Assuntos
Janus Quinase 2/genética , Prurido/patologia , Água/efeitos adversos , Programas de Triagem Diagnóstica , Eritropoetina/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Transtornos Mieloproliferativos/complicações , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/genética , Transtornos Mieloproliferativos/patologia , Policitemia Vera/complicações , Policitemia Vera/diagnóstico , Policitemia Vera/genética , Policitemia Vera/patologia , Prurido/etiologia , Estudos RetrospectivosAssuntos
Consenso , Técnicas de Diagnóstico Molecular/métodos , Transtornos Mieloproliferativos/genética , Análise Mutacional de DNA , Marcadores Genéticos/genética , Humanos , Irlanda , Janus Quinase 2/genética , Técnicas de Diagnóstico Molecular/normas , Transtornos Mieloproliferativos/diagnóstico , Transtornos Mieloproliferativos/enzimologia , Guias de Prática Clínica como AssuntoRESUMO
PURPOSE: Chronic neutrophilic leukemia is a rare form of myeloproliferative neoplasm characterized by mature neutrophil hyperleukocytosis. The majority of patients harbor somatic mutations of CSF3R gene and are potentially amenable to targeted therapy with JAK inhibitors. The incidence and clinical significance of additional mutations requires clarification. MATERIALS AND METHODS: A next-generation sequencing approach for myeloid malignancy-associated mutations was applied to diagnostic and matched blast crisis samples from four chronic neutrophilic leukemia patients. RESULTS: Next-generation sequencing confirmed the CSF3R T618I in all patients with identification of concurrent SRSF2, SETBP1, NRAS and CBL mutations at diagnosis. At blast crisis, clonal evolution was evidenced by an increased CSF3R T618I allele frequency and by loss or acquisition of CBL and NRAS mutations. CONCLUSION: The diagnostic utility of a targeted next-generation sequencing approach was clearly demonstrated with the identification of additional mutations providing the potential for therapeutic stratification of chronic neutrophilic leukemia patients.
Assuntos
Crise Blástica/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Leucemia Neutrofílica Crônica/genética , Leucemia Neutrofílica Crônica/patologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , MutaçãoRESUMO
Detection of BCR-ABL transcripts in chronic myeloid leukaemia (CML) is used to confirm the diagnosis and to monitor residual disease. Quantitative techniques are required to predict response to therapy or early relapse. We have evaluated an assay in which transcription-mediated amplification (TMA) of BCR-ABL and ABL transcripts is achieved using reverse transcriptase and RNA polymerase. The products are quantified in the hybridisation protection assay (HPA) using acridinium ester-labelled DNA probes and chemiluminescence. The method is a single tube procedure which uses small amounts of RNA (<500 ng/triplicate analysis), is technically simple (requiring just two waterbaths and a luminometer), rapid (total assay time <4 h) and sensitive (capable of detecting one BCR-ABL-positive K562 cell in the presence of 10(4)-10(5) BCR-ABL-negative cells). BCR-ABL signals from patient RNA samples were quantified relative to known amounts of K562 RNA and normalised to levels of ABL. BCR-ABL/ABL ratios ranged from 0.15 to 1.59 (median 0.65) in RNA from diagnostic blood or bone marrow of 18 CML patients and were < or =0.0001 in 20 normal controls. Sequential samples analysed from six CML patients post-allogeneic bone marrow transplantation who relapsed and received donor lymphocyte infusions showed BCR-ABL/ABL ratios which reflected patient status or treatment. A BCR-ABL/ABL ratio of 0.01 served as a useful arbitrary indicator value, with results above and below this value generally correlating with relapse or remission, respectively.
Assuntos
Proteínas de Fusão bcr-abl/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Recidiva Local de Neoplasia/genética , RNA Mensageiro/análise , RNA Neoplásico/análise , Acridinas , Transplante de Medula Óssea , Sondas de DNA , Proteínas de Fusão bcr-abl/metabolismo , Amplificação de Genes , Facilitação Imunológica de Enxerto , Humanos , Hibridização Genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Medições Luminescentes , Recidiva Local de Neoplasia/diagnóstico , Recidiva Local de Neoplasia/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , RNA Neoplásico/biossíntese , Fatores de Risco , Succinimidas , Transcrição GênicaAssuntos
Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Mutação Puntual , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Proto-Oncogênicas c-abl/genética , Adulto , Resistencia a Medicamentos Antineoplásicos , Proteínas de Fusão bcr-abl/genética , Humanos , Masculino , Inibidores de Proteínas Quinases/farmacologiaAssuntos
Calreticulina/genética , Mutação , Inibidores da Agregação Plaquetária/uso terapêutico , Quinazolinas/uso terapêutico , Trombocitemia Essencial/tratamento farmacológico , Trombocitemia Essencial/genética , Idoso , Alelos , Humanos , Masculino , Contagem de Plaquetas , Trombocitemia Essencial/diagnósticoRESUMO
Hairy cell leukaemia (HCL) has distinct clinical, morphological and immunophenotypic features with no recurrent cytogenetic or molecular abnormalities reported until the recent description of the BRAF V600E mutation in patients with classical HCL. The incidence of this mutation was sought in 27 patients with either classical HCL or HCL variant by an allele-specific PCR approach and findings related to morphology, cytochemistry and immunophenotype. A high degree of correlation was noted between the presence of BRAF V600E and established diagnostic criteria in 26/27 patients with HCL/HCL variant. Detection of the BRAF V600E mutation is therefore a useful adjunct in the differential diagnosis of HCL and HCL variant and highlights the value of a multifaceted approach to the diagnosis of this malignancy.
Assuntos
Leucemia de Células Pilosas/diagnóstico , Leucemia de Células Pilosas/genética , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Células da Medula Óssea/patologia , Diagnóstico Diferencial , Feminino , Variação Genética , Histocitoquímica , Humanos , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosAssuntos
Cromossomos Humanos Par 21/genética , Cromossomos Humanos Par 8/genética , Leucemia Mieloide/genética , Mutação , Proteínas Proto-Oncogênicas c-kit/genética , Trissomia , Doença Aguda , Humanos , Proteínas de Fusão Oncogênica/genética , Proteínas de Fusão Oncogênica/metabolismo , Translocação GenéticaRESUMO
BACKGROUND: Acute myeloid leukaemia (AML) is a heterogeneous clonal disorder of haematopoietic progenitor cells. Approximately half of all adult AML patients have a normal karyotype (NK-AML) and an intermediate risk prognosis. AIMS: To determine the incidence and prognostic significance of NPM1 and FLT3-ITD mutations in a population of patients with NK-AML. METHODS: FLT3-ITD and NPM1 mutation status was retrospectively sought in presentation samples from 44 NK-AML patients. RESULTS: FLT3-ITD and NPM1 mutations were detected in 45.5 and 54.5% of patients, respectively, allowing stratification according to genotype. CONCLUSIONS: FLT3-ITD and NPM1 mutation status can be defined in NK-AML. Prospective screening for these mutations is advocated in all NK-AML patients, as the genotype is of clinical importance when considering treatment options including stem cell transplantation.
Assuntos
Leucemia Mieloide Aguda/genética , Proteínas Nucleares/genética , Fosfoproteínas/genética , Tirosina Quinase 3 Semelhante a fms/genética , Adolescente , Adulto , Idoso , Eletroforese em Gel de Ágar , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Nucleofosmina , Prognóstico , Estudos Retrospectivos , Estudos Soroepidemiológicos , Cariótipo XYY , Adulto JovemAssuntos
Biomarcadores Tumorais/genética , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/genética , Leucemia Mieloide Crônica Atípica BCR-ABL Negativa/terapia , Mutação de Sentido Incorreto , Receptores de Fator Estimulador de Colônias/genética , Aloenxertos , Sequência de Bases , DNA de Neoplasias/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transplante de Células-Tronco , Fatores de TempoRESUMO
BACKGROUND: The chronic myeloproliferative disorders (MPD) are clonal haemopoietic stem cell disorders. AIMS: The incidence of JAK2 V617F mutation was sought in a population of patients with MPD. METHODS: The JAK2 V617 mutation status was determined in 79 patients with known MPD and 59 patients with features suggestive of MPD. RESULTS: The mutation was found in patients with polycythaemia vera, essential thrombocythaemia, idiopathic myelofibrosis and in patients with other chronic myeloproliferative disorders. Eight JAK2 V617F positive cases were identified amongst those patients with features suggestive of MPD. CONCLUSIONS: The incidence of the JAK2 V617F mutation in MPD patients is similar to that reported by other groups. The assay confirmed and refined the diagnosis of several patients with features indicative of MPD. We suggest screening for this mutation in all patients with known and suspected MPD as identification is valuable in classification and is a potential target for signal transduction therapy.
Assuntos
Janus Quinase 2/genética , Transtornos Mieloproliferativos/genética , Doença Crônica , Humanos , Mutação , Policitemia Vera/genética , Reação em Cadeia da Polimerase , Mielofibrose Primária/genética , Trombocitemia Essencial/genéticaAssuntos
Proteínas de Fusão bcr-abl/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Sequência de Aminoácidos , Sequência de Bases , Feminino , Humanos , Íntrons , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase em Tempo Real , Deleção de SequênciaAssuntos
Regulação Leucêmica da Expressão Gênica , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Receptores de Citocinas/genética , Adolescente , Adulto , Idoso , Feminino , Testes Genéticos , Humanos , Hibridização in Situ Fluorescente , Cariótipo , Masculino , Pessoa de Meia-Idade , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto JovemRESUMO
Acute myeloid leukaemia (AML) may not only occur as a de novo disease but may evolve from a preceding myelodysplastic syndrome (MDS) or may result from therapy for a previous malignancy. These secondary acute myeloid leukaemias (sAML) possess some common biological and clinical features of the corresponding de novo disorders. The cytokine interleukin-1 (IL-1) is known to have a role in haematopoiesis, and modulation of its action might contribute to the deregulation of proliferation seen in leukaemia. It has recently been reported that a variable number tandem repeat (VNTR) polymorphism in the IL-1 receptor antagonist (IL-1ra) gene is closely associated with the severity of many inflammatory and autoimmune diseases, and may also play a role in the pathogenesis of sAML. We sought to confirm this finding in a large group of patients classified as having sAML. We found no differences in either the genotypic or allele frequencies of the polymorphism studied when compared with those of normal controls or other haematological disorders. No differences were observed in allele frequencies between younger and older patients, or between those patients who had an antecedent myelodysplasia and those who had received prior chemotherapy or radiotherapy. We conclude that the described polymorphism in the IL-1ra gene is not associated with the development of sAML.
Assuntos
Leucemia Mieloide , Polimorfismo Genético , Sialoglicoproteínas/genética , Doença Aguda , Fatores Etários , Frequência do Gene , Humanos , Proteína Antagonista do Receptor de Interleucina 1 , Síndromes Mielodisplásicas/complicaçõesRESUMO
Acute myeloid leukaemia (AML) patients with either a t(15;17), t(8;21) or inv(16) at diagnosis have 'good-risk' disease with a favourable response to therapy and improved survival. Detection of cryptic fusion genes created by these translocations has been reported where there is no cytogenetic evidence of the corresponding abnormality. It is likely that these cases share the same favourable prognosis. Secondary cytogenetic changes commonly associated with these rearrangements are +8 with t(15;17), del(9q) with t(8;21) and +22 with inv(16). These secondary abnormalities are also observed alone, raising the possibility that they may be markers of underlying cryptic rearrangements. In order to determine the frequency of these rearrangements in AML cases with +8, del(9q) or +22 we have performed an analysis of 63 such patients in whom there was no evidence of a t(15;17), t(8;21) or inv(16) by cytogenetics. No disease-related fusion transcripts were identified, indicating that the secondary changes are rarely markers for cryptic rearrangements.
Assuntos
Biomarcadores Tumorais/genética , Cromossomos Humanos Par 22/genética , Cromossomos Humanos Par 8/genética , Rearranjo Gênico , Leucemia Mieloide/genética , Proteínas de Fusão Oncogênica/genética , Trissomia , Doença Aguda , Adulto , Inversão Cromossômica , Cromossomos Humanos Par 15/genética , Cromossomos Humanos Par 16/genética , Humanos , Translocação GenéticaRESUMO
It has been established that cytogenetic findings at diagnosis of acute myeloid leukaemia (AML) are a powerful prognostic indicator. Patients who have the inv(16)(p13q22), closely associated with the FAB subtype M4Eo. are deemed to have good-risk disease. This subtle translocation may be difficult to detect in poor-quality metaphase preparations and if missed could lead to the incorrect assignment of risk group and influence further treatment strategies. We studied 321 patients with AML at diagnosis for the presence of inv(16)(p13q22) and CBF beta/MYH11 fusion transcripts by cytogenetic and RT-PCR techniques respectively. Karyotypic analysis detected 21 cases of inv(16) (p13q22), all of which were PCR positive. A further 12 cases were detected at the molecular level only, in FAB types other than M4Eo. The observed frequencies of CBF beta/MYH11 fusion transcripts in our study have been adjusted for the reported incidence of each FAB subtype and we calculate that 10.1% of all new cases of AMLs have molecular evidence of inv(16)(p13q22). only half of which are of the M4Eo subtype. We conclude that molecular screening for the presence of CBF beta/MYH11 fusion transcripts should be mandatory in all case of AML at diagnosis.