RESUMEN
We evaluated the depletion of TCR-alpha/beta cells from the graft of children with high-risk AML, who received transplantation from unrelated (n=20) and haploidentical donors (n=13). The preparative regimen included treosulfan, melphalan, fludarabine and anti-thymocyte globulin. Grafts were PBSC engineered by TCR-alpha/beta and CD19 depletion. The graft contained a median of 9 × 10(6)/kg of CD34+ and 20 × 10(3)/kg of αß-T cells. Post-transplant immune suppression included tacrolimus till day +30 and Mtx in 21 patients, tacrolimus in 5, Mtx in 2 and no prophylaxis in 5 patients. Sixteen patients received native or TCR-alpha/beta-depleted donor lymphocytes at a median of 47 (40-204) days. Median follow-up is 1.76 years. Primary engraftment was achieved in 33 patients (100%). Cumulative incidence of acute GvHD (aGvHD) grade 2-3 was 39 (26-60)%, half of them had skin-only aGvHD. Cumulative incidence of chronic GvHD was 30(18-50)%. Transplant-related mortality is 10(4-26)%. Event-free survival (EFS) is 60(43-76)% and overall survival (OS) is 67(50-84)% at 2 years. In a subgroup of patients, who received transplantation in CR, EFS is 66(48-84)% and OS-72(53-90)% at 2 years. Our data suggest that TCR-alpha/beta and CD19 depletion is a robust method of graft manipulation, which can be used to engineer grafts for children with AML.
Asunto(s)
Antígenos CD19/análisis , Busulfano/análogos & derivados , Leucemia Mieloide Aguda/terapia , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Acondicionamiento Pretrasplante/métodos , Trasplante Haploidéntico/métodos , Adolescente , Antígenos CD19/aislamiento & purificación , Busulfano/uso terapéutico , Niño , Preescolar , Supervivencia sin Enfermedad , Femenino , Supervivencia de Injerto , Enfermedad Injerto contra Huésped , Humanos , Lactante , Leucemia Mieloide Aguda/complicaciones , Leucemia Mieloide Aguda/mortalidad , Masculino , Receptores de Antígenos de Linfocitos T alfa-beta/aislamiento & purificación , Trasplante Haploidéntico/mortalidad , Donante no Emparentado , Adulto JovenRESUMEN
We used the Rapid Identification of Genomic Splits technique to get a detailed methylation landscape of a 1-megabase-long human genome region (FXYD5-COX7A1, chromosome 19) in normal and tumor lung tissues and in the A549 lung cancer cell line. All three samples were characterized by an essentially uneven density of unmethylated sites along the fragment. Strikingly enough, the distribution of hypomethylated regions did not correlate with gene locations within the fragment. We also demonstrated that the methylation pattern of this long genomic DNA fragment was rather stable and practically unchanged in human lung cancer tissue as compared with its normal counterpart. On the other hand, the methylation landscape obtained for the A549 cell line (human lung carcinoma) in the USF2-MAG locus showed clear differences from that of the tissues mentioned above. A comparative analysis of transcriptional activity of the genes in this region demonstrated the general absence of direct correlation between methylation and expression, although some data suggest a possible role of methylation in the regulation of MAG expression through cis-regulatory elements. In total, our data provide new evidence for the necessity of revising currently prevailing views on the functional significance of methyl groups in genomic DNA.
Asunto(s)
Cromosomas Humanos Par 19/genética , Metilación de ADN , Complejo IV de Transporte de Electrones/genética , Glicoproteínas de Membrana/genética , Proteínas de Neoplasias/genética , Línea Celular Tumoral , Cromosomas Humanos Par 19/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Humanos , Canales Iónicos , Pulmón/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Microfilamentos , Proteínas de Neoplasias/metabolismoRESUMEN
Individual differences in drug efficacy or toxicity can be influenced by genetic factors. We investigated whether polymorphisms of pharmacogenes that interfere with metabolism of drugs used in conditioning regimen and graft-versus-host disease (GvHD) prophylaxis could be associated with outcomes after HLA-identical hematopoietic stem cell transplantation (HSCT). Pharmacogenes and their polymorphisms were studied in 107 donors and patients with leukemia receiving HSCT. Candidate genes were: P450 cytochrome family (CYP2B6), glutathione-S-transferase family (GST), multidrug-resistance gene, methylenetetrahydrofolate reductase (MTHFR) and vitamin D receptor (VDR). The end points studied were oral mucositis (OM), hemorrhagic cystitis (HC), toxicity and venoocclusive disease of the liver (VOD), GvHD, transplantation-related mortality (TRM) and survival. Multivariate analyses, using death as a competing event, were performed adjusting for clinical factors. Among other clinical and genetic factors, polymorphisms of CYP2B6 genes that interfere with cyclophosphamide metabolism were associated with OM (recipient CYP2B6(*)4; P=0.0067), HC (recipient CYP2B6(*)2; P=0.03) and VOD (donor CYP2B6(*)6; P=0.03). Recipient MTHFR polymorphisms (C677T) were associated with acute GvHD (P=0.03), and recipient VDR TaqI with TRM and overall survival (P=0.006 and P=0.04, respectively).Genetic factors that interfere with drug metabolisms are associated with treatment-related toxicities, GvHD and survival after HLA-identical HSCT in patients with leukemia and should be investigated prospectively.
Asunto(s)
Antineoplásicos/efectos adversos , Biotransformación/genética , Enfermedad Injerto contra Huésped/genética , Trasplante de Células Madre Hematopoyéticas , Inmunosupresores/efectos adversos , Leucemia/cirugía , Agonistas Mieloablativos/efectos adversos , Polimorfismo Genético , Subfamilia B de Transportador de Casetes de Unión a ATP , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Adolescente , Adulto , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapéutico , Hidrocarburo de Aril Hidroxilasas/genética , Enfermedad Hepática Inducida por Sustancias y Drogas , Niño , Preescolar , Citocromo P-450 CYP2B6 , Femenino , Genes MDR , Predisposición Genética a la Enfermedad , Glutatión Transferasa/genética , Antígenos HLA/genética , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Inmunosupresores/farmacocinética , Inmunosupresores/uso terapéutico , Inflamación/inducido químicamente , Inflamación/genética , Leucemia/genética , Leucemia/mortalidad , Hepatopatías/genética , Masculino , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Persona de Mediana Edad , Agonistas Mieloablativos/farmacocinética , Agonistas Mieloablativos/uso terapéutico , Proteínas de Neoplasias/genética , Oxidorreductasas N-Desmetilantes/genética , Receptores de Calcitriol/genética , Hermanos , Acondicionamiento Pretrasplante/efectos adversos , Trasplante Homólogo/mortalidad , Adulto JovenRESUMEN
Allogeneic stem cell transplantation remains the best option for young patients with SAA. With genetically identical twin as an ideal donor, the majority of SAA patients require appropriate immunosuppression before and after stem cell transplantation to obtain long-term hematopoietic reconstitution. Alkylating agents, used during conditioning, are associated with short- and long-term toxic effects that lead to poor compliance of treatment and could compromise the quality of future life. Three SAA patients, transplanted from genetically identical twins without using alkylating agents during conditioning, showed rapid and sustained hematological reconstitution without any evidence of conditioning-related toxicity.
Asunto(s)
Alquilantes , Anemia Aplásica/cirugía , Antineoplásicos/farmacología , Trasplante de Células Madre , Acondicionamiento Pretrasplante/métodos , Gemelos Monocigóticos/genética , Vidarabina/análogos & derivados , Contraindicaciones , Femenino , Estudios de Seguimiento , Humanos , Masculino , Agonistas Mieloablativos , Estudios Retrospectivos , Trasplante Homólogo , Resultado del Tratamiento , Vidarabina/farmacologíaRESUMEN
Allogeneic hematopoietic stem cell transplantation (SCT) from unaffected donors remains the only modality for the correction of hematological abnormalities in Fanconi anemia (FA) patients. We performed four HLA-matched related donor SCT using a novel irradiation and cyclophosphamide-free conditioning regimen. The protocol included fludarabine 150 mg/m(2), busulfan 4 mg/kg, and antithymocyte globulin 90 mg/kg. Graft-versus-host disease (GVHD) prophylaxis was cyclosporin A, MTX, and daclizumab. The engraftment and occurrence of full stable donor hemopoiesis was rapid in all cases with minimal short-term toxic complications. There were no infections or febrile episodes during the inpatient phase. Three patients developed acute GVHD grade I-II involving gut and skin and one patient progressed to extensive chronic GVHD. The preparative conditioning regimen is safe and associated with low organ toxicity and effective immunosupression for the stable engraftment in FA patients undergoing SCT with matched related donors.
Asunto(s)
Suero Antilinfocítico/uso terapéutico , Trasplante de Médula Ósea , Busulfano/uso terapéutico , Anemia de Fanconi/cirugía , Acondicionamiento Pretrasplante/métodos , Vidarabina/análogos & derivados , Vidarabina/uso terapéutico , Trasplante de Médula Ósea/inmunología , Niño , Preescolar , Infecciones por Citomegalovirus/diagnóstico , Quimioterapia Combinada , Anemia de Fanconi/inmunología , Femenino , Prueba de Histocompatibilidad , Humanos , Inmunosupresores/uso terapéutico , Donadores Vivos , Masculino , Complicaciones Posoperatorias/virología , Trasplante de Células Madre , Trasplante HomólogoRESUMEN
Pure red cell aplasia (PRCA) is a well-known although infrequent hematologic complication after allogeneic bone marrow transplantation. PRCA occurs in cases of major ABO-mismatch between donor and recipient and is believed to be due to inhibition of donor erythroid progenitors by residual host isohemagglutinins. We report a 10-year-old boy with post-hepatitis aplastic anemia (AA) who developed PRCA after HLA-matched familial peripheral blood stem cell transplantation (SCT) following conditioning with Cph 200 mg/kg + ATG 90 mg/kg. Granulocyte engraftment occurred on day +18, platelet engrafted on day +40, while reticulocytopenia at 0% persisted until day +118, and erythroid precursors were totally absent from bone marrow. After a single dose of rituximab 200 mg/m(2)administered on day +118 PRCA resolved and on day +132 the reticulocytes rose to 5.7%. On day +139 the Hb reached 137 g/l and the erythroid lineage in BM increased to 21%. We conclude that due to the rapid recovery from PRCA and lack of side effects, rituximab should be tried as first-line treatment of PRCA after allo-SCT.