Fusion Proteins, bcr-abl ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Male ; Middle Aged

Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Mutation

Fusion Proteins, bcr-abl ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive

BACKGROUND: STI571, which is a selective inhibitor of the BCR-ABL tyrosine kinase, is a promising new drug for the treatment of patients with chronic myelogenous leukemia. But it has been noted that this drug is frequently associated with an adverse cutaneous reaction. OBJECTIVE: The purpose of this study was to find the clinical and histopathological characteristics of drug eruptions caused by STI571. METHOD: We reviewed the clinical records of 10 patients diagnosed as drug erupions caused by STI571. RESULT: The mean age was 36.4 years and it was observed predominantly in females as the sex ratio of 7: 3. The most common clinical type was exanthematous eruption(70%), and followed by erythema multiforme-like eruption(20%), urticarial eruption(10%). In most cases(90%), the distribution was generalized, which involved trunk and extremities. The mean latent period was 17.1 days and peak incidence(70%) was noted between 1 and 2 weeks. Commonly associated adverse effects included fever(60%) and diarrhea(30%). Histopathologically, common findings included perivascular inflammatory cell infiltration(100%), eosinophil(80%), exocytosis(80%). CONCLUSION: Because drug eruptions caused by STI571 are dose-related and develop with a high frequency, we need a careful monitoring of patients who are treated with STI571, especially with a high dose.
Drug Eruptions* ; Erythema ; Extremities ; Female ; Fusion Proteins, bcr-abl ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Sex Ratio ; Imatinib Mesylate

BACKGROUND: STI571, which is a selective inhibitor of the BCR-ABL tyrosine kinase, is a promising new drug for the treatment of patients with chronic myelogenous leukemia. But it has been noted that this drug is frequently associated with an adverse cutaneous reaction. OBJECTIVE: The purpose of this study was to find the clinical and histopathological characteristics of drug eruptions caused by STI571. METHOD: We reviewed the clinical records of 10 patients diagnosed as drug erupions caused by STI571. RESULT: The mean age was 36.4 years and it was observed predominantly in females as the sex ratio of 7: 3. The most common clinical type was exanthematous eruption(70%), and followed by erythema multiforme-like eruption(20%), urticarial eruption(10%). In most cases(90%), the distribution was generalized, which involved trunk and extremities. The mean latent period was 17.1 days and peak incidence(70%) was noted between 1 and 2 weeks. Commonly associated adverse effects included fever(60%) and diarrhea(30%). Histopathologically, common findings included perivascular inflammatory cell infiltration(100%), eosinophil(80%), exocytosis(80%). CONCLUSION: Because drug eruptions caused by STI571 are dose-related and develop with a high frequency, we need a careful monitoring of patients who are treated with STI571, especially with a high dose.
Drug Eruptions* ; Erythema ; Extremities ; Female ; Fusion Proteins, bcr-abl ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Sex Ratio ; Imatinib Mesylate

OBJECTIVETo construct the P210(T315I-BCR/ABL) transgenic mice model.

METHODSThe transgenic vector in which the P210(T315I-BCR/ABL) gene and eGFP gene was derived by APN/CD13 promoter was constructed and microinjected into the single-cell fertilized eggs of C57 mice. Transgene integration was conformed by PCR genotyping and P210(T315I-BCR/ABL) expression levels was evaluated by RT-PCR. The CML phenotype was confirmed by blood routine examination, Wright's staining for peripheral blood and bone marrow smears, HE staining for organs of transgenic mice.

RESULTSThree transgenic mice lines with high expression of P210(T315I-BCR/ABL) gene and eGFP gene was selected. Compared with the wild type mice, the levels of WBC, platelet and neutrophil granulocyte of transgenic mice began to increase gradually at 2 months, and increase to 23.9×10⁹/L, 4 136×10⁹/L, and 74.6% respectively at 6 months. The remarkable hyperplasia of granulocytes was seen in the peripheral blood and bone marrow smears with splenomegaly infiltrated by leukemic cells.

CONCLUSIONThe P210(T315I-BCR/ABL) transgenic mice was constructed and provided a model to explore the mechanism of T315I CML and screen out the drug for T315 CML patient.

OBJECTIVETo revalidate the conversion factor（CF）for the conversion of BCR-ABL （P210）transcript levels to the international scale（BCR- ABLIS）in chronic myeloid leukemia（CML） which validated before.

METHODSPeking University People's Hospital（PKUPH）prepared the exchange samples for revalidation of CFs of 15 laboratories which validated nine or eighteen months ago. The fresh BCR-ABL（P210）（+）bone morrow or peripheral blood nucleated cells were diluted with BCR-ABL （P210）（-）cells to achieve different BCR- ABL levels, totally 16 sets and 24 samples per set were prepared. TRIzol reagent was added in each tube. Each laboratory tested BCR-ABL transcript levels of one set of samples. Agreement between BCR-ABLIS of each laboratory and PKUPH was assessed by the Bland- Altman method. For laboratories which did not meet the criteria of revalidation, linear regression equation was derived after the samples with maximum BCR-ABL deviation were removed until R²>0.98, then new CF was calculated.

RESULTS10 laboratories met the revalidation criteria with both bias within ±1.4 fold and 95% limits of agreement within ±6 folds, and their CFs still could be used for accurately conversion of BCR-ABLIS. New CFs were recalculated as of 1.8-6.3 folds of their previous CFs in 5 laboratories not met the criteria.

CONCLUSIONRevalidation of CF by sample exchange among laboratories was necessary for accurate and continuous application of BCR-ABLIS, which not only tested the validity of CF acquired before but also calculated new available CFs for those with invalid CFs.

Transgenic animal model provide a good platform to research the pathogenesis and therapy of chronic myelogenous leukemia (CML). To date, a number of BCR/ABL transgenic animal models have been established using different promoter or tetracycline-controlling system. Some of them appear the characteristics of human CML, which have contributed greatly to research the pathogenesis and therapy of CML. In this article, the researching progress, advantage and drawback, application of BCR/ABL transgenic animal model are reviewed.
Animals ; Animals, Genetically Modified ; Disease Models, Animal ; Fusion Proteins, bcr-abl ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive

OBJECTIVE: To analyze the clinical features of chronic myeloid leukemia (CML) with T315 I mutation (CML-T315I) and compare the effectiveness of different treatments. METHODS: We retrospectively analyzed the clinical data and outcomes of 19 patients with CML-T315I receiving different treatments. The T315 I mutations in these patients were detected by examination of BCR-ABL kinase domain (KD) mutation by RTQ-PCR and Sanger sequencing. The relapse following the treatments, defined as hematological, cytogenetic and molecular biological recurrences, were analyzed in these patients. RESULTS: Of the 19 patients with CML-T315I, 14 (73.7%) were in CML-CP stage at the initial diagnosis, and 13 (81.2%) were high-risk patients based on the Sokal scores. All the 19 patients were treated with TKI after the initial diagnosis, and during the treatment, 15 (78.9%) patients were found to have additional chromosomal aberrations, and 10 (52.6%) had multiple mutations; 13 (68.4%) of the patients experienced disease progression (accelerated phase/blast crisis) before the detection of T315I mutation, with a median time of 40 months (5-120 months) from the initial diagnosis to the mutation detection. After detection of the mutation, 12 patients were treated with ponatinib and 7 were managed with the conventional chemotherapy regimen, and their overall survival rates at 3 years were 83.3% and 14.2%, respectively ( < 0.001). CONCLUSIONS CML patients resistant to TKI are more likely to have T315I mutations, whose detection rate is significantly higher in the progressive phase than in the chronic phase. These patients often have additional chromosomal aberrations and multiple gene mutations with poor prognoses and a high recurrence rate even after hematopoietic stem cell transplantation. Long-term maintenance therapy with ponatinib may improve the prognosis and prolong the survival time of the patients.
Drug Resistance, Neoplasm ; Fusion Proteins, bcr-abl ; Humans ; Imidazoles ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Mutation ; Pyridazines ; Retrospective Studies

OBJECTIVE: To investigate the pro-apoptotic effect and mechanism of miR-30a overexpression on chronic myeloid leukemia K562 cells. METHODS: The k562 cells were transfected with the recombinant plasmid pEGFP-pre-miR-30a, the real-time quantitative PCR was used to detect the level of miR-30a and BCR/ABL, and then the cell apoptosis was assessed by flow cytometry with AnnexinV-FITC/PI double staining. Western blot was used to detect the expression of BCR/ABL protein,apoptosis-related protein BCL-2 and BAX, PTEN, AKT and p-AKT. RESULTS: Sequencing and digestion map indicated that the recombinant plasmid was constructed successfully. Compared with 2 control groups, the miR-30a expression in k562 cells transfected with recombinant plasmid pEGFP-pre-miR-30a was obviously up-regulated. The expression of BCR/ABL mRNA and BCR/ABL protein was both significantly down-regulated. Apoptotic rate was significantly enhanced (both P<0.05),and the expression of anti-apoptotic protein BCL-2 was down-regulated while the expression of pro-apoptotic protein BAX was up-regulated. The level of PTEN was significantly up-regulated in omparison with control groups,no variation was found in total AKT, but the expression of p-AKT was down-regulated. CONCLUSION The overexpression of miR-30a is abled to down-regulate the level of BCR/ABL mRNA and BCR/ABL protein, and increase apoptotic rate, its mechanism may be related with inhibition of the activity of BCR/ABL-PTEN/AKT signaling pathway.
Apoptosis ; Cell Proliferation ; Fusion Proteins, bcr-abl ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; MicroRNAs ; genetics