RESUMEN

We aimed to test the efficacy and toxicity of autologous hematopoietic cell transplant (HCT) in Multiple Myeloma (MM) patients aged ≥65 years compared to patients aged 60-64. Two hundred twenty consecutive patients (age ≥65, n = 87) with MM aged 60 and above, who underwent HCT as part of an upfront MM treatment, at four Israeli centers between 2000 and 2014 were included. A melphalan dose of 200 mg/m2 was more frequent in the 60-64 age group vs. the ≥65 age group (77 vs. 57%, p = 0.002). There were no differences between groups in median day of neutrophil engraftment, incidence of infections, grades 3-4 mucositis, cardiovascular events, or non-relapse mortality at 100 days post HCT (4.7, vs. 5%, p = 0.9). A similar rate of improvement in response level was observed (36, vs. 35%, p = 0.87). At 3 years post HCT progression-free survival (PFS) was higher in the 60-64 age group (42 vs. 29%, p = 0.04); however, it was no longer so after adjustment for disease status prior to HCT (p = 0.49). In a Multivariate analysis, melphalan doses and age did not predict PFS. There was no difference in overall survival (OS) between age groups (p = 0.2). We conclude that toxicity profile, response, PFS, and OS of HCT in aged ≥65 patients with myeloma is similar to patients aged 60-64.

Asunto(s)

Trasplante de Células Madre Hematopoyéticas/tendencias , Mieloma Múltiple/diagnóstico , Mieloma Múltiple/terapia , Acondicionamiento Pretrasplante/tendencias , Anciano , Estudios de Cohortes , Femenino , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Trasplante de Células Madre Hematopoyéticas/mortalidad , Humanos , Israel/epidemiología , Masculino , Persona de Mediana Edad , Mieloma Múltiple/mortalidad , Estudios Retrospectivos , Tasa de Supervivencia/tendencias , Acondicionamiento Pretrasplante/efectos adversos , Acondicionamiento Pretrasplante/mortalidad , Trasplante Autólogo/efectos adversos , Trasplante Autólogo/mortalidad , Trasplante Autólogo/tendencias , Resultado del Tratamiento

RESUMEN

Automated morphological assessment of peripheral blood slides has become an important modality facilitating characterization and quantification of cells in a uniform, fast and robust manner. In this study, we evaluated the morphological diversity in peripheral blood films of 94 chronic lymphocytic leukemia (CLL) patients using the DM1200 CellaVision automated microscopy system. Aberrant lymphocytes and smudge cells were enumerated and correlated with CLL immunophenotype, chromosomal aberrations and prognostic parameters. Herein, we show that the percentages of aberrant and smudge cells was highly variable between patients and did not correlate with each other. Increased aberrant lymphocytes and fewer smudge cells were associated with an atypical immunophenotype including low expression of CD23, higher levels of FMC7 and bright surface levels of CD20. High fraction of aberrant lymphocytes also was associated with trisomy 12. These cells were predominantly of small/medium size, sometimes with cleft nuclei. No correlation was noted between aberrant or smudge cells and clinical stage, CD38, ZA70 or time to first treatment. Taken together, automated morphological analysis of peripheral blood leukocytes emerged as a powerful and robust tool for the quantitative morphological stratification of CLL. Integration of the automated morphological features discriminates between different CLL phenotypes and distinct chromosomal aberrations.

Asunto(s)

Aberraciones Cromosómicas , Leucemia Linfocítica Crónica de Células B/genética , Leucemia Linfocítica Crónica de Células B/patología , Leucocitos Mononucleares/patología , Automatización de Laboratorios , Estudios de Casos y Controles , Forma de la Célula , Diagnóstico Diferencial , Citometría de Flujo , Humanos , Inmunofenotipificación , Leucemia Linfocítica Crónica de Células B/clasificación , Linfocitos/patología , Pronóstico

RESUMEN

Peripheral absolute monocyte count (AMC) has been reported to correlate with clinical outcome in different types of cancers. This association may relate to alteration in circulating monocytic subpopulations and tumor infiltrating macrophages. In this study we evaluated the clinical significance of peripheral AMC in 80 treatment naive patients with CLL. Measurement of AMC was based on direct morphological enumeration, due to our findings that complete blood count data may yield incorrect monocytes enumeration values in CLL. The median AMC in patients with CLL was within normal limits, however the AMC range exceeded the values of healthy individuals. The AMC trichotomized patients into 3 distinct sub-groups with different characteristics and outcomes. High AMC patients were younger and had higher absolute lymphocytes count, while patients with low AMC had prominent immune dysregulation (lower serum IgA levels, susceptibility to infections and a tendency for positive direct anti-globulin test). The low and high AMC patients had a shorter time to treatment compared to the intermediates AMC subgroups, whereas low AMC was associated with increased mortality caused by infectious complications. In conclusion, AMC quantification during the disease course classifies CLL patients into subgroups with unique clinical features and outcomes.

Asunto(s)

Leucemia Linfocítica Crónica de Células B/diagnóstico , Leucemia Linfocítica Crónica de Células B/inmunología , Monocitos/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Progresión de la Enfermedad , Femenino , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina A/inmunología , Leucemia Linfocítica Crónica de Células B/mortalidad , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias

Asunto(s)

Anticuerpos Monoclonales de Origen Murino/inmunología , Síndrome Inflamatorio de Reconstitución Inmune/inmunología , Linfoma/inmunología , Anticuerpos Monoclonales de Origen Murino/efectos adversos , Antineoplásicos/efectos adversos , Antineoplásicos/inmunología , Antituberculosos/efectos adversos , Antituberculosos/inmunología , Diagnóstico Diferencial , Humanos , Síndrome Inflamatorio de Reconstitución Inmune/inducido químicamente , Síndrome Inflamatorio de Reconstitución Inmune/diagnóstico , Linfoma/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/inmunología , Mycobacterium tuberculosis/aislamiento & purificación , Rituximab , Tuberculosis Pulmonar/tratamiento farmacológico , Tuberculosis Pulmonar/inmunología , Tuberculosis Pulmonar/microbiología

RESUMEN

Emerging data on intraclonal diversity imply that this phenomenon may play a role in the clinical outcome of patients with chronic lymphocytic leukemia (CLL), where subsets of the CLL clone responding more robustly to external stimuli may gain a growth and survival advantage. In this study, we report intraclonal diversity resolved by responses to CD19 engagement in CLL cells, which can be classified into CD19-responsive (CD19-R) and -nonresponive subpopulations. Engagement of CD19 by anti-CD19 Ab rapidly induced cellular aggregation in the CD19-R CLL cells. The CD19-R CLL cells expressed higher surface levels of CD19 and c-myc mRNA, exhibited distinct morphological features, and were preferentially abolished in rituximab-treated patients. Both subpopulations reacted to sIgM stimulation in a similar manner and exhibited similar levels of Akt and Erk phosphorylation, pointing to functional signaling divergence within the BCR. CD19 unresponsiveness was partially reversible, where nonresponding CD19 cells spontaneously recover their signaling capacity following incubation in vitro, pointing to possible in vivo CD19-signaling attenuating mechanisms. This concept was supported by the lower CD19-R occurrence in bone marrow-derived samples compared with cells derived from the peripheral blood of the same patients. CLL patients with >15.25% of the CD19-R cell fraction had a shorter median time to treatment compared with patients with <15.25% of CD19-R cell fraction. In conclusion, divergence in CD19-mediated signaling unfolds both interpatient and intraclonal diversity in CLL. This signaling diversity is associated with physiological implications, including the location of the cells, their responses to anti-CLL therapeutics, and disease progression.

Asunto(s)

Antígenos CD19/metabolismo , Leucemia Linfocítica Crónica de Células B/metabolismo , Transducción de Señal , Anticuerpos Monoclonales de Origen Murino/farmacología , Antineoplásicos/farmacología , Células Cultivadas , Colesterol/metabolismo , Progresión de la Enfermedad , Humanos , Leucemia Linfocítica Crónica de Células B/genética , Microdominios de Membrana/química , Microdominios de Membrana/efectos de los fármacos , Microdominios de Membrana/metabolismo , Rituximab , Transducción de Señal/efectos de los fármacos

RESUMEN

UNLABELLED: Recent studies have reevaluated whether gemtuzumab ozogamicin (GO) improves the outcome of acute myeloid leukemia (AML) in elderly patients. Over 5 years, we treated 16 elderly patients with AML with GO and cytarabine. A high response rate, prolonged survival, and low toxicity were observed in the favorable and intermediate-I genetic groups of AML. Our study raises the issue about the optimal protocol for these patients. BACKGROUND: The benefit of gemtuzumab ozogamicin (GO) in combination with chemotherapy as frontline therapy in patients with acute myeloid leukemia (AML) is still debated. PATIENTS AND METHODS: We evaluated the safety and efficacy of low-dose GO with cytarabine in elderly patients with newly diagnosed AML. Over the past 5 years, we have treated 16 elderly patients with AML (64-82 years) with GO (3 mg/m(2)) followed by continuous infusion of cytarabine (100 mg/m(2)) for 7 days. RESULTS: Complete remission (CR) was achieved in 68.8% of patients; however, this was true only in patients in the favorable or intermediate-I cytogenetic risk groups. Of the 12 patients with AML in the favorable and intermediate-I genetic groups, 11 (91.7%) achieved CR. By comparison, of all 4 patients in the intermediate-II or adverse genetic groups, none of the patients achieved CR (P = .003). The median disease-free survival and overall survival (OS) was 10.9 and 18.8 months, respectively, for patients who achieved CR. The estimated median survival was 15 months in the favorable and intermediate-I cytogenetic groups and only 4.4 months in the intermediate-II and unfavorable risk groups (P = .008). The toxicity profile was also manageable in patients with AML who were mainly older than 70 years with good performance status (PS). The 8-week mortality rate was 6.25%, which is relatively low in this high-risk group of patients. These data are in line with results from 2 randomized trials suggesting that the addition of low-dose GO should be further investigated to reevaluate its role in selected elderly patients with AML and raises the issue of the optimal protocol.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Leucemia Mieloide Aguda/tratamiento farmacológico , Anciano , Anciano de 80 o más Años , Aminoglicósidos/administración & dosificación , Aminoglicósidos/efectos adversos , Anticuerpos Monoclonales Humanizados/administración & dosificación , Anticuerpos Monoclonales Humanizados/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Citarabina/administración & dosificación , Citarabina/efectos adversos , Citogenética , Supervivencia sin Enfermedad , Femenino , Gemtuzumab , Humanos , Masculino , Persona de Mediana Edad , Inducción de Remisión , Estudios Retrospectivos , Factores de Riesgo

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Cromosomas Humanos Par 11/genética , Leucemia Linfocítica Crónica de Células B/tratamiento farmacológico , Leucemia Mieloide Aguda/etiología , Leucemia Mieloide Aguda/genética , Proteína de la Leucemia Mieloide-Linfoide/genética , Neoplasias Primarias Secundarias/etiología , Neoplasias Primarias Secundarias/genética , Anticuerpos Monoclonales de Origen Murino/administración & dosificación , Anticuerpos Monoclonales de Origen Murino/efectos adversos , Secuencia de Bases , Cromosomas Humanos Par 9/genética , Ciclofosfamida/administración & dosificación , Ciclofosfamida/efectos adversos , Sondas de ADN/genética , Reordenamiento Génico , N-Metiltransferasa de Histona-Lisina , Humanos , Hibridación Fluorescente in Situ , Masculino , Persona de Mediana Edad , Rituximab , Translocación Genética , Vidarabina/administración & dosificación , Vidarabina/efectos adversos , Vidarabina/análogos & derivados

Asunto(s)

Leucemia Linfocítica Crónica de Células B , Regresión Neoplásica Espontánea , Biopsia , Examen de la Médula Ósea , Humanos , Leucemia Linfocítica Crónica de Células B/diagnóstico por imagen , Leucemia Linfocítica Crónica de Células B/patología , Masculino , Persona de Mediana Edad , Tomografía Computarizada por Rayos X

RESUMEN

Chronic lymphocytic leukemia (CLL) cells depend on their microenvironment for proliferation and survival. Ectonucleotidase CD39 has anti-inflammatory properties as it hydrolyzes proinflammatory extracellular ATP, generates anti-inflammatory adenosine, and also protects regulatory T cells from ATP-induced cell death. In this study, we investigated the clinical significance of CD39 expression on CD4(+) T cells in 62 patients with CLL as well as its compartmental regulation and explored the possible mechanisms for its induction. Compared to healthy individuals, CD4(+)CD39(+) lymphocytes were increased in the peripheral blood of patients with CLL and correlated with the advanced stage of disease. CD4(+)CD39(+) cells were also higher in patients with CLL, who needed therapeutic intervention, and in those who had unmutated immunoglobulin heavy chain variable region gene, were ZAP70(+) or had ß2-microglobulin levels of >3 g/L. There were more CD4(+)CD39(+) lymphocytes in the bone marrow compartment than in the peripheral blood, and in vitro studies showed that CD39 can be induced on CD4(+) cells by exposure to ATP or indirectly, following B cell receptor engagement. This may support the notion that the leukemic cells contribute to create an immune-subversive environment, and perhaps to a poorer prognosis. CD39(+) may also serve as a future target for the development of novel therapies with immune-modulating antitumor agents in CLL.

Asunto(s)

Antígenos CD/metabolismo , Apirasa/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Leucemia Linfocítica Crónica de Células B/diagnóstico , Anciano , Antígenos CD/genética , Antígenos de Superficie/genética , Antígenos de Superficie/metabolismo , Apirasa/genética , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos/patología , Estudios de Casos y Controles , Progresión de la Enfermedad , Femenino , Humanos , Leucemia Linfocítica Crónica de Células B/inmunología , Leucemia Linfocítica Crónica de Células B/metabolismo , Leucemia Linfocítica Crónica de Células B/patología , Masculino , Persona de Mediana Edad , Pronóstico , Regulación hacia Arriba/genética

RESUMEN

Imatinib has been accepted as frontline treatment for patients with chronic myeloid leukaemia (CML), and patients generally receive doses ranging from 400 to 800 mg/day. Previous studies have demonstrated that maintaining imatinib plasma levels (IMPLs) >1000 ng/mL leads to improved responses and long-term outcomes. However, IMPLs vary among patients because of factors such as drug-drug interactions, adherence, toxicity and differential levels of expression of cellular efflux/influx proteins. In this study, IMPLs were analysed in 191 patients with CML and were compared with achievement of molecular and cytogenetic responses (CyR). IMPLs were also correlated with renal and hepatic dysfunction. Additionally, self-reported adherence was monitored. The median and mean IMPLs were 994 ng/mL and 1070 ± 686 ng/mL, respectively, with 96 patients (50%) achieving plasma levels >1000 ng/mL. Self-reported patient compliance was 98%. Patients who achieved a complete CyR (CCyR) had significantly higher IMPLs (1078 ± 545 ng/mL) than those without CyR (827 ± 323 ng/mL, p = 0.045). When grouped together, patients who achieved a CCyR or partial CyR had significantly higher IMPLs than patients who achieved a minimal CyR or did not achieve a CyR (1066 ng/mL vs 814 ng/mL, p = 0.002). There was no significant difference observed in the IMPLs between patients who achieved molecular responses (n = 177) on treatment (major molecular response, 976 ± 385 ng/mL versus complete molecular response, 1138 ± 809 ng/mL, p = 0.387). Mean IMPLs were similar in patients with or without renal or hepatic impairment. Overall, this study confirmed previous reports that higher IMPLs correlate with clinical responses and demonstrated that imatinib exposure did not differ in patients with or without liver and/or renal dysfunction. The use of IMPL testing and patient diaries may be practical tools for the management of imatinib therapy in patients with CML.

Asunto(s)

Antineoplásicos/sangre , Antineoplásicos/uso terapéutico , Crisis Blástica/tratamiento farmacológico , Leucemia Mieloide de Fase Crónica/tratamiento farmacológico , Piperazinas/sangre , Piperazinas/uso terapéutico , Pirimidinas/sangre , Pirimidinas/uso terapéutico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos/farmacocinética , Benzamidas , Femenino , Estudios de Seguimiento , Humanos , Mesilato de Imatinib , Riñón/efectos de los fármacos , Riñón/fisiopatología , Hígado/efectos de los fármacos , Hígado/fisiopatología , Masculino , Persona de Mediana Edad , Cooperación del Paciente , Piperazinas/farmacocinética , Pronóstico , Pirimidinas/farmacocinética , Distribución Tisular , Adulto Joven

Asunto(s)

Ejercicio Físico , Leucemia Linfocítica Crónica de Células B/inmunología , Linfocitos T Reguladores/inmunología , Células Th17/inmunología , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Interleucina-2/sangre , Interleucina-2/inmunología , Interleucina-6/sangre , Interleucina-6/inmunología , Leucemia Linfocítica Crónica de Células B/sangre , Leucemia Linfocítica Crónica de Células B/patología , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Linfocitos T Reguladores/metabolismo , Linfocitos T Reguladores/patología , Células Th17/metabolismo , Células Th17/patología , Factor de Crecimiento Transformador beta/sangre , Factor de Crecimiento Transformador beta/inmunología

RESUMEN

OBJECTIVE: To determine the role of bone marrow biopsy (BMBX), performed in association with comprehensive blood and imaging tests, in the evaluation of patients with fever of unknown origin (FUO). PATIENTS AND METHODS: We reviewed the medical records of 475 hospitalized patients who underwent BMBX in our medical center from January 1, 2005, to April 30, 2010. We identified 75 patients who fulfilled the accepted classic Petersdorf criteria for FUO. All patients underwent in-hospital investigation for fever, including chest and abdominal computed tomography. RESULTS: In 20 patients (26.7%), BMBX established the final diagnosis. Sixteen patients had hematologic disorders, including 8 patients with non-Hodgkin lymphoma, 2 with acute leukemia, 1 with multiple myeloma, 1 with myelodysplastic syndrome, and 4 with myeloproliferative disorders. The remaining patients with diagnostic BMBX specimens had solid tumors (2 patients), granulomatous disease (1 patient), and hemophagocytic syndrome (1 patient). Multivariate analysis revealed the following as the significant positive predictive parameters for a diagnostic BMBX specimen: male sex (odds ratio [OR], 7.35; 95% confidence interval [CI], 1.19-45.45), clinical lymphadenopathy (OR, 21.98; 95% CI, 1.97-245.66), anemia (OR, 2.21; 95% CI, 1.28-3.80), and increased lactate dehydrogenase levels (OR, 1.003; 95% CI, 1.001-1.006). CONCLUSION: Bone marrow biopsy is still a useful ancillary procedure for establishing the diagnosis of FUO, particularly if used in the appropriate clinical setting. Clinical and laboratory parameters associated with hematologic disease are predictive of a diagnostic BMBX specimen in patients with FUO.

Asunto(s)

Médula Ósea/patología , Fiebre de Origen Desconocido/diagnóstico , Enfermedades Hematológicas/diagnóstico , Linfohistiocitosis Hemofagocítica/diagnóstico , Linfoma no Hodgkin/diagnóstico , Mieloma Múltiple/diagnóstico , Neoplasias/diagnóstico , Adulto , Anciano , Biopsia , Femenino , Fiebre de Origen Desconocido/etiología , Enfermedades Hematológicas/complicaciones , Enfermedades Hematológicas/patología , Humanos , Leucemia/complicaciones , Leucemia/diagnóstico , Leucemia/patología , Linfohistiocitosis Hemofagocítica/complicaciones , Linfohistiocitosis Hemofagocítica/patología , Linfoma no Hodgkin/complicaciones , Linfoma no Hodgkin/patología , Masculino , Persona de Mediana Edad , Mieloma Múltiple/complicaciones , Mieloma Múltiple/patología , Neoplasias/complicaciones , Neoplasias/patología , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad

RESUMEN

Acute myeloid leukemia (AML) with a complex karyotype (CK) has frequent alterations in TP53 and a very poor prognosis. We examined whether a prompt and simple fluorescence in situ hybridization (FISH) analysis for 17p13 deletion at diagnosis has a predictive value for response to therapy and overall survival in subgroups of AML. In 15 patients with a normal karyotype the TP53 FISH analysis was normal, whereas in 16 patients with CK 75% had only one copy of the TP53 allele. The deletion was also detected in 33% of six patients with monosomy or partial monosomy of chromosome 5, 7, 9, or 12. This loss of TP53 correlated significantly with a poor response to chemotherapy, and the median survival time of these patients was shorter. TP53 FISH analysis carried out at diagnosis has a predictive value with respect to chemotherapy response and can therefore facilitate a rapid decision on treatment strategies.

Asunto(s)

Hibridación Fluorescente in Situ , Leucemia Mieloide Aguda/diagnóstico , Proteína p53 Supresora de Tumor/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos/uso terapéutico , Femenino , Eliminación de Gen , Humanos , Cariotipificación , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Análisis de Supervivencia , Resultado del Tratamiento , Proteína p53 Supresora de Tumor/genética

RESUMEN

The outcome and quality of life of chronic myeloid leukemia (CML) patients has remarkably changed with the treatment of tyrosine kinase inhibitors (TKIs). Currently, hematopoietic stem cell transplantation (HSCT) is considered mainly as a third line salvage therapy in cases of TKIs resistance or intolerance. Here we describe a patient with chronic phase CML who developed both resistance and late occurrence of s severe thrombo-cytopenia on first and second generation TKIs and eventually underwent HSCT. Although the mechanism of the myelosuppression is not fully understood, we showed for the first time the development of dose dependent platelet antibodies in the presence of TKIs, suggesting the possibility of TKIs induced thrombocytopenia. Our case emphasizes that late development of severe myelosuppression during imatinib treatment is probably an important indication for consideration of early HSCT.

RESUMEN

We have earlier shown an advantage in overall survival for malignant lymphoma (ML) patients who received combined Interleukin-2 (rIL-2) and interferon alpha (IFN-alpha) immunotherapy after autologous stem cell transplantation (AutoSCT) in comparison with historic controls. On the basis of these results, we initiated a control prospective randomized multicenter 2-arm study, comparing the same combined immunotherapy treatment versus control for patients with malignant lymphoma after AutoSCT. One hundred nine patients were included in this study. After AutoSCT, patients were randomized either to the treatment group or to the control group. Patients in the treatment group received daily subcutaneous injections of rIL-2 6x10 IU/m/d for 5 consecutive days followed by 2-weeks rest period. Subsequently, they were treated daily with rIL-2 6x10 IU/m/d combined with INF-alpha 3x10 U/d for 5 consecutive days per week for 4 consecutive weeks. After 4 weeks of rest, IFN-alpha 3x10 U was administered for the next 6 months 3 times per week. Patients in the control group were followed up at the outpatient clinic. There was a significant enhancement of survival (P=0.05) and a clear trend in disease-free survival in favor of NHL patients receiving post-AutoSCT immunotherapy, in comparison with the control group. Eighty-nine percent of patients with NHL treated with immunotherapy were alive and 64% were disease free. In the control group, 66% of patients survived and 46% were disease free. Among the HL patients no significant differences were observed regarding survival, disease-free survival, and relapse rate. No serious toxicity events were observed. These results suggest that outpatient administered immunotherapy with IFN-alpha and rIL-2 is relatively well tolerated and may intensify remission in NHL patients, but not HL patients after AutoSCT.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Enfermedad de Hodgkin/terapia , Linfoma no Hodgkin/terapia , Trasplante de Células Madre/métodos , Adolescente , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Terapia Combinada , Femenino , Fiebre/inducido químicamente , Enfermedad de Hodgkin/patología , Humanos , Inmunoterapia , Interferón Tipo I/administración & dosificación , Interleucina-2/administración & dosificación , Interleucina-2/efectos adversos , Interleucina-2/genética , Linfoma no Hodgkin/patología , Masculino , Persona de Mediana Edad , Análisis Multivariante , Náusea/inducido químicamente , Estudios Prospectivos , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/efectos adversos , Análisis de Supervivencia , Trasplante Autólogo , Resultado del Tratamiento , Vómitos/inducido químicamente , Adulto Joven

RESUMEN

The genetic profiling of B-cell malignancies is rapidly expanding, providing important information on the tumorigenic potential, response to treatment, and clinical outcome of these diseases. However, the relative contributions of inherent gene expression versus microenvironmental effects are poorly understood. The regulation of gene expression programs by means of adhesive interactions was studied here in ARH-77 human malignant B-cell variants, derived from the same cell line by selective adhesion to a fibronectin matrix. The populations included cells that adhere to fibronectin and are highly tumorigenic (designated "type A" cells) and cells that fail to adhere to fibronectin and fail to develop tumors in vivo ("type F" cells). To identify genes directly affected by cell adhesion to fibronectin, type A cells deprived of an adhesive substrate (designated "AF cells") were also examined. Bioinformatic analyses revealed a remarkable correlation between cell adhesion and both B-cell differentiation state and the expression of multiple myeloma (MM)-associated genes. The highly adherent type A cells expressed higher levels of NFkappaB-regulated genes, many of them associated with MM. Moreover, we found that the transcription of several MM-related proto-oncogenes is stimulated by adhesion to fibronectin. In contrast, type F cells, which display poor adhesive and tumorigenic properties, expressed genes associated with higher levels of B-cell differentiation. Our findings indicate that B-cell differentiation, as manifested by gene expression profiles, is attenuated by cell adhesion to fibronectin, leading to upregulation of specific genes known to be associated with the pathogenesis of MM.

Asunto(s)

Linfocitos B/metabolismo , Regulación Neoplásica de la Expresión Génica/genética , Trastornos Linfoproliferativos/genética , Trastornos Linfoproliferativos/metabolismo , Activación Transcripcional/genética , Linfocitos B/patología , Adhesión Celular/genética , Diferenciación Celular/genética , Línea Celular Tumoral , Transformación Celular Neoplásica/genética , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Perfilación de la Expresión Génica/métodos , Humanos , Trastornos Linfoproliferativos/fisiopatología , Mieloma Múltiple/genética , Mieloma Múltiple/metabolismo , FN-kappa B/genética , Invasividad Neoplásica/genética , Invasividad Neoplásica/fisiopatología , Proto-Oncogenes/genética

Asunto(s)

Leucemia Linfocítica Crónica de Células B/terapia , Linfocitosis/diagnóstico , Trasplante de Células Madre de Sangre Periférica , Donantes de Tejidos , Anciano , Femenino , Histocompatibilidad , Prueba de Histocompatibilidad , Humanos , Masculino , Persona de Mediana Edad , Hermanos , Resultado del Tratamiento

RESUMEN

Severe neutropenia and protracted thrombocytopenia remain serious clinical problems following cord blood transplantation (CBT) due to the paucity of stem and progenitor cells in the grafts. Administration of ex-vivo expanded megakaryocyte progenitor cells may facilitate platelet production. We propose a novel strategy to expand these rare cells ex-vivo, from a small portion of the cord blood (CB) unit, using fibronectin (FN), a major component of hematopoietic niches, combined with cytokines, including thrombopoietin and the hematopoietic stress-associated acetylcholinesterase readthrough peptide (ARP). Application of multiple gates and high definition flow cytometry enabled clear resolution of expanded hematopoietic stem/precursor cells (HSPC) and megakaryocyte progenitors (Mk-p) and their early subsets while eliminating positively stained non-relevant cells. FN increased viability, expansion of all CD34(+) HSPC populations and Mk-p. The combination of FN + thrombopoietin + ARP maintained and expanded very early myeloid and thrombopoietic precursors, increased the proliferation of megakaryocyte, granulocyte-macrophage and multilineage colony-forming progenitors and supported Mk maturation as measured by ploidy and glycoprotein IIb/IIIa expression by quantiative reverse transcription polymerase chain reaction. This approach, which involves expanding HSPC and Mk precursors from a small portion of the CB unit, without sacrificing the coveted stem cells, may lead to improved cell therapy modalities to facilitate earlier myelopoiesis and platelet production post-CBT.

Asunto(s)

Sangre Fetal/citología , Células Madre Hematopoyéticas/citología , Células Progenitoras de Megacariocitos/citología , Acetilcolinesterasa/farmacología , Antígenos CD34/análisis , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias , Citocinas/farmacología , Fibronectinas/farmacología , Citometría de Flujo/métodos , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Células Progenitoras de Megacariocitos/efectos de los fármacos , Fragmentos de Péptidos/farmacología

Asunto(s)

Trasplante de Médula Ósea , Talasemia beta/cirugía , Adolescente , Preescolar , Humanos , Masculino

RESUMEN

Multiple myeloma is characterized by the malignant growth of immunoglobulin producing plasma cells, predominantly in the bone marrow. The effects of primary human mesenchymal stromal cells on the differentiation phenotype of multiple myeloma cells were studied by co-culture experiments. The incubation of multiple myeloma cells with bone marrow-derived mesenchymal stromal cells resulted in significant reduction of the expression of the predominant plasma cell differentiation markers CD38 and CD138, and cell surface immunoglobulin light chain. While the down-regulation of CD138 by stromal cells was completely dependent on their adhesive interactions with the multiple myeloma cells, interleukin-6 induced specific down-regulation of CD38. Mesenchymal stromal cells or their conditioned media inhibited the growth of multiple myeloma cell line, thereby reducing the overall amounts of secreted light chains. Analysis of primary multiple myeloma bone marrow samples reveled that the expression of CD38 on multiple myeloma cells was not affected by adhesive interactions. The ex vivo propagation of primary multiple myeloma cells resulted in significant increase in their differentiation markers. Overall, the data indicate that the bone marrow-derived mesenchymal stromal cells revert multiple myeloma cells to less differentiated phenotype by the combined activities of adhesive interactions and interleukin-6.

Asunto(s)

Interleucina-6/fisiología , Mesodermo/citología , Mesodermo/fisiología , Mieloma Múltiple/patología , Mieloma Múltiple/fisiopatología , Células del Estroma/citología , Células del Estroma/fisiología , ADP-Ribosil Ciclasa 1/metabolismo , Anticuerpos Monoclonales/farmacología , Biomarcadores de Tumor/metabolismo , Células de la Médula Ósea/citología , Células de la Médula Ósea/fisiología , Adhesión Celular , Diferenciación Celular/efectos de los fármacos , Línea Celular , Línea Celular Tumoral , Técnicas de Cocultivo , Regulación hacia Abajo , Humanos , Cadenas kappa de Inmunoglobulina/metabolismo , Interleucina-6/antagonistas & inhibidores , Interleucina-6/farmacología , Glicoproteínas de Membrana/metabolismo , Mesodermo/efectos de los fármacos , Pruebas de Neutralización , Fenotipo , Células Plasmáticas/citología , Células Plasmáticas/fisiología , Proteínas Recombinantes/farmacología , Células del Estroma/efectos de los fármacos , Sindecano-1/metabolismo