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

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

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

OBJECTIVE: To investigate the curative efficacy of first generation TKI in the treatment of CML-CP combined with vPh and genetic characteristics. METHODS: 60 patients with CML-CP combined with vPh from January 2010 to May 2017 in the First Affiliated Hospital of Kunming Medical University were chosen as CML-CP-vPh group, and 107 patients with CML-CP combined with typical Ph chromosome at the same time were chosen as control group. The patients in two groups were treated with imatinib; The curative efficacy, karyotype and FISH signal type were compared between 2 groups, and the factors influencing long-term survival of patients were analyzed by Cox risk model. RESULTS: There was no significant difference in the demographic and hematological baseline data between 2 groups (P＞0.05), and there was no significant difference in the incidence of drug-resistance between 2 groups (P＞0.05). The incidence of primary drug-resistance and primary hematological drug-resistance in the CML-CP-vPh group were significantly higher than that in control group (P＜0.05). There was no significant difference in the accumulative CCyR rate, accumulative MMR rate, OS and EFS between 2 groups (P＞0.05). Multivariate Cox model analysis showed that vPh not correlated with OS and EFS of patients with CML-CP (P＞0.05). The factors influencing OS in CML-vPh patients included high risk of Sokal scores, peripheral blood basophils proportion ≥10%, BCR-ABL in 3 months after treatment＜10%, achieviag CCyR in 6 months after treatment and achieviag MMR in 12 months after treatment (P＜0.05). The factors influencing EFS included BCR-ABL＜10% in 3 months after treatment and achieving MMR in 12 months after treatment (P＜0.05). The regions with high frequency of heterotopic involvement included 12q1, 12q2 [9 cases (15.00%)] and 1p3 [8 cases (13.33%)]. The percentage of 2G2R1Y, 1G1R2F, 1G2R1Y, 2G1R1Y and 1G1R1Y in FISH signal types were 73.33%, 10.00%, 1.67%, 1.67% and 1.67% respectively. CONCLUSION Patients with CML-CP combined with vPh possess higher primary drug-resistance rate and primary hematological drug-resistance rate for the first generation TKI, while second-generation TKI can efficiently improve long-term survival, its efficacy is similar to efficacy for patients with typical Ph chromosomes. CML-CP combined with vPh does not display special demographic and hematological characteristics. The main involved regions of heterotopic variants include 12q1, 12q2 and 1p3, while 2G2R1Y type is the most common type of FISH signal.
Fusion Proteins, bcr-abl ; Humans ; Imatinib Mesylate ; Karyotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Protein Kinase Inhibitors ; Treatment Outcome

OBJECTIVE: To analyze the significance of various abnormal signal patterns appreared in CML and B-ALL patients by using BCR/ABL/ASS1 tricolor dual-fusion probe, and to explore its application value in detecting BCR/ABL fusion gene and ASS1 gene deletion. METHODS: 50 newly diagnosed CML patients and 50 newly diagnosed B-ALL patients were detected by fluorescence in situ hybridization (FISH) with BCR/ABL/ASS1 tricolor dual-fusion probe. Meanwhile, karyotype analysis was performed on all the patients using the 24 hours short-term culture and R-banding. RESULTS: Among the 50 CML patients, Ph was found in 49 cases, 5 normal interphase karyotype was observed in 1 case. FISH detection showed that BCR/ABL fusion gene existed in all patients (100%), while the positive signal pathway showed that 1R1G2B2F was observed in 39 cases (78%), 2R1G2B1F in 2 cases (4%) and 1R1G2B1F in 6 cases (12%), simultaneous existence of 1R1G1B1F and 1R1G2B3F in 1 case (2%), 2R1G1B1F in 1 case (2%) 1R1G3B3F in 1 case (2%). FISH detection also showed that the karyotype of 6 case at ASS1 gene deletion (1R1G1B1F) all were simple t (9; 22) translocation, and other abnormalities not were observed. Among 50 cases of B-ALL, Ph was found in 13 cases, the numerical aberration and structural aberration of non t (9; 22) in 16 cases, normal karyotype in 20 cases, absence of mitotic phase in 1 case. FISH detection showed that 16 cases (32%) had BCR/ABL fusion gene including 13 cases (26%) of 1R1G2B2F, 1 case (2%) of stimultaneous exitance of 1R1G2B2F and 1R1G3B3F 1 case (2%) of 2R1G1B1F, 1 case (2%) of 1R1G3B2F. FISH detection also showed that 3 cases had BCR/ABL fusion gene, including 1 case with ASS1 gene deletion (2R1G1B1F), 1 case with classical t (9; 22) translocation (1R1G2B2F) and 1 case with BCR/ABL fusion gene and increase of ASS1 gene copy (1R1G3B3F). CONCLUSION Tricolor dual-fusion FISH probe for detecting BCR/ABL fusion gene and ASS1 gene deletion is simple, rapid, sensitive and stable. It can detect various forms of molecular fusion and avoid the false positive results due to coincidental overlap of signals generated by D-FISH probe and ES-FISH probe. In addition, this detection method not only can directly observe the presence or absence of ASS1 gene deletion, but also improve the reliability of the positive results of newly diagnosed BCR/ABL fusion gene and accuracy of monitoring results of minimal residual disease for the subsequent visit.
Fusion Proteins, bcr-abl ; genetics ; Gene Deletion ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Reproducibility of Results