC2 Protein Kinase antagonists & inhibitors genetics metabolism MeSH
- Flow Cytometry MeSH
- Purines pharmacology MeSH
- DNA Replication drug effects MeSH
- Cell Survival drug effects MeSH
- Blotting, Western MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
