[Show abstract][Hide abstract] ABSTRACT: Focal adhesions have been associated with poor prognosis in multiple cancer types, but their prognostic value in diffuse large B-cell lymphoma (DLBCL) has not been evaluated. The aim of this study was to investigate the expression patterns and the prognostic value of focal adhesion proteins FAK, Pyk2, p130Cas and HEF1 in DLBCL.
Focal adhesion proteins expression was examined using immunohistochemistry in normal lymphoid tissues and in 60 DLBCL patient sample sections. Kaplan-Meier survival and Cox regression analysis were performed to evaluate focal adhesion proteins correlation with patient prognosis.
FAK, Pyk2, p130Cas and HEF1 expression was mostly found in the germinal centers of normal human lymphoid tissues. When assessed in DLBCL samples, FAK, Pyk2, p130Cas and HEF1 were highly expressed in 45, 34, 42, and 45% of the samples, respectively. The multivariate COX analysis revealed that decreased FAK expression was a significant independent factor predictive of poor disease outcome.
FAK expression is an independent prognostic factor in DLBCL. Our results suggest that the addition of FAK immunostaining to the current immunohistochemical algorithms may facilitate risk stratification of diffuse large B cell lymphoma patients. This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: We analyze the impact of cyclosporine (CsA) levels in the development of acute graft-versus-host disease (aGVHD) after reduced intensity conditioning allogeneic hematopoietic transplantation (allo-RIC). We retrospectively evaluated 156 consecutive patients who underwent HLA-identical sibling allo-RIC at our institution. CsA median blood levels in the 1st, 2nd, 3rd and 4th weeks after allo-RIC were 134 (range: 10-444), 219 (54-656), 253 (53-910) and 224 (30-699) ng/mL; 60%, 16%, 11% and 17% of the patients had median CsA blood levels below 150 ng/mL during these weeks. 53 patients developed grade 2-4 aGVHD for a cumulative incidence of 45% (95% CI 34-50%) at a median of 42 days. Low CsA levels on the 3rd week and sex-mismatch were associated with the development of GVHD. Risk factors for 1-year NRM and OS were advanced disease status (HR: 2.2, P = 0.02) and development of grade 2-4 aGVHD (HR: 2.5, P < 0.01), while there was a trend for higher NRM in patients with a low median CsA concentration on the 3rd week (P = 0.06). These results emphasize the relevance of sustaining adequate levels of blood CsA by close monitoring and dose adjustments, particularly when engraftment becomes evident. CsA adequate management will impact on long-term outcomes in the allo-RIC setting.
Mediators of Inflammation 01/2014; 2014:620682. · 3.88 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The non-Hodgkin lymphomas are 12(th) most prevalent cancers in Europe. No recent update in the epidemiology of these lymphomas has been performed in our country. We diagnosed 701 new lymphomas during the period beginning January 1, 2000 and ending December 31, 2009 in our center.
The most frequent lymphoma was diffuse large B cell lymphoma, followed by follicular lymphoma and then classic Hodgkin's disease. The male:female ratio is 1.2:1. Diagnosis by age showed that non-Hodgkin's lymphoma is by far more frequent in the 61-80 years old patients. On the other hand, classic Hodgkin's lymphoma is more frequent in the 20-40 years old population.
Our results are very similar to those published by other centers in Europe and United States.
[Show abstract][Hide abstract] ABSTRACT: In the rituximab era, lymphoma patients with persistent disease receiving autologous transplantation have a very poor outcome. The addition of radioimmunotherapy to the conditioning regimen may improve outcome for these patients. We have evaluated, in a prospective phase 2 study, the safety and efficacy of the addition of 90Y-Ibritumomab tiuxetan to the conditioning chemotherapy in refractory diffuse large B cell lymphoma patients. Thirty patients with induction failure (primary refractory; n=18) or refractory to salvage immunochemotherapy at relapse (n=12) were included in the study. Patients with a median age of 53 years (range, 25-67) received 90Y-Ibritumomab tiuxetan at a fixed dose of 0.4mCi/kg (maximum dose 32mCi) 14 days prior to the preparative chemotherapy regimen. Histology included de novo diffuse large B cell lymphoma (22) and transformed diffuse large B cell lymphoma (8). All patients had persistent disease at the time of transplantation, with 25 patients considered to be chemorefractory. Median time to neutrophil recovery (>500/ml) was 11 days (9-21), and to platelet recovery (>20.000/ml) was 13 days (11-35). Overall response at day +100 was 70% (95% CI, 53.6-86.4) with 60% (95% CI, 42.5-77.5) complete responses. After a median follow-up of 31 months for alive patients (range, 16-54), estimated 3-year overall and progression-free survival is 63% (95% CI, 48-82) and 61% (95% CI, 45-80), respectively. We conclude that autologous transplantation with conditioning including 90Y-Ibritumomab tiuxetan is safe, and results in a very high response rate with promising survival in this very poor prognosis group of refractory diffuse large B cell lymphoma patients. Study registered at European Union Drug Regulating Authorities Clinical Trials (EudraCT) No. 2007-003198-22.
[Show abstract][Hide abstract] ABSTRACT: We have previously reported that LITAF is silenced by promoter hypermethylation in germinal centre-derived B-cell lymphomas, but beyond these data the regulation and function of lipopolysaccharide-induced tumour necrosis factor (TNF) factor (LITAF) in B cells are unknown. Gene expression and immunohistochemical studies revealed that LITAF and BCL6 show opposite expression in tonsil B-cell subpopulations and B-cell lymphomas, suggesting that BCL6 may regulate LITAF expression. Accordingly, BCL6 silencing increased LITAF expression, and chromatin immunoprecipitation and luciferase reporter assays demonstrated a direct transcriptional repression of LITAF by BCL6. Gain- and loss-of-function experiments in different B-cell lymphoma cell lines revealed that, in contrast to its function in monocytes, LITAF does not induce lipopolysaccharide-mediated TNF secretion in B cells. However, gene expression microarrays defined a LITAF-related transcriptional signature containing genes regulating autophagy, including MAP1LC3B (LC3B). In addition, immunofluorescence analysis co-localized LITAF with autophagosomes, further suggesting a possible role in autophagy modulation. Accordingly, ectopic LITAF expression in B-cell lymphoma cells enhanced autophagy responses to starvation, which were impaired upon LITAF silencing. Our results indicate that the BCL6-mediated transcriptional repression of LITAF may inhibit autophagy in B cells during the germinal centre reaction, and suggest that the constitutive repression of autophagy responses in BCL6-driven lymphomas may contribute to lymphomagenesis.
British Journal of Haematology 06/2013; · 4.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: MYC alterations influence the survival of patients with diffuse large B-cell lymphoma. Most studies have focused on MYC translocations but there is little information regarding the impact of numerical alterations and protein expression. We analyzed the genetic alterations and protein expression of MYC, BCL2, BCL6, and MALT1 in 219 diffuse large B-cell lymphomas. MYC rearrangement occurred as the sole abnormality (MYC single-hit) in 3% of cases, MYC and concurrent BCL2 and/or BCL6 rearrangements (MYC double/triple-hit) in 4%, MYC amplifications in 2% and MYC gains in 19%. MYC single-hit, MYC double/triple-hit and MYC amplifications, but not MYC gains or other gene rearrangements, were associated with unfavorable progression free survival and overall survival. MYC protein expression, evaluated using a computerized image analysis, captured the unfavorable prognosis of MYC translocations/amplifications and identified an additional subset of patients without gene alterations but with similar poor prognosis. Tumors expressing both MYC/BCL2 had the worst prognosis, whereas double negative tumors had the best outcome. High MYC expression was associated with shorter overall survival irrespective of the International Prognostic Index and BCL2 expression. In conclusion, MYC protein expression identifies a subset of diffuse large B-cell lymphoma with very poor prognosis independently of gene alterations and other prognostic parameters.
[Show abstract][Hide abstract] ABSTRACT: Strongyloides stercoralis is an intestinal nematode that causes strongyloidiasis, which affects 30 to 100 million people worldwide. Risk factors for hyperinfection and disseminated disease include immunosuppressive drug therapy, human T-lymphotropic virus-1 (HTLV-1) infection, solid organ and bone marrow transplantation, hematologic malignant diseases, hypogammaglobulinemia, and severe malnutrition and associated conditions. The diagnosis can be difficult because a single stool examination fails to detect larvae in up to 70% of the cases, and the symptoms are nonspecific. Although eosinophilia is a common finding in patients with chronic Strongyloides infection, it is an unreliable predictor of hyperinfection. Furthermore, the lack of eosinophilia while receiving immunosuppressive therapy cannot reliably exclude the underlying chronic Strongyloides infection. We report here a fatal Strongyloides hyperinfection in a patient receiving allogeneic stem cell transplantation; risk factors and outcome in this clinical setting are discussed.
[Show abstract][Hide abstract] ABSTRACT: Refractory acute graft-versus-host disease (aGVHD) remains an important cause of mortality after allogeneic stem cell transplantation. No standard therapy exists once steroids fail to obtain a good response. In 2006, our group published a series of patients who received inolimomab, an anti-interleukin-2 receptor monoclonal antibody, as salvage therapy with initial encouraging results. In this update, we have analysed a larger group of patients with prolonged follow-up. Ninety-two consecutive patients were treated with inolimomab in our center between April 1999 and December 2011. Overall response rate was 42% (complete response [CR] in 14%) on day +30. Predictors of failure to respond in the multivariate analysis were overall aGVHD grade IV, instauration of inolimomab before day 15 of aGVHD diagnosis and severe lymphopenia. Patients without gastrointestinal (GI) involvement appeared to do better, with a 70% response rate compared with 39% in patients with GI involvement (p=0.06). However, the 2-year overall survival (OS) was 18% (95% CI 10-26%) for the entire cohort and 33% (95% CI 25-40%) for day-30 responders. Acute GVHD was the main cause of death (49%) followed by opportunistic infections (27%). In conclusion, results of this update show that although inolimomab is a well tolerated drug with a moderate number of short term responses, it is associated with long-term survival in only one third of responding patients. These data highlight the need of investigating new rescue treatments with sustained effect and theimportance of reportinglong-term outcomesin GVHD studies.
Biology of blood and marrow transplantation: journal of the American Society for Blood and Marrow Transplantation 11/2012; · 3.15 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background. This phase II clinical trial evaluated the efficacy, safety and pharmacokinetics of plitidepsin 3.2 mg/m2 administered as a 1-hour intravenous infusion weekly on Days 1, 8 and 15 every 4 weeks in 67 adult patients with relapsed/refractory aggressive non-Hodgkin's lymphoma. Design and Methods. Patients were divided into 2 cohorts: non-cutaneous peripheral T-cell lymphoma (n=34) and other lymphomas (n=33). Efficacy was evaluated using the International Working Group criteria (1999). Results. Response occurred in 6 of 29 evaluable patients with non-cutaneous peripheral T-cell lymphoma (overall response rate 20.7%; 95% confidence interval, 8.0%-39.7%), including 2 complete responses and 4 partial responses. No responses occurred in 30 evaluable patients with other lymphomas (including 27 B-cell lymphomas). Most common plitidepsin-related adverse events were nausea, fatigue and myalgia (grade 3 in <10% of cases). Severe laboratory abnormalities (lymphopenia, anemia, thrombocytopenia, and transaminase and creatine phosphokinase increases) were transient and easily managed by plitidepsin dose adjustments. Pharmacokinetic profile did not differ from that previously reported in patients with solid tumors. Conclusions. Plitidepsin monotherapy has clinical activity in relapsed/refractory T -cell lymphomas. Combinations of plitidepsin with other chemotherapeutic drugs deserve further evaluation in non-cutaneous peripheral T-cell lymphoma. (clinicaltrials.gov identifier: NCT00884286).
[Show abstract][Hide abstract] ABSTRACT: Pretransplant pulmonary function tests (PFTs) have been checked mostly in myeloablative allogeneic stem cell transplantation (Allo-SCT). Their value in the setting of reduced intensity conditioning Allo-SCT (Allo-RIC) has been less explored. We retrospectively evaluated the predictive value of PFTs on posttransplant pulmonary complications (PPC) and outcomes in 195 consecutive Allo-RIC patients, based on fludarabine plus busulphan or melphalan. PFT parameters included forced vital capacity (FVC), forced expiratory volume in the first second (FEV1), FEV1/FVC ratio, total lung capacity (TLC), residual volume, and diffusion capacity for carbon monoxide (DLCo) corrected for the hemoglobin levels. Pretransplant PFTs abnormalities were observed in 130 patients (66%). The most frequent abnormalities were abnormal DLCO (n = 83, 44%), followed by FEV1/FVC (n = 75, 38%) and FVC (n = 47, 24%). The abnormalities were severe in 25 (13%) patients, moderate in 65 (33%) and mild in 40 patients (21%). Multivariate analysis showed that TLC was significantly associated with PPC, nonrelapse mortality and overall survival (OS), (HR 4.2, 95% CI. 2-8.5; HR 3.8, 95% CI. 1.7-8.5; HR 2.3, 95% CI. 1.3-4.1, respectively, P = 0.01), while abnormal FVC had a negative impact on PPC and OS (HR 1.8, 95% CI. 0.98-3.6, P = 0.06 and HR 1.7, 95% CI. 1.1-2.6, P = 0.008). This study emphasizes the valuable role of PFTs in identifying patients at risk for PPC, NRM, and lower OS in the Allo-RIC setting.
American Journal of Hematology 01/2012; 87(1):9-14. · 4.00 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Whether the intensity of the conditioning regimen affects febrile neutropenia (FN) and severe bacterial infections (SBIs) is not well established. We analyzed the risk factors (RFs) for the development of FN and SBI in the first 100d post-transplant in 195 consecutive adult recipients of a reduced-intensity conditioning allogeneic hematopoietic stem cell transplantation (RIC-allo).
The RIC regimens consisted of fludarabine plus melphalan (62%) or busulphan (38%) (FluMel or FluBu). SBIs include pneumonia, urinary tract infections, and bacteremia.
FN occurred in 141 patients (72%), always in the first 30d post-allo-RIC. However, a SBI occurred in only 27 patients (14%) during this early post-transplant period (<day +30), while 29 evaluable patients (15%) developed a SBI in the intermediate post-transplant period (days +31 to +100). In multivariate analysis, RFs for the development of FN included onset of neutropenia before day +5 after allo-RIC (P<0.02) and NCI CTC grade III-IV mucosal damage in the first 10d post-transplantation (P=0.03). RFs identified to SBI by multivariate analysis included corticosteroid therapy before day +100 (P<0.01), mycophenolate mofetil-based graft-versus-host disease (GVHD) prophylaxis (P<0.01), and previous SBI before day +30 (P<0.01). The rate of SBI from day +30 to +100 varied according to the number of RFs; thus, the rate of SBI was 1% in patients without any RF, 17% in patients with one RF, 29% with one RFs, and 53% in those with all three RFs.
After an RIC-allo, FN and early SBI occurred mostly in patients with severe mucositis and early-onset neutropenia, while postengraftment high-dose steroid therapy for acute GVHD was the major RF.
European Journal Of Haematology 01/2012; 88(1):46-51. · 2.55 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Chemoimmunotherapy with anti-CD20 monoclonal antibody rituximab is increasingly used for the treatment of patients with chronic lymphocytic leukemia (CLL). Antibody-dependent cytotoxicity (ADCC) is one of the most important mechanisms of action of rituximab against B-cell malignancies. We studied ways to increase the cytotoxic effect of rituximab on CLL cells by enhancing ADCC. Peripheral blood mononuclear cell (PBMC) or purified natural killer (NK) cells from healthy donors were activated with interleukin-15 (IL-15) and cultured with rituximab-coated CLL cells, and ADCC was evaluated using a (51)chromium release assay. The IL-15 significantly enhanced in vitro ADCC against CLL cells, and this effect was mainly mediated by NK cells. The IL-15 treated effector cells with the low affinity FcγRIIIA receptor (158FF) had an ADCC comparable to those with the high affinity FcγRIIIA form (158VF). In addition, IL-15 enhanced rituximab-mediated ADCC of CLL cells in the presence of transforming growth factor-beta. The IL-15 increases rituximab-mediated ADCC against CLL, and supports the use of such agents with the goal of improving clinical response to chemoimmunotherapy in patients with CLL.
[Show abstract][Hide abstract] ABSTRACT: Diffuse large B-cell lymphoma (DLBCL) heterogeneity has prompted investigations for new biomarkers that can accurately predict survival. A previously reported 6-gene model combined with the International Prognostic Index (IPI) could predict patients' outcome. However, even these predictors are not capable of unambiguously identifying outcome, suggesting that additional biomarkers might improve their predictive power.
We studied expression of 11 microRNAs (miRNA) that had previously been reported to have variable expression in DLBCL tumors. We measured the expression of each miRNA by quantitative real-time PCR analyses in 176 samples from uniformly treated DLBCL patients and correlated the results to survival.
In a univariate analysis, the expression of miR-18a correlated with overall survival (OS), whereas the expression of miR-181a and miR-222 correlated with progression-free survival (PFS). A multivariate Cox regression analysis including the IPI, the 6-gene model-derived mortality predictor score and expression of the miR-18a, miR-181a, and miR-222, revealed that all variables were independent predictors of survival except the expression of miR-222 for OS and the expression of miR-18a for PFS.
The expression of specific miRNAs may be useful for DLBCL survival prediction and their role in the pathogenesis of this disease should be examined further.
Clinical Cancer Research 06/2011; 17(12):4125-35. · 7.84 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Several gene-expression signatures predict survival in diffuse large B-cell lymphoma (DLBCL), but the lack of practical methods for genome-scale analysis has limited translation to clinical practice. We built and validated a simple model using one gene expressed by tumor cells and another expressed by host immune cells, assessing added prognostic value to the clinical International Prognostic Index (IPI). LIM domain only 2 (LMO2) was validated as an independent predictor of survival and the "germinal center B cell-like" subtype. Expression of tumor necrosis factor receptor superfamily member 9 (TNFRSF9) from the DLBCL microenvironment was the best gene in bivariate combination with LMO2. Study of TNFRSF9 tissue expression in 95 patients with DLBCL showed expression limited to infiltrating T cells. A model integrating these 2 genes was independent of "cell-of-origin" classification, "stromal signatures," IPI, and added to the predictive power of the IPI. A composite score integrating these genes with IPI performed well in 3 independent cohorts of 545 DLBCL patients, as well as in a simple assay of routine formalin-fixed specimens from a new validation cohort of 147 patients with DLBCL. We conclude that the measurement of a single gene expressed by tumor cells (LMO2) and a single gene expressed by the immune microenvironment (TNFRSF9) powerfully predicts overall survival in patients with DLBCL.
[Show abstract][Hide abstract] ABSTRACT: Infection-related mortality (IRM) is responsible for a major proportion of all cases of non-relapse mortality (NRM) after allogeneic PBSCT (alloPBSCT). We analyzed 580 consecutive adults who received a first alloPBSCT from an HLA-identical sibling from 1994 to 2008 at a single institution to describe the severe infections and report the incidence, causes and risk factors for IRM and NRM. Both IRM and NRM decreased with time; within the period of 1994-2000, the 2-year incidence of IRM and NRM was 22 and 31%, respectively, vs 11 and 16% within the period of 2001-2008 (P<0.05 for both comparisons). In multivariate analysis, the variables that increased IRM were within the earlier period of 1994-2000 (P<0.01), poor performance status (P<0.01), grade II-IV acute GVHD (P<0.001) and invasive fungal infection (IFI) (P<0.001) or CMV disease (P<0.001) after transplant. With respect to NRM, earlier time period was also identified as a risk factor (P<0.001), as well as IFIs (P<0.001) and CMV disease (P<0.001). The intensity of the conditioning regimen had no effect on IRM and NRM. These results showed a significant reduction in IRM and NRM over a period of 15 years. The development of IFIs and CMV disease continue to have an impact on NRM.
Bone marrow transplantation 05/2011; 46(5):690-701. · 3.00 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Acute Graft-versus-host disease (GVHD) is a major complication after allogeneic hematopoietic stem cell transplantation. Although this process is thought to consist of several phases, T-cell activation plays a critical role in the pathogenesis of acute GVHD. To become efficient effectors, T-cells require additional costimulation after T-cell receptor signaling. A number of molecules are involved in costimulation of T-cells such as CD28, CD40L, CD30, OX40, 4-1BB, ICOS, and LIGHT. The system is regulated by inhibitory molecules, CTLA-4, and PD-1. There is experimental evidence that those molecules are implicated in the pathogenesis of GHVD. We describe how these molecules are involved in acute GVHD and how the blockade of costimulatory molecules may have potential implications for the treatment of patients with acute GVHD.
[Show abstract][Hide abstract] ABSTRACT: Monoclonal antibodies are standard therapeutics for several cancers including the anti-CD20 antibody rituximab for B cell non-Hodgkin lymphoma (NHL). Rituximab and other antibodies are not curative and must be combined with cytotoxic chemotherapy for clinical benefit. Here we report the eradication of human NHL solely with a monoclonal antibody therapy combining rituximab with a blocking anti-CD47 antibody. We identified increased expression of CD47 on human NHL cells and determined that higher CD47 expression independently predicted adverse clinical outcomes in multiple NHL subtypes. Blocking anti-CD47 antibodies preferentially enabled phagocytosis of NHL cells and synergized with rituximab. Treatment of human NHL-engrafted mice with anti-CD47 antibody reduced lymphoma burden and improved survival, while combination treatment with rituximab led to elimination of lymphoma and cure. These antibodies synergized through a mechanism combining Fc receptor (FcR)-dependent and FcR-independent stimulation of phagocytosis that might be applicable to many other cancers.
[Show abstract][Hide abstract] ABSTRACT: Interphase fluorescence in situ hybridization (I-FISH) studies have a remarkable prognostic value in patients with chronic lymphocytic leukemia (CLL). I-FISH studies can be performed either on tetradecanoylphorbol acetate stimulated peripheral blood cells (I-FISH-TPA) or unstimulated peripheral blood mononuclear cells (I-FISH-PBMC). The aim of the study was to evaluate whether this finding was clinically relevant in a group of 235 patients with CLL. Fifty-six patients had both I-FISH-TPA and I-FISH-PBMC results. Compared with uncultured cells, the cytogenetic detection rate rose from 57 to 80% with the use of TPA-stimulated cells (P = 0.014). I-FISH-TPA provided a better prediction of treatment-free survival compared with I-FISH-PBMC (P = 0.031 vs. 0.166). Then, I-FISH-PBMC results from 93 historical patients were compared with 86 recent patients with I-FISH-TPA results. Genomic aberrations were detected in 46 and 67% of patients from the I-FISH-PBMC and I-FISH-TPA cohorts, respectively. The detection rate of 13q deletion as the only aberration increased from 10% with I-FISH-PBMC to 37% with I-FISH-TPA (P = 0.006). In conclusion, I-FISH-TPA increased the detection rate of 13q deletion and had an improved prognostic value compared with I-FISH-PBMC.
Genes Chromosomes and Cancer 04/2010; 49(4):327-32. · 3.55 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Heat-shock protein (HSP)90 is a molecular chaperone involved in the proper folding and cellular transportation of many signaling proteins that are deregulated in lymphoma. HSP90 inhibition results in proteasomal degradation of these proteins, leading to antitumoral activity. Recent studies have focused on the use of HSP90 inhibitors as potential therapies for non-Hodgkin lymphoma. BCL6 plays a critical role in the pathogenesis of most diffuse large B-cell lymphoma (DLBCL), the most frequent non-Hodgkin lymphoma. The current study demonstrates that HSP90 forms a complex with BCL6, and inhibition of HSP90 with the drug PU-H71 selectively kills BCL6-positive DLBCL in animal models. These data support the use of HSP90 inhibitor PU-H71 for treating patients with BCL6-positive DLBCL.
Expert Review of Hematology 04/2010; 3(2):157-9. · 2.38 Impact Factor