Assuntos
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
Fatores de Transcrição Kruppel-Like/genética , Linfoma de Zona Marginal Tipo Células B/genética , Mutação , Neoplasias Esplênicas/genética , Alelos , Transformação Celular Neoplásica , Análise Mutacional de DNA , Deleção de Genes , Dosagem de Genes , Células HEK293 , Humanos , Hibridização in Situ Fluorescente , Células Jurkat , Polimorfismo de Nucleotídeo ÚnicoRESUMO
We conducted a phase II, noncomparative, open-label, multicenter GIMEMA (Gruppo Italiano Malattie EMatologiche dell'Adulto) study (CLL0809) to assess the efficacy and safety of bendamustine in combination with ofatumumab (BendOfa) in relapsed/refractory chronic lymphocytic leukemia (CLL). Forty-seven patients from 14 centers were evaluated. Therapy consisted of bendamustine (70 mg/m(2)) for 2 consecutive days every 28 days, and ofatumumab 300 mg on day 1 and 1000 mg on day 8 during the first cycle, and 1000 mg on day 1 subsequently. Treatment was administered up to six cycles. The overall response rate (ORR), as per intention-to-treat analysis, was 72.3% (95% confidence of interval (CI), 57-84%), with 17% complete responses. After a median follow-up of 24.2 months, the overall survival was 83.6% (95% CI, 73.0-95.7%) and the progression-free survival (PFS) was 49.6% (95% CI, 35.9-68.6%). The median PFS was 23.6 months. Univariate and multivariate analyses were used to identify clinical and biological characteristics associated with ORR and PFS. Myelosuppression was the most common toxicity; grade ≥3 neutropenia was observed in 61.7% of patients; however, grade ≥3 infections occurred in 6% of patients. BendOfa is feasible and effective in relapsed/refractory CLL patients, including patients with high-risk clinical and biological features.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Idoso , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais Humanizados , Cloridrato de Bendamustina , Feminino , Humanos , Leucemia Linfocítica Crônica de Células B/patologia , Masculino , Compostos de Mostarda Nitrogenada/administração & dosagem , RecidivaRESUMO
BACKGROUND: Genomic complexity can predict the clinical course of patients affected by chronic lymphocytic leukemia (CLL) with a normal FISH. However, large studies are still lacking. Here, we analyzed a large series of CLL patients and also carried out the so far largest comparison of FISH versus single-nucleotide polymorphism (SNP) array in this disease. PATIENTS AND METHODS: SNP-array data were derived from a previously reported dataset. RESULTS: Seventy-seven of 329 CLL patients (23%) presented with a normal FISH. At least one large (>5 Mb) genomic aberration was detected by SNP array in 17 of 77 patients (22%); this finding significantly affected TTT. There was no correlation with the presence of TP53 mutations. In multivariate analysis, including age, Binet stage, IGHV genes mutational status and large genomic lesion, the latter three factors emerged as independent prognosticators. The concordance between FISH and SNP array varied between 84 and 97%, depending on the specific genomic locus investigated. CONCLUSIONS: SNP array detected additional large genomic aberrations not covered by the standard FISH panel predicting the outcome of CLL patients.
Assuntos
Aberrações Cromossômicas , Leucemia Linfocítica Crônica de Células B/genética , Feminino , Genótipo , Humanos , Cadeias Pesadas de Imunoglobulinas/genética , Região Variável de Imunoglobulina/genética , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Prognóstico , Proteína Supressora de Tumor p53/genéticaAssuntos
Antineoplásicos/farmacologia , Ácidos Hidroxâmicos/farmacologia , Leucemia Megacarioblástica Aguda/tratamento farmacológico , Leucemia Megacarioblástica Aguda/metabolismo , NF-kappa B/metabolismo , Proteínas Wnt/metabolismo , Apoptose/efeitos dos fármacos , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ensaio de Desvio de Mobilidade Eletroforética , Citometria de Fluxo , Perfilação da Expressão Gênica , Inibidores de Histona Desacetilases , Humanos , Leucemia Megacarioblástica Aguda/patologia , NF-kappa B/genética , Análise de Sequência com Séries de Oligonucleotídeos , Transdução de Sinais , Células Tumorais Cultivadas , VorinostatAssuntos
Antineoplásicos/farmacologia , Ácidos Borônicos/farmacologia , Inibidores Enzimáticos/farmacologia , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Genes do Tumor de Wilms/efeitos dos fármacos , Ácidos Hidroxâmicos/farmacologia , Proteínas de Neoplasias/biossíntese , Pirazinas/farmacologia , Proteínas WT1/biossíntese , Bortezomib , Linhagem Celular Tumoral/metabolismo , Regulação para Baixo/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Sinergismo Farmacológico , Humanos , Leucemia Megacarioblástica Aguda/patologia , Leucemia Mielomonocítica Crônica/patologia , Megacariócitos/efeitos dos fármacos , Megacariócitos/metabolismo , VorinostatAssuntos
Angiopoietina-2/análise , Cadeias Pesadas de Imunoglobulinas/genética , Leucemia Linfocítica Crônica de Células B/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Prognóstico , Resultado do TratamentoRESUMO
Acute promyelocytic leukaemia (APL) is a well-defined disease characterized by a typical morphology of leukaemic cells, the presence of t(15;17) translocation and the unique sensitivity to the differentiating effect of all-trans retinoic acid. Nevertheless, some aspects are variable among APL patients, with differences substantially related to morphological variants, peripheral leukocytes count, the presence of a disseminated intravascular coagulopathy, different PML/RARalpha isoforms (long, variable or short) and Fms-like tyrosine kinase 3 (Flt3) mutations. In order to better define this variability, we investigated the gene expression profiles of 18 APL cases revealing, besides a high uniformity in gene expression pattern, the presence of few robust differences among patients able to identify, by an unsupervised analysis, two major clusters of patients characterized by different phenotypes (hypogranular M3v vs classical M3) and by the presence or absence of Flt3 internal tandem duplications (ITDs). Further supervised analysis confirmed that Flt3 status was the APL parameter best associated with these two subgroups. We identified, between Flt3 wild-type and Flt3-ITDs subsets, 147 differentially expressed genes that were involved in the cytoskeleton organization, in the cell adhesion and migration, in the proliferation and the coagulation/inflammation pathways as well as in differentiation and myeloid granules constitution suggesting a role of Flt3 mutations in the pathogenesis and clinical manifestations of APL.
Assuntos
Perfilação da Expressão Gênica , Leucemia Promielocítica Aguda/genética , Família Multigênica , Tirosina Quinase 3 Semelhante a fms/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise por Conglomerados , Éxons , Feminino , Humanos , Leucemia Promielocítica Aguda/classificação , Leucemia Promielocítica Aguda/diagnóstico , Masculino , Pessoa de Meia-Idade , Mutação , FenótipoAssuntos
Transformação Celular Neoplásica/patologia , Leucemia Linfocítica Crônica de Células B/patologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Segunda Neoplasia Primária/etiologia , Linhagem da Célula/genética , Linhagem da Célula/imunologia , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/imunologia , Células Clonais/patologia , Análise Citogenética , Humanos , Imunofenotipagem , Leucemia Linfocítica Crônica de Células B/etiologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/etiologia , Linfócitos/patologia , Pessoa de Meia-Idade , Células Mieloides/patologia , Segunda Neoplasia Primária/patologiaRESUMO
Seventy-two patients with non-Hodgkin's lymphoma were evaluated for the presence of molecular markers (IgH, bcl-1, bcl-2 rearrangement) on bone marrow, at diagnosis and after PBSCT, and on harvests in order to find a possible predictive role of minimal residual disease on treatment outcome. At diagnosis, 41 (59%) out of 69 available bone marrows showed molecular involvement. Fifty-six percent of leukaphereses were involved, mainly indolent lymphoma (P = 0.001) or advanced disease (P = 0.01). Ex vivo purging cleared only one stem collection out of 31 PCR-positive leukaphereses. Aggressive lymphomas showed both a longer overall survival (OS) (P = 0.03) and relapse-free survival RFS (P = 0.02) when transplanted with unpurged stem cells, whereas indolent NHL survival was not influenced by ex vivo purging. Twenty out of 26 samples taken during follow-up had bone marrow involvement at diagnosis. Of these, 15 cleared their bone marrow; both OS and RFS were significantly longer in the PCR-negative cases (P = 0.05 and P = 0.005). At 1 year after PBSCT, 75% of patients were PCR negative, with 50% molecular remissions; the relapse rate was 55% for patients still PCR positive vs 29% for those who were PCR negative. Thus, after high-dose chemotherapy, close molecular monitoring of MRD using qualitative PCR techniques seems to represent a reliable prognostic indicator.
Assuntos
Biomarcadores Tumorais/análise , Medula Óssea/química , Ciclina D1/análise , Cadeias Pesadas de Imunoglobulinas/análise , Linfoma não Hodgkin/química , Proteínas de Neoplasias/análise , Proteínas Proto-Oncogênicas c-bcl-2/análise , Adolescente , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Purging da Medula Óssea , Terapia Combinada , Ciclina D1/genética , Ciclofosfamida/administração & dosagem , Intervalo Livre de Doença , Doxorrubicina/administração & dosagem , Feminino , Seguimentos , Genes bcl-2 , Humanos , Cadeias Pesadas de Imunoglobulinas/genética , Leucaférese , Tábuas de Vida , Linfoma não Hodgkin/tratamento farmacológico , Linfoma não Hodgkin/mortalidade , Linfoma não Hodgkin/patologia , Linfoma não Hodgkin/terapia , Masculino , Pessoa de Meia-Idade , Neoplasia Residual , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Prednisona/administração & dosagem , Proteínas Proto-Oncogênicas c-bcl-2/genética , Estudos Retrospectivos , Análise de Sobrevida , Resultado do Tratamento , Vincristina/administração & dosagemRESUMO
Nodal marginal zone B-cell lymphoma (NMZL) is actually considered as a distinct entity that must be distinguished from extra-nodal and splenic marginal zone lymphomas. To define the cell origin and the role of antigen stimulation we determined the nucleotide sequence of the tumor-related immunoglobulin heavy chain variable genes in 10 cases of NMZL. The results were also evaluated on the basis of the presence of chronic hepatitis C virus (HCV) infection. All 10 cases harbored VH somatic mutations with a sequence homology compared to the closest germline gene, ranging from 83.33 to 98.28%. Interestingly, different VH segments were preferentially used in HCV-positive and HCV-negative patients: three of five HCV-negative NMZLs used a VH4-34 segment joined with different D and JH segments whereas three of five HCV-positive NMZLs used a VH1-69 gene joined with a D3-22 and a JH4 segment, with very strong similarities in the CDR3s among the three different cases. These data indicate: 1) NMZL is derived from B cells that have experienced the germinal center reaction; 2) the preferential usage of a VH1-69 segment in the majority of the HCV-positive NMZL cases with similar CDR3s suggests the presence of a common antigen, probably a HCV antigen epitope, involved in the B-cell selection; and 3) the use of a VH4-34 segment suggests a role of yet unknown B-cell superantigen(s) in the selection of tumor B-cell precursors in HCV-negative NMZL.
Assuntos
Hepacivirus/isolamento & purificação , Cadeias Pesadas de Imunoglobulinas/genética , Região Variável de Imunoglobulina/genética , Imunoglobulinas/genética , Linfoma de Células B/genética , Linfoma de Células B/virologia , Mutação/fisiologia , Sequência de Bases/genética , Rearranjo Gênico , Humanos , Dados de Sequência Molecular , Mutação/genética , Reação em Cadeia da PolimeraseAssuntos
Proteínas de Fusão bcr-abl/genética , Trombocitemia Essencial/genética , Doença Aguda , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Itália/epidemiologia , Leucemia Mieloide/genética , Masculino , Pessoa de Meia-Idade , Cromossomo Filadélfia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Trombocitemia Essencial/sangue , Trombocitemia Essencial/epidemiologiaRESUMO
BACKGROUND: Human herpesvirus 8 (HHV-8) infection has been linked to the development of Kaposi's sarcoma and to rare lymphoproliferative disorders. METHODS: We used molecular methods, serologic methods, in situ hybridization, and immunohistochemical analyses to study HHV-8 infection in association with nonmalignant illnesses in three patients after transplantation. RESULTS: Primary HHV-8 infections developed in two patients four months after each received a kidney from the same HHV-8-seropositive cadaveric donor. Seroconversion and viremia occurred coincidentally with disseminated Kaposi's sarcoma in one patient and with an acute syndrome of fever, splenomegaly, cytopenia, and marrow failure with plasmacytosis in the other patient. HHV-8 latent nuclear antigen was present in immature progenitor cells from the aplastic marrow of the latter patient. Identification of the highly variable K1 gene sequence of the HHV-8 genome in both the donor's peripheral-blood cells and the recipients' serum confirmed that transmission had occurred. HHV-8 viremia also occurred after autologous peripheral-blood stem-cell transplantation in an HHV-8-seropositive patient with non-Hodgkin's lymphoma. Reactivation of the infection was associated with the development of fever and marrow aplasia with plasmacytosis; there was no evidence of other infections. HHV-8 transcripts and latent nuclear antigen were expressed in the aplastic marrow but not in two normal marrow samples obtained before transplantation. CONCLUSIONS: Primary HHV-8 infection and reactivation of infection may be associated with nonneoplastic complications in immunosuppressed patients.
Assuntos
Doenças da Medula Óssea/etiologia , Transmissão de Doença Infecciosa , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Infecções por Herpesviridae/transmissão , Herpesvirus Humano 8/isolamento & purificação , Transplante de Rim/efeitos adversos , Sarcoma de Kaposi/etiologia , Adulto , Anticorpos Antivirais/sangue , Contagem de Células Sanguíneas , Medula Óssea/virologia , Doenças da Medula Óssea/sangue , Doenças da Medula Óssea/virologia , Evolução Fatal , Genoma Viral , Infecções por Herpesviridae/etiologia , Infecções por Herpesviridae/virologia , Herpesvirus Humano 8/genética , Herpesvirus Humano 8/imunologia , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Sarcoma de Kaposi/virologia , Viremia/etiologia , Ativação ViralRESUMO
In transplant patients, Kaposi sarcoma (KS)-associated herpesvirus or human herpesvirus-8 (HHV-8) infection is associated with the development of KS, primary effusion lymphoma and Castleman disease. Whether HHV-8 is either reactivated in the recipient or transmitted by the donor has been investigated so far only by serologic studies. Thus, we addressed the issue of HHV-8 transmission in the transplantation setting by molecular methods. We exploited the high level variability of the orf-K1 gene and the polymorphism of the orf-73 gene of the HHV-8 genome to assess the genetic relatedness of the HHV-8 strains identified in the posttransplant KS lesions that developed, simultaneously, 20 months after transplantation, in 2 recipients of twin kidneys from the same cadaver donor. The 100% identity of nucleotide sequence of the most variable viral region and the presence of the same, single orf-73 type in both patients provides strong molecular evidence of organ-related transmission of HHV-8 in the setting of transplantation.
Assuntos
Herpesvirus Humano 8/isolamento & purificação , Transplante de Rim , Complicações Pós-Operatórias , Sarcoma de Kaposi/virologia , Sequência de Bases , Feminino , Genótipo , Herpesvirus Humano 8/genética , Herpesvirus Humano 8/fisiologia , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Alinhamento de Sequência , Ativação ViralRESUMO
Fever, cutaneous rash, and hepatitis-for which an infectious cause was suspected-developed in an Italian patient with non-Hodgkin lymphoma after autologous peripheral blood stem cell (PBSC) transplantation. Polymerase chain reaction (PCR) with degenerate primers for the highly conserved DNA polymerase gene of herpesviruses detected herpesvirus sequences 100% identical to human herpesvirus-8 (HHV-8) in serial cell-free serum samples, collected immediately before or concomitant with the occurrence of clinical symptoms; no other common infections were documented. The presence of the HHV-8 genome (clade C) was confirmed by PCR with HHV-8-specific primers for orf 26 and orf-K1. HHV-8 viremia was undetectable either before transplantation or when the patient was clinically asymptomatic. Semiquantitative PCR analysis showed variations of the viral load correlating with the clinical status. Anti-HHV-8 antibodies were detected before and after transplantation by an immunofluorescence assay for lytic antigens. Active HHV-8 infection may be associated with nonmalignant illness after PBSC/bone marrow transplantation.
Assuntos
Linfoma de Burkitt/virologia , Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 8 , Sarcoma de Kaposi/diagnóstico , Adulto , Sequência de Aminoácidos , Anticorpos Antivirais/sangue , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Linfoma de Burkitt/tratamento farmacológico , Linfoma de Burkitt/cirurgia , DNA Viral/sangue , DNA Polimerase Dirigida por DNA/química , DNA Polimerase Dirigida por DNA/genética , Imunofluorescência , Herpesvirus Humano 8/enzimologia , Herpesvirus Humano 8/genética , Herpesvirus Humano 8/imunologia , Humanos , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Sarcoma de Kaposi/etiologia , Sarcoma de Kaposi/virologia , Transplante Autólogo , Carga ViralRESUMO
The late appearance of a cytogenetic/molecular hallmark in human leukemias is a rare event. We report on a case of acute myeloid leukemia with morphology, immunophenotype and clinical features typical of promyelocytic subtype (APL), in which the specific PML/RARalpha gene rearrangement was molecularly detected only at second relapse of disease, without cytogenetic evidence of the t(15;17). The emergence of the PML/RARalpha gene may be therapy-related or may represent the exceptional result of a clonal evolution during progression of neoplasia. At second relapse, a novel cell clone bearing a t(12;13)(p13.2;q14) was also observed and a molecular deletion and rearrangement of a locus at 13q14, distinct from retinoblastoma (Rb1) locus, was found. In this unusual case, the PML/RARalpha product seems to be not essential for the expression of the promyelocytic phenotype at diagnosis and, when detectable, it is not the sole genetic defect.
Assuntos
Cromossomos Humanos Par 12/ultraestrutura , Cromossomos Humanos Par 13/ultraestrutura , Leucemia Promielocítica Aguda/genética , Proteínas de Neoplasias/genética , Proteínas de Fusão Oncogênica/genética , RNA Mensageiro/biossíntese , RNA Neoplásico/biossíntese , Translocação Genética , Cromossomos Humanos Par 15/ultraestrutura , Cromossomos Humanos Par 17/ultraestrutura , Células Clonais/patologia , Progressão da Doença , Evolução Fatal , Feminino , Humanos , Leucemia Promielocítica Aguda/patologia , Pessoa de Meia-Idade , Recidiva , Fatores de TempoRESUMO
BACKGROUND AND OBJECTIVE: Acute promyelocytic leukemia is characterized by the chromosomal translocation t(15;17) which yields the fusion product PML/RARa. All-trans retinoic acid probably induces differentiation of atypical promyelocytes and clinical remission in APL patients by binding to the ligand binding domain (LBD) of the RARa portion of the PML-RARa chimeric protein. Structural alterations of the LBD of the PML/RARa have been revealed in ATRA-resistant APL cell lines and in a few APL patients with acquired clinical resistance to ATRA therapy. Two APL relapsed patients with clinical resistance to ATRA therapy were evaluated for the presence of nucleotide mutations in the LBD of PML/RARa gene and then treated with arsenic trioxide (As2O3). DESIGN AND METHODS: DNA fragments from the LBD of the PML/RARa chimeric transcript were obtained by reverse-transcribed polymerase chain reaction. Direct sequencing was performed by an unambiguous bi-directional automatic analysis. Samples representative of APL onset and relapse were analyzed from both patients. RESULTS: In both patients, at the ATRA-resistant relapse, a missense point mutation in the LBD of the PML/RARa gene was found. The mutations, absent at APL onset, led to an Arg272Gln and to an Arg276Trp amino acid substitution, according to the sequence of the RARa protein. Both patients had complete clinical and hematologic remission after treatment with As2O3. INTERPRETATION AND CONCLUSIONS: LBD missense mutations appear to be a significant mechanism of acquired ATRA-resistance in vivo, closely related to clinical APL relapse. The two cases reported here provide the first in vivo evidence of APL relapsed patients, who have become ATRA-resistant for molecular reasons, being sensitive to arsenic trioxide.