-
Elena Sebastián,
Miguel Alcoceba,
Ana Balanzategui,
Luis Marín,
Santiago Montes-Moreno,
Teresa Flores,
David González,
M Eugenia Sarasquete,
M Carmen Chillón, Noemí Puig,
Rocío Corral,
Emilia Pardal,
Alejandro Martín,
Eva González-Barca,
M Dolores Caballero,
Jesús F San Miguel,
Ramón García-Sanz,
Marcos González
[show abstract]
[hide abstract]
ABSTRACT: The pathogenesis of diffuse large B-cell lymphoma (DLBCL) remains partially unknown. The analysis of the B-cell receptor of the malignant cells could contribute to a better understanding of the DLBCL biology. We studied the molecular features of the immunoglobulin heavy chain (IGH) rearrangements in 165 patients diagnosed with DLBCL not otherwise specified. Clonal IGH rearrangements were amplified according to the BIOMED-2 protocol and PCR products were sequenced directly. We also analyzed the criteria for stereotyped patterns in all complete IGHV-IGHD-IGHJ (V-D-J) sequences. Complete V-D-J rearrangements were identified in 130 of 165 patients. Most cases (89%) were highly mutated, but 12 sequences were truly unmutated or minimally mutated. Three genes, IGHV4-34, IGHV3-23, and IGHV4-39, accounted for one third of the whole cohort, including an overrepresentation of IGHV4-34 (15.5% overall). Interestingly, all IGHV4-34 rearrangements and all unmutated sequences belonged to the nongerminal center B-cell-like (non-GCB) subtype. Overall, we found three cases following the current criteria for stereotyped heavy chain VH CDR3 sequences, two of them belonging to subsets previously described in CLL. IGHV gene repertoire is remarkably biased, implying an antigen-driven origin in DLBCL. The particular features in the sequence of the immunoglobulins suggest the existence of particular subgroups within the non-GCB subtype.
American Journal Of Pathology 09/2012; 181(5):1879-88. · 4.89 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We have evaluated the use of CD138+ positively selected bone marrow samples to identify a molecular target for minimal residual disease assessment by polymerase chain reaction (PCR) in 25 untreated patients with multiple myeloma. A fraction of each sample was used for CD138+ selection, and the rest served as a reference control. VDJH, DJH, and Kde gene rearrangements were tested for amplification according to the BIOMED-2 Concerted Action. PCR products were directly sequenced in an automated ABI 3130 DNA sequencer using Big-Dye terminators. Within the CD138+ selected group, VDJH rearrangements were detected in all cases (100 %), DJH in 16 (64 %), and Kde in 18 (72 %) cases; whereas in the control samples, VDJH, DJH, and Kde rearrangements were detected in 19 (76 %), 11 (44 %), and 12 (48 %) cases, respectively. After sequencing, 24 (96 %) cases within the CD138+ group had a PCR target for MRD detection compared with 15 (60 %) cases in the control group. We conclude that the use of CD138+ positively selected bone marrow samples increases the applicability of minimal residual disease studies by PCR in patients with multiple myeloma.
Annals of Hematology 09/2012; · 2.62 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Minimal residual disease (MRD) assessment by PCR in multiple myeloma (MM) has several shortcomings, including the lack of a suitable target. Kappa deleting element (KDE) rearrangements occur in virtually all Ig-lambda B-cell malignancies and in 1/3 of Ig-kappa are not affected by somatic hypermutation and, as in ALL, could be used as PCR targets.
We have first investigated the incidence, gene segment usage, and CDR3 composition of IGK-KDE rearrangements in 96 untreated myeloma patients. Second, we tested 16 KDE gene rearrangements as molecular targets for MRD assessment by RQ-PCR using a germline reverse primer and a germline Taqman probe in combination with allele-specific oligonucleotides (ASO) as forward primers.
Monoclonal KDE rearrangements were amplified in 45% (43/96) of cases, monoallelic in 2/3 of them (29 cases), and biallellic in the remaining 14 cases. Overall, 88% of cases were successfully sequenced, KDE being equally frequently rearranged with VK and with intron-Recombination signal sequence (RSS). Median numbers of inserted and deleted nucleotides in the junctional region were one and five, respectively.
Using KDE rearrangements as additional PCR target for MRD assessment in MM improves the applicability of these studies in 9% of cases overall and in 20% of lambda cases. Its use in the latter subset could represent a significant advance.
European Journal Of Haematology 07/2012; 89(4):328-35. · 2.61 Impact Factor
-
Annals of Hematology 02/2011; 90(2):227-9. · 2.62 Impact Factor
-
Carlos Santamaría,
María C Chillón,
Ramón García-Sanz,
Cristina Pérez,
María D Caballero,
María V Mateos,
Fernando Ramos,
Alfonso García de Coca,
José M Alonso,
Pilar Giraldo,
Teresa Bernal,
José A Queizán,
Juan N Rodríguez, Noemí Puig,
Ana Balanzategui,
María E Sarasquete,
Miguel Alcoceba,
Joaquín Díaz-Mediavilla,
Jesús San Miguel,
Marcos González
[show abstract]
[hide abstract]
ABSTRACT: We have analyzed brain and acute leukemia, cytoplasmic (BAALC) gene expression and other genetic markers (ERG, EVI1, MN1, PRAME, WT1, FLT3, and NPM1 mutations) in 127 intermediate-risk acute myeloid leukemia (AML) patients: 98 cytogenetically normal and 29 with intermediate-risk cytogenetic alterations. High versus low BAALC expressers showed a higher refractoriness to induction treatment (31% vs 10%; p = .005), lower complete remission rate after salvage therapy (82% vs 97%; p = .010), and lower 3-year overall (23% vs 58%, p < .001) and relapse-free survival (26% vs 52%, p = .006). Similar results were found when cytogenetic subgroups were analyzed separately. Multivariate models confirmed the unfavorable prognosis of this marker. In conclusion, BAALC is a relevant prognostic marker in intermediate-risk AML.
Annals of Hematology 11/2009; 89(5):453-8. · 2.62 Impact Factor
-
Noemí Puig,
Javier De La Rubia,
Isidro Jarque,
Miguel Salavert,
Federico Moscardó,
Jaime Sanz,
Ignacio Lorenzo,
Pau Montesinos,
Guillermo Martín,
Jesús Martínez,
Guillermo Sanz,
Margarita Blanes,
Miguel Sanz
[show abstract]
[hide abstract]
ABSTRACT: We analyzed the incidence, etiology, risk factors and outcomes of 49 episodes of pneumonia that developed in 326 adult patients undergoing autologous stem-cell transplantation (ASCT) from January 1990 to December 2005. The median time for the onset of pneumonia after transplantation was 11 days (range 0-148). Empirical antibiotic therapy in patients with pneumonia consisted of piperacillin-tazobactam (20 cases, 49%), third-generation cephalosporin (11 cases, 27%) and carbapenem (8 cases, 19%). Multivariate analysis showed that a higher risk of pneumonia could be predicted for patients with myeloma (P = 0.006) and for patients with an absolute neutrophil count <0.5 x 10(9)/L >7 days (P = 0.008). Cumulative incidence of transplant-related mortality at 6 months was 51% versus 8% for patients with or without pneumonia, respectively (P = 0.001). Pneumonia after ASCT is a severe complication more commonly observed in patients with myeloma and with prolonged duration of neutropenia.
Leukemia and Lymphoma 01/2008; 48(12):2367-74. · 2.58 Impact Factor
-
Noemí Puig,
Javier de la Rubia,
Isidro Jarque,
Miguel Salavert,
Pau Montesinos,
Jaime Sanz,
Guillermo Martín,
Guillermo Sanz,
Susana Cantero,
Ignacio Lorenzo,
Miguel A Sanz
[show abstract]
[hide abstract]
ABSTRACT: Infectious complications are a major cause of morbidity and mortality in patients who undergo autologous stem cell transplantation (ASCT). We examined 476 patients with hematologic malignancies (401) or solid tumors (75) who underwent ASCT between February 1990 and May 2005. Anti-infectious prophylaxis consisted of different combinations of ciprofloxacin, cotrimoxazole, fluconazole, aerosolized amphotericin B, acyclovir, and intravenous immunoglobulins. Overall, 454 patients (95%) developed fever in the first 60 days after ASCT. In the majority of patients, initial antibiotic therapy consisted of broad-spectrum beta-lactamic with or without amikacin. A glycopeptide was administered as initial therapy in 86 cases. Overall, there were 132 (29%) clinically documented infections (37 pneumonias), 79 (17%) microbiologically documented infections (65 bacteremias), and 243 (54%) fevers of unknown origin. Coagulase-negative staphylococci (18, 25%) and E coli (18, 25%) were the organisms most frequently isolated. The pattern of infection did not change throughout the study except for a significantly higher incidence of bacteremia due to gram-positive bacteria in the first 5 years of the study. Infection-related mortality was 5% (21 cases), with pneumonia the most frequent cause of death. ASCT should be considered a low-risk procedure, although new therapeutic approaches for patients developing severe respiratory infections are still needed.
International Journal of Hematology 09/2007; 86(2):186-92. · 1.27 Impact Factor
-
Noemí Puig,
Javier de la Rubia,
María J Remigia,
Isidro Jarque,
Guillermo Martín,
Luca Cupelli,
Guillermo F Sanz,
Ignacio Lorenzo,
Jaime Sanz,
Jesús A Martínez,
Carmen Jiménez,
Miguel A Sanz
[show abstract]
[hide abstract]
ABSTRACT: CBV and BEAM are the two most frequently used regimens for patients with lymphoma undergoing autologous hematopoietic stem-cell transplantation (ASCT). This study compared their morbidity and transplant-related mortality (TRM) in 113 patients with non-Hodgkin's lymphoma (69) and Hodgkin's disease (44) undergoing ASCT between 1990 - 2004. CBV (cyclophosphamide, 6000 mg m(-2); VP-16, 750 mg m(-2); and high-dose BCNU, 800 mg m(-2)) was administered to 75 patients and 38 received BEAM (BCNU, 300 mg m(-2); VP-16, 800 mg m(-2); cytarabine, 800 mg m(-2); melphalan, 140 mg m(-2)). Patients in the BEAM group had a significantly higher median age (p = 0.002) and were more heavily treated before ASCT (p = 0.003). More patients showed active disease at transplant in the BEAM group (p = 0.04). Sinusoidal obstruction syndrome (SOS) was more frequent in the CBV group (11% vs 0%, p = 0.048). There were 20 (18%) transplant-related deaths, 18 in the CBV and two in the BEAM group. Infectious complications (12 patients, seven with pneumonia) and SOS (four) were the most frequent causes of death. The cumulative incidences of TRM were 25% in the CBV and 7% in the BEAM group (p = 0.02). CBV thus produced a higher incidence of SOS and TRM than BEAM in this series.
Leukemia and Lymphoma 09/2006; 47(8):1488-94. · 2.58 Impact Factor
