Adult ; Blast Crisis ; Female ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; complications ; diagnosis ; Ulcer ; diagnosis ; etiology

This study was aimed to investigate the immunologic characteristics of refractory anemia with excess blasts-II (RAEB-II) which belongs to a new subtype of World Health Organization (WHO) classification of myelodysplastic syndrome (MDS) and to screen out the independent immunologic prognostic factors of MDS. 35 cases of adult patients with de novo MDS were investigated. The immunofluorescent analysis by multiparameter flow cytometry was performed at the double gating of CD45/SSC to determine the immunophenotype of MDS cells in all cases. All patients were followed up. 47 cases of acute myeloid leukemia (AML) M1, 51 cases of AML-M(2) and 38 cases of acute lymphocytic leukemia (ALL) were selected as control. Software SPSS 13.0 was applied to analyze all the related data. The results showed that the positive expression rate of HLA-DR in RAEB-II was 100%, which was high in sensitivity and specificity. CD13 (94.74%), CD33 (84.21%) and CD117 (78.95%) were also highly expressed in RAEB-II. CD13 in RAEB-II was significantly higher than that in refractory cytopenia with or without multilineage dysplasia (RA/RCMD) (p < 0.01) and REAB-I (p < 0.05); CD33, CD117 (p < 0.05) and stem cell antigen CD34 (p < 0.01) in RAEB-II were significantly higher than that in RCMD (p < 0.01), but no statistically significant difference was found as compared with RAEB-I (p > 0.05). Compared with AML-M(1) and AML-M(2), no significant difference of CD13 and CD117 in RAEB-II was found (p > 0.05). CD33 (p < 0.01) and CD34 (p < 0.05) were significantly lower than that in AML-M(1), but no significant difference was found as compared with AML-M(2) (p > 0.05); CD15 (p < 0.01) and CD11b (p < 0.05) was significantly lower than that in M(2), but no significant difference was found as compared with AML-M(1) (p > 0.05); MPO was significantly lower than that in AML-M(1) and M(2) (p < 0.05); HLA-DR was significantly higher than that on AML-M(2) (p < 0.05), but no significant difference was found as compared with AML-M(1) (p > 0.05). RAEB-II did not express CD2, CD3, CD5 and CD8 (positive rate 0%, p < 0.01) when compared with T-ALL; CD4 (p < 0.05) and CD7 (p < 0.01) were significantly lower than that in T-ALL. RAEB-II did not express CD19 and CD20 (positive rate 0%, p < 0.01) as compared with B-ALL; CD10, CD22 and cCD79a were significantly lower than that in B-ALL (p < 0.05). CD117 (p = 0.0197) and MPO (p = 0.0085) were the two prognostic immunological antigens as regards the overall survival (OS) of MDS; CD117 (p = 0.003) was the single parameter in Cox regression. It is concluded that RAEB-II expresses mainly myeloid antigen without or with little expression of lymphoid antigen. Unique individual immunophenotypic features can be detected in patients with RAEB-II. HLA-DR can be a specific parameter to distinguish the other subtypes of MDS. CD117 may be an independent prognostic immunological antigen as regards OS of MDS.
Adolescent ; Adult ; Aged ; Anemia, Refractory ; complications ; diagnosis ; immunology ; Blast Crisis ; complications ; diagnosis ; immunology ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; diagnosis ; immunology ; Prognosis ; Young Adult

BACKGROUND: Philadelphia(Ph) chromosome is found in about 95 percent of chronic myelogenous leukemia(CML) patients. Ph chromosome results from a reciprocal translocation between the long arms of chromosomes 9 and 22, and the fusion gene, BCR-ABL contribute to oncogenesis. Three to five years after first diagnosis, CML progresses to the blast crisis, and is accompanied by secondary cytogenetic changes in about 85% of cases. In this study, we investigated the incidence of ABL deletion of derivative 9 chromosome in CML and evaluated the association between this deletion and progression to the blast crisis by interphase fluorescence in situ hybridization(FISH). METHOD: The subjects included in this study were a consecutive series of 58 patients who were diagnosed as CML at Seoul National University Hospital between January 1997 and April 2000. On 90 archival bone marrow aspirate samples from these 58 CML patients, interphase FISH was performed with a commercially available probe. RESULTS: The ABL deletion of derivative 9 chromosome was detected in 17(29.3%) of 58 patients with CML. Eighteen of 58 patients progressed to blast crisis in this period. ABL deletion was found in 7 of 18 patients with blast crisis, and not in 11 remainders. The mean duration from the diagnosis to blast crisis was 37.1 months in 7 patients with the ABL deletion, while the mean duration was 74.2 months in 11 patients without the ABL deletion. The mean duration from the diagnosis to blast crisis in patients with ABL deletion was significantly shorter than in patients without ABL deletion(P=0.043). CONCLUSIONS: We found that 29.3% of patients with CML had the ABL deletion on derivative 9 chromosome. In these patients, the time taken for evolution to blast crisis was significantly shorter than that of the patients without ABL deletion.
Arm ; Blast Crisis* ; Bone Marrow ; Carcinogenesis ; Cytogenetics ; Diagnosis ; Fluorescence* ; Humans ; Hydrogen-Ion Concentration ; In Situ Hybridization* ; Incidence* ; Interphase* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Philadelphia Chromosome ; Seoul

BACKGROUND: Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder characterized by a progressive course outlined by the transition from the chronic phase (CP) to blastic crisis (BC). The course of CML-BC is still fatal, and in spite of various efforts in treatment, median survival remains short. The aim of the present study was to analyze the prognostic factors having an impact on response to treatment and survival in patients with CML-BC. We also investigated prognostic influence on survival in myeloid BC according to FAB classification. METHODS: All patients (N=35) with CML with onset of CML-BC between January 1992 and May 2002 were reviewed. RESULTS: The median survival for all patients after onset of CML-BC was 7 weeks, and probable survival rate at 24 months was 5.9%. The adverse prognostic factors for survival of CML-BC were high and intermediate risk of Hasford score at diagnosis of CML (P=0.05), normal serum LDH (P=0.016), bone marrow blasts > or =60% (P=0.092), no treatment at CML-BC (P=0.0056), platelet count <20x10(9)/L (P=0.13). Clonal evolution at diagnosis of CML-BC was associated with a shorter survival. Especially in our study, FAB subtype M4-7 (median survival, 4 weeks; 95% CI, 2~6) had shorter survival duration than M0-2 (median survival, 16 weeks; 95% CI, 5~27) in patients with myeloid CML-BC. CONCLUSION: The management of patients with CML-BC remains highly unsatisfactory. Once blast crisis has occurred, there are useful parameters to assess the prognosis for the individual patient and these may be of interest in planning therapy. Our experience suggests that FAB classification M4-7 are poor prognostic factor in patients with myeloid CML-BC.
Blast Crisis ; Bone Marrow ; Classification* ; Clonal Evolution ; Diagnosis ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Myeloproliferative Disorders ; Platelet Count ; Prognosis ; Retrospective Studies* ; Survival Rate

The purpose of this study was to investigate the risk factors in prognosis of patients with myelodysplastic syndrome (MDS) to emphatically study the clinical significance of different scoring systems such as Bournemouth-, Spanish-, Düsseldorf-, Lille, and the International prognostic scoring systems (IPSS), French-American-British (FAB) and World Health Organization (WHO) classifications as well as abnormal clone of chromosome, bone marrow and hematological indicators in evaluating prognosis of MDS patients, and to identify the independent factor related to prognosis. All clinical data of 69 consecutive patients diagnosed as primary MDS over a period of 5 years were collected and stored by Epi Data 3.0, 63 patients out of which were followed up. SPSS 13.0 software was applied to deal with all data. The statistical methods included life table, Kaplan-Meier, Log-rank test and Cox regression. The results indicated that the median age of 63 patients was 38 years. 26 out of 63 patients had karyotype aberration (41.27%). Median OS was 30.63 months, and 33 patients (52.38%) died. All five prognostic scoring systems could successfully discriminate risk groups as regards overall survival. IPSS, Lille and Spanish prognostic scoring system were more effective (p<0.0001). Multivariate Cox regression analysis indicated that IPSS (p<0.0001) and Lille chromosome classifications (p<0.0001) were most important factors for OS followed by bone marrow blasts (p=0.00062), Spanish prognostic scoring system (p=0.00064) and Lille prognostic system (p=0.008). The WHO classifications also successfully discriminated between risk groups (p<0.0001). The new WHO subgroups [refractory cytopenia with multilineage dysplasia (RCMD), with or without ringed sideroblasts] showed a significantly different prognosis (p=0.003) for OS, in comparison to the subgroups having erythroid dysplasia only (RA/RARS) and 5q-syndrome. All patients were reclassified to FAB classification, and the low risk group (RA/RAS) and high risk group (RAEB) also had significant difference (p=0.00012) as regards OS. It is concluded that the major independent prognostic variables for OS are percentage of bone marrow blasts and karyotype aberration. The use of WHO classification have more significance for improving predictive value than that of the FAB classification, and the IPSS can be used for clinical decision-making in patients with cytogenetic results. In the hospitals which cannot carry out the cytogenetic examination, Spanish prognostic system can be applied to the patients without cytogenetic results.
Adolescent ; Adult ; Aged ; Blast Crisis ; Child ; Child, Preschool ; Cytogenetic Analysis ; Female ; Humans ; Infant ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; classification ; diagnosis ; genetics ; Prognosis ; Risk Factors ; Severity of Illness Index ; Survival Analysis ; World Health Organization ; Young Adult

Erythroleukemic blast crisis of chronic myeloid leukemia (CML) is very rare. We report two cases of erythroleukemic blast crisis of CML resistant to imatinib treatment. Both patients made a rapid progression to blast crisis 6 and 4 months after diagnosis while being treated with imatinib 400 mg/day. Bone marrow aspiration revealed predominant erythroid precursors with 65.4% and 54.8% each. There were significant proportions (more than 20%) of myeloblasts among non-erythroid cells. Immunophenotyping revealed expression of glycophorin A confirming erythroleukemic blast crisis. The karyotyping result of patient 1 was 46,XX,t(9;22)(q34;q11.2)[3]/52,idem,+8,+12,+18,+21,+22,+der(22)t(9;22)[17] and that of patient 2 was 46,XX,inv(3)(q21q26.2),t(9;22)(q34;q11.2)[20]. Patient 1 showed no response to imatinib and BMS-354825 in the following bone marrow study. She died of septic shock as a complication of an infection after 69 days of blast crisis. Patient 2 received allogeneic bone marrow transplantation (BMT) in the cytogenetically no response state, but she also died of graft-versus-host disease 9 weeks after BMT. The poor prognosis and rapid progression of disease in both cases were correspondent to most of the reported cases. During the course of the disease of the two patients, we monitored the BCR-ABL chimeric mRNA with real-time quantitative polymerase chain reaction (RT-PCR), and it was found useful in predicting the imatinib response and progression to blast crisis of CML. Although both of our cases showed the typical bad prognosis and findings of erythroleukemic blast crisis of CML, the karyotypes were different from the expected type of t(3;21)(q26;q22). But the relationship between additional changes of EVI1 on chromosome 3q26 shown in case 2, and progression to the erythroleukemic blast crisis need further investigation.
Blast Crisis* ; Bone Marrow ; Bone Marrow Transplantation ; Diagnosis ; Glycophorin ; Graft vs Host Disease ; Granulocyte Precursor Cells ; Humans ; Immunophenotyping ; Karyotype ; Karyotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Polymerase Chain Reaction ; Prognosis ; RNA, Messenger ; Shock, Septic ; Dasatinib ; Imatinib Mesylate

BACKGROUND: In most cases of chronic myelogenous leukemia (CML), one of the two abl promotors (Pa) is nested within the bcr-abl hybrid gene and should be able to transcribe the 6-kb normal abl mRNA from the Philadelphia chromosome. However, 6-kb transcript is present only in CML cell lines containing a normal abl allele. Since the nested Pa in the bcr-abl hybrid gene is contained within a CpG island, de novo methylation of Pa CpG islands may cause silencing of the c-abl gene. In this study, using a polymerase chain reaction (PCR) and methylation-sensitive restriction enzymes, we investigated the methylation status of the nested Pa CpG islands in the bcr-abl hybrid gene at two stages of CML and the usefulness of it as a disease progression marker. METHODS: Genomic DNA was extracted from the preserved bone marrow smear slides of ten CML patients, at chronic phase and blast crisis respectively. K-562 cell line and normal peripheral lymphocytes as a negative control were also used. The extracted genomic DNA was digested with methylation-sensitive restriction enzymes and methylation-insensitive restriction enzymes, respectively, followed by PCR amplification for Pa promotor and electrophoresed for detection of 171 bp PCR products. RESULTS: The patients were transformed from chronic phase to blast crisis during 11-60 months after initial diagnosis. The patients displayed the following three types of methylation patterns. A, no methylation; B, partial methylation; C, complete methylation. All cases of chronic phase showed pattern A, but three cases of blast crisis were pattern B and seven cases, pattern C. CONCLUSIONS: De novo DNA Methylation at the Pa promotor of the abl gene in the bcr-abl hybrid gene is a distinct and common molecular event in blast crisis of CML. Since the process of Pa CpG methylation is of a progressive nature, Pa methylation pattern may be used as a molecular marker for progression of CML to blast crisis.
Alleles ; Blast Crisis ; Bone Marrow ; Cell Line ; CpG Islands ; Diagnosis ; Disease Progression ; DNA Methylation* ; DNA* ; Genes, abl ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Lymphocytes ; Methylation ; Philadelphia Chromosome ; Polymerase Chain Reaction ; RNA, Messenger

The t(3;3)(q21;q26.2) is known to be mainly observed in hematologic myeloid malignancies, as a form of 3q21q26 syndrome. Cytogenetic abnormalities of 3q21q26 syndrome result in RPN1-EVI1 fusion transcripts involving ecotropic viral integration site-1 (EVI1) at 3q26.2 and ribophorin I (RPN1) at 3q21, and the fusion transcripts play an important role in leukemogenesis and disease progression. They are usually associated with dysplasia, especially of megakaryocytes. Patients with these cytogenetic abnormalities show extremely poor prognosis even with aggressive anti-leukemic therapy. We report a case of blastic crisis of CML with both t(3;3)(q21;q26.2) and t(9;22)(q34;q11.2) and associated severe multilineage dysplasia. The patient showed a poor response to imatinib, dasatinib and aggressive induction therapy. When both t(3;3)(q21;q26.2) and t(9;22)(q34;q11.2) are observed in cases of leukemia with increased blasts, they are best considered as aggressive phases of CML with t(3;3)(q21;q26.2), rather than AML with t(9;22)(q34;q11.2) by 2008 WHO classification.
Adolescent ; Antineoplastic Agents/therapeutic use ; Blast Crisis/*diagnosis ; Bone Marrow Cells/pathology ; *Chromosomes, Human, Pair 3 ; Drug Resistance, Neoplasm ; Humans ; Karyotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/*diagnosis/drug therapy/genetics ; Male ; Piperazines/therapeutic use ; Pyrimidines/therapeutic use ; Thiazoles/therapeutic use ; *Translocation, Genetic

OBJECTIVETo explore the genetic and clinical characteristics of isodicentric Ph chromosomes [idic(Ph)] in lymphoid blast crisis of chronic myeloid leukemia (CML-BLC).

METHODSBone marrow aspirates of 2 patients with CML-BLC were analyzed by R banding after 24 hours of culturing. Genomic copy number variations (CNV) were analyzed by single nucleotide polymorphism array (SNP array) in case 1. The results were confirmed with fluorescence in situ hybridization (FISH). Variations of acute lymphoblastic leukemia-related genes including CDKN2A/AB and PAX5 were detected by multiplex ligation-dependent probe amplication (MLPA).

RESULTSDeletions and duplications on derivative chromosome 9 detected by FISH were confirmed by SNP array analysis. The distances between the BCR/ABL fusion signals on the idic(Ph) chromosomes in the two patients have differed greatly. The idic(Ph) in the second patient was supposed to be formed by two Ph chromosomes joined at their q terminals, where as the idic(Ph) in the first patient have been shown to be fused at the satellite regions of their p arms.

CONCLUSIONThe idic(Ph) chromosomes presented in CML-BLC may predict resistance to Imatinib and response to Dasatinib.

Blast Crisis ; diagnosis ; genetics ; therapy ; Chromosome Aberrations ; Chromosome Banding ; Chromosome Deletion ; Chromosome Duplication ; Chromosomes, Human, Pair 9 ; genetics ; DNA Copy Number Variations ; Fatal Outcome ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; genetics ; therapy ; Male ; Middle Aged ; Philadelphia Chromosome