Asunto(s)
Células Dendríticas/patología , Leucemia Mielomonocítica Crónica/patología , Neoplasias de Células Plasmáticas/patología , Anciano , Anciano de 80 o más Años , Células Dendríticas/inmunología , Humanos , Inmunofenotipificación , Leucemia Mielomonocítica Crónica/genética , Leucemia Mielomonocítica Crónica/inmunología , Masculino , Neoplasias de Células Plasmáticas/genética , Neoplasias de Células Plasmáticas/inmunologíaAsunto(s)
Proteínas de Unión al ADN/genética , Cariotipo , Trastornos Mieloproliferativos/diagnóstico , Trastornos Mieloproliferativos/genética , Proteínas de Fusión Oncogénica/genética , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biopsia , Médula Ósea/patología , Análisis Mutacional de ADN , Humanos , Recuento de Leucocitos , Masculino , Trastornos Mieloproliferativos/tratamiento farmacológico , UltrasonografíaAsunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Proteínas de Complejo Poro Nuclear/genética , Asparaginasa/uso terapéutico , Biomarcadores de Tumor/genética , Ciclofosfamida/uso terapéutico , Citarabina/uso terapéutico , Daunorrubicina/uso terapéutico , Regulación hacia Abajo/genética , Humanos , Mercaptopurina/uso terapéutico , Metotrexato/uso terapéutico , Mutación/genética , Proteínas de Fusión Oncogénica/genética , Prednisona/uso terapéutico , Pronóstico , Translocación Genética/genética , Regulación hacia Arriba/genética , Vincristina/uso terapéuticoAsunto(s)
Mutación , Síndromes Mielodisplásicos/genética , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Progresión de la Enfermedad , GTP Fosfohidrolasas/genética , Genes p53 , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Proteínas de la Membrana/genética , Proteínas Nucleares/genética , Proteínas Represoras/genética , Ribonucleoproteínas/genéticaAsunto(s)
Anemia Macrocítica/genética , Anemia Macrocítica/metabolismo , Eritroblastos/metabolismo , Leucina/farmacología , Proteínas Ribosómicas/deficiencia , Transducción de Señal/efectos de los fármacos , Serina-Treonina Quinasas TOR/metabolismo , Deleción Cromosómica , Cromosomas Humanos Par 5/genética , Cromosomas Humanos Par 5/metabolismo , Activación Enzimática/efectos de los fármacos , Humanos , Proteínas Ribosómicas/genéticaRESUMEN
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.
Asunto(s)
Ensamble y Desensamble de Cromatina/genética , Sitios Frágiles del Cromosoma , Cromosomas Humanos Par 10 , Reparación del ADN/genética , Leucemia/genética , Proteínas de Ensamble de Clatrina Monoméricas/genética , Factores de Transcripción/genética , Regulación hacia Arriba , Animales , Humanos , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Translocación GenéticaAsunto(s)
Azacitidina/uso terapéutico , Metilación de ADN , Proteínas de Unión al ADN/genética , Mutación/genética , Síndromes Mielodisplásicos/tratamiento farmacológico , Síndromes Mielodisplásicos/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteínas Proto-Oncogénicas/genética , Adulto , Anciano , Antimetabolitos Antineoplásicos/uso terapéutico , Dioxigenasas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Retrospectivos , Factores de Riesgo , Tasa de SupervivenciaRESUMEN
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.
Asunto(s)
Análisis Citogenético/métodos , Análisis Citogenético/normas , Pruebas Genéticas , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Neoplasias/diagnóstico , Garantía de la Calidad de Atención de Salud , Genotipo , Humanos , Italia , Neoplasias/genética , Diagnóstico Prenatal , Factores de TiempoAsunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Cromosomas Humanos Par 12/genética , Cromosomas Humanos Par 5/genética , Leucemia Monocítica Aguda/genética , Proteínas del Tejido Nervioso/genética , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Translocación Genética , Adulto , Humanos , Hibridación Fluorescente in Situ , Leucemia Monocítica Aguda/diagnóstico , Leucemia Monocítica Aguda/terapia , MasculinoAsunto(s)
Inestabilidad Genómica , Hepatitis C/complicaciones , Linfoma no Hodgkin/genética , Adulto , Anciano , Anciano de 80 o más Años , Aberraciones Cromosómicas , Femenino , Humanos , Hibridación Fluorescente in Situ , Linfoma no Hodgkin/virología , Masculino , Persona de Mediana Edad , Hibridación de Ácido NucleicoAsunto(s)
Cromosomas Humanos Par 16 , Cromosomas Humanos Par 5 , Fusión Génica , Leucemia Mielomonocítica Crónica/genética , Síndrome de Noonan/genética , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Metiltransferasa de ADN de Sitio Específico (Adenina Especifica)/genética , Transcripción Genética , Adulto , Secuencia de Bases , Femenino , Humanos , Hibridación Fluorescente in Situ , Translocación GenéticaRESUMEN
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.
Asunto(s)
Proteínas de Fusión bcr-abl/análisis , Síndrome Hipereosinofílico/patología , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Células Madre Neoplásicas/enzimología , Proteínas de Fusión Oncogénica/análisis , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/análisis , Factores de Escisión y Poliadenilación de ARNm/análisis , Antígeno AC133 , Antígenos CD/análisis , Antígenos CD34/análisis , Antineoplásicos/uso terapéutico , Benzamidas , Linaje de la Célula , Enfermedad Crónica , Células Clonales/enzimología , Resistencia a Medicamentos , Eosinófilos/enzimología , Eritrocitos/enzimología , Proteínas de Fusión bcr-abl/antagonistas & inhibidores , Glicoforinas/análisis , Glicoproteínas/análisis , Granulocitos/enzimología , Células Madre Hematopoyéticas/enzimología , Humanos , Síndrome Hipereosinofílico/tratamiento farmacológico , Síndrome Hipereosinofílico/enzimología , Síndrome Hipereosinofílico/genética , Mesilato de Imatinib , Inmunofenotipificación , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/enzimología , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Subgrupos Linfocitarios/enzimología , Megacariocitos/enzimología , Monocitos/enzimología , Células Mieloides/enzimología , Proteínas de Fusión Oncogénica/antagonistas & inhibidores , Péptidos/análisis , Piperazinas/uso terapéutico , Inhibidores de Proteínas Quinasas/uso terapéutico , Pirimidinas/uso terapéutico , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/antagonistas & inhibidores , Ensayo de Tumor de Célula Madre , Inactivación del Cromosoma X , Factores de Escisión y Poliadenilación de ARNm/antagonistas & inhibidoresAsunto(s)
Antígenos CD1/análisis , Antígenos CD34/análisis , Leucemia-Linfoma de Células T del Adulto/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Linfocitos T/inmunología , Adolescente , Línea Celular , Femenino , Genes Codificadores de la Cadena beta de los Receptores de Linfocito T , Humanos , Inmunofenotipificación , Leucemia-Linfoma de Células T del Adulto/genética , Leucemia-Linfoma de Células T del Adulto/patologíaAsunto(s)
Fusión Génica , Síndrome Hipereosinofílico/genética , ARN Mensajero/genética , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Tropomiosina/genética , Cromosomas Humanos Par 1 , Cromosomas Humanos Par 10 , Cromosomas Humanos Par 5 , Enfermedad Crónica , Cartilla de ADN , Humanos , Síndrome Hipereosinofílico/patología , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Neoplasias de la Tiroides/genéticaRESUMEN
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.
Asunto(s)
Cromosomas Humanos Par 2/genética , Cromosomas Humanos Par 3/genética , Leucemia Mieloide/genética , Translocación Genética/genética , Enfermedad Aguda , Adulto , Análisis Citogenético/métodos , Femenino , Humanos , Hibridación Fluorescente in Situ , Cariotipificación , Leucemia Mieloide/diagnóstico , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Valor Predictivo de las Pruebas , Pronóstico , TrisomíaRESUMEN
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.
Asunto(s)
Cromosomas Humanos Par 6/genética , Leucemia Mieloide/genética , Síndromes Mielodisplásicos/genética , Neoplasias Primarias Secundarias/genética , Translocación Genética/genética , Enfermedad Aguda , Adulto , Anciano , Anciano de 80 o más Años , Análisis Citogenético , Femenino , Humanos , Hibridación Fluorescente in Situ , Leucemia Mieloide/diagnóstico , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/diagnóstico , Neoplasias Primarias Secundarias/diagnóstico , Sensibilidad y EspecificidadRESUMEN
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.
