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Marta Garcia-Recio,1,2 Jordi Martinez-Serra,1 Francesc Mestre,2,3 Leyre Bento,1,2 Jordi Gines,4 Rafael Ramos,2,5 Jaime Daumal,2,6 Paloma López,2,3 Antonia Sampol,1 Antonio Gutierrez1,2 1Hematology Department, 2Lymphoma Unit, 3Radiotherapy Department, 4Pharmacy Department, 5Pathology Department, 6Nuclear Medicine Department, Son Espases University Hospital, IdISBa, Palma, Spain Abstract: Hodgkin lymphoma (HL) represents ~11% of all lymphoma cases. This disease occurs in young adults, but also affects people over 55 years of age. Despite the fact that >80% of all newly diagnosed patients under 60 will achieve a sustained complete response (CR), 5%–10% of HL patients are refractory to initial treatment and 10%–30% of patients will eventually relapse after an initial CR. The treatment recommendation for primary refractory or relapsed HL patients is salvage therapy followed by high-dose chemotherapy and autologous stem cell transplantation. Following this approach, a significant part will still relapse at any moment. Thus, further research and new drugs or combinations are required. Overexpression of COX-2 has been associated with poor prognosis in relapse/refractory HL patients, so it could be a potential therapeutic target in HL. For this purpose, several drugs may have a role: specific COX-2 inhibitors such as celecoxib or other anti-inflammatory drugs such as lenalidomide may further inhibit lipopolysaccharide-mediated induction of COX-2. Moreover, lenalidomide and COX-2 inhibitors (celecoxib) have been tested in solid tumors with encouraging results. We present a case of a young female diagnosed with a heavily pretreated HL nodular sclerosis subtype who, after failing six treatment lines, only achieved clinical and radiological CR after six cycles of lenalidomide/celecoxib that resulted in an event-free survival of 22 months. We explain the rationale of using this chemotherapy regimen and our patient follow-up. Keywords: Hodgkin lymphoma, relapse, celecoxib, lenalidomide, COX-2

The International Prognostic Index (IPI) is the most widely used score for non-Hodgkin lymphoma but lacks the ability to identify a high-risk population in diffuse large B-cell lymphoma (DLBCL). Low absolute lymphocyte count and high monocytes have proved to be unfavourable factors. Red-cell distribution width (RDW) has been associated with inflammation and beta-2 microglobulin (B2M) with tumour load. The retrospective study included 992 patients with DLBCL treated with R-CHOP. In the multivariate analysis, age, Eastern Cooperative Oncology Group performance status (ECOG-PS), stage, bulky mass, B2M, RDW, and lymphocyte/monocyte ratio (LMR) were independently related to progression-free survival (PFS). A new prognosis score was generated with these variables including age categorized into three groups (0, 1, 2 points); ECOG ≥ 3-4 with two; stage III/IV, bulky mass, high B2M, LMR 2·25 and RDW 0·96 with one each; for a maximum of 9. This score could improve the discrimination of a very high-risk subgroup with five-year PFS and overall survival (OS) of 19% and 24% versus 45% and 59% of R (revised)-IPI respectively. This score also showed greater predictive ability than IPI. A new score is presented including complete blood cell count variables and B2M, which are readily available in real-life practice without additional tests. Compared to R-IPI, it shows a more precise high-risk assessment and risk discrimination for both PFS and OS.

Introduction: Micro-RNAs (miRNAs), have been shown to be one of the main regulators of gene expression and other biological processes. Recently, attention is focused on their potential role as prognostic predictors in different types of cancer, including diffuse large B cell lymphoma (DLBCL). We previously reported several miRNAs associated to progression/refractoriness in DLBCL (miR-1244, miR-193b-5p and miR-1231). The aim of this study was to validate these resultsin vitro, exploring the molecular pathways involved. Material and methods: The DLBCL cell line, U-2932 (activated B-cell (ABC) subtype) (ACC 633) was obtained from DSMZ (Germany). Transient reverse transfection of DLBCL cells for 48 or 72 h with mirVana miRNA mimics or inhibitors (miR-1244, miR-193b-5p and miR-1231) or with a miRNA negative control (100 nM) was performed using Viromer®GREEN reagent (Lipocalyx). In some experiments, after 24 h of transfection cells were treated with vehicle (1% DMSO) or with increasing concentrations of CHOP (cyclophosphamide, adriamycin, vincristine sulfate, and prednisone) (Sigma-Aldrich) for 48 h. The ratio of the four drugs was 80 mg/ 5.33 mg/ 0.16 mg/ 5.77 mg. After 48 h of transfection or treatment, cell viability was measured using the CellTiter 96®AQueous One Solution and total RNA was isolated using the RNeasy Mini Kit from Qiagen. Gene expression profiling (GEP) was performed using GeneChip human Clarion S of Affymetrix-ThermoFisher. Results: Reverse transfection of U-2932 with the miRNA mimic negative control (100 nM) for 48 h did not alter the cell viability compared to non-transfected (NT) cells, showing that the transfection method did not affect viability in these DLBCL cells. Transfection of mimic miR-1244, miR-193b-5p or miR-1231 (100 nM) did not significantly alter the viability compared to cells transfected with the miRNA mimic negative control. However, inhibition of these endogenous miRNA molecules for 48 h significantly decreased by 24.1 ± 5.7%, 28.9 ± 6.5% and 30.9 ± 3.3% cell viability, respectively, suggesting that these miRNAs could be involved in the survival of these tumor cells. In these experiments, the percentage of cell transfection was 89.6%. The effect of miRNA 1244, 193b-5p or 1231 on the response of DLBCL cells to CHOP treatment was also evaluated. Transfection of U-2932 cells with miRNA mimic negative control did not alter its sensibility to CHOP, which dose-dependently decreased its viability with an IC50 of 1.45 μg/ml in NT or transfected cells. Transfection of U-2932 cells with mimic miR-1244 or miR-193b-5p (100 nM) increased cell viability in the presence of vehicle (1% DMSO) and also reverted the inhibitory effect of CHOP (0.3 μg/ml) after 48 h of treatment. However, transfection of U-2932 cells with mimic miR-1231 (100 nM) did not alter the inhibitory effect of CHOP (0.3, 1 and 3 μg/ml) on cell viability at any of the concentrations studied. GEP studies showed that these miRNAs reduced the expression of genes associated to chemosensitivity (MCTP1) in the case of miR-1244 and miR-193-5p or tumoral suppressor genes (NRN1) in the case of miR-1231. Conclusions: We validatedin vitroour previously reported miRNAs about the role of miR-1244, miR-193b-5p and miR-1231 associated to treatment failure. The inhibition of endogenous miR-1244, miR-193b-5p or miR-1231 molecules significantly decreased cell viability. Transfection of U-2932 cells with mimic miR-1244 or miR-193b-5p (100 nM) increased cell viability of cells treated with CHOP. GEP studies showed that these miRNAs were linked to chemoresistance and inhibition of tumoral suppressor genes. Future works will have to translate these results to clinical practice in DLBCL. Disclosures Salar: Celgene:Speakers Bureau;Janssen:Speakers Bureau;Roche:Speakers Bureau.

Introduction: Micro-RNAs (miRNAs) are 19-24 nucleotide non-coding RNAs that regulate gene expression through the inactivation of their messenger RNA. Previous studies have demonstrated the important role of some miRNAs in the development of cancer. Specifically, in diffuse large B cell lymphoma (DLBCL), miRNAs involved in lymphomagenesis were identified but understanding their biological function continues to be a challenge. Nevertheless, the pathogenesis effects of some miRNAs such as miR-125a, miR-17-92 cluster or mi-R-155 are well characterized. Some studies suggest that miRNAs also possess a prognostic potential role in DLBCL. For this reason, our objective was to analyze the different miRNAs involved in the chemo-sensitivity or resistance to the first line treatment, their correlation with standard prognostic factors at diagnosis and the role of these miRNAs in survival in patients with DLBCL. Material and methods: Patients homogeneously treated with R-CHOP from 1999-2013 were reviewed from 3 Spanish centers. They were retrospectively obtained from Pathology Department registry to avoid selection bias. We included those patients with valid genomic material in formalin-fixed-paraffin-embedded tissue and with available clinical data. Samples were processed using the miRNA 4 Affymetrix microarrays kit in a discovery group that included 2 cohorts (patients with durable complete remission (CR) versus refractory or early relapsing patients). Those miRNAs with differential expression were validated in the whole series with quantitative RT-PCR. We also analyzed the role of these miRNAs as predictors of event-free survival (EFS) and overall survival (OS) and their relationship with standard prognostic factors in DLBCL. Results: We identified 156 patients homogeneously treated with R-CHOP. Finally, 96 samples were obtained with valid material for RNA extraction. To identify those miRNAs with prognostic implication, a discovery cohort of 12 patients was used in which all the cases had poor prognosis with high tumor load and advance disease (III-IV stage and unfavorable R-IPI). On this basis of poor prognosis, 2 groups were defined totally opposed from the point of view of the treatment response and evolution: chemo-resistance group (n=6) including refractory or relapsed (RR) patients (first 12 months) and chemo-sensitive group (n=6) including patients with at least 3 years of CR. A hierarchical clustering was performed in which 26 miRNAs differentially expressed were identified. A screening of miRNAs was carried out based on the fold-change and pathways involved and finally we obtained 10 miRNAs differentially expressed in RR group. The validation of these miRNAs was performed with quantitative RT-PCR in the whole series which finally included 68 samples with valid material. A univariate survival analysis including clinical prognostic factors and the selected 10 miRNAs was performed. We confirmed that 7 of them (miR-20b-5p, miR-1244, miR-6840-3p, miR-1231, miR-193b-5p, miR-6860-5p y miR-199a-5p) significantly influenced EFS and 6 of them (miR-1244, miR-1231, miR-193b-5p, miR-885-3p, miR-182-5p y miR-199a-5p) on OS. From these 10 miRNAs, only 3 had a significant prognosis role not only for EFS but also for OS and progression-free survival (PFS): miR-1244, miR-193b-5p and miR-1231. The overexpression of miR-1244 and miR-193-5p were associated with advance stage and worse clinical prognosis factors (p15% were independently associated with worse SG and EFS (Figure 1). In previously reported studies, these 3 miRNAs have been related with proliferative events such as the overexpression of myc or anti-apoptosis. Also, the miR-1231 may be related with new lymphomagenesis pathways associated to viral infections. Conclusions: Through a discovery group focused on progression/refractoriness, a group of new miRNAs differentially expressed on chemo-resistant patients with DLBCL was identified. The overexpression of miR-1244 and miR-193-5p was associated with more extensive disease and worse clinical prognostic factors. The high R-IPI, reduction of >15% RDI and the overexpression of miR-1231 were independently associated with worse EFS and OS. Disclosures Sánchez-González: Amgen: Consultancy, Speakers Bureau; Gilead: Speakers Bureau; Navartis: Consultancy, Speakers Bureau; Shire: Speakers Bureau; Takeda: Consultancy, Speakers Bureau. Salar:Roche: Research Funding, Speakers Bureau; Janssen Pharmaceuticals: Consultancy, Speakers Bureau; Gilead: Consultancy, Speakers Bureau; Celgene: Consultancy.

Introduction DLBCL is the most common non-Hodgkin lymphoma and a heterogeneous subtype from the biological and clinical point of view. 20-40% of the patients relapse so it is important to identify a high risk population that could benefit from different therapeutic approaches. For this purpose, several scores have been developed. The IPI has been the most widely used but lacks the ability to identify this very high risk population in Rituximab era. Low absolute lymphocyte count and high levels of blood monocytes have shown to be unfavorable risk factors. The red cell distribution width (RDW) has been associated with aging and active inflammatory processes and beta-2-microglobulin (B2M) with tumor load and proliferation as well as comorbidities such as kidney failure. All these variables are easily obtainable at the time of diagnosis. Patients and methods From the national database of GELTAMO-IPI Project, we selected those patients with DLBCL homogeneously treated with R-CHOP ± radiotherapy. The complete blood count (CBC) values were standardized using the normal reference values of each centre. The survival analysis was estimated with Kaplan-Meier method. The comparison between variables was performed through Log-Rank test and multivariate analysis with Cox regression. The CBC quantitative variables cut points were calculated through MAXSTAT. A new prognosis score was generated taking into account the results of multivariate analysis for progression free survival (PFS), spliting the sample in two cohorts (discovery and validation). Results Nine hundred and ninety-two patients with DLBCL were retrospectively analyzed with a median follow up of 55 months (12-185). The characteristic of the patients are summarized in table 1. In the multivariate analysis, age, ECOG, stage, bulky mass, B2M, RDW and lymphocytes/monocytes ratio (LMR) were significantly independent variables for PFS. A new prognosis score was generated with these variables including age categorized in 3 groups (18-64, 65-80 y >80) with 0, 1 y 2 points, ECOG>3-4 with 2 points, stage III-IV, bulky mass, high B2M, LMR0.96 with 1 point each for a maximum of 9. This score could improve the discrimination of a very high risk subgroup with a 5-year PFS of 17% (Figure 1) versus 45%, 52%, 32% or 29% of R-IPI, NCCN-IPI, R-TS and GELTAMO-IPI, respectively and 5-year overall survival (OS) of 20% (Figure 2) versus 59%, 46%, 45% and 40% for R-IPI, NCCN-IPI, R-TS and GELTAMO-IPI. Conclusion A new prognosis score including easily obtainable CBC variables (LMR and RDW) and B2M is presented. This score identifies a high risk population both for PFS and OS in comparison with other scores for DLBCL. The addition of other biological or image markers could improve these results. Disclosures Martín: Roche: Consultancy, Honoraria, Other: Travel expenses; Janssen: Honoraria, Other: Travel expenses; Celgene: Consultancy, Honoraria, Other: Travel expenses; Servier: Honoraria, Other: Travel expenses. Sancho:SERVIER: Honoraria; JANSSEN: Honoraria, Speakers Bureau; MUNDIPHARMA: Honoraria; SANOFI: Honoraria; GILEAD: Honoraria, Research Funding; KERN FHARMA: Honoraria, Speakers Bureau; CELGENE: Honoraria; ROCHE: Honoraria, Speakers Bureau.

Introduction: Six percent of all non-Hodgkin lymphomas (NHL) cases are mantle cell lymphoma (MCL). MCL has a poor prognosis and is considered incurable with current strategies. Intensive approaches are not applicable to most these patients considering the median age at diagnosis (70 years old). Due to the molecular events that drive MCL to be a lymphoma with an aggressive and indolent behavior as well as a tendency to clonal evolution (dysfunction of cell cycle, response to cellular damage and apoptosis), we present a global therapeutic approach which includes an effective induction with a low toxic profile, as well as consolidation and maintenance with the final aim of increasing and maintaining responses. Methods: From December-2008 to January-2018 all MCL patients treated in first line in our center were included. The therapeutic approach was based on an induction with R-GemOx (rituximab, gemcitabine and oxaliplatin) followed by consolidation with autologous stem cell transplant or ibritumomab tiuxetan followed by rituximab maintenance (Rm) (every 2 months for 2-3 years). Since 2016, all patients with less than complete response after induction/consolidation, received Ibrutinib 560 mg daily added to Rm. Standard prognostic variables were collected at diagnosis including MIPI; the response evaluation and follow-up was made considering Cheson criteria; for toxicity assessment, we used OMS grading scales of toxicity criteria. Overall survival (OS) and progression free survival (PFS) were estimated from the beginning of the treatment with the Kaplan-Meier method. Results: Thirteen treatment-naïve MCL patients were included from December-2008 to January-2018. Main characteristics of patients are shown in Table 1. Briefly, this is an old high risk series with a median age at diagnosis of 71 years old and poor prognosis (31% and 69% belong to intermediate and high risk MIPI groups, respectively). Two cases were refractory to R-GemOx induction (a blastic MCL and another case with high MIPI (7.7)). Four cases relapsed between 30 and 66 months after treatment was started. All cases with CR or PR after induction (n=11; 85%), obtained a CR at the end of the maintenance. One of the refractory cases after a second line followed by rituximab and ibrutinib maintenance also reached CR. Only 2 patients died (15%): one disease progression and another non-related pulmonary thromboembolism. The median follow-up was 51 months. Four-years OS was of 83% with a median PFS of 58 months (Figure 1). R-GemOx toxicity profile was manageable: most of them grade 1-2 neutropenia (43%), vomits (100%), diarrhea (30%) and anemia (30%). Grade 3-4 toxicity was scarce (neutropenia 14% and thrombocytopenia 36%). Conclusions: R-GemOx followed by consolidation and Rm should was an effective approach with manageable toxicity and excellent survival considering the median age and the high MIPI risk of the series. We plan to test this global approach in a prospective clinical trial. Disclosures No relevant conflicts of interest to declare.

Introduction : Follicular lymphoma (FL) is an indolent non-Hodgkin lymphoma that represents 15-30% of all newly diagnosed lymphomas. It is characterized by a high rate of response followed by continuous relapses being considered incurable with standard immunochemotherapy regimens (Ardeshna et al. Lancet. 2003). The combination of immunochemotherapy and rituximab maintenance (Rm) improves progression free survival (PFS) when compared with a cohort without Rm (51% vs 35%) (Salles G. et al. ASH, 2017), being the standard for many centers. This improvement may have changed the risk assessment in FL. However, there is a need for identifying those patients with a poor prognosis with standard Rm-based therapy and candidates to receive alternative approaches using new drugs such as obinutuzumab with better reported results in this frontline setting (Gallium trial) (Marcus et al., 2017). Methods : From February-2002 to October-2016, all newly diagnosed FL patients with the intention to treat with immunochemotherapy and Rm where included. The participant centers were Son Espases University Hospital and Hospital del Mar of Barcelona. Clinical data were retrospectively collected from clinical histories and standard prognostic variables were evaluated at diagnosis including FLIPI and FLIPI-2. Another prognostic markers were defined: POD12 (progression of disease at 12 months), POD24 (progression of disease at 24 months) and POD30 (progression of disease at 30 months). The regimen effectiveness was evaluated based on the CT or PET/CT findings. Overall survival (OS) and PFS were determined from the date of the first cycle and they were estimated through Kaplan-Meier. Results: A total of 96 patients were included in the study with a median follow-up of 68 months. Main characteristics of the sample are shown in Tables 1. Briefly, median age was 61 years old, 88% where diagnosed in an advanced stage (III-IV), 36% and 38% presented with high risk (3-5) FLIPI and FLIPI2. Five-year OS and PFS was of 94% and 78% respectively. Univariate survival analysis proved that ECOG and response rate affected PFS and that stage and response influenced OS. In this analysis, no significant relation was found between the studied variables and POD12, POD24 or POD30 except beta2-microglobuline in POD30. Conclusions: Frontline Rm significantly improves outcomes of FL patients. It seems that it is not possible to predict only with clinical variables which patients are in high risk of failure to a standard approach with immunochemotherapy followed by Rm. Additional prognostic factors such as molecular markers may help to identify these patients at risk of failure at the beginning of a first line of immunochemotherapy followed by Rm, and candidates for alternative approaches. Disclosures No relevant conflicts of interest to declare.

INTRODUCTION The standard first line treatment for patients with diffuse large B-cell lymphoma (DLBCL) is rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) independently of cell origin subtype. Retrospective studies have shown that non-germinal center (NGC) subtype is associated with a worse progression free survival (PFS) and overall survival (OS) compared to germinal center subtype. In phase II studies the combination of lenalidomide (lena) or bortezomib (bor) with R-CHOP in this lymphoma subtype has shown better results in comparison to R-CHOP alone. For this reason, the aim of our study is to analyze the efficacy and toxicity of R-CHOP plus lena/bor in comparison with R-CHOP alone in patients with NGC DLBCL. PATIENTS AND METHODS In order to avoid selection bias, all patients with NGC DLBCL (confirmed by immunohistochemistry with Hans algorithm) who received R-CHOP or R-CHOP-lena/bor (25mg lenalidomide daily from day 1 to 10 / bortezomib 1.3mg/m2 days 1 and 4) were retrospectively selected from the Pharmacy Registry. Patients in our center received R-CHOP from March 2002 to June 2013, R-CHOP-bor from June 2013 to July 2015 and R-CHOP-lena from November 2014 until now. Cheson and Lugano criteria were used to confirm response rates. Toxicity was checked with CTCAE v4-5 criteria. RESULTS Fifty-five patients with NGC DLBCL were identified from 2002 to 2018, 18 of whom were treated with R-CHOP-lena/bor and 33 with R-CHOP. Baseline characteristics are specified in Table 1. Overall response rate was similar in both treatment regimens, 94% versus 87%, respectively. 12.5% progression/stable disease cases were identified with R-CHOP alone compared to 5.3% with R-CHOP-lena/bor. With a median follow-up of 45 months (6-126) for R-CHOP and 22 months (6-59) for R-CHOP-lena/bor, the median PFS was 65 months for R-CHOP and was not reached for R-CHOP-lena/bor. 47% versus 9% of progressions were observed in R-CHOP and R-CHOP-lena/bor, respectively (p=0.006). Regarding OS, the death rate in the R-CHOP group was 45% in comparison with 23% in R-CHOP-lena/bor group (p=0.015). Higher hematological toxicity was observed in the R-CHOP-lena/bor group comparing with R-CHOP: neutropenia (95% versus 68%), grade 3-4 neutropenia (81% versus 37%) and anemia (81% vs 56%) (p CONCLUSIONS The addition of lenalidomide or bortezomib to R-CHOP seems to be a good option in NGC DLBCL, with a lower progression rate, refractoriness and less mortality in comparison with R-CHOP alone. R-CHOP-lena/bor is associated to higher rates of hematological toxicity but manageable in all cases. These retrospective data should be confirmed in ongoing phase III clinical trials. Disclosures No relevant conflicts of interest to declare.

15 Páginas; 6 Figuras, Several important biological activities have been attributed to the pentacyclic triterpene ursolic acid (UA), being its antitumoral effect extensively studied in human adenocarcinomas. In this work, we focused on the efficacy and molecular mechanisms involved in the antitumoral effects of UA, as single agent or combined with doxorubicin (DXR), in human soft tissue sarcoma cells. UA (5–50 μM) strongly inhibited (up to 80%) the viability of STS cells at 24 h and its proliferation in soft agar, with higher concentrations increasing apoptotic death up to 30%. UA treatment (6–9 h) strongly blocked the survival AKT/GSK3β/β-catenin signalling pathway, which led to a concomitant reduction of the anti-apoptotic proteins c-Myc and p21, altogether resulting in the activation of intrinsic apoptosis. Interestingly, UA at low concentrations (10–15 μM) enhanced the antitumoral effects of DXR by up to 2-fold, while in parallel inhibiting DXR-induced AKT activation and p21 expression, two proteins implicated in antitumoral drug resistance and cell survival. In conclusion, UA is able to induce intrinsic apoptosis in human STS cells and also to sensitize these cells to DXR by blocking the AKT signalling pathway. Therefore, UA may have beneficial effects, if used as nutraceutical adjuvant during standard chemotherapy treatment of STS., Research was supported by the Institute of Health Carlos III through the subprogramme “CIBERobn (Centro de Investigación Biomédica en Red de la Fisiopatología de la Obesidad y Nutrición)”. VHV was supported by a predoctoral fellowship from ‘Govern de les Illes Balears, Conselleria d’Educació, Cultura i Universitats’ under a program of joint financing with the European Social Fund.

Marta Garcia-Recio,1,2 Jordi Martinez-Serra,1,2 Leyre Bento,1,2 Rafael Ramos,2,3 Jordi Gines,4 Jaime Daumal,5 Antonia Sampol,1,2 Antonio Gutierrez1,2 1Service of Hematology, 2Instituto de Investigación Sanitaria de Palma (IdISPa), 3Service of Pathology, 4Pharmacy Department, 5Service of Nuclear Medicine, Son Espases University Hospital, Palma de Mallorca, Spain Abstract: Blastic plasmocytoid dendritic cell neoplasm is characterized by aggressive behavior with a tendency for systemic dissemination and a predilection for skin, lymph nodes, soft tissues, peripheral blood, or bone marrow. It usually occurs in elderly patients with a mean age between 60 and 70 years. Despite initial response to chemotherapy, the disease regularly relapses with a short median overall survival. Better outcomes have been reported with high-dose acute leukemia-like induction chemotherapy followed by consolidation with allogeneic hematopoietic stem cell transplantation. However, elderly patients are not candidates for intensive therapy or allogeneic stem cell transplantation. So, new active and tolerable drugs are needed. Our case illustrates that one cycle of lenalidomide and celecoxib provides at least a partial cutaneous and hematologic response, but this regimen was discontinued due to toxicity and followed by a consolidation/maintenance phase with azacitidine, thus achieving a final complete response with a much higher than expected progression-free and overall survival in an elderly patient with comorbidities. This information may be useful in the design of treatment approaches for elderly patients with blastic plasmocytoid dendritic cell neoplasm. However, it should be confirmed in clinical trials as well as by optimizing the induction and extending the consolidation/maintenance period to avoid early relapses after discontinuation and improve progression-free survival. Keywords: blastic plasmocytoid dendritic cell neoplasm, elderly, immunomodulatory, COX-2, hypomethylating

Purpose Doxorubicin and trabectedin are considered active drugs in soft tissue sarcoma (STS). The combination of both drugs was hypothesized to be advantageous and safe on the basis of preclinical evidence and a previous phase I trial, respectively. The aim of this study was to compare the clinical outcome of trabectedin plus doxorubicin with doxorubicin as first-line treatment of advanced STS patients. Patients and Methods In this open-label randomized phase II trial, the main end point was progression-free survival (PFS). Trabectedin 1.1 mg/m2 in a 3-hour infusion plus doxorubicin 60 mg/m2 as the experimental arm and doxorubicin 75 mg/m2 as the control arm were administered for up to six cycles. Translational research was planned to correlate the expression of apoptotic and DNA repair genes with clinical outcome. Results In 115 randomly assigned patients, the median PFS was 5.5 months in the control arm and 5.7 months in the experimental arm (hazard ratio, 1.16; 95% CI, 0.79 to 1.71; P = .45) in the intent-to-treat analysis. The trial was stopped for futility after the interim analysis, because the results in the experimental arm showed the risk reduction for the main end point to be < 9.64%. The proportion of patients with grade 3 or 4 thrombocytopenia, asthenia, and liver toxicity was significantly higher in the experimental arm. FAS and p53 were shown to be prognostic factors for PFS (7.0 months if FAS+ and p53−; 3.4 months if FAS+/p53+ or FAS−/p53−; and 0.7 months if FAS− and p53+; P < .001) and for overall survival. Conclusion Trabectedin plus doxorubicin did not show superiority over doxorubicin alone as first-line treatment of advanced STS. The prognostic role of apoptotic key genes, FAS and p53, was shown to be robust enough to continue this research line.

Objectives Within the laboratory protocols, used for the study of BCR-ABL resistance mutations in chronic myeloid leukemia patients treated with Imatinib, direct sequencing remains the reference method. Since the incidence of patients with a mutation-related loss of response is not very high, it is very useful in the routine laboratory to perform a fast pre-screening method. Design and methods With this in mind, we have designed a new technique, based on a single Real-Time FRET-based PCR, followed by a study of melting peaks. This new tool, developed in a LightCycler 2.0, combines four different fluorescence channels for the simultaneous detection, in a single close tube, of critical mutations within the ABL kinase domain. Results Assay evaluation performed on 33 samples, previously genotyped by sequentiation, resulted in full concordance of results. Conclusions This new methodology detects in a few steps the presence of critical mutations associated to Imatinib resistance.

Trabectedin, a naturally occurring substance isolated from the Caribbean marine invertebrate Ecteinascidia turbinata, is the active compound of the antitumor drug Yondelis®. The mechanism of action of Trabectedin has been attributed to interactions with the minor groove of the DNA double helix, thereby affecting transcription of different genes involved in DNA repair and thus facilitating lethal DNA strand breaks. Nevertheless, the existence of other clinically important molecular mechanisms has not yet been fully explored. In this paper we demonstrate how Yondelis®, apart from activating the caspase-8-dependent cascade of apoptosis, sensitizes cancer cells to Fas-mediated cell death at achievable concentrations similar to those found in the plasma of patients. In addition we show that the facilitated apoptosis activated through the Fas death receptor, is associated with a significant increase of membrane Fas/FasL, as well as the modulation of accessory proteins regulating this route, such as FLIP (L) or Akt. Thus, our results propose that the sensitization of the death receptor pathway is an essential mechanism amplifying the cytotoxic properties of Yondelis® that could explain the hepatotoxicity observed in patients treated with this drug. Finally, we also show how the use of dexamethasone as a prophylactic agent that protects against hepatotoxicity induced by Yondelis® may also inhibit some of the cytotoxic properties described in this work. The study of this important mechanism of action should set up the basis for reassessing clinical therapy with Yondelis® in order to improve antitumor treatment outcome.

Summary CA125, a tumor marker normally used to follow the clinical course of ovarian cancer, also may have a role in lymphoma. All available series were analyzed using the standard reference value 35 U/ml, but age and sex may influence serum CA125 (sCA125) levels. We aim to study the prognostic value of serum CA125 (sCA125) levels in diffuse large B-cell lymphoma (DLBCL), considering the influence of age and sex on sCA125 levels. We investigated the relationship between sCA125 and clinical outcome after treatment in 42 patients with DLBCL, comparing both the standard (35 U/ml) and a new age and sex adjusted (sex/age-adjusted) reference value proposed by our group. We found that patients with elevated sCA125 levels had significantly more adverse prognostic factors at diagnosis, lower CR rates, higher relapse rates and worse survival. In the low-risk IPI categories, the presence of elevated sCA125 defined a particularly high-risk subgroup with poorer 3-year PFS when compared with patients with normal sCA125 levels. The use of a sex/age-adjusted reference value for sCA125 may increase the sensitivity to identify those patients with elevated sCA125 levels truly related to DLBCL activity.

Several new Cu–hippurate derivative–phenanthroline ternary complexes have been prepared. The X-ray structure of one of them, [Cu(hip)(phen)2]+·(hip−) (2) (where hip is hippurate and phen is 1,10-phenanthroline) has been solved. The structure of this new compound shows important differences (3D-pattern) to other similar related complexes (2D-pattern). A study of the biological activity of [Cu(hip)(phen)2]+·(hip−)·2H2O (2), [Cu(BGG)(phen)2]+·(BGG−)·6H2O (3), [Cu(BIGG)2(phen)](H2O) (4) and [Cu(I-hip)(bpy)2]+·(I-hip−)·3.5H2O (5) (where I-hip is ortho-iodohippurate, BGG corresponds to benzoylglycilglycine, and BIGG is ortho-iodobenzoylglycilglycine) is included and compared with the anti-proliferative activity of [Cu(I-hip)(phen)2]+·(I-hip−)·7H2O (1) previously described, resulting in a greater cytotoxic activity of the compounds with 1,10-phenanthroline instead of those with 2,2′-bipyridyl, in the same way that removing iodine substitution or lengthening the peptidic chain diminishes the activity of compounds compared with 1. The presence of an ortho-iodine group and the direct bond between Ar–CO and glycine moieties yield to the best results.

DLBCL is an aggressive lymphoma treated with R-CHOP. Recently, attempts have been made to improve the outcome by increasing both dose-density and intensity but there have been no benefits in terms of survival. When treating malignancies RDI is important to consider but there is little published information on DLBCL. The purpose of this study was to analyze the differential prognostic impact of RDI in two cohorts of DLBCL patients treated with R-CHOP21 or R-CHOP14. From January 2001 to August 2013 we included DLBCL patients homogenously treated with R-CHOP21 or R-CHOP14, with or without radiotherapy, at University Hospital Son Espases, Hospital Son Llatzer of Palma and Hospital del Mar of Barcelona (N = 157). In order to avoid selection bias the patients were retrospectively identified from the Pathology Department and Pharmacy registries. Median follow-up was 68 months. There was no difference in the response or survival between the two cohorts. In the R-CHOP21 group, both a reduction higher than 15% in RDI (RR 7.41) and R-IPI (RR 2.99) were independently associated with OS. However, a reduction higher than 15% in RDI (RR 4.41) was only noted for PFS. In the R-CHOP14 group, NCCN-IPI (RR 7.09) and B-symptoms (RR 5.37) for OS; AA stage III-IV (RR 6.26) and bulky disease (RR 4.05) for PFS. There was a trend towards a higher rate of RDI reduction observed in the R-CHOP14 group but it only made an impact in the R-CHOP21 group. We conclude that R-CHOP21 and R-CHOP14 are equivalent regimens in terms of response and survival, but only if RDI reductions are avoided. For patients receiving R-CHOP21 we recommend using clinical and support measures in order to avoid RDI reductions.

Jordi Martinez-Serra,1 Juan Robles,2 Antoni Nicolàs,3 Antonio Gutierrez,1 Teresa Ros,1 Juan Carlos Amat,1 Regina Alemany,4 Oliver Vögler,4 Aina Abelló,2 Aina Noguera,2 Joan Besalduch1 1Department of Hematology, 2Department of Clinical Analysis, Hospital Universitary Son Espases, Palma de Mallorca, Spain; 3ECOGEN, Barcelona, 4Department of Cell Biology, University of the Balearic Islands, Palma de Mallorca, Spain Abstract: Blood samples are extensively used for the molecular diagnosis of many hematological diseases. The daily practice in a clinical laboratory of molecular diagnosis in hematology involves using a variety of techniques, based on the amplification of nucleic acids. Current methods for polymerase chain reaction (PCR) use purified genomic DNA, mostly isolated from total peripheral blood cells or white blood cells (WBC). In this paper we describe a real-time fluorescence resonance energy transfer-based method for genotyping directly from blood cells. Our strategy is based on an initial isolation of the WBCs, allowing the removal of PCR inhibitors, such as the heme group, present in the erythrocytes. Once the erythrocytes have been lysed, in the LightCycler® 2.0 Instrument, we perform a real-time PCR followed by a melting curve analysis for different genes (Factors 2, 5, 12, MTHFR, and HFE). After testing 34 samples comparing the real-time crossing point (CP) values between WBC (5×106 WBC/mL) and purified DNA (20 ng/µL), the results for F5 Leiden were as follows: CP mean value for WBC was 29.26±0.566 versus purified DNA 24.79±0.56. Thus, when PCR was performed from WBC (5×106 WBC/mL) instead of DNA (20 ng/µL), we observed a delay of about 4 cycles. These small differences in CP values were similar for all genes tested and did not significantly affect the subsequent analysis by melting curves. In both cases the fluorescence values were high enough, allowing a robust genotyping of all these genes without a previous DNA purification/extraction. Keywords: real-time PCR, LightCycler® 2.0 Instrument, melting peak, FRET, white blood cells, lysis, erythrocytes

Purpose To analyze the role of radiation therapy (RT) on the adverse prognostic influence of cyclooxygenase-2 (COX-2) expression on Reed-Sternberg (RS) cells, in the setting of early Hodgkin lymphoma (HL) treated with ABVD (adriamycin, vinblastine, bleomycin, dacarbazine). Methods and Materials In the present study we retrospectively investigated the prognostic value of COX-2 expression in a large (n=143), uniformly treated early HL population from the Spanish Network of HL using tissue microarrays. Univariate and multivariate analyses were done, including the most recognized clinical variables and the potential role of administration of adjuvant RT. Results Median age was 31 years; the expression of COX-2 defined a subgroup with significantly worse prognosis. Considering COX-2 + patients, those who received RT had significantly better 5-year progression-free survival (PFS) (80% vs 54% if no RT; P =.008). In contrast, COX-2 − patients only had a modest, nonsignificant benefit from RT in terms of 5-year PFS (90% vs 79%; P =.13). When we compared the outcome of patients receiving RT considering the expression of COX-2 on RS cells, we found a nonsignificant 10% difference in terms of PFS between COX-2 + and COX-2 − patients ( P =.09), whereas the difference between the 2 groups was important (25%) in patients not receiving RT ( P =.04). Conclusions Cyclooxygenase-2 RS cell expression is an adverse independent prognostic factor in early HL. Radiation therapy overcomes the worse prognosis associated with COX-2 expression on RS cells, acting in a chemotherapy-independent way. Cyclooxygenase-2 RS cell expression may be useful for determining patient candidates with early HL to receive consolidation with RT.

The xCELLigence system is a new technological approach that allows the real-time cell analysis of adherent tumor cells. To date, xCELLigence has not been able to monitor the growth or cytotoxicity of nonadherent cells derived from hematological malignancies. The basis of its technology relies on the use of culture plates with gold microelectrodes located in their base. We have adapted the methodology described by others to xCELLigence, based on the pre-coating of the cell culture surface with specific substrates, some of which are known to facilitate cell adhesion in the extracellular matrix. Pre-coating of the culture plates with fibronectin, compared to laminin, collagen, or gelatin, significantly induced the adhesion of most of the leukemia/lymphoma cells assayed (Jurkat, L1236, KMH2, and K562). With a fibronectin substrate, nonadherent cells deposited in a monolayer configuration, and consequently, the cell growth and viability were robustly monitored. We further demonstrate the feasibility of xCELLigence for the real-time monitoring of the cytotoxic properties of several antineoplastic agents. In order to validate this technology, the data obtained through real-time cell analysis was compared with that obtained from using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. This provides an excellent label-free tool for the screening of drug efficacy in nonadherent cells and discriminates optimal time points for further molecular analysis of cellular events associated with treatments, reducing both time and costs., The authors thank A Font and J Ortiz de Zarate (Roche Diagnostics, Basel, Switzerland) for their valuable technical support. This work was supported by a grant from Mundipharma (Cambridge, UK).

Chronic myelogenous leukemia (CML) results from the neoplastic transformation of a hematopoietic stem cell. CML is cytogenetically characterized by the presence of the Philadelphia chromosome (Ph’). Most patients with CML express e13a2 or e14a2 mRNAs that result from a rearrangement of the major breakpoint cluster regions (M-BCR) generating the 210-kDa (p210BCR-ABL) fusion proteins b2a2 or b3a2 respectively. The e1a3 CML-related atypical translocation has been reported with an indolent clinical course, low leukocyte count, long chronic phase even without treatment and good response to therapy. We report the case of a patient initially diagnosed as CML in chronic phase whose cells expressed the e1a3 variant. The patient readily responded to imatinib 400 mg with the achievement of a rapid complete cytogenetic response and the normalization of the blood count values, but after 5 months transformed into lymphoid blast crisis.

Red cell distribution width (RDW) is an indicator of the variability in the size of circulating erythrocytes (anisocytosis); different conditions can increase the RDW levels; such as hemolysis, ineffective erythropoiesis and blood transfusions. Recently, different studies have shown an association between increased levels of RDW and inflammation in different diseases, being proposed as a surrogate marker of inflammation and a strong predictor of adverse outcome. The proposed mechanism of this association departs from the finding that Inflammatory cytokines like TNFand IL-6 (part of the classic inflammatory cascade), have been found to inhibit erythropoietin-induced erythrocyte maturation, which is reflected in the RDW increase. Some reports have found a relationship between RDW and mortality related to age or several malignant or non-malignant conditions. However, there is no information about the role of RDW in overall survival (OS) of patients with DLBCL. We aim to evaluate the prognostic role of RDW levels in DLBCL patients at diagnosis. METHODS We retrospectively evaluated 83 patients with DLBCL homogenously treated in frontline with R-CHOP from 2002 to 2013 in the Son Espases University Hospital. To avoid selection bias patients were obtained from Pharmacy and Pathology Departments registries. Main clinical and prognostic factors at diagnosis were obtained from medical records. Cheson criteria were used for response assessment. The RDW was collected from the hemogram at diagnosis. The IBM SPSS STADISTICS program was used for all statistical analyses. PFS (time to progression/relapse) and overall survival (OS) (time to death) were measured from the date of ABVD onset, and were estimated according to the Kaplan-Meier method. We performed the comparisons between those interest variables with the log-rank test. A comparison between categorical variables was made with the chi-square of Fisher's exact test, as appropriate. All reported P-values were two-sided, and statistical significance was defined at P RESULTS: Main characteristics of patients were as follows: median age was 62 (20-86) years, 24% had ECOG PS>1, 64% advanced III-IV Ann Arbor (AA) stage, 39% B-symptoms, 51% adjusted-International Prognostic Index (a-IPI) and 39% belong to the high risk (3-5) subgroups of R-IPI Median RDW was 14.6 (11.1-21.1). Using ROC curves we selected the cutoff 14.05 for the death event. We evaluated the association of increased RDW with main prognostic factors at diagnosis. RDW >14.05 at diagnosis was associated with a more advanced age, worse ECOG PS, a more advanced AA stage, higher incidence of B symptoms and IPI>2. However, RDW was not related to disease control in terms of response to therapy (p=0.39) or relapse/progression (p=0.21) rates. Inversely, RDW>14.05 was in fact associated to a higher mortality (47%) compared to only 17% in patients with RDW≤14.05 (p=0.008). Median follow-up was 77 (20-137) months. Univariate survival analysis showed age>60 years (p=0.001), ECOG PS>1 (p=0.036), high risk R-IPI (p=0.005), a higher than 15% reduction in relative dose-intensity (RDI) (p=0.026) and RDW>14.05 (p=0.008) were significantly related to worse OS. By contrast, RDW did not significantly influence progression-free survival (p=0.19). CONCLUSIONS: Higher RDW at diagnosis in this series of DLBCL patients was related with older age, worse ECOG PS and more advanced disease but this was not translated into a worse control of disease in terms of only a small non statistically significant impact in response or PFS. By contrast higher RDW was linked to a significantly higher mortality and worse OS possibly related to a higher proinflammatory basal status and comorbidities. Patients with higher RDW may be at risk of reduction in RDI. These findings could justify including RDW in scores of comorbidities in DLBCL as well as in other malignant and non-malignant conditions. Disclosures No relevant conflicts of interest to declare.

The pentacyclic triterpenes oleanolic acid (OLA) and maslinic acid (MLA) are natural compounds present in many plants and dietary products consumed in the Mediterranean diet (e.g., pomace and virgin olive oils). Several nutraceutical activities have been attributed to OLA and MLA, whose antitumoral effects have been extensively evaluated in human adenocarcinomas, but little is known regarding their effectiveness in soft tissue sarcomas (STS). We assessed efficacy and molecular mechanisms involved in the antiproliferative effects of OLA and MLA as single agents or in combination with doxorubicin (DXR) in human synovial sarcoma SW982 and leiomyosarcoma SK-UT-1 cells. As single compound, MLA (10-100 μM) was more potent than OLA, inhibiting the growth of SW982 and SK-UT-1 cells by 70.3±1.11% and 68.8±1.52% at 80 μM, respectively. Importantly, OLA (80 μM) or MLA (30 μM) enhanced the antitumoral effect of DXR (0.5-10 μM) by up to 2.3-fold. On the molecular level, efflux activity of the multidrug resistance protein MRP-1, but not of the P-glycoprotein, was inhibited. Most probably as a consequence, DXR accumulated in these cells. Kinetic studies showed that OLA behaved as a competitive inhibitor of substrate-mediated MRP-1 transport, whereas MLA acted as a non-competitive one. Moreover, none of both triterpenes induced a compensatory increase in MRP-1 expression. In summary, OLA or MLA sensitized cellular models of STS to DXR and selectively inhibited MRP-1 activity, but not its expression, leading to a higher antitumoral effect possibly relevant for clinical treatment. © 2014 Elsevier Inc.

Cyclooxygenase 2 (COX-2) is an inflammatory enzyme involved in the pathogenesis and prognosis of several malignancies. In the present study, we investigated the prognostic value of COX-2 expression in a large (N = 242), uniformly treated Hodgkin lymphoma (HL) population from the Spanish Network of HL using tissue microarrays. Univariate and multivariate analysis was done, including comparing the most recognized clinical variables: the early- and advanced-stage subgroups. COX-2 was expressed on Reed-Sternberg cells in 37% of patients. There were no differences in the distribution of clinical variables according to COX-2 expression. With a median follow-up time of 58 months, PFS at 5 years was 60% and 79% for COX-2+ and COX-2− patients, respectively (P = .003). The overall survival was 73% and 91%, respectively (P < .001). The major impact on prognosis was observed in the early AA stage (I-II) group. In fact, in these low-risk groups the expression of COX-2 defined a group with significantly worse progression-free and overall survival. In conclusion, COX-2 was expressed on Reed-Sternberg cells in one-third of HL patients and was a major independent, unfavorable prognostic factor in early-stage HL. We conclude that COX-2 may be a major prognostic variable in HL and a potential therapeutic target.

The therapeutic effect of doxorubicin (DXR) in the treatment of soft tissue sarcomas (STS) is limited by its toxicity and the development of multidrug resistance (MDR), the latter mainly induced by high expression of efflux pumps (e. g., P-glycoprotein [P-gp]). Therefore, the search for alternative therapies, which sensitize these tumors to chemotherapy while maintaining a low toxicity profile, is a rational approach. We assessed efficacy and molecular mechanisms involved in the antiproliferative effects of the tyrosine kinase inhibitors, nilotinib and imatinib, as single agents or in combination with DXR, in human synovial sarcoma SW982 and leiomyosarcoma SK-UT-1 cells. As single compound nilotinib (1-10 mu M) was more potent than imatinib inhibiting the growth of SK-UT-1 and SW982 cells by 33.5-59.6%, respectively. Importantly, only nilotinib synergized the antitumoral effect of DXR (0.05-0.5 mu M) by at least 2-fold, which clearly surpassed the mere sum of effects according to isobolographic analysis. Moreover, nilotinib in combination with DXR had a sustained effect on cell number (-70.3+/-5.8%) even 12 days after withdrawal of drugs compared to DXR alone. On the molecular level, only nilotinib fully blocked FBS-induced ERK1 and p38 MAPK activation, hence, reducing basal and DXR-induced up-regulation of P-gp levels. Moreover, efflux activity of the MDR-related proteins P-gp and MRP-1 was inhibited, altogether resulting in intracellular DXR retention. In high-risk STS tumors 53.8% and 15.4% were positive for P-gp and MRP-1 expression, respectively, with high incidence of P-gp in synovial sarcoma (72.7%). In summary, nilotinib exhibits antiproliferative effects on cellular models of STS and sensitizes them to DXR by reverting DXR-induced P-gp-mediated MDR and inhibiting MRP-1 activity, leading to a synergistic effect with potential for clinical treatment., Research was supported by grants Jose Ma Buesa del grupo GEIS and the Asociacion Espanola contra el Cancer (Junta de Baleares) and by Grand EC10-150 awarded by the Spanish Ministry of Health in 2010 (Independent Clinical Research Call). V. H. V. is supported by a PhD fellowship from the Conselleria d'Economia, Hisenda i Innovacio (Govern de les Illes Balears). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Introduction: DLBCL is the most common non-Hodgkin lymphoma. There is increasing evidence that tumor microenvironment and host immunity play an important role in lymphoma progression. The absolute lymphocyte count (ALC), calculated from the complete blood count, is considered a surrogate for host immunity and has been proposed as a part of a score in combination with IPI. In contrast to a high ALC being associated with a good prognosis, high levels of blood monocytes (AMC) have just the opposite effect. The absolute lymphocytes/monocytes ratio (LMR) is under investigation and could be a useful prognostic marker in DLBCL. The aim of this study is to analyze the prognostic role or LMR in patients with DLBCL and definite a new score with the combination of RLM and R-IPI that could improve its risk definition. Patients and methods: All patients diagnosed of DLBCL from January-1999 to June-2013 at University Hospital Son Espases were retrospectively identified from Pathology and Pharmacy Departments registries to avoid selection bias. Only patients treated with R-CHOP +/- radiotherapy were included (N=79). Other regimes, consolidations or maintenance were excluded. The ALC and AMC were derived from pre-treatment CBC counts. LMR at diagnosis was calculated using ROC curves. Also, other standard prognostic factors including R-IPI and NCCN-IPI were obtained. Results: The cutoff for LMR was determined as 2.55. Main characteristics of patients are summarized in Table 1. Patients with LMR≤2.55 were associated with ECOG PS>1 (p=0.019), high LDH (p=0.025), a-IPI>1 (p=0.043) and NCCN-IPI intermediate-high (p=0.007). The complete response rate was similar in both groups: 82% for the LMR≤2.55cohort and 95% for LMR>2.55cohort(p=0.15). With a median follow up of 73 months (8-137) the progression free survival (PFS) was influenced by LMR, R-IPI, stage and B-symptoms. However, the multivariate analysis confirmed only LMR as independent significant predictor for PFS [HR 4,7; CI95% 1.5-14.1 (p=0.006)]. In the univariate analysis, only age, B-symptoms and NCCN-IPI were predictors for overall survival (OS). A prognostic score with the combination of LMR and R-IPI was stablished (LMR-R-IPI): patients with very good/good R-IPI and LMR>2.55 in one group and patients with poor R-IPI and/or LMR≤2.55 in other group with 6y-PFS of 96% and 59% (p=0.001), as well as 6y-OS of 87% and 64% (p=0.031), respectively. The multivariate analysis confirmed this new prognosis score as the only independent significant predictor for PFS and OS in our series. Conclusions: The addition of LMR improves the R-IPI in patients with DLBCL treated with R-CHOP. In our series, LMR-R-IPI was the only independent significant predictor for PFS and OS. This score identifies a high risk population that could benefit from different therapeutic approaches. Table 1. Overall group LMR ≤ 2.55(n=40) LMR > 2.55(n=39) P Median age (range) 62 (20-87) 64 (20-87) 60 (29-79) 0.19 Sex M/F (%) 44 (56%) / 35 (44%) 20 (50%) / 20 (50%) 24 (61%) / 15 (38%) 0.37 ECOG PS > 1 20 (25%) 15 (37%) 5 (13%) 0.019 Stage III-IV 49 (62%) 26 (65%) 23 (59%) 0.65 B symptoms 32 (40%) 19 (47%) 13 (33%) 0.25 High LDH 41 (52%) 19 (53%) 15 (38%) 0.025 > 1 extranodal site 12 (15%) 8 (20%) 4 (10%) 0.35 Bulky mass 25 (32%) 16 (40%) 9 (23%) 0.15 a-IPI > 1 38 (48%) 24 (60%) 14 (36%) 0.043 Poor R-IPI 29 (37%) 19 (47%) 10 (26%) 0.062 NCCN-IPI:- Low- Low-intermediate- High-intermediate- High 9 (11%)34 (44%)30 (38%)5 (6%) 3 (8%)11 (28%)22 (56%)3 (8%) 6 (15%)23 (59%)8 (20%)2 (5%) 0.007 High Beta-2-microglobulin 36 (49%) 19 (53%) 17 (46%) 0.64 Disclosures No relevant conflicts of interest to declare.

Introduction The use of maintenance treatment with rituximab in B lymphomas has improved the results in terms of progression free survival in follicular lymphoma (PRIMA trial). This improvement has been seen in other types of indolent lymphomas including chronic lymphatic leukemia or specific cases of aggressive lymphomas with or without low grade associated component. However, the rituximab maintenance, even though is a well-tolerated strategy, is not free of toxicity. Our objective is reporting the rituximab toxicity associated to the maintenance with rituximab in a series of B-cell lymphomas treated with immunochemotherapy in first line in Hospital Son Espases (HUSE) and Hospital del Mar (HM). Methods and materials To avoid selection bias, we retrospectively identified from the Pharmacy registries of HUSE and HM those patients who had been treated with rituximab maintenance between 2002 and 2014 in first line of induction treatment with conventional immunotherapy or immunochemotherapy. Second lines or more intensive treatments were excluded. We obtained information about diagnosis, prognostic and outcome in all the patients, including levels of immunoglobulins as well as the seriousness of infections or other related side effects. Polymorphisms FcgRIIIa were analyzed using RT-PCR and sequencing. Results With the previous criteria 82 patients were included. Table 1 shows the diagnostic characteristics and the treatment of the patients. The median of age was 65 years; most cases were follicular lymphoma treated with R-CHOP or R-B and receiving maintenance with rituximab every 2 months (66%). Table 2 describes toxicity related to rituximab maintenance: hypogammaglobulinaemia IgG, IgA or both in the 27%, 21% and 11% of the cases respectively. When we analyzed the role of polymorphisms of FcgRIIIa we found that the patients with homozigous polymorphism VV were exempt of hypogammaglobulinaemia IgG or IgA. Moreover the polymorphism FF was associated meaningfully with a greater incidence of hypogammaglobulinaemia IgA (p=0.022). Only the 8% of the cases of hypogammaglobulinaemia needed treatment with intravenous immunoglobulins and only 6% had grade 3-4 infections. 21% of the patients treated with rituximab maintenance developed grade 3-4 neutropenia, generally reversible with the administration of G-CSF and without serious infections. Increase of mortality was not observed related with the presence of hypogamaglobulinaemia IgG (p=0.34), IgA (p=0.59) or both (p=0.59) or grade 3-4 neutropenia secondary to rituximab (p=0.38). Conclusions Between one-quarter or one-fifth of the patients who received maintenance therapy with rituximab during 2 years associate hypogammaglobulinaemia IgG, IgA and/or neutropenia as toxicity. Nevertheless these complications are generally mild, without related severe infections or increase of mortality. Table 1. Clinical characteristics of the series Median of age 65 (27-85) Sex 41 (50%) / 41 (50%) ECOG PS >1 7 (9%) Type of lymphoma:- Folicular lymphoma- DLBCL- CLL/SLL- Marginal lymphoma- Mantel cell lymphoma- Other 46 (56%)13 (16%)8 (10%)6 (7%)5 (6%)4 (5%) Stage III-IV 71 (87%) B symptoms 29 (35%) Induction treatment:- R-CHOP/R-CVP- R-B- Rituximab alone- Schema with Fludarabine-R- Intensive schemas- Others 50 (61%)13 (16%)9 (11%)4 (5%)3 (4%)3 (4%) Type of maintenance (2 years):- Every 2 months- Every 3 months- Every 6 months (4 weeks) 54 (66%)15 (18%)1 3 (16%) Table 2. Rituximab maintenance attributable toxicity Polimorphism FcgRIIIA Type of toxicity Total FF FV VV p Hypogammaglobulinaemia- IgG- IgA- Double IgG + IgA 22 (27%)17 (21%)9 (11%) 7 (23%)11 (37%)3 (10%) 15 (31%)6 (12%)6 (12%) 0 (0%)0 (0%)0 (0%) 0.340.0220.74 Treatment with immunoglobulins iv 7 (8%) 3 (10%) 4 (8%) 0 (0%) 0.79 Grade 3-4 infections: 5 (6%) 2 (7%) 3 (6%) 0 (0%) 0.87 Grade 3-4 neutropenia: 17 (21%) 8 (27%) 8 (17%) 1 (25%) 0.59 Disclosures No relevant conflicts of interest to declare.

Background During the last years the appearance of the acquired V617F mutation of the Janus Kinase 2 gene (JAK2) in patients suffering different thrombotic events has been described. We decided to develop a new and rapid multiplex real-time Polymerase Chain Reaction (PCR) in order to detect the V617F mutation together with the inherited prothrombotic mutations of factors F5 and F2. Design and methods The method was carried out on the LightCycler 2.0 (Roche Diagnostics, Mannheim, Germany) and consisted in a first step of simultaneous amplification by real-time PCR of the three genes to be genotyped, in a 20 μl closed tube, using a primer pair together with the correspondent FRET-hybridization probes for each gene. Results We assayed 41 samples in the multiplex PCR reaction, 19 were positive (46.34%) for V617F mutation. From the V617F positive samples we found 1 sample heterozygous for F2 (5.26%) and 1 sample heterozygous for F5 (5.26%), so a 10.52% of the samples tested combine V617F mutation with inherited thrombophilic mutations. Results were clear, rapid and reliable allowing a significant time saving. Conclusions The technique presented in this manuscript is a new achievement in the field of the molecular diagnosis that combines the genotyping of F5 and F2 with the assessment of the JAK2 (V617F) mutation load.

Introduction: DLBCL is the more common non Hodgkin lymphoma. This is an aggressive lymphoma that is treated with a standard chemotherapy regimen: R-CHOP. In the last years attempts have been done to improve the outcome both increasing dose-density (CHOP14) or intensity (CHOEP, ACVBP, autologous stem cell transplantation) without obtaining benefit in terms of survival. This has allowed setting R-CHOP administered every 21 days (R-CHOP21) as the standard treatment for DLBCL patients. RDI is an important issue to consider when treating malignancies. Although this a well-known prognostic factor in Hodgkin lymphoma, scarce information has been published in DLBCL. Objective: The purpose of this study is further analyzing the prognostic impact of RDI in two cohorts of DLBCL patients treated with R-CHOP21 or R-CHOP14, to evaluate its differential impact when increasing dose density. Material and methods: All patients diagnosed of DLBCL from January-1999 to June-2013 at University Hospital Son Espases were retrospectively identified from Pathology Department registry to avoid selection bias. Only patients treated with R-CHOP21 or R-CHOP14 +/- radiotherapy were included (N=115). To increase the R-CHOP14 cohort we added also all patients treated with R-CHOP14 in the same time period in two additional hospitals (Hospital Son Llatzer of Palma and Hospital del Mar of Barcelona) identified from their Pharmacy registries to avoid selection bias (N=42). Other regimes, consolidations or maintenance were excluded. Table 1 shows main characteristics of the global series (N=157). RDI represents the ratio of the amount of a drug actually administered to the amount planned for a fixed time period. RDI was calculated as previously described. Briefly, RDI of each drug was obtained followed by an average of RDI in CHOP consisting in the sume of RDI of the 3 drugs divided by 3. Main prognostic factors at diagnosis in DLBCL were obtained, including international prognostic index (IPI) factors. Evaluations were carried out following standard guidelines. Results: Overall response and complete response rates were similar in both groups:86% and 76% for R-CHOP21 and 94% and 74% for R-CHOP14(p=0.17 and p=0.85, respectively). Median follow-up for alive patients was 68 months (4-156). There were no differences between the two cohorts in terms of either OS or PFS (Figure 1). In the R-CHOP21, both a reduction higher than 15% in RDI [RR 7.41 (2.51-21.83); (p Conclusions: Overall in our series there were no differences in terms of response or survival between patients treated with R-CHOP21 or R-CHOP14. A higher rate of RDI reduction was observed in the R-CHOP14 group. However, the impact of RDI reductions on response and survival was only observed in the R-CHOP21 group but not in patients treated with R-CHOP14. We can conclude that R-CHOP21 and R-CHOP14 are equivalent regimens in terms of response and survival only if RDI reductions are avoided. For patients receiving R-CHOP21 we recommend using clinical and support measures in order to avoid RDI reductions. Table 1. Main clinical characteristics of the patients. R-CHOP21 group(n=74) R-CHOP14 group(n=83) P Age (median & range) 65 (25-88) 55 (15-79) 0.005 Sex (M/F) 34 (46%) / 40 (54%) 51 (61%) / 32 (39%) 0.056 ECOG PS > 1 17 (23%) 18 (22%) 0.85 Ann Arbor stage III-IV 40 (54%) 48 (58%) 0.75 B-symptoms 25 (34%) 26 (31%) 0.86 Elevated LDH 33 (46%) 40 (49%) 0.75 > 1 extranodal site 8 (11%) 19 (23%) 0.057 Bulky disease 23 (31%) 34 (41%) 0.24 R-IPI unfavorable 24 (32%) 26 (32%) 1 NCCN-IPI: - Low - Low-intermediate - High-intermediate - High 9 (13%) 27 (39%) 26 (38%) 7 (10%) 17 (21%) 35 (44%) 24 (30%) 4 (5%) 0.31 Elevated Beta-2-microglobulin 32 (49%) 31 (39%) 0.24 Radiotherapy 27 (36%) 27 (32%) 0.62 Figure 1. Figure 1. Figure 2. Figure 2. Disclosures No relevant conflicts of interest to declare.

Antonio Gutierrez,1,* Jose Rodriguez,1,* Jordi Martinez-Serra,1 Jordi Gines,2 Pilar Paredes,1 Florencia Garcia,3 Javier Vercher,4 Josep Balanzat,4 Raquel del Campo,5 Pilar Galan,6 Miguel Morey,1 Antonia Sampol,1,7 Andres Novo,1 Leyre Bento,1 Lucia García,1 Joan Bargay,5 Joan Besalduch1,7 1Service of Hematology, 2Service of Pharmacy, 3Service of Oncology, Son Espases University Hospital, Palma de Mallorca, Spain; 4Service of Hematology, Can Misses Hospital, Ibiza, Spain; 5Service of Hematology, Son Llatzer Hospital, Palma, Spain; 6Service of Hematology, Mateu Orfila Hospital, Menorca, Spain; 7Service of Hematology, Policlínica Miramar, Instituto de Investigación Sanitaria de Palma (IdISPa), Palma, Spain *These authors contributed equally to this work Background: Most Hodgkin lymphomas (HL) can be cured with current strategies. However, one-third of the cases do not respond or relapse and need salvage regimens. We report the results of a retrospective study using the gemcitabine and oxaliplatinum (GemOx) regimen.Methods: Patients who relapsed or failed to achieve complete response were eligible and received GemOx salvage therapy. To avoid selection bias and thus to overcome the retrospective nature of the study, all treated patients were included from the pharmacy database.Results: Between 2003–2013, 24 HL patients – relapsing (number [n]=12) or refractory (n=12) – were included, receiving a total of 26 induction treatments with GemOx. Mean previous regimens were 2.38 (42% relapsing after autologous transplantation). Median follow-up was 37 months, and 71% responded (38% of patients achieved complete response). The factors related to better progression-free survival were: B symptoms; response to GemOx; and consolidation with stem cell transplantation. Grades 1 and 2 neurological toxicity was present in 17% of patients. Hematological toxicity was common, with grades 3 and 4 neutropenia (25%) and thrombocytopenia (34%) observed. Progression-free survival was better in patients consolidated with stem cell transplantation. The peripheral blood stem cell collection after GemOx was successful for all candidates.Conclusion: 1) The GemOx regimen is effective in relapsed or refractory HL with manageable toxicity. 2) No mobilization failures were observed. 3) Consolidation after response is needed. 4) Its efficacy and favorable toxicity profile might make multiple administrations possible in several recurrences in HL.Keywords: gemcitabine, oxaliplatinum, GemOx, Hodgkin lymphoma, treatment

Background DLBCL is the more common non Hodgkin lymphoma. This is an aggressive lymphoma that is treated with a standard chemotherapy regimen: R-CHOP. In the last years attempts have been done to improve the outcome both increasing dose-density (DD) (CHOP14) or intensity (CHOEP, ACVBP, frontline high dose therapy followed by autologous stem cell transplantation). Although phase 2 studies of these interventions suggested promising results, when randomized phase 3 trials have been conducted, there is no demonstrated benefit of these higher toxicity approaches when compared with R-CHOP alone. Only addition of rituximab to CHOP has proved a survival advantage. This has allowed setting R-CHOP administered every 21 days (R-CHOP21) as the standard treatment for DLBCL patients. The purpose of this study is further analyzing the prognostic impact of DD delays in two cohorts of DLBCL patients treated with R-CHOP21 or R-CHOP14. Methods All patients diagnosed between 1999 and 2013 of DLBCL in University Hospital Son Espases were identified from Pathology Department registry. Only patients treated with R-CHOP21 or R-CHOP14 +/- radiotherapy were included. Patients receiving other chemotherapy regimens or consolidations were excluded. DD delay was calculated as follows: DD delay = real number of days from first to last cycle of chemotherapy / expected number of days from first to last cycle in every regimen. Results A total of 166 cases were identified: considering inclusion and exclusion criteria finally 111 cases were selected (71 in the R-CHOP21 cohort and 40 in the R-CHOP14 cohort). Respectively for R-CHOP21 and R-CHOP14, 61% and 37% were older than 60 years (p=0.02), 26% and 35% had an ECOG PS higher than 1 (p=0.3), 49% and 62% had an Ann Arbor stage III-IV (p=0.09), 44% and 51% an a-IPI higher than 1 (p=0.47). Median DD delay was 2% versus 14% for R-CHOP21 and R-CHOP14 groups (p Conclusions Overall in our series there were no differences in terms of response or survival between patients treated with R-CHOP21 or R-CHOP14. Significantly higher rates of DD delay were observed in the R-CHOP14 group. However, the impact of DD delays on response and survival was only observed in the R-CHOP21 group but not in patients treated with R-CHOP14. We can conclude that R-CHOP21 and R-CHOP14 are equivalent regimens in terms of response and survival only if DD delays are avoided. For patients receiving R-CHOP21 we recommend using clinical and support measures in order to avoid DD delays. Disclosures: No relevant conflicts of interest to declare.

Background Cyclooxygenase-2 (COX-2) is an inflammatory enzyme involved in the pathogenesis and prognostic of Hodgkin lymphoma. We previously reported that expression of COX-2 on Reed-Sternberg cell is a major independent, unfavorable prognostic factor in patients with early Hodgkin lymphoma treated with ABVD (Blood 2012; 119: 6072). The relationship between COX-2 expression and poor prognosis has been associated with pathogenic phenomena such as induction of angiogenesis, chemoresistance through induction of antiapoptotic mechanisms such as up-regulation of bcl-2 or resistance to Fas, or promotion of invasion through induction of some metalloproteinases. As patients in our series were treated with ABVD +/- radiotherapy, we aim to analyze the role of radiotherapy on the adverse prognostic factor of COX-2 RS expression. Methods In the present study, we investigated the prognostic value of COX-2 expression in a large (N = 143), uniformly treated early Hodgkin Iymphoma (HL) population from the Spanish Network of HL using tissue microarrays. Univariate and multivariate analysis was done, including the most recognized clinical variables and the potential role of administration of adjuvant radiotherapy. Results Median age was 31 years; there was B-symptoms or bulky disease in 30% of patients; ECOG PS > 1 in 6%, EORTC score > 1 in 84% and 66% received radiotherapy. The expression of COX-2 defined a subgroup with significantly worse prognosis. Accordingly, the 46 (32%) COX-2+ patients had a PFS at 5 years of 72% versus 86% of COX-2- patients (P = .017). Similarly, the OS at 5 years also differed for both groups, with 82% and 94% for COX-2+ and COX-2-, respectively (P = .004). By multivariate analysis, COX-2 expression (HR = 2.76), B symptoms (HR = 2.34), and administration of RT (HR = 0.32) were independently associated with PFS, whereas only ECOG PS (HR = 9) and COX-2 RS expression (HR = 4.81) were independently associated with OS. Considering patients COX-2+, those that received RT had a significantly better 5y-PFS (80% versus 54% if no RT; p = .008). In contrast, COX-2- patients only had a modest non-significant benefit from RT in terms of 5y-PFS (90% versus 79%; p=0.13). When compared the outcome of patients receiving RT considering the expression of COX-2 on RS we found a non-significant 10% difference in terms of PFS between COX-2+ and COX-2- patients (p=0.09) while this difference between the two groups was important (25%) in patients non receiving RT (p=0.04). Conclusions COX-2 RS expression is an adverse independent prognostic factor in early HL. Radiotherapy overcome the worse prognosis associated to COX-2 expression on RS acting in a chemotherapy-independent way. COX-2 RS expression may be useful to determine patients with early HL candidates to receive consolidation with RT. Disclosures: No relevant conflicts of interest to declare.

Abstract 4845 Hodgkin lymphoma (HL) represents 10–15% of all types of lymphoma. At present, more than 70% of patients can be cured with current strategies based on chemotherapy with or without radiotherapy. However, one third of the cases finally relapses and needs salvage regimens usually consolidated with high dose chemotherapy and autologous stem cell transplantation. The number of regimens and drugs available are limited and new protocols that increase the efficacy with manageable toxicity are needed. In the present communication we report the results of a retrospective study using the GemOx schema that combines the efficacy of gemcitabine in Hodgkin lymphoma with oxaliplatin, a less toxic and effective platinum-based drug. Patients and methods: Patients were eligible for this retrospective study, according to the following criteria: diagnosis of HL, which relapsed or failed to achieve complete remission after induction treatment. They received GemOx regimen that consisted of gemcitabine 1000mg/m2 and oxaliplatin 100mg/m2 on day 1. Treatment was given every 15 days if feasible or every 21 days. To evaluate response and toxicity Cheson criteria and OMS toxicity scale were used respectively. Results: Between 2003 and 2012, 29 patients with Hodgkin lymphoma were retrospectively included in this study. All patients had recurrent (n=17) or refractory (n=12) disease. Median age was 24 (14–76) years and 50% had an International Prognosis Score (IPS) higher than 2. Patients received a mean of 2.79 previous regimens and 79% more than 1 regimen before GemOx with 48% relapsing after a prior autologous stem cell transplant (ASCT). Median follow-up was 41 months. 76% of patients responded (31% complete responses; CR). Responses were better in the relapsed setting or partial response (PR) (85% with a 45% of CR) compared to the truly refractory cases (55% PR) (p=0.037). Main prognostic factors for HL were assessed to view their impact on survival. Factors related with progression- free survival (PFS) and overall survival (OS) were age lower than 45 years, response to GemOx and consolidation with stem cell transplantation (p=0.001). Presence of B-symptoms at diagnosis also influenced OS. Neurologic toxicity was present in 9% of patients, all of them grade I or II. Hematologic toxicity was also common, including grade 3 or 4 neutropenia in 23% of patients, and grade 3 or 4 thrombocytopenia in 33%. Nausea and vomiting occurred in all the patients at grade 2, or lower. At last follow-up, 13 patients (45%) are alive and remain free of progression. However, 16 patients (54%) had died: 12 (41%) due to progression of disease, 3 (10%) due to complications due to a subsequent allogenic transplant (graft versus host disease, thrombotic thrombocytopenic purpura and bleeding) and 1 due to pneumonia. PFS was better in patients consolidated with autologous or allogeneic transplantation (100%) compared with patients not consolidated (14%) (p=0.009). PBSC collection after GemOx and G-CSF was successful for all of candidates. Conclusions: 1) GemOx regimen is effective in relapsed or refractory Hodgkin lymphoma with manageable toxicity; 2) Results are better in relapsed or chemosensitive disease compared to truly refractory cases; 3) No mobilization failures were observed; 4) Consolidation after response is needed. Disclosures: No relevant conflicts of interest to declare.

Abstract 1560 Poster Board I-583 Background Hodgkin lymphoma is defined by Reed Stemberg (RS) tumor cells in a microenvironment composed of a deregulated cytokine network and diverse cell populations, associated with their respective extracellular matrix, defining an inflammatory microenvironment. COX-2 (cyclooxygenase 2) is an inflammatory induced enzyme involved in the pathogenesis and prognosis of several solid tumors (colorectal, breast, ovarian, lung, etc). Its role in hematological malignancies is being recognized and specifically its expression in Hodgkin lymphoma has been associated with higher proliferation and angiogenesis. Aim To investigate the prognostic value of COX-2 expression in a large uniformly treated population with Hodgkin lymphoma. Methods We examined tissue microarrays for COX-2 expression in 242 patients of the Hodgkin Lymphoma Spanish Network treated with ABVD with or without Radiotherapy (XRT) according to AA (Ann Arbor) staging. Tissue sections were stained with monoclonal antibodies to COX-2 and the expression was scored and validated by a pathology review. Cox-2 was dichotomized as either negative (no RS staining) or positive. Plots of Kaplan Meier were constructed for survival analysis and most recognized clinical variables (age, sex, presence of bulky disease, B symptoms, AA stage, International prognostic system for Hodgkin lymphoma (IPS), ECOG performance status (PS) were investigated for their prognostic importance along with COX-2 expression by univariate and multivariate analysis. Results Median age was 33 (10 to 83). COX-2 was expressed in 89 pts (37%), 99 pts (41%) presented with AA stage III-IV, 98 pts presented B symptoms (40%) and 124 pts (51%) received XRT added to ABVD chemotherapy, 66% of patients with AA stage I-II and 30% of pts presenting with AA stage III-IV. There were no differences in the distribution of clinical variables according to the COX-2 expression, except for a trend to increased positivity in nodular sclerosis and lymphocyte-depleted subtypes and higher AA stage. With a median follow up for alive patients of 59 months, the PFS at 5 years were 61% and 79% for COX-2 + and negative respectively (p=0.004). OS were 73% and 91% respectively (p=2, IPS>2, no XRT treatment and COX-2 expression were associated with an unfavourable PFS and OS. By multivariate analysis only the IPS (HR 3.72) and COX-2 (HR 1.72) expression were independently associated with the outcome. Interestingly the major impact in the prognosis was observed in the favourable AA stage (I-II) and IPS (0-2) groups. In fact in these low risk groups the expression of COX-2 define a group of significant worse prognosis. Accordingly, the 46 patients (32%) of the 143 patients presenting an AA stage of I-II had a PFS at 5 years of 73% versus 86% of those COX-2 negative (p=0.031). Similarly the OS at 5 years of both groups also differ with 82 and 94% for COX + and negative respectively (p=0.004). Conclusion COX-2 is expressed in approximately a third of RS cells and is a major independent unfavourable prognostic factor. Of main relevance is the fact that in the favourable groups of the AA and IPS the expression of COX-2 identify a group of significant worse prognosis that need different approach. According to our data COX-2 can be a major prognostic variable in this disease and potentially could be a therapeutic target. Disclosures No relevant conflicts of interest to declare.

Abstract 1561 Poster Board I-584 Vascular endothelial growth factor (VEGF) is a key regulator of tumor angiogenesis as well as a growth and metastatic inducer in several malignancies. Numerous studies have demonstrated up-regulation of this protein at both mRNA and protein level in various tumours and a correlation with advanced stage and prognosis has been demonstrated in several solid tumors and hematologic malignancies such as multiple myeloma or non-Hodgkin lymphomas. There is evidence that also the neoplastic Hodgkin-Reed-Stenberg cells express VEGF and their receptors and that VEGF expression both in macrophages and the extracellular matrix might facilitate tumour progression, contributing to the pathophysiology of this malignancy. We aim to investigate the prognostic value of VEGF expression in a large uniformly treated population with Hodgkin lymphoma. In this immunohistochemical study we retrospectively included 252 patients of the Hodgkin Lymphoma Spanish Network uniformly treated with ABVD (3-6 cycles, in function of clinical factors) with or without radiotherapy. Tissue sections were stained with monoclonal antibodies to VEGF-A and the expression was scored and validated by a pathology review. VEGF expression was scored as negative (no RS staining), middle or strong positive staining. Overall survival (OS) and progression-free survival (PFS) were measured from the diagnosis and were estimated according to the Kaplan-Meier method. Comparisons among those variables of interest were performed by the log-rank test. Multivariate analysis with the variables that proved to be significant in univariate analysis was performed according to the Cox proportional hazard regression model. Most recognized clinical variables (age, sex, presence of bulky disease, B symptoms, AA stage, international prognostic score for Hodgkin lymphoma (IPS) and ECOG performance status (PS) were investigated for their prognostic importance along with VEGF expression. Median age was 33 years (10-84). Prognostic factors were as follows: 41% Ann Arbor stage III-IV, 40% B-symptoms, 9% ECOG PS >1 and 26% IPS > 2. One hundred and thirty patients (52%) of cases received ABVD plus radiotherapy. Median follow-up was 59 months (8-199). Relapses were significantly higher in patients with positive VEGF staining in a stain-dependent manner (no stain: 10%; middle stain: 44% and strong stain: 65%) (p 60 years, III-IV AA stage, ECOG PS > 1, B-symptoms, IPS > 2 and non receiving radiotherapy). By multivariate analysis only the IPS (HR 2.19) and VEGF middle (HR 3.2) or strong (HR 7.24) expression were independently associated with the outcome. Conclusion Middle and strong VEGF staining on neoplastic Hodgkin-Reed-Stenberg cells is associated with a greater risk for relapse and a worst PFS. However in this series this fact only affected OS in the strong staining cases. This could imply that these VEGF-positive cases may need a more intensive induction therapy and justify the development of trials including anti-VEGF or antiangiogenic drugs as part of therapeutic regimens in this malignancy. Disclosures No relevant conflicts of interest to declare.

B Acute Lymphoblastic leukemia (B-ALL) with Philadelphia chromosome (Ph') is a neoplasm of lymphoblast committed to the B cell lineage. The clinical presentation of B-ALL Ph'+ is similar to B-ALL, but is more common in adults than in children. The e1a3 rare variant is produced by the fusion of BCR exon 1 to ABL exon 3. The presence of this translocation has been associated with good disease outcome for chronic myeloid leukemia in a very small series of only 5 cases; there is no such evidence for B-ALL. We report two new cases of B-ALL Ph+ with the rare e1a3 fusion transcript. The e1a3 and e1a2 (p190) transcripts have been reported to have a similar molecular weight and probably a similar clinical profile, thus in these cases the presence of e1a3 was associated with extramedullary infiltration and disease acceleration.