Genetic variability profiling of the p53 signaling pathway in chronic lymphocytic leukemia. Individual and combined analysis of TP53, MDM2 and NQO1 gene variants.

TP53 gene disruption, including 17p13 deletion [del(17p)] and/or TP53 mutations, is a negative prognostic biomarker in chronic lymphocytic leukemia (CLL) associated with disease progression, treatment failure and shorter survival. Germline variants in p53 signaling pathway genes could also lead to p53 dysfunction, but their involvement in CLL has not been thoroughly evaluated. The aim of this study was to determine the association of TP53, MDM2 and NQO1 gene variability with clinical and genetic data of CLL patients. Individual genotype and haplotype data of CLL patients were compared with clinical prognostic factors, cytogenetic and molecular cytogenetic findings as well as IGHV and TP53 mutational status. The study included 116 CLL patients and 161 healthy blood donors. TP53 (rs1042522, rs59758982, rs1625895), NQO1 (rs1800566) and MDM2 (rs2279744, rs150550023) variants were genotyped using different PCR approaches. Analysis of genotype frequencies revealed no association with the risk of CLL. TP53 rs1042522, rs1625895 and MDM2 rs2279744 variants were significantly associated with abnormal karyotype and the presence of del(17p). Similarly, these two TP53 variants were associated with TP53 disruption. Moreover, TP53 C-A-nondel and G-A-del haplotypes (rs1042522-rs1625895-rs59758982) were associated with an increased likelihood of carrying del(17p) and TP53 disruptions. MDM2 T-nondel haplotype (rs2279744-rs150550023) was found to be a low risk factor for del(17p) (OR = 0.32; CI: 0.12-0.82; p = 0.02) and TP53 disruptions (OR = 0.41; CI: 0.18-0.95; p = 0.04). Our findings suggest that TP53 and MDM2 variants may modulate the risk to have chromosome alterations and TP53 disruptions, particularly del(17p). To our knowledge this is the first study of several germline variants in p53 pathway genes in Argentine patients with CLL.

Expression dynamics of the immune mediators ARG1, TBET, CIITA, IL10 and TGFB1 in chronic myeloid leukaemia patients during the first year of imatinib therapy.

The use of tyrosine kinase inhibitors seems to restore the broadly compromised immune system described in chronic myeloid leukaemia (CML) patients at diagnosis leading to a re-activation of the effector-mediated immune surveillance. Here, we describe the expression dynamics of immune factors during the first year on imatinib therapy. Gene expression was evaluated in 132 peripheral blood samples from 79 CML patients, including 34 who were serially followed. An aliquot of the stored sample used to monitor BCR-ABL1 levels was retro-transcribed to cDNA and gene expression was quantified by real-time PCR. An elevated expression of ARG1 was observed at diagnosis, while TBET, CIITA, IL10 and TGFB1 were significantly decreased. Once on therapy, each gene displayed a particular behaviour. ARG1 normalized to control levels at 3 months only in optimal molecular responders and was identified as the major contributor to the difference among patients. TBET reached normal levels after 12 months in optimal responders and non-responders, regardless the Th1-response previously associated, and CIITA continued downregulated. IL10 and TGFB1 achieved normal levels early at 3 months in both groups, afterwards IL10 was sustained while TGFB1 was slightly increased after 1 year in responders. Our findings are in agreement with an immune re-activation after imatinib initiation; however, some immune mediators may require a longer exposition. The follow-up of novel and reliable biomarkers, such as ARG1, one of the principal mechanisms of myeloid-derived-suppressor cells to inhibit immune system, may be useful to deepen the characterization of early responder patients.

Haploidentical transplant in adult patients with acute lymphoblastic leukemia in Argentina: a comparison with matched related and unrelated donors.

We aimed at analyzing the outcome of allogeneic stem cell transplant (ASCT) in adult patients with acute lymphoblastic leukemia (ALL), comparing Haploidentical (Haplo) with HLA-matched (sibling and unrelated) donors. Between 2008 and 2017, we collected data from 236 patients (median age 31 years; range 16-64; 90% HCT-CI 0-1) who underwent unmanipulated ASCT in first complete remission and subsequent remissions in 15 Argentinian centers. Donors were HLA-matched (n = 175; 74%) and Haplo (n = 61; 26%). Two-year overall survival (OS) was 55% (95% CI 47-63) for the HLA-matched group and 49% (95% CI 34-62) for the Haplo group (p = 0.351). For OS, crude HR, adjusted HR for covariates (HR 1.24; 95% CI 0.77-1.99; p = 0.363) and HR including a propensity score in the model (HR 1.22; 95% CI 0.71-2.08; p = 0.414) showed no impact of donor category on the OS. No difference was found in terms of nonrelapse mortality, relapse, leukemia-free survival, and grade 3-4 acute graft-versus-host disease (GVHD); 2-year incidence of chronic GVHD was higher in HLA-matched vs Haplo group (p = 0.028). Patients with ALL who underwent ASCT were young subjects with low HCT-CI. In this setting, a Haplo donor represents an alternative widely available in the absence of an HLA-matched donor. Relapse remains a challenge for all donor categories.

A der(11)t(4;11)(q21;p15) in a T-ALL/LBL patient.

Translocation t(4;11)(q21;p15) is a rare recurrent change associated to T-cell acute leukemia. In most cases, this alteration appears as the only abnormality or as part of a simple karyotype. In this report, we present the first case of T acute lymphoblastic leukemia/lymphoma (T-ALL/LBL) with the unbalanced translocation der(11)t(4;11)(q21;p15) as part of a very complex karyotype with multiple chromosome abnormalities, most of them not previously described in the literature. FISH (fluorescence in situ hybridization) and spectral karyotype (HiSKY) analysis confirmed the presence of complex alterations. The patient, a 16-year-old male, showed poor response to treatment and short survival (11 months). A detailed review of previously reported cases with t(4;11)(q21;p15) is also provided. The description of this type of alterations may contribute to the identification of new molecular mechanism associated to neoplastic development.

Perfil de seguridad de la angioplastia pulmonar con balón bajo anticoagulación con antagonistas de vitamina K / Safety profile of balloon pulmonary angioplasty under anticoagulation with vitamin K antagonists

La hipertensión pulmonar tromboembólica crónica se trata con angioplastia pulmonar con balón (APB) cuando la enfermedad es distal o la clase funcional cardíaca no permite la tromboendarectomía. La anticoagulación con antagonistas de vitamina k es uno de los pilares del tratamiento. Se describe la experiencia de 25 procedimientos de ABP bajo tratamiento anticoagulante con AVK y sus complicaciones de sangrado (8% de las ABP). Realizar APB, con RIN mayor igual a 2 no presenta un riesgo significativo de sangrado en nuestra serie.

Células madre en el corazón chagásico / Células tronco no coraçâo chagásico / Stem cells in the Chagas heart

Objetivo: El objetivo de la presente investigación es valorar la seguridad y factibilidad, como así mismo analizar los cambios clínicos y de la función ventricular en pacientes portadores de IC secundaria a miocardiopatía dilatada de origen chagásico, sometidos a cardioimplante de células mononucleares de médula ósea por cateterismo intracoronario. Material y Métodos: De los ocho pacientes tratados por enfermedad de Chagas-Mazza, estamuestra analizada comprende 5 pacientes consecutivos con el seguimiento necesario para las conclusiones (3 masculinos), edad promedio de 55.8 ± 8.1 años, que presentaban IC secundaria a cardiomiopatía chagásica. Todos ellos tenían tres reacciones serológicas positivas para enfermedad de Chagas. Dos pacientes tenían colocados marcapasos definitivos. Los datos preoperatorios promedio de estos pacientes indicaban una clase funcional (NYHA) de 2.6 ± 0.5 en promedio, mientras que la fracción del ventrículo izquierdo correspondía al 26.4 ± 5.7%; el diámetro diastólico ventricular izquierdo era de 61.6 ± 5.6 mm y el diámetro sistólico del ventrículo izquierdo de 46.9 ± 6.3 mm. La suspensión celular obtenida de células mononucleares de la médula ósea contenía (valores promedio) 10.6 ± 7 ml con un conteo de 1.43 ± 0.6 (E+08) de células con una viabilidad mayor al 95%. Correspondían a la fracción CD34+, 5.07 ± 9.51 (E+06) y a la CD133+, 5.11 ± 4.3 (E+06). El tiempo de isquemia inducida fue de unos 20 minutos. No hubo complicaciones ni óbitos hospitalarios. Resultados: A los 22.8 ± 13.3 meses de seguimiento promedio, todos los enfermos se hallaban vivos en clase funcional I. Un paciente falleció a los 17 meses de muerte súbita luego de completadoeste análisis. En este tiempo de evolución, no se observaron efectos adversos en ninguno de los pacientes referidos al implante celular. Los estudios realizados en el control postoperatorio fueron los mismos que los llevados a cabo en el preoperatorio. Con relación a la fracción de eyección...

Objetivo: O objetivo deste trabalho de pesquisa é valorizar a segurança e factibilidade, além da análise de mudanças clínicas e da função ventricular em pacientes portadores de IC secundária à miocardiopatia dilatada de origem chagásica, submetidos a cardioimplante de células mononucleares de medula óssea por cateterismo intracoronariano. Material e Métodos: Dos oito pacientes tratados por doença de Chagas-Mazza, esta amostra analisada compreende 5 pacientes consecutivos com o acompanhamento necessário para asconclusões (3 masculinos), média de idade, de 55.8 ± 8.1 anos, que apresentavam IC secundária à cardiomiopatia chagásica. Todos tiveram três reações serológicas positivas para a doença de Chagas. Dois pacientes eram portadores de marcapassos definitivos. A média dos dados pré operatórios destes pacientes indicava uma classe funcional de, em média, (NYHA) de 2.6 ± 0.5, enquanto a fração do ventrículo esquerdo correspondia a 26.4 ± 5.7%; o diâmetro diastólico ventricular esquerdo era de 61.6 ± 5.6 mm e o diâmetro sistólico do ventrículo esquerdo de 46.9 ± 6.3 mm. A suspensão celular obtida de células mononucleares da medula óssea continha (valores médios) 10.6 ± 7 ml com uma contagem de 1.43 ± 0.6 (E+08) de células com uma viabilidade superior a 95%. Correspondiam à fração CD34+, 5.07 ± 9.51 (E+06) e à CD133+, 5.11 ± 4.3 (E+06). O tempode isquemia induzida foi de 20 minutos aproximadamente. Não ocorreram complicações nem óbitos hospitalares.Resultados: Após 22.8 ± 13.3 meses de seguimento médio, todos os doentes se encontravam em classe funcional I. Um paciente faleceu 17 meses depois, de morte súbita, após o término destaanálise. Durante este tempo de evolução, não foram observados efeitos adversos em nenhum dos pacientes referidos ao implante celular. Os estudos realizados no controle pós operátorio foram os mesmos que os realizados no período pré operatório. Com relação à fração de ejeção, esta apresentou um aumento de...

Purpose: The purpose of this research is to assess the safety and feasibility, as well as the clinical and ventricular function changes, in patients with heart failure secondary to Chagas dilatedcardiomyopathy, of cardiac bone marrow mononuclear cell transplantation carried out by intracoronary catheterization. Material and Methods: Out of eight patients with Chagas-Mazza disease, the sample analyzed includes 5 consecutive patients with the controls necessary to reach the conclusions (3 males), average age 55.8 ± 8.1 years, with heart failure secondary to Chagas cardiomyopathy. They all presented three positive serological reactions for Chagas disease. Two patients had a definitive pacemaker placed. Average preoperative data of these patients indicated an average functionalclass (NYHA) of 2.6 ± 0.5 while the left ventricular ejection fraction corresponded to 26.4 ± 5.7 %;the left ventricular diastolic diameter was 61.6 ± 5.6 mm and the left ventricular systolic diameter was 46.9 ± 6.3 mm. The cell suspension obtained from bone marrow mononuclear cells contained (average values) 10.6 ± 7 ml, counting 1.43 ± 0.6 (E+08) cells with a viability over 95%. 5.07 ± 9.51 (E+06) corresponded to the CD34+ fraction, 5.11 ± 4.3 (E+ CD133+, 06) to theCD133+. The induced ischemia time was around 20 minutes. There were no hospital complications nor deaths.Results: At 22.8 ± 13.3 months of average follow-up all patients were alive and in functional Class I. One patient died after 17 months due to a sudden death after completing this analysis. During the follow-up there were no side effects in any of the patients related to the cell transplantation. The tests carried out in the postoperative period were the same as in the preoperative one. Inrelation to the ejection fraction, it increased from 26.4 ± 5.7% before the transplantation to 35.6 ± 5.3% (p<0.05). In turn, the functional class improved from 2.6 ± 0.5 to 1 ± 0 (p<0.005). The data collected about the...

Autologous stem cell transplantation for patients with chronic myeloid leukemia. The Argentine Group of Bone Marrow Transplantation (GATMO) experience.

BACKGROUND: The objective of this analysis was to evaluate the role of autologous stem cell transplantation (ASCT) in prolonging disease free survival (DFS) and overall survival (OS) in patients with chronic myeloid leukemia (CML) who received autografts of Philadelphia chromosome (Ph) positive or Ph negative cell harvests. METHODS: Over a 4-year period (1994-1999), 53 patients who underwent ASCT for CML were reported to the Argentine Group of Bone Marrow Transplantation (GATMO) Registry. RESULTS: Ph negative cell products were harvested in only 18 patients (34%). Comparison of disease status at the time of autograft, duration of neutropenia, thrombocytopenia, days of antibiotics, and transfusional requirements of red blood cells and platelets did not reveal statistical significant differences between the Ph positive group and the Ph negative group. Only days of hospitalization were increased significantly in patients who received Ph positive autografts. Although DFS at 36 months was significantly longer after infusion of Ph negative cell products (54% vs. 14%; P

Resultados alejados del implante miocárdico de células madre en la miocardiopatía chagásica / Late Results in Myocardial Implantation of Stem Cells in Chagasic Myocardial Disease

Objetivo La presente investigación clínica persigue evaluar la seguridad y la factibilidad del cardioimplante de células mononucleares de médula ósea, así como analizar los cambios clínicos y de la función ventricular, en pacientes portadores de insuficiencia cardíaca secundaria a miocardiopatía dilatada de origen chagásico sometidos a este procedimiento. Material y métodos La muestra comprende 5 pacientes consecutivos (3 masculinos) con una edad promedio de 55,8 ± 8,1 años, que presentaban insuficiencia cardíaca secundaria a miocardiopatía chagásica. Los datos preoperatorios promedio de estos pacientes indicaban una clase funcional (NYHA) de 2,6 ± 0,5, mientras que la fracción del ventrículo izquierdo correspondía al 27,6% ± 5,9%, el diámetro diastólico ventricular izquierdo era de 62,9 ± 6,2 mm y el diámetro sistólico del ventrículo izquierdo era de 49,7 ± 7,6 mm. La suspensión celular obtenida de células mononucleares de la médula ósea contenía 10,6 ± 7 ml con un conteo de 1,43 ± 0,6 (E+08) células con una viabilidad mayor del 95%. Correspondían a la fracción CD34+, 5,07 ± 9,51 (E+06) y a la CD133+, 5,11 ± 4,3 (E+06). La suspensión se inyectó por pulsos en cuatro pacientes por vía intracoronaria y en el quinto por vía transepicárdica en el transcurso de una cirugía (reemplazo valvular mitral, revascularización y colocación de matriz bioartificial degradable). Resultados A los 17,2 ± 8,8 meses de seguimiento promedio, 4 enfermos se hallaban vivos y en clase funcional I (p < 0,005). Un paciente falleció de muerte súbita a los 17 meses de seguimiento. En este tiempo de evolución no se observaron efectos adversos en ninguno de los pacientes referidos al implante celular. En relación con la fracción de eyección, se notó un incremento a 36,6% ± 2,3% (p < 0,05). Los datos obtenidos en la medición de los diámetros sistólico y diastólico del ventrículo izquierdo no mostraron cambios estadísticos significativos. Conclusiones Este trabajo representa la segunda comunicación mundial sobre el uso de stem cells en cardiopatía de Chagas-Mazza. El procedimiento resultó factible y seguro y se asoció con mejoría de la capacidad funcional y de la función ventricular en el seguimiento. Estos hallazgos provisorios deberán ser corroborados por estudios clínicos controlados más amplios.

Objective The present clinical study is aimed at assessing the safety and feasibility of cardio implantation of mononuclear bone marrow cells, as well as analyzing the clinical and ventricular function changes in patients with cardiac failure secondary to dilated myocardial disease of chagasic origin that underwent this procedure. Material and Methods The sample included 5 consecutive patients (3 male), mean age 55, 8 ± 8, 1 years that showed cardiac failure secondary to chagasic myocardial disease. Average pre-surgery data in these patients were 2, 6 ± 0, 5 functional class (NYHA), whereas the left ventricle fraction corresponded to 27, 6% ± 5, 9%, the left ventricle diastolic diameter was 49, 7 ± 7, 6 mm. The cellular suspension obtained from mononuclear bone marrow cells contained 10, 6 ± 7 ml with 1, 43 ± 0, 6 (E+08) cell counting with a viability higher than 95%. Corresponding to CD34+, 5, 07 ± 9, 51 (E+06) fraction and CD133+, 5, 11 ± 4, 3 (E+06). Suspension was intracoronary injected in pulses to four patients and transepicardially to the fifth during the course of a surgery (mitral valve replacement, revascularization and degradable bioartificial matrix). Results At 17,2 ± 8,8 average months follow up, 4 patients were alive and with functional class I (p < 0,005). One patient died of sudden death after 17 months of follow up. During this evolution time period no adverse events were observed in none of the patients referred for cellular implantation. In regards to the ejection fraction, a 36,6% ± 2,3% (p < 0,05) increase was observed. Data obtained from the assessment of the left ventricle systolic and diastolic diameters did not show significant statistical changes. Conclusions This study represents the second world communication on the use of stem cells in Chagas-Mazza heart disease. The procedure resulted feasible and safe and was associated to improvement of the functional capacity and ventricular function during follow up. These conditional results should be corroborated by broader controlled clinical trials.

Banco de células progenitoras hematopoyéticas de cordón umbilical / Umbilical cord hematopoietic progenitor cells bank

Transplantation of hematopoietic progenitor cells (HPC) from bone marrow and mobilized peripheral blood is a standard therapy in malignant and non malignant diseases. The lack of suitable donors is an important limitation. The discovery that umbilical cord blood (CB) contains high numbers of HPC that can be used as an alternative source for allogeneic stem cell transplantation led ITMO to establish BANCEL, the first Argentine and Latinoamerican experience of its kind. The blood remaining in the umbilical cord and in the placenta was requested from women who were in the last quarter of pregnancy. An informed consent together with a medical record focused on family disease was completed. Out of 65 donations, 55 (85 per cent) were collected and 51 (78 per cent) were cryopreserved. Mean collected volume was 110 ml with 68 per cent (75 ml) reduction and mean cryopreservation of 35 ml; ABO and Rh blood group systems were determined, HLA, class I, A and B loci, and class II, DR locus were typed by molecular biology methods using PCR-SSOP. Infectious disease screening was carried out for brucellosis, syphilis, Chagas, hepatitis B and C, HIV I and II, HTLV I and II, toxoplasmosis and cytomegalovirus. Two positive units for hepatitis B (anticore) and two positive units for Chagas were discarded. The quantity of total nucleated cells (TNC), CD34+ cells and the clonogenic capacity were determined twice at the collection and after the procedures of volume reduction previous to cryopreservation. A 5 per cent reduction in both TNC and CD34 cells and a 10 per cent in the colony forming units (CFU) were detected. A good correlation coefficient between TNC and CFU was obtained.

Trasplante de medula osea en leucemia mieloide cronica / Bone marrow transplantation in chronic myelogenous leukemia

La Leucemia Mieloide Crónica (LCM) es un proceso oncohematológico caracterizado por una proliferación clonal que afecta a la célula hematopoyética primitiva. Un 95 por ciento de pacientes presenta una alteración citogenética característica conocida como Cromosoma Philadelphia (Ph1), producto de una traslocación cromosómica 9:22, que da lugar a un gen hídrido BCR/ABL. Diecinueve pacientes con LMC recibieron Trasplante Alogeneico de Médula Osea (TMO), 9 fueron de sexo femenino y 10 de sexo masculino. La média de edad fue de 32 años (rango 9-47); 15 de los pacientes estaban en fase crónica (FC) y 4 en fase acelerada (FA). Todos los pacientes al momento del diagnóstico fueron Ph1+, BCR/ABL+. El régimen de acondicionamiento consistió en Busulflán y Ciclofosfamida, con el agregado de Etoposido en los pacientes en FA. La profilaxis de EICH se efectuó con Ciclosporina A, Metotrexato y Metilprednisona en 17 pacientes y con las 2 primeras drogas en 2 pacientes. La traslocación 922 se estudió mediante técnica de RT-PCR, con empleo de las sondas NB1+, Abl3, B2A, CA3 y A2. La sensibilidad del método fue de 1 x 10(-6). De 19 pacientes que ingresaron al protocolo, 14 permanecen vivos y en remisión clínica, hematológica y citogenética (Ph, negativo); 3 pacientes fallecieron de EICH agudo, 1 de fallo de "angraftment" y 1 de síndrome urémico hemolítico. De 4 pacientes trasplantados en FA, 3 están vivos y en remisión completa. Los pacientes con LMC trasplantados presentaron una sobrevida del 74 por ciento con un seguimiento medio de 655 días. El quimerismo hematopoyético completo se demostró en 16 pacientes, mediante el estudio de 3 loci, D1S80, APO B y D17S30. No se encontró ninguna relación entre la presencia post TMO del híbrido BCR/ABL (RT.PCR) y la recaída de la enfermedad; la presencia de EICH agudo y/o crónico no tuvo influencia entre la positividad del BCR/ABL. El TMO ha demostrado se en nuestra experiencia la única alternativa terapéutica para la LMC con obtención de remisión completa, clínica, hematológica y citogenética, con una sobrevida media del 74 por ciento comparable a la de centros internacionales de trasplante.