ALBERT

Description: Background The use of combination antiretroviral therapy (CART) has improved the prognosis of HIV-related diffuse large B cell lymphoma (DLBCL). In the pre-CART era, HIV-infected individuals usually presented DLBCL of primary extranodal origin and advanced stage, unlike non-HIV-infected individuals. In the CART era, studies conducted before the introduction of immunochemotherapy pointed outcomes of HIV-infected patients to be approaching those of non-HIV-infected patients. Recently, studies have reported the lack of influence of HIV-infection in the prognosis of Burkitt’s lymphoma treated with immunochemotherapy. Nevertheless, there is scarce information comparing the clinical presentation and outcome of DLBCL treated with immunochemotherapy between non-HIV-infected patients and HIV-infected patients in CART era. Methods We retrospectively studied two series of DLBCL patients treated with RCHOP; 81 HIV-infected patients included in a Spanish multicentre trial and 84 non-HIV-infected patients diagnosed in our institution between 2002 and 2010. Demographic, HIV-infection, and DLBCL data on each case were collected. DLBCL cases were classified in nodal and extranodal according to a previously reported definition (JCO 2005). Continuous and categorical variables are presented using descriptive statistics. Survival analyses were performed using the Kaplan-Meier method, and compared using the log-rank test. P-values of less than 0.05 were considered statistically significant. Results The median follow-up of HIV-infected patients was 6.5 years and of non-HIV-infected patients was 4.6 years. Non-HIV-infected patients were older at the time of DLBCL diagnosis than HIV-infected patients, median age (range) 62 years (24-80) and 44 years (24-74) respectively; (P

Description: Background: Follicular lymphoma (FL) is the most frequent indolent lymphoma and is characterized by a high response to immunochemotherapy (ICT). However, patients refractory to first-line ICT have a worse prognosis. The objective of this study was to determine the prevalence of refractory FL, the factors that predict refractoriness as well as the salvage treatment and outcome. Patients and methods: This is a retrospective analysis including stage II-IV FL patients treated with first-line ICT in 3 Spanish institutions. The cohort was divided into ICT-refractory patients (less than partial response after induction or maintenance/consolidation therapy, as well as relapse or progression within 6 months of the last dose of therapy) and ICT-sensitive. Baseline features, therapy received and outcome were analyzed. Results: 283 patients were included, the median age was 58 years-old (range 28 to 85) and 53% were female. 200/231 (87%) had a good performance status (ECOG 〈 2), 260/295 (88%) presented with stages III and IV and 163/284 (57%) had bone marrow involvement. High-risk FLIPI score was seen in 108/256 (42%), high serum LDH in 78/263 (28%) and high serum B2-microglobulin in 138/253 (54%). RCHOP was administered to 226 (80%), RCVP to 36 (13%) and rituximab in combination with fludarabine or bendamustine-based therapy to 21 (7%). Seventeen patients received consolidation with radioimmunotherapy and 140 received maintenance with rituximab (n=137) or interferon (n=3). Sixteen patients received complementary radiotherapy. Forty-three (16%) patients were ICT-refractory (37 within 6 months of the completion of induction and 6 during or within 6 months of the completion of maintenance/consolidation therapy). On univariate analysis, high-risk FLIPI (OR 5.4, [95% CI 2.3-12.6]), high-risk FLIPI2 (5.4, [2.4-12.4]), B symptoms (3.2, [1.6-6.6]), ECOG ≥ 2 (4.6, [2-10.9]), involvement of 〉 4 nodal regions (2.3, [1.02-5.3]), hepatomegaly (7.5, [2.6-21.5]), splenomegaly (2.8, [1.4-5.9]), high B2-microglobulin (4, [1.7-9.5]), high serum LDH (3.9, [1.8-8]) and treatment with RCVP (compared with RCHOP, 2.8, [1.2-6.2]) were correlated with refractoriness. On multivariate analysis, high-risk FLIPI score (4.9, [2.1-11.7]) and treatment with RCVP (3.4, [1.2-9.4]) were the only variables associated with refractoriness. After exclusion of FLIPI, ECOG ≥ 2 (3, [1.1-8.4]) and high serum LDH (4.7, [2-11]) were correlated with refractoriness, in addition to RCVP therapy (4.5, [1.5-13.2]). Ten-year OS probabilities in ICT-sensitive and ICT-refractory patients were 83% (95% CI 76%-90%) and 33% (12%-54%), respectively (p

Description: Abstract 1437 JAK2 mutation testing and karyotye are routinely used for diagnosis of myeloproliferative neoplasms (MPNs) but they have not been incorporated into risk stratification. Although JAK2V617F mutation in MPNs has been one of the most seminal medical discoveries in recent years, it is not clear the importance of the amount of JAK2V617F allele. Some studies correlate the JAK2 allele burden with a higher hemoglobin level, leukocyte count, splenomegaly and thrombosis and more probability of transformation to MF or AML. The aim of this study was to determine whether cytogenetic data, JAK2 mutation status and allele burden correlates with cytological subtypes, clinical complications or provide prognostic information. Methods A retrospective study was conducted with samples centralized in a unique laboratory since 2005. A total of 526 patients were included (median age 63; 243 males) with classic MPNs (348 ET, 135 PV, 43 PMF) fulfilling 2008 WHO criteria and in accordance with the Declaration of Helsinki. Conventional cytogenetic was performed in bone marrow samples obtained at diagnosis (n=205) and at progression to MF or AML (n=46). DNA was extracted from peripheral blood using QIAamp DNA mini kit (Qiagen). All samples were coded and assayed blindly in duplicate to detect JAK2V617F mutation with an allele-specific PCR using TaqMan allelic discrimination, with 2 specific probes to measure the respective fluorescence of each allele. Then, JAK2 MutaQuant assay (Ipsogen, Luminy Biotech) was used to detect the JAK2V617F quantity by real-time PCR, detecting fluorescent signals using double-dye hyrolysis oligonucleotide probes with calibration standards at 4 different concentrations. Homozygous (HOZ) ratio was considered when percentage was higher than 50. Laboratory (hemoglobin, WBC and platelet counts) and clinical data (constitutional symptoms, splenomegaly, complications, OS and DFS) were collected. For continuous variables parametric and non parametric statistics were used. Survival analysis was performed using Kaplan-Meier estimate and log-rank tests were used for comparisons. The χ2 and Fisher's exact tests were used to analyze differences in the distribution of variables among patient subsets. p-value less than 0.05 were considered statistically significant. Results Aberrant karyotypes were seen in 15/205 (7%) cases at diagnosis (4% in ET and PV and 40% in PMF). At progression to MF or AML we have cytogenetic studies in 22 patients, and 10 (45%) harbor alterations. A total of 283 patients (64%) were JAK2V617F, 61% ET (4% HOZ), 75% PV (28% HOZ) and 55% PMF (16% HOZ). The median value of JAK2V617F was 26% (range, 1–99.9%). No correlation was seen between JAK2 and karyotype at diagnosis, but 7/9 patients with aberrant karyotype at progression had JAK2V617F mutation. JAK2 correlations with laboratory and clinical data are summarized in Table 1 and 2. Conclusions JAK2V617F is associated with a more pronounced myeloproliferative phenotype (higher hemoglobin level, platelet or leukocyte count). Patients with JAK2V617F HOZ have a higher probability of splenomegaly. Hematological complications do not depend on JAK2 ratio but mutated patients have more probability of thrombosis or hemorrhage. OS and PFS do not depend on JAK2 status, but patients with JAK2V617F heterozygous seem to have a slightly better outcome. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 4897 Objective. To study the frequency, type and prognostic significance of clonal chromosomal abnormalities following SCT in patients with AML and MDS. Patients and methods. One hundred thirty patients were studied between 2000 and 2010. Karyotypes were analysed by G-banded chromosomes obtained from 24 hours bone marrow cultures, and were described according to ISCN 2009. Results. Clonal abnormalities were observed in 36/130 patients (28%) with a median follow-up was 11 months (range 3–131). Initial diagnosis (OMS 2008): AML with maduration (8 patients), acute erythroid leukaemia (4), acute monocytic leukaemia (3), AML with multilineage dysplasia (3), AML with myelodysplasia-related changes (3), AML with inv(16)(p13q22) (2), acute monoblastic leukaemia (2), AML with minimal differentation (2), AML without maduration (1), acute myelomonocytic leukaemia (1), refractory anaemia with excess blasts (4), chronic myelomonocytic leukaemia (2) and refractory cytopenia with multilineage dysplasia (1). Treatment before SCT: idarubicin, cytarabine, etoposide and mitoxantrone (25), idarubicin, cytarabine and etoposide (6), idarubicin, cytarabine, etoposide and gemtuzumab (2), FLAG-ida (2) and azacitidine (1). SCT type: autologous (23 patients), allogeneic of reduced intensity (7), allogenic (4), umbilical cord blood (1) and syngeneic transplant (1). Conditioning regimen: TBI and cyclophosphamide (24 cases), fludarabine and busulfan (7), busulfan and cyclophosphamide (4) and thymoglobulin, thiotepa, fludarabine and busulfan (1). The median time between SCT and the appearance of clonal abnormalities was 6.5 months (range 2–51). At the time of clonal abnormality detection, 32 patients were in cytological and/or clinical relapse and 4 in cytological remission. In 15 cases the initial clone reappeared, 2 showed the initial abnormalities with an acquired abnormality and 19 presented de novo clonal abnormalities (53%). The most frequent clonal chromosomal abnormalities observed were: complex karyotype (22%), estructural abnormalities with afection of chromosomes 1, 4, 6, 7, 10, 11, 12, 16 and 17 (22%), +21 (5%) and +11 (3%). The median survival from the appearance of clonal abnormalities was 2.5 months (range 1–100). There were no differences when we compared the survival of patients with de novo clonal abnormalities with that of patients with initial abnormalities (including those with acquired abnormality). Furtheremore there were no differences when we compared the median survival of relapsed patients with normal Karyotype with that of patients with clonal chromosomal abnormalities following SCT (2.5 months [range 1–25] and 2.5 months [range 1–100] respectively). At the time of the analysis only 1 patients are alive and in complete remission (100 months). Conclusions. 1. The appearance of clonal abnormalities following SCT in patients with AML and MDS is frequent and the majority of cases are detected at the time of relapse. Half of cases presented de novo clonal abnormalities and the majority were not related with prior irradiation exposition, alkylating agents neither topoisomerase II inhibitors. The prognosis of patients with acquired clonal abnormalities after SCT is poor. Disclosures: No relevant conflicts of interest to declare.

Description: Introduction Monoclonal B-Cell Lymphocytosis (MBL) is a frequent condition characterized by a clonal B-cell population detected in the blood or bone marrow, in otherwise asymptomatic individuals.A small proportion of patients with MBL need clinical attention. Its prevalence is around 3-5% in healthy adults over age 40 and rises with age. The largest studies of MBL features are focused on healthy population, therefore additional data regarding the population referred to hospital are necessary. Genotype and immunophenotype data are well described in the literature so far, but a comprehensive framework of the condition is still required. Data of features not well described heretofore like bone marrow pattern of infiltration would help in the differential diagnosis. The aim of this work was to analyze the clinical and laboratory findings of MBL reclassified from CLL or other B-cell lymphoproliferative disorders (LPD) with peripheral involvement, referred to a centre. Patients and methods Three hundred seventy-one LPD patients with peripheral blood involvement diagnosed between years 2000 and 2009 in a single centre were revised to reclassify them according to the World Health Organization consensus cut-off of 5x109/L circulating B cells. From those reclassified as MBL, blood count, biochemistry, immunophenotype, cytogenetics, bone marrow trepine biopsy and results of CT screening were registered, as well as treatment and outcome data. Only those patients who were followed in our centre were included. Results Seventy-three LPD (20%) were reclassified as MBL, with a median age of 72 years (range: 42-94) and a 52% of males. Proportions of CLL-like LPD, CD5 negative non CLL-like and CD5 positive non CLL-like switched from 65%, 21% and 14% to 84%, 7% and 9% after the reclassification. In only seven out of 41 patients the bone marrow was not infiltrated, while the majority had an interstitial and a mixed nodular and interstitial infiltration (44% and 36%, respectively). Only 2 out of 49 patients had a concomitant infection with hepatitis C virus. Fifty-nine % of patients had a FISH alteration, predominantly chromosome 13 deletion. Thirty-seven out of 69 patients (54%) fulfilled criteria of LPD progression, with 10 patients (15%) requiring treatment (7 out of them because of progression of CLL). The mean (SD) B-lymphocyte count at diagnosis of patients with LPD progression was 3.58x10^9/L (0.9x10^9/L) versus 3.03x10^9/L (1.1x10^9/L) in those who did not progress, p=0.023. The 12-year progression free survival of the whole series was 38% (CI 95% 23-53) with a median of 4.08 years (range: 2.3-5.8). The 12-year overall survival was 54% (29-79), with a median follow-up of 7.73 years (1.7 – 12.9). Conclusions An important proportion of LPD with peripheral involvement can be reclassified to MBL, with a higher prevalence of CLL-like LPD than the global set of patients. The pattern of bone marrow infiltration was predominantly interstitial, followed by a mixed pattern of infiltration. A high percentage of patients in this series evolved to LPD, but a small subset required specific treatment. The patients with LPD progression had a significantly higher B-lymphocyte count than those who did not. Disclosures: No relevant conflicts of interest to declare.

Description: Introduction Epstein-Barr virus (EBV) viral load and serum free light chains (sFLC) are easily measurable and are possible biomarkers for screening and prognosis in HIV-related lymphomas. EBV load and sFLC levels have been shown to be increased at the diagnosis of HIV-related lymphomas but there is a lack of data on their levels at complete response (CR). We aimed to determine the clinical relevance and the potential use of combining EBV load and sFLC measurements as tumor markers in patients with HIV-related lymphomas. Methods Retrospective study of HIV-infected patients diagnosed with non-Hodgkin (NHL) and Hodgkin’s lymphoma (HL) treated in a single institution from 1998 to 2012. EBV loads were determined in plasma by means of a commercial real-time PCR technique (EBV PCR kit, Qiagen GmbH, Hilden, Germany). Levels of κ and λ sFLC were measured in serum using a standardized assay (FREELITE, The Binding Site, Birmingham, UK). sFLC were considered elevated when κ, λ, or the sum of κ and λ were above the upper normal limit. Clinical and biological data were collected from the clinical records. Results Sixty-eight patients were studied (53 NHL and 15 HL). At diagnosis (N=47), levels of κ, λ, and sum of κ and λ sFLC were elevated in 87%, 70%, and 81% of patients, respectively. At CR (N=29), the percentage of patients with elevated κ, λ, and κ+λ was 79%, 45%, and 62%, respectively. EBV load was detectable in 60% of the patients at diagnosis (N=50) and in 7.4% at CR (N=27). Median levels of κ, λ, and κ+λ sFLC at diagnosis were higher than at CR (6.06 mg/dL vs 3.19 mg/dL, P=0.006; 3.94 mg/dL vs 2.21 mg/dL, P=0.021; 11.38 mg/dL vs 5.43 mg/dL, P=0.004; respectively). Median levels of EBV loads at diagnosis were also higher than at CR (121.5 copies/mL vs. 0 copies/mL, P

Description: Histone deacetylase 6 (HDAC6) is a protein modifier that is an increasingly attractive pharmacological target. Interestingly, the observation that the HDAC6 knock-out mouse is not lethal, in contrast to those undergoing complete loss of class I, II and III HDACs, suggests that specific HDAC6 inhibitors may be better tolerated than pan-HDAC inhibitors or drugs that target the other HDAC classes. In this regard, the compound ACY-1215 (Rocilinostat), the described selective HDAC6 inhibitors, is undergoing clinical trials for the treatment of multiple myeloma. Taking into account the previous information about HDAC6 inhibitor structures, the structural differences between HDAC6 and other HDAC isoforms and also the structural information of other developed HDAC inhibitors, we have previously designed and synthesized a new potential HDAC6 selective inhibitor, QTX125 with growth inhibitory effects in mantle cell lymphoma (MCL) cell lines, mouse models and ex vivo treatment of primary samples obtained from patients with MCL. Herein, we have extended these findings to show that the newly identified HDAC6 inhibitor QTX125 is also able to inhibit the growth of preclinical models of other B-cell lymphomas such as follicular lymphoma and Burkitt's cell lymphoma, but also of acute acute myeloid leukemia. In addition beyond a-tubulin, a well known HDAC6 target, we have developed a pharmacological and proteomic screening to identify other proteins modified by HDAC6 that can contribute to the described lymphoma and leukemia phenotypes. Disclosures Eneko: Quimatryx: Employment. Yosu:Quimatryx: Employment. Myriam:Oncomatryx: Employment. Cristina:Oncomatryx: Employment. González-Barca:Roche: Speakers Bureau; Celtrion: Consultancy; Gilead: Consultancy; janssen: Consultancy, Speakers Bureau. Fernando:Quimatryx: Consultancy.

Description: Abstract 3682 Background: Non-Hodgkin Lymphoma (NHL) remains the most common malignancy in patients with HIV infection, but there are few randomized phase II or III trials that provide level 1 evidence regarding the optimal treatment. We previously reported a pooled analysis of 16 trials including 1,144 patients with HIV–associated NHL (ASCO 2012, abstract 8005), and herein report the updated final analysis of 1,546 patients enrolled on 19 trials. Our objective was to assess lymphoma-, HIV-, and treatment-specific factors and their influence on response and survival. Methods: We performed a systematic review to search for prospective clinical phase II or III trials that assessed therapeutic interventions for HIV-associated NHL and assembled individual patient data from 19 trials published between 1993 and 2011 including 1,546 patients (median N=61/trial, range 14–467). Treatment factors included rituximab use (N=542) and type of chemotherapy (CHOP, N=632; EPOCH, N=166; CDE, N=191; dose-intense (intensive) regimens, N=155; ABCVP-based regimens, N=158; low-dose CHOP, N=165; VS, N=41; oral regimens, N=38). Endpoints included complete response (CR), progression-free survival (PFS), and overall survival (OS). We used logistic regression and Cox proportional hazard models adjusted for age, sex, histological subtype, age-adjusted international prognostic index (aaIPI), CD4 count at baseline, prior history of AIDS, type of chemotherapy, use of rituximab, concurrent use of GCSF and combination antiretroviral therapy (cART) with chemotherapy, and enrollment period. Results: The majority of patients were enrolled in the cART era (75%), were male (84%), had DLBLC (69%; BL/BLL 26%; other 6%), had high risk disease (aaIPI≥2; 59%) and a baseline CD4 count ≥50 cells/μl (86%). CHOP was the most commonly prescribed regimen (41%). Among the lymphoma- and HIV-specific covariates evaluated, lower baseline CD4 count and higher aaIPI score were associated with inferior CR rates (CD4: p

Description: Background: In the combination antiretroviral therapy (cART) era the prognosis of HIV-infected patients with Hodgkin’s lymphoma (HL) approaches to that of the HIV-negative population. Treatment of advanced stage HL with ABVD has been shown feasible and effective in HIV patients. However, there is scarce information about the impact of cART on clinical and biological characteristics of HIV-related HL as well as on its prognosis. Aim: To compare the clinical and biological characteristics and outcome of patients with advanced stage HL treated with ABVD according to HIV-status in the cART era. Methods: Two groups of patients with advanced stage HL treated with ABVD were studied. The first group consisted of 63 HIV-infected patients diagnosed with HL in the cART era in 15 Spanish hospitals, and the control group included 31 HIV-negative patients diagnosed in the same period in a single institution. Demographic, clinical and biological data were collected, and complete response rate (CRR), disease-free survival (DFS) and overall survival (OS) were calculated. Then, HIV-infected patients were stratified according to whether they started cART therapy before or concomitantly with chemotherapy. Survival analyses were performed using the Kaplan-Meier method and compared using the log-rank test. The Cox regression proportional hazards model was used for multivariate analyses. Significance level was established at p

Description: Abstract 4442 Introduction: Chronic myeloid leukemia (CML) is a model of disease in the development of targeted therapies. Tyrosine kinase inhibitors (TKIs) have transformed the approach to management of CML and have dramatically improved patients' outcome. Clinical response is obtained in the majority of patients. However, a significant proportion of patients do not achieve the optimal desirable outcome or are completely resistant to this treatment. ABL kinase domain mutations have been extensively implicated in the pathogenesis of TKI resistance. Treatment with second-generation TKIs has produced high rates of hematologic and cytogenetic response in mutated ABL patients. The aim of this study was analyzed the presence of ABL mutations in imatinib resistant patients and determine the importance of changing to second-generation TKIs treatment as soon as failure or suboptimal response is recognized. Patients and methods: From 420 CML patients diagnosed in 6 centers between 2004 and 2010, we have amplified and sequenced the ABL1 domain from BCR-ABL1 amplicon of 45 imatinib resistant patients (23 patients with suboptimal response, 14 with treatment failure, 4 who lost the molecular response and 4 patients who progressed to blast phase). The obtained sequences were compared with the published ABL1 sequence, GenBank U07563, using BLAST 2 software. Results: We have detected mutations in 15 of 45 patients (33%), some of them with more than one mutation (Table 1). Seven of these patients were treated with second-generation TKIs as a single treatment. Three of them achieve a major molecular response (MMR), one patient is in complete cytogenetic response (CCyR) and the other two patients are in major (MCyR) and partial (PCyR) cytogenetic response. Another patient received nilotinib followed by hematological stem cell transplantation (HSCT) and is in MMR. Two patients were submitted to a HSCT and achieve MMR. Only one patient treated with nilotinib as second option has not reach a cytogenetic response one year after detection of the mutation. Two of the patients with the T315I mutation were treated with IFN and nilotinib achieving PCyR and MCyR, respectively, and are still alive. The other T315I patient, and two patients in blast-crisis (BC) disease with the F317L mutation who received dasatinib prior to the study of ABL mutations, died before a change of treatment could have been performed. Conclusions: Disclosures: No relevant conflicts of interest to declare.