Assuntos
Células Dendríticas/patologia , Leucemia Mielomonocítica Crônica/patologia , Neoplasias de Plasmócitos/patologia , Idoso , Idoso de 80 Anos ou mais , Células Dendríticas/imunologia , Humanos , Imunofenotipagem , Leucemia Mielomonocítica Crônica/genética , Leucemia Mielomonocítica Crônica/imunologia , Masculino , Neoplasias de Plasmócitos/genética , Neoplasias de Plasmócitos/imunologiaAssuntos
Proteínas de Ligação a DNA/genética , Cariótipo , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/genética , Proteínas de Fusão Oncogênica/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biópsia , Medula Óssea/patologia , Análise Mutacional de DNA , Humanos , Contagem de Leucócitos , Masculino , Transtornos Mieloproliferativos/tratamento farmacológico , UltrassonografiaAssuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/genética , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Asparaginase/uso terapêutico , Biomarcadores Tumorais/genética , Ciclofosfamida/uso terapêutico , Citarabina/uso terapêutico , Daunorrubicina/uso terapêutico , Regulação para Baixo/genética , Humanos , Mercaptopurina/uso terapêutico , Metotrexato/uso terapêutico , Mutação/genética , Proteínas de Fusão Oncogênica/genética , Prednisona/uso terapêutico , Prognóstico , Translocação Genética/genética , Regulação para Cima/genética , Vincristina/uso terapêuticoAssuntos
Mutação , Síndromes Mielodisplásicas/genética , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Progressão da Doença , GTP Fosfo-Hidrolases/genética , Genes p53 , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Proteínas de Membrana/genética , Proteínas Nucleares/genética , Proteínas Repressoras/genética , Ribonucleoproteínas/genéticaAssuntos
Anemia Macrocítica/genética , Anemia Macrocítica/metabolismo , Eritroblastos/metabolismo , Leucina/farmacologia , Proteínas Ribossômicas/deficiência , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Deleção Cromossômica , Cromossomos Humanos Par 5/genética , Cromossomos Humanos Par 5/metabolismo , Ativação Enzimática/efeitos dos fármacos , Humanos , Proteínas Ribossômicas/genéticaRESUMO
The t(10;11)(p12;q14) is a recurring chromosomal translocation that gives rise to the CALM/AF10 fusion gene, which is found in acute myeloid leukemia, acute lymphoblastic leukemia and malignant lymphoma. We analyzed the fusion transcripts in 20 new cases of CALM/AF10-positive leukemias, and compared the gene expression profile of 10 of these to 125 patients with other types of leukemia and 10 normal bone marrow samples. Based on gene set enrichment analyses, the CALM/AF10-positive samples showed significant upregulation of genes involved in chromatin assembly and maintenance and DNA repair process, and downregulation of angiogenesis and cell communication genes. Interestingly, we observed a striking upregulation of four genes located immediately centromeric to the break point of the t(10;11)(p12;q14) on 10p12 (COMMD3 (COMM domain containing 3), BMI1 (B lymphoma Mo-MLV insertion region 1 homolog), DNAJC1 (DnaJ (Hsp40) homolog subfamily C member 1) and SPAG6 (sperm associated antigen 6)). We also conducted semiquantitative reverse transcriptase-PCR analysis on leukemic blasts from a murine CALM/AF10 transplantation model that does not have the translocation. Commd3, Bmi1 and Dnajc1, but not Spag6 were upregulated in these samples. These results strongly indicate that the differential regulation of these three genes is not due to the break point effect but as a consequence of the CALM/AF10 fusion gene expression, though the mechanism of regulation is not well understood.
Assuntos
Montagem e Desmontagem da Cromatina/genética , Sítios Frágeis do Cromossomo , Cromossomos Humanos Par 10 , Reparo do DNA/genética , Leucemia/genética , Proteínas Monoméricas de Montagem de Clatrina/genética , Fatores de Transcrição/genética , Regulação para Cima , Animais , Humanos , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Translocação GenéticaAssuntos
Azacitidina/uso terapêutico , Metilação de DNA , Proteínas de Ligação a DNA/genética , Mutação/genética , Síndromes Mielodisplásicas/tratamento farmacológico , Síndromes Mielodisplásicas/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas/genética , Adulto , Idoso , Antimetabólitos Antineoplásicos/uso terapêutico , Dioxigenases , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Taxa de SobrevidaRESUMO
BACKGROUND: The Italian external quality assessment scheme in classical cytogenetics was started in 2001 as an activity funded by the National Health System and coordinated by the Italian Public Institute of Health. OBJECTIVES: The aim of our work is to present data from the first 4 years of activity, 2001-2004. METHODS: Italian cytogenetics public laboratories were enrolled on a voluntary basis, and this nationwide program covered prenatal, postnatal and oncological diagnosis. The scheme is annual and retrospective; a panel of experts reviewed the quality of images and reports in order to assess technical, analytical and interpretative performance. RESULTS: Over the 4-year period, the number of participating laboratories increased: from 36 in 2001, 46 in 2002, 49 in 2003 to 51 in 2004. The overall technical performance was satisfactory. Inadequacy or lack of information in reporting was the most frequent analytical inaccuracy identified in all parts of the scheme. However, the percentage of complete reports increased significantly during the period: by 36% in postnatal diagnosis between 2001 and 2004 (p < 0.001) and by 42% in oncological diagnosis between 2002 and 2004 (p = 0.003). CONCLUSIONS: Our experience reveals that participation in external quality assessment programs has significant advantages, helping to standardize and to assure quality in cytogenetic testing.
Assuntos
Análise Citogenética/métodos , Análise Citogenética/normas , Testes Genéticos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Neoplasias/diagnóstico , Garantia da Qualidade dos Cuidados de Saúde , Genótipo , Humanos , Itália , Neoplasias/genética , Diagnóstico Pré-Natal , Fatores de TempoAssuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Cromossomos Humanos Par 12/genética , Cromossomos Humanos Par 5/genética , Leucemia Monocítica Aguda/genética , Proteínas do Tecido Nervoso/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Translocação Genética , Adulto , Humanos , Hibridização in Situ Fluorescente , Leucemia Monocítica Aguda/diagnóstico , Leucemia Monocítica Aguda/terapia , MasculinoAssuntos
Instabilidade Genômica , Hepatite C/complicações , Linfoma não Hodgkin/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Aberrações Cromossômicas , Feminino , Humanos , Hibridização in Situ Fluorescente , Linfoma não Hodgkin/virologia , Masculino , Pessoa de Meia-Idade , Hibridização de Ácido NucleicoAssuntos
Cromossomos Humanos Par 16 , Cromossomos Humanos Par 5 , Fusão Gênica , Leucemia Mielomonocítica Crônica/genética , Síndrome de Noonan/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , DNA Metiltransferases Sítio Específica (Adenina-Específica)/genética , Transcrição Gênica , Adulto , Sequência de Bases , Feminino , Humanos , Hibridização in Situ Fluorescente , Translocação GenéticaRESUMO
We investigated genetically affected leukemic cells in FIP1L1-PDGFRA+ chronic eosinophilic leukemia (CEL) and in BCR-ABL1+ chronic myeloid leukemia (CML), two myeloproliferative disorders responsive to imatinib. Fluorescence in situ hybridization specific for BCR-ABL1 and for FIP1L1-PDGFRA was combined with cytomorphology or with lineage-restricted monoclonal antibodies and applied in CML and CEL, respectively. In CEL the amount of FIP1L1-PDGFRA+ cells among CD34+ and CD133+ cells, B and T lymphocytes, and megakaryocytes were within normal ranges. Positivity was found in eosinophils, granulo-monocytes and varying percentages of erythrocytes. In vitro assays with imatinib showed reduced survival of peripheral blood mononuclear cells but no reduction in colony-forming unit growth medium (CFU-GM) growth. In CML the BCR-ABL1 fusion gene was detected in CD34+/CD133+ cells, granulo-monocytes, eosinophils, erythrocytes, megakaryocytes and B-lymphocytes. Growth of both peripheral blood mononuclear cells and CFU-GM was inhibited by imatinib. This study provided evidence for marked differences in the leukemic masses which are targeted by imatinib in CEL or CML, as harboring FIP1L1-PDGFRA or BCR-ABL1.
Assuntos
Proteínas de Fusão bcr-abl/análise , Síndrome Hipereosinofílica/patologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Células-Tronco Neoplásicas/enzimologia , Proteínas de Fusão Oncogênica/análise , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/análise , Fatores de Poliadenilação e Clivagem de mRNA/análise , Antígeno AC133 , Antígenos CD/análise , Antígenos CD34/análise , Antineoplásicos/uso terapêutico , Benzamidas , Linhagem da Célula , Doença Crônica , Células Clonais/enzimologia , Resistência a Medicamentos , Eosinófilos/enzimologia , Eritrócitos/enzimologia , Proteínas de Fusão bcr-abl/antagonistas & inibidores , Glicoforinas/análise , Glicoproteínas/análise , Granulócitos/enzimologia , Células-Tronco Hematopoéticas/enzimologia , Humanos , Síndrome Hipereosinofílica/tratamento farmacológico , Síndrome Hipereosinofílica/enzimologia , Síndrome Hipereosinofílica/genética , Mesilato de Imatinib , Imunofenotipagem , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/enzimologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Subpopulações de Linfócitos/enzimologia , Megacariócitos/enzimologia , Monócitos/enzimologia , Células Mieloides/enzimologia , Proteínas de Fusão Oncogênica/antagonistas & inibidores , Peptídeos/análise , Piperazinas/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêutico , Pirimidinas/uso terapêutico , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/antagonistas & inibidores , Ensaio Tumoral de Célula-Tronco , Inativação do Cromossomo X , Fatores de Poliadenilação e Clivagem de mRNA/antagonistas & inibidoresAssuntos
Antígenos CD1/análise , Antígenos CD34/análise , Leucemia-Linfoma de Células T do Adulto/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/análise , Linfócitos T/imunologia , Adolescente , Linhagem Celular , Feminino , Genes Codificadores da Cadeia beta de Receptores de Linfócitos T , Humanos , Imunofenotipagem , Leucemia-Linfoma de Células T do Adulto/genética , Leucemia-Linfoma de Células T do Adulto/patologiaAssuntos
Fusão Gênica , Síndrome Hipereosinofílica/genética , RNA Mensageiro/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Tropomiosina/genética , Cromossomos Humanos Par 1 , Cromossomos Humanos Par 10 , Cromossomos Humanos Par 5 , Doença Crônica , Primers do DNA , Humanos , Síndrome Hipereosinofílica/patologia , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Neoplasias da Glândula Tireoide/genéticaRESUMO
Six patients with de novo acute myeloid leukemia (AML) and a t(2;3)(p15-21;q26-27) were identified among approximately 1000 cases enrolled in the GIMEMA trial. The t(2;3) was the sole anomaly in three patients, whereas in three cases monosomy 7, trisomy 15 and 22, and trisomy 14 represented additional aberrations. No cryptic chromosome deletions at 5q, 7q, 12p, and 20q were observed. One patient carried a FLT3 D835 mutation; FLT3 internal tandem duplication (ITD) was not detected in three patients tested. Characterization of the translocation breakpoints using a 3q26 BAC contig specific for the PRDM3 locus showed that the breakpoints were located 5' to EVIl as follows: within myelodysplatic syndrome (MDS) intron 1 (# 3), between MDS1 exons 2 and 3 in three patients (# 1, 2, 4) with a 170bp cryptic deletion distal to the breakpoint in one (# 2), and in a more centromeric position spanning from intron 2 to the 5' region of EVI1 (# 6, 5). A set of 2p16-21 BAC probes showed that the breakpoints on chromosome 2p were located within BCL11A in two separate regions (# 1, 4 and # 2-5), within the thyroid adenoma-associated (THADA) gene (# 6) or distal to the ZFP36L2 locus (# 3). Regulatory elements were present in proximity of these breakpoints. RACE PCR studies revealed a chimeric transcript in 1/6 patient analyzed, but no fusion protein. Quantitative PCR showed a 21-58-fold over-expression of the EVIl gene in all cases analyzed. The patients showed dysplasia of at least two myeloid cell lineages in all cases; they had a low-to-normal platelet count and displayed an immature CD34+ CD117+ immunophenotype. Despite intensive chemotherapy and a median age of 43 years (range 36-59), only two patients attained a short-lived response; one patient is alive with active disease at 12 months, five died at 4-14 months. We arrived at the following conclusions: (a) the t(2;3) is a recurrent translocation having an approximate 0.5% incidence in adult AML; (b) breakpoints involve the 5' region of EVIl at 3q26, and the BCL11A, the THADA gene or other regions at 2p16.1-21; (c) cryptic deletions distal to the 3q26 breakpoint may occur in some cases; (d) the juxtaposition of the 5' region of EVIl with regulatory elements normally located on chromosome 2 brings about EVI1 overexpression; (e) clinical outcome in these cases is severe.
Assuntos
Cromossomos Humanos Par 2/genética , Cromossomos Humanos Par 3/genética , Leucemia Mieloide/genética , Translocação Genética/genética , Doença Aguda , Adulto , Análise Citogenética/métodos , Feminino , Humanos , Hibridização in Situ Fluorescente , Cariotipagem , Leucemia Mieloide/diagnóstico , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Prognóstico , TrissomiaRESUMO
Fluorescence in situ hybridization and comparative genomic hybridization characterized 6p rearrangements in eight primary and in 10 secondary myeloid disorders (including one patient with Fanconi anemia) and found different molecular lesions in each group. In primary disorders, 6p abnormalities, isolated in six patients, were highly heterogeneous with different breakpoints along the 6p arm. Reciprocal translocations were found in seven. In the 10 patients with secondary acute myeloid leukemia/myelodysplastic syndrome (AML/MDS), the short arm of chromosome 6 was involved in unbalanced translocations in 7. The other three patients showed full or partial trisomy of the 6p arm, that is, i(6)(p10) (one patient) and dup(6)(p) (two patients). In 5/7 patients with unbalanced translocations, DNA sequences were overrepresented at band 6p21 as either cryptic duplications (three patients) or cryptic low-copy gains (two patients). In the eight patients with cytogenetic or cryptic 6p gains, we identified a common overrepresented region extending for 5-6 megabases from the TNF gene to the ETV-7 gene. 6p abnormalities were isolated karyotype changes in four patients. Consequently, in secondary AML/MDS, we hypothesize that 6p gains are major pathogenetic events arising from acquired and/or congenital genomic instability.
Assuntos
Cromossomos Humanos Par 6/genética , Leucemia Mieloide/genética , Síndromes Mielodisplásicas/genética , Segunda Neoplasia Primária/genética , Translocação Genética/genética , Doença Aguda , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise Citogenética , Feminino , Humanos , Hibridização in Situ Fluorescente , Leucemia Mieloide/diagnóstico , Masculino , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/diagnóstico , Segunda Neoplasia Primária/diagnóstico , Sensibilidade e EspecificidadeRESUMO
Mutations in exon 12 of the nucleophosmin (NPM1) gene occur in about 60% of adult AML with normal karyotype. By exploiting a specific feature of NPM1 mutants, that is insertion at residue 956 or deletion/insertion at residue 960, we developed highly sensitive, real-time quantitative (RQ) polymerase chain reaction (PCR) assays, either in DNA or RNA, that are specific for various NPM1 mutations. In all 13 AML patients carrying NPM1 mutations at diagnosis, cDNA RQ-PCR showed >30 000 copies of NPM1-mutated transcript. A small or no decrease in copies was observed in three patients showing partial or no response to induction therapy. The number of NPM1-mutated copies was markedly reduced in 10 patients achieving complete hematological remission (five cases: <100 copies; five cases: 580-5046 copies). In four patients studied at different time intervals, the number of NPM1 copies closely correlated with clinical status and predicted impending hematological relapse in two. Thus, reliable, sensitive RQ-PCR assays for NPM1 mutations can now monitor and quantify MRD in AML patients with normal karyotype and NPM1 gene mutations.
