bluuebunny/crossref_metadata_2025 huggingface dataset:
PF317 ELEVATION OF SERUM IL‐10 IS ASSOCIATED WITH ADVERSE CLINICAL OUTCOMES IN PATIENTS WITH DIFFUSE LARGE B‐CELL LYMPHOMA WHO WERE TREATED WITH R‐CHOP
Authors: Yi J.H. , Yoon S.E. , Ko Y.H. , Kim W.S. , Kim S.J.
Published: June 2019

Background:

Cytokines are one of the major non‐cellular component of tumor microenvironment, and they act as messengers between malignant cells and the cellular component of tumor microenvironment to promote cancer cell proliferation and to evade immune surveillance. While the role of cytokine had been widely investigated in patients with DLBLC, their impacts on secondary CNS involvement had rarely been reported.

Aims:

To address prognostic roles of serum cytokines regarding the OS and CNS events in DLBCL patients.

Methods:

Patients out of our prospective cohort study (NCT#00822731) who met the following criteria were included in the present study. 1) Patients with newly diagnosed, histologically confirmed DLBCL, other than primary CNS lymphoma; 2) Patients who were treated with R‐CHOP as the first‐line treatment; 3) Patients whose serum samples were collected. The clinical data of patients were collected. Serum levels of IL‐2, IL‐6, IL‐10, IL‐21, CXCL10, CCL3, CCL5, PAI‐1, survivin, and TNFα were measured. The primary end‐point of the current study was estimated 3‐year overall survival (OS) rate, and the secondary end‐point was time to secondary CNS involvement

Results:

A total of 313 patients were eligible for the analysis. The median age was 56 years (range, 16 to 86 years) and 177 patients (57%) were male. The 3‐year OS rate was 73.6% ( ±  2.5%), and all the factors in IPI significantly discriminated the OS. Six cytokines were correlated with the 3‐year OS rate (FIGURE). Time to CNS involvement was assess in 294 patients, and 16 events (5.4%) of secondary CNS involvement were detected. Only one cytokine could predict it, as patients with elevated IL‐10 was associated with significantly short time to CNS involvement compared to patients without IL‐10 elevation (p = 0.012). While only 1 out of 99 patients (1.0%) without IL‐10 elevation developed secondary CNS involvement, 15 out of 195 patients (7.7%) with IL‐10 elevation developed secondary CNS involvement (p = 0.017). Of note, among 19 patients who initially had CNS involvement, 14 patients (74%) had elevated serum IL‐10.

Summary/Conclusion:

We have found that elevation of CCL3, CCL5, CXCL10, IL‐6, IL‐10, and survivin were associated with inferior 3‐year OS rate, and elevation of IL‐10 was associated with short time to secondary CNS involvement. Considering their biologic roles in lymphomagenesis, further studies are required to incorporate these results to conventional prognostic factors to better predict outcomes of patients with DLBCL

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PF326 VITAMIN D INSUFFICIENCY AND PROGNOSIS IN LYMPHOMA
Authors: Sanchez‐Gonzalez B. , Platero J. , Ortuño A. , Garcia‐Pallarols F. , Perez S. , Torres E. , Bento L. , Parraga I. , Gimeno E. , Gutierrez A. , Salar A.
Published: June 2019

Background:

Vitamin D deficiency has been recently associated, as an independent risk factor, with lower rates of progression‐free survival (PFS) and overall survival (OS) in patients with Hodgkin (HL) and non‐Hodgkin's lymphoma (NHL).

Aims:

To investigated the impact of vitamin D status on the outcome of lymphoma patients in two Institutions.

Methods:

From 2015 to 2018, we tested circulating 25‐hydroxyvitamin D3 (25[OH]D3) levels in a prospective cohort of 135 newly diagnosed patients with NHL and HL in two Institutions in Spain. 25[OH]D3 levels were measured by chemoluminescent immunoassay. Determinations were based on guidelines from the Food and Nutrition Board of the Institute of Medicine. According to current guidelines, we considered the following thresholds: vitamin D deficient (<10 ng/ml), vitamin D insufficiency (10 to 30 ng/ml) and normal vitamin D level (>30 ng/ml). Patients with <20 ng/ml from one Institution had vitamin D substitution. OS and PFS were calculated using Kaplan‐Meier estimators, comparison between categories performed by log‐rank test and Cox PH regression, and the effect of covariates reported with 95% confidence interval (95 CI).

Results:

135 newly diagnosed patients with NHL and HL were included. Median age at diagnosis was 67 years (range, 20 to 91 years). NHL subtypes: 47% diffuse large b cell lymphoma (DBCL), 30% Indolent lymphoma, 5% mantle cell lymphoma, 6% Tcell lymphoma, 10% Hodgkin lymphoma and 3% others NHL 34% had advanced stage and and 39% of NHL patient had ≥ 3score of International Pronostic Index (IPI).55% of the patients were diagnosed in spring or summer. Median 25(OH)D levels at diagnosis were 18 ng/ml. 11% of patients had normal 25(OH)D levels, 77% had insufficient 25(OH)D levels and 12% were deficient at lymphoma diagnosis. Median follow‐up was 12 months and 22 deaths were observed. The three groups of vitamin D levels were associated with OS, but this difference was not statistically significant (Figure 1).

In the DLBCL subset, we inspected in a Cox Model the effect of Vitamin D levels for OS/PFS, adjusting for IPI Score and vitamin D substitution. Vitamin D levels did not achieve statistical significance for PFS (hazard ratio [HR], 1.02; 95% CI, 0.99 to 1.04) and OS (HR: 0.99; 95% CI, 0.94 to 1.06). Remarkably, there was a tendency towards a better statistically significant outcome in patients supplemented (HR for PFS: 0.36; 95% CI, 0.10 to 1.29; p = 0.10).

Summary/Conclusion:

Most of our patients present with low levels of vitamin D[JPD1]. In our limited series, with the current follow‐up time, baseline vitamin D values did not significantly impact PFS and OS. Further studies are needed to confirm our data.

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PF341 LUSUTROMBOPAG FOR TREATMENT OF THROMBOCYTOPENIA IN PATIENTS WITH CHRONIC LIVER DISEASE WHO ARE UNDERGOING PLANNED INVASIVE PROCEDURES: POOLED SAFETY ANALYSIS OF BLEEDING‐RELATED ADVERSE EVENTS
Authors: Giannini E.G. , Kano T. , Ochiai T. , Bentley R. , Shrestha P.
Published: June 2019

Background:

Thrombocytopenia is a common complication in patients with chronic liver disease. Furthermore, a low platelet count in chronic liver disease patients who require an invasive procedure is a serious condition that may be associated with an increased risk of bleeding during or after the procedure. Lusutrombopag is a thrombopoietin receptor agonist approved in Japan and US for improvement of thrombocytopenia, and in EU for severe thrombocytopenia, associated with chronic liver disease in patients undergoing planned invasive procedures.

Aims:

A pooled safety analysis was performed to assess differences in bleeding events following treatment with lusutrombopag without platelet transfusion or placebo with platelet transfusion.

Methods:

Safety data were pooled from 3 double‐blind, placebo‐controlled studies, in patients with thrombocytopenia associated with chronic liver disease undergoing a planned invasive procedure (Phase 2b: M0626 [Japan; JapicCTI‐121944]; Phase 3: L‐PLUS 1 [Japan; JapicCTI‐132323], L‐PLUS 2 [global; NCT02389621]). Patients enrolled in these studies were randomized to lusutrombopag 3 mg or placebo for up to 7 days prior to procedure. Platelet transfusion was administered if platelet count was <50x109/L no more than 2 days prior to procedure. Patients were classified into 1 of 4 subgroups: lusutrombopag with or without platelet transfusion or placebo with or without platelet transfusion. This analysis focused on patients in the lusutrombopag without platelet transfusion and placebo with platelet transfusion subgroups. Bleeding events were summarized pre‐, during, and post‐procedure.

Results:

Lower bleeding event rates, regardless of severity and timing of onset, were observed for lusutrombopag alone vs placebo+platelet transfusion (Table). Percent of patients with bleeding events was numerically higher with placebo+platelet transfusion vs lusutrombopag alone, both during (4.8% vs 3.2%) and post‐procedure (7.1% vs 4.0%). Percent of patients with bleeding events in liver‐related procedures was greater in the placebo+platelet transfusion arm (16.1%) vs lusutrombopag alone (11.1%), and a larger difference was seen with gastrointestinal‐related procedures in placebo+platelet transfusion (8.9%) vs lusutrombopag alone (2.0%).

Summary/Conclusion:

In this pooled analysis, a trend toward lower bleeding event rates was observed with lusutrombopag alone vs placebo+platelet transfusion, regardless of type of invasive procedure.

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PF346 ZEBRAFISH AS A NOVEL MODEL TO STUDY GATA2 HAPLOINSUFFICIENCY SYNDROMES
Authors: Gioacchino E. , Koyunlar C. , de looper H. , Peulen J. , Bosch D. , hoogenboezem R. , van strien P. , Bindels E. , Touw I. , de Pater E.
Published: June 2019

Background:

Patients with GATA2 mutations have up to 80% chance to develop a myeloid malignancy. MonoMAC and Emberger syndromes are GATA2 deficiency syndromes and are characterized by monocytopenia, B, NK cell lymphopenia and a predisposition to acute myeloid leukemia (AML). Although the disease initiating mutation is known, it is unclear how these patients develop hematological malignancies.

Aims:

In our research we use both mouse and zebrafish models of Gata2 haploinsufficiency to describe their phenotypes and study the mechanisms of malignant transformation.

Methods:

In mice we analyzed the transcriptome and functionality of LSK SLAM in Gata2 mutants. In zebrafish, two orthologues of Gata2 exist: gata2a and gata2b of which gata2b was shown to be predominantly expressed in hematopoietic cells. We used CRISPR‐Cas9 to generate zebrafish gata2b mutants and evaluate their phenotype using flow cytometry, RNAseq, histology and in situ hybridization. Using single‐cell RNA sequencing we aim to identify early onset markers of dysplasia and unveil the mechanism of malignant transformation.

Results:

Gata2 heterozygous mutant mice survive into adulthood without developing signs of alterations in the myeloid lineage. Phenotypic HSCs of these mutant adult mice have unchanged Gata2 expression but are lower in frequency, more proliferative and transcriptionally more committed to differentiation compared to WT. Additionally, they display increased DNA damage indicating these HSCs experience proliferative stress. gata2b

+/− zebrafish show reduced myeloid differentiation and the kidney marrow smears of this aged fish show dysplastic myeloid cells.

Summary/Conclusion:

Our mouse model doesn’t incur a malignancy even though it acquires an HSCs phenotype. Our zebrafish model develops a dysplastic kidney marrow and recapitulates characteristics of the GATA2 deficiency syndromes. Both models can give new insights into the physiopathology of these diseases.
PF373 IMPACT OF ATM ALTERATIONS ON CHRONIC LYMPHOCYTIC LEUKEMIA IN GENERAL PRACTICE IN THE AREA OF NEXT GENERATION SEQUENCING
Authors: Ganard M. , Houot R. , Henry C. , Boulland M.‐L. , Luycx O. , Norwood J. , Bareau B. , Grulois I. , Launay V. , Jacomy D. , Roussel M. , De Tayrac M. , De Guibert S. , Fest T. , Lamy T. , Pastoret C.
Published: June 2019

Background:

Chronic Lymphocytic Leukemia (CLL) is characterized by a very heterogeneous clinical course. Del11q22–23 affecting ATM gene is the most common cytogenetic abnormality associated with poor prognosis. Clinical significance of ATM mutations, isolated or associated with del11q22–23, remains unclear specifically since the widespread use of new therapeutic agents.

Aims:

Evaluate frequency, associations and clinical significance of ATM alterations in a daily clinical practice cohort.

Methods:

We retrospectively analyzed a cohort of 268 CLL patients tested by NGS (Next Generation Sequencing) including ATM mutations in daily clinical practice.

Results:

We identified 50 different ATM mutations among 42 patients (15.7%) by NGS, including a majority of missense mutations and 35 del11q using FISH. Bi‐allelic ATM alterations (ATM mutation and del11q) were found in only 13 patients (37.5% of the del11q patients and 31% of the ATM mutated patients). All but 11 patients with ATM alterations received anti‐CD20 combined chemotherapy directly after NGS realization. ATM alterations had no significant impact on overall survival (OS) but significantly affected time to first treatment (TTFT) in univariate analysis (median of 2.4 years versus 6.1 years (p = 0.004)) (Figure 1A) as well as in multivariate analysis (HR 1.6 IC 95% 1–2.6 ‐ p = 0.023). ATM alterations also affected significantly the time to next treatment (TTNT) with a median time of 2.0 years versus non‐reached in the absence of ATM alteration (p = 0.003) (Figure 1B).Outcome in the bi‐allelic ATM alterations subgroup was particularly pejorative with a median TTNT of 5.5 months (IC 95% 4.8–6.2 months). In multivariate analysis, only ATM mutation had a significant impact on TTNT (HR 3.98 IC 95% 1.45–10.9 ‐ p = 0.008) even when BCR‐inhibitors were used.

Summary/Conclusion:

ATM alterations are frequent in CLL either at diagnosis or at relapse. They confer worse disease evolutionary profile and earlier relapses, especially in case of bi‐allelic inactivation or multiple‐hit profile. These findings emphasize the importance of integrating ATM gene in a targeted NGS panel to better characterize CLL patients while waiting availability ofnew therapeutic agents targeting ATM deficiency.

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PF404 TUMOR‐DERIVED EXOSOMES MAY BE USED AS NEW INFORMATIVE TOOL FOR THE DETECTION OF BONE MARROW RESIDUAL CML LEUKEMIC CELLS ACTIVITY
Authors: Bernardi S. , Foroni C. , Zanaglio C. , Re F. , Polverelli N. , Turra A. , Morello E. , Farina M. , Cattina F. , Gandolfi L. , Zollner T. , Accorsi Buttini E. , Malagola M. , Russo D.
Published: June 2019

Background:

Ph+ Chronic myeloid leukemia (CML) is a clonal myeloproliferative neoplasia characterized by BCR‐ABL1 fusion‐gene, derived from the t (9;22) translocation. Tyrosine Kinase Inhibitors (TKIs) target BCR‐ABL1 protein, induce molecular response in the most of patients, prolong the survival and allow the treatment free remission. Many studies have demonstrated the persistence of leukemic cells in the bone marrow (BM) niche, even after treatment. The conventional minimal residual disease monitoring for CML patients is based on the RT‐qPCR quantification of BCR‐ABL1 transcript in the peripheral blood (PB) cells. However, it is not able to appreciate the activity of residual CML leukemic cells in the BM, responsible of the molecular relapse in the patients undergoing TKIs discontinuation. Exosomes, carrying different information of the cell of origin, have garnered considerable attention in cancer research thanks to the discovery of their messenger role in cellular communication. Moreover, there are some evidences regarding the possibility to isolate the different exosomes sub‐populations targeting the antigens on their surface.

Aims:

In this study we aimed to explore the feasibility of the tumor‐derived exosomes fraction enrichment in CML patients in chronic phase (CP) and treated with TKIs. Ph‐ patients and healthy subjects served as controls.

Methods:

BCR‐ABL1 transcript was quantified following on PB cells both conventionally, according to the International Scale, and by dPCR, given the last published evidences of its accuracy and sensitivity. The exosomes enrichment was performed by a specific kit based on immuno‐selection for a pan‐cancer antigen. The quantification of BCR‐ABL1exosomal transcript was performed by dPCR. In addition, Y4 transctript, an exosomal housekeeper, was quantified in order to assess the exosomal RNA quantity.

Results:

The possibility of the tumor‐derived exosomes enrichmentwas confirmed by the quantification of Y4 and the immuno‐selection of the exosomes resulted specific thanks to the negative results obtained in healthy controls. In Ph‐controls and in CML patients we were able to isolate tumor exosomes (Y4 positive). Moreover, the presence of BCR‐ABL1 transcript in all CML exosomal samples highlighted, for the first time, the presence of active leukemic cells in CML patients in CP.

Summary/Conclusion:

These results suggest that tumor derived exosomes could be consider as a new toolfor the identification of active leukemic cells and for the assessment of an innovative monitoringfocused on the biological meaning of exosomes in CML.

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PF444 IDENTIFICATION OF T‐CELL RECEPTORS FROM THE ALLOGENEIC‐HLA REPERTOIRE SPECIFIC FOR MELANOMA‐ASSOCIATED ANTIGENS
Authors: de Rooij M. , van der Meent M. , van der Steen D. , Remst D. , Hagedoorn R. , Wouters A. , Kester M. , van Veelen P. , Falkenburg F. , Heemskerk M.
Published: June 2019

Background:

Melanoma‐associated antigens (MAGE) are highly expressed in multiple myelomas as well as solid tumors, but silent in normal cells except for male germ line cells. This tumor‐restricted expression makes it an excellent target for adoptive T cell therapy. However, due to thymic selection high‐affinity T cells against self‐antigens presented in the context of self‐HLA are deleted from the T‐cell repertoire of a patient. To overcome this problem, we identify T‐cell receptors (TCRs) from the allogeneic‐HLA repertoire instead of the autologous‐HLA repertoire.

Aims:

To identify high‐affinity T‐cell receptors from the allogeneic‐HLA repertoire specific for melanoma‐associated antigens.

Methods:

In this study we selected MAGE peptides by peptide elution experiments from HLA class I and subsequent mass spectrometric analysis of tumor cells. Only peptides binding in the most frequently occurring HLA‐alleles were selected. From the identified peptides, HLA‐tetramers were formed to isolate peptide specific CD8+ T cells from healthy allogeneic donors. Avidity of the selected T‐cell clones was determined by stimulation assays including a broad range of MAGE expressing tumor cell lines. To examine off‐target recognition, T‐cell clones were stimulated with MAGE negative cell lines and EBV‐transformed B lymphoblastic cells expressing all prevalent HLA molecules.

Results:

MAGE peptides were eluted from multiple myelomas, EBV‐transformed lymphoblastic cells, acute myeloid leukemia and ovarium carcinomas. For MAGE‐A1, T‐cell clones specific for four different epitopes presented in the context of HLA‐A∗01:01, HLA‐A∗03:01, HLA‐B∗07:02 and HLA‐C∗07:02 were selected. For MAGE‐A3, T‐cell clones specific for one epitope presented in the context of HLA‐A∗01:01 and HLA‐B∗35:01 were identified. The selected T‐cell clones demonstrated efficient recognition of MAGE‐A1 or MAGE‐A3 positive multiple myeloma and solid tumor cell lines without detectable cross‐reactivity.

Summary/Conclusion:

MAGE‐A1 and MAGE‐A3 specific T‐cell clones were isolated from the allogeneic‐HLA repertoire of healthy individuals. The identified clones were activated upon exposure to MAGE‐A1 or MAGE‐A3 positive tumor cell lines and showed limited off‐target recognition. These newly identified TCRs could potentially be used for TCR‐gene therapy strategies targeting MAGE‐A1 or MAGE‐A3 positive tumors.
PF508 AUTOCHTHONOUS MOUSE MODELS OF MYD88 P.L265P ‐ AND BCL2‐DRIVEN DIFFUSE LARGE B CELL LYMPHOMA
Authors: Fluemann R. , Knittel G. , Nieper P. , Rehkämper T. , Pfeiffer P. , Holzem A. , Peifer M. , Büttner R. , Persigehl T. , Kashkar H. , Reinhardt C.
Published: June 2019

Background:

There are several recurrent mutations in DLBCL leading to the constitutive activation of NF‐kB, frequently by activating the BCR and TLR pathways (mostly via mutations in CD79B and MYD88, respectively), or by the loss of a negative feedback loop mediated by the loss of TNFAIP3.

Aims:

Here, we aimed to generate novel mouse models of human DLBCL to investigate the role of the most recurrent genetic alterations affecting MYD88, CD79B, TNFAIP3 and BCL2 in lymphomagenesis in vivo.

Methods:

We generated a novel mouse model, in which Cre‐mediated recombination leads to the conditional expression of Myd88 p.L252P (the orthologue of the human MYD88 p.L265P mutation) from the endogenous locus. We combined this Myd88 allele (abbr. M) with a BCL2 overexpression allele (abbr. B) and both alleles were activated B cell specifically with CD19:Cre (abbr. C). The effects of these mutations in vivo were carefully characterized. Additionally, we generated a conditional Cd79b p.Y195H allele and also obtained a previously published Tnfaip3 fl allele to model the loss of Tnfaip3 in DLBCL.

Results:

MBC animals showed elevated levels of self‐reactive antibodies. Immunization experiments revealed that Myd88 p.L252P and BCL2 overexpression cooperate in generating exaggerated antibody responses to exogenous antigen. Ultimately, these animals develop clonal tumors which resemble features of ABC DLBCL histologically and transcriptionally. They were exquisitely sensitive to inhibition of BCL2 in vitro and in vivo. MBC tumors expressed PD‐L1 at elevated levels compared to Eμ:Myc driven lesions. Anti‐PD1 antibody treatment had a stabilizing effect on tumor growth and resulted in a survival benefit. We successfully crossed our novel Cd79b p.Y195H allele into our MBC background to generate a mouse model of human DLBCL with oncogenic CD79B. Additionally, we generated a mouse model of TNFAIP3‐deficient lymhpoma by crossing the Tnfaip3 fl allele into our MBC background.

Summary/Conclusion:

Taken together, we established a Myd88 p.L252P driven mouse model for B cell lymphoma which shares many features with human ABC DLBCL. These tumors are sensitive to inhibition of BCL2 by ABT‐199 and respond to immune checkpoint blockade by anti‐PD1 treatment, suggesting these treatments as potential new strategies in the therapy of a subset of DLBCL patients. Additionally, we are currently investigating a novel conditional Cd79b p.Y195H allele and are modelling the loss of Tnfaip3 in DLBCL using a previously published allele.
PF521 CHOP TREATMENT ENHANCES DEPENDENCY ON BCL‐2, MCL‐1 AND BCL‐XL IN DIFFUSE LARGE B‐CELL LYMPHOMA
Authors: De Jong M. , Langendonk M. , Reitsma B. , Nijland M. , Huls G. , van den Berg A. , Ammatuna E. , Visser L. , van Meerten T.
Published: June 2019

Background:

Deregulation of anti‐apoptotic proteins BCL‐2, MCL‐1, BCL‐XL or BCL‐W is often a contributing factor to oncogenic transformation or progression, including in diffuse large B‐cell lymphoma (DLBCL). Gene expression profiling and immunohistochemistry analysis has shown overexpression of BCL‐2 in DLBCL is an important adverse prognostic factor, and has been correlated to resistance of standard therapy composed of cyclophosphamide, vincristine, doxorubicin, prednisolone and the anti‐CD20 antibody rituximab (R‐CHOP). Although these studies show a clear relation between BCL‐2 and therapy response, at the moment it remains unclear what the effect is of CHOP therapy on the intracellular anti‐apoptotic dependency in DLBCL.

Aims:

Here, we aim to determine the effect of CHOP treatment on the anti‐apoptotic proteins by employing BH3 profiling and investigate whether CHOP treatment enhances sensitivity for BH3 mimetic drugs.

Methods:

We first investigated the predictive value of anti‐apoptotic gene expression in 1800 DLBCL patients treated with CHOP or R‐CHOP. Next, dynamic BH3 profiles were acquired for 8 DLBCL cell lines treated with CHOP for 18 hours. Validation experiments were performed for CHOP combined with BH3 mimetic drugs on selected DLBCL cell lines. Finally, individual CHOP components were tested using BH3 profiling.

Results:

Gene expression analysis of anti‐apoptotic proteins revealed BCL‐2 had a significant negative association with survival in DLBCL patients, which was not observed for MCL‐1, BCL‐XL or BCL‐W. Analysis of anti‐apoptotic proteins expression in DLBCL cell lines demonstrated a strong adverse correlation for BCL‐2 protein expression and venetoclax (BCL‐2i) sensitivity, and a strong positive correlation for BCL‐2 protein expression and CHOP sensitivity. Dynamic BH3 profiling of CHOP‐treated DLBCL cell lines showed CHOP was able to enhance dependency on MCL‐1, BCL‐XL and BCL‐2. Validation experiments with BH3 mimetic drugs confirmed cell line specific enhanced sensitivity for S63845 (MCL‐1i), venetoclax (BCL‐2i) and/or navitoclax (BCL‐2/BCL‐XL/BCL‐Wi). No sensitivity‐related changes were observed on anti‐apoptotic protein expression after CHOP treatment. Dynamic BH3 analysis of single CHOP components revealed doxorubicin and vincristine were able to induce changes that were similar to the dynamic CHOP‐treated profiles.

Summary/Conclusion:

Gene expression analysis confirm that BCL‐2 status is a treatment‐independent predictive factor for DLBCL survival. However, our results demonstrate that CHOP treatment also induces cell‐specific increased anti‐apoptotic dependency on MCL‐1 or BCL‐XL, and not just BCL‐2. These results provide new perspective for the treatment of CHOP‐resistant DLBCL and underline the potential of BH3 profiling in predicting therapy outcome.

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PF548 PROGNOSTIC MUTATIONS IN PATIENTS WITH MYELODYSPLASTIC SYNDROME TREATED WITH HEMATOPOIETIC STEM‐CELL TRANSPLANTATION
Authors: Liu Z. , Cao S. , Lou F. , Sun Y. , Jiang E. , Han M.
Published: June 2019

Background:

Genetic mutations in MDS patients are closely related with clinical phenotypes and prognosis in MDS patients. But whether mutations are prognostic for outcomes after allogeneic hematopoietic stem‐cell transplantation(allo‐HSCT) remains to be elaborated.

Aims:

This research was done to figure out whether genetic mutations help the risk‐stratification in MDS patients received transplantation.

Methods:

Targeted mutational analysis were performed on samples obtained before transplantation from 151 patients underwent HSCT. Overall‐survival(OS) was calculated from the date of transplantation to date of death, and surviving patients were censored at the date on which they were last known to be alive. Diease‐free survival(DFS) was calculated from the time of transplantation to date of relapse or death and was censored at the last date known to alive without relapse. We analyzed the relationship of mutations and clinical outcomes. Curves for OS and DFS were generated using the Kaplan‐Meier method and compared using log‐rank test.Cumulative incidence of nonrelapse death and relapse was assessed using competing risks from time of HSCT to nonrelapse death or relapse by the Gray test.

Results:

All 151 patients carried more than one mutations, most frequently in DNAH2(47.02%), KDM6B(42.38%), USH2A(41.06%), KMT2D(38.41%), PCLO(33.11%), CCDC168(32.45%), U2AF1(29.14%), TET2(27.81%), ASXL1(25.83%), ARID1B(23.18%). In univariable analyses, TP53 or PPM1D mutations, ANKRD26 mutations, FLT3 abnormalities(including FLT3‐ITD), PDGFRB mutations were associated with shorter OS(TP53 or PPM1D, P = 0.064; ANKRD26, P = 0.025, FLT3 abnormalities: P = 0.046; PDGFRB, P = 0.044) and DFS(TP53 or PPM1D, P = 0.019; ANKRD26, P = 0.058, FLT3 abnormalities: P = 0.047; PDGFRB, P = 0.048). In multivariable analysis including clinical variables, monosomal karyotype status, HCT‐CI, and candidate genes, PDGFRB mutations and FLT3 abnormalities were each independently associated with shorter OS(PDGFRB mutations, HR = 2.979, P = 0.021; FLT3 abnormalities, HR = 2.677, P = 0.018) and DFS(PDGFRB mutations, HR = 2.779, P = 0.019; FLT3 abnormalities, HR = 2.288, P = 0.03). PDGFRB mutations(P = 0.042), ANKRD26 mutations (P = 0.004) were associated with higher non‐relapse mortality rate. FLT3 abnormalities was associated with higher relapse rate in RAEB patients(P = 0.004).

Summary/Conclusion:

PDGFR mutations and FLT3 abnormalities are independently associated with shorter survival in MDS patients treated with allo‐HSCT.
PF549 AUTOIMMUNE AND INFLAMMATORY DISEASES ASSOCIATE TO R‐IPSS SCORE BUT NOT TO OVERALL SURVIVAL OR LEUKEMIC EVOLUTION IN MYELODYSPLASTIC SYNDROMES
Authors: Brunet T. , Roy‐Peaud F. , Landron C. , Gallego‐Hernanz M.P. , Cayssials E. , Puyade M. , Torregrosa‐Díaz J.M.
Published: June 2019

Background:

Myelodysplastic syndromes (MDS) are clonal hemopathies with a relatively heterogeneous spectrum of presentation and a variable risk of transformation into acute myeloid leukemia (AML). Autoimmune diseases (AID) are present in 10 to 30% of MDS, with uncertain prognostic significance in this context. To date, no MDS‐related factor has been clearly identified as a predictor for the occurrence of AIDs.

Aims:

To study the association between MDS characteristics and the occurrence of an AID and its impact on overall survival and leukemia‐free survival.

Methods:

We retrospectively collected data from primary MDS adult patients followed‐up at the University Hospital of Poitiers between January 2010 and December 2017. Immunosuppressive therapy secondary‐MDS were excluded. The date of onset and type of AID as established by the standard criteria were collected. Steroid dependence was defined as a prednisone equivalent amount > 10 mg/day during at least 3 months. Simultaneous diagnosis of MDS and AID were considered if established in a 3‐months interval from the MDS one. Baseline characteristics at diagnosis of the MDS were registered and compared between those with and without AIDs.

Results:

AIDs were identified in 44 of 179 MDS patients (24.6%). MDS‐AID patients were more frequently male (29/44, p < 0.05). AIDs occurred before MDS in 63.6% of cases. Associated AIDs were: systemic vasculitis (23%), inflammatory rheumatism (18%), neutrophilic dermatosis (12.5%), connective tissue diseases (12.5%) and autoimmune cytopenia (11%). At the time of AID diagnosis, 48% of patients had general signs, 37.5% had cutaneous and/or joint signs. AID was steroid‐dependent in 39% of cases (9/23). No association between AID and MDS subtypes was observed. A significant linear tendency was found between the R‐IPSS and the presence of AID (p = 0.02): that is, the lower the IPSS‐R score, the higher the risk of association with AID. The presence of AID did not affect survival or progression to acute leukemia.

Summary/Conclusion:

The association of MDS to AIDs is frequent and of clinical heterogeneous expression. It occurs mainly in low‐risk MDS and does not seem to impact overall survival, that seems logical. The pathogenic mechanisms are complex and remain still completely unknown, but the hypothesis of a common pathogenic pathway seems strongly supported. The association MDS‐AID does not seem to impact overall survival in our cohort and occurs mainly in low‐risk MDS with a significant linear augmentation of the risk by the creasing categories of R‐IPSS. That raises the question of the role of different physio‐pathological processes underlying low‐risk and high‐risk MDS in the predisposition of AIDs. Larger prospective ongoing studies will better clarify our results.
PF639 IMMUNE ELECTRON MICROSCOPY AND MASS SPECTROMETRY FOR CLASSIFICATION OF AMYLOID DEPOSITS
Authors: Abildgaard N. , Rojek A.M. , Møller H.E. , Hansen C.T. , Palstrøm N.B. , Rasmussen L.M. , Beck H.C. , Marcussen N.
Published: June 2019

Background:

Amyloidosis is a shared name for a number of rare, complex and serious disease entities with an annual incidence in Denmark of about 2 per 100,000. Amyloidosis is caused by extra‐cellular deposits of amyloid substance created by accumulation of different misfolded proteins. Accurate characterization of the amyloid protein is essential. Amyloid light chain (AL) amyloidosis is a plasma cell dyscrasia that can be treated with chemotherapy, whereas transthyretin (ATTR) and secondary Amyloid A (AA) amyloidosis are other entities that are not treated within haematology. It is generally recognized that conventional immunohistochemistry has limitations, particular due to unspecific staining. Immune electron microscopy (IEM) and laser dissection microscopy followed by tandem mass spectrometry (LDM‐MS) are the new gold standards. It has not been established if one of the methods is superior; each method have some strengths and weaknesses. At Odense Amyloidosis Center, IEM and LDM‐MS have been established for standard diagnostics in 2017.

Aims:

To compare the sensitivity and specificity of IEM and MS for sybtyping amyloid deposits in different organs

Methods:

We performed a retrospective study of 106 Congo red positive biopsies from a number of different involved organs; heart, kidney, lung, gut mucosa, subcutaneous fat, bone marrow. IEM was performed with gold‐labeled antibodies against kappa, lambda, transthyretin and amyloid A.

Results:

IEM identified specific staining of amyloid fibrils in 91.6% of the biopsies; in 6 biopsies amyloid fibrils were not identified, and in 2 biopsies the subtype of the amyloid fibrils could not be established. MS identified an amyloigenic protein signature in 104/106 biopsies but in 9 of the biopsies the amyloid subtype could not be clearly identified. Protein subtype was established in 95 biopsies (89.6%). The concordance of the results ranged from 89.6–100% for the different subtypes. Importantly, the combined use of both methods increased the diagnostic sensitivity to 100%. Some variety in performances of the methods was observed on organ level.

Summary/Conclusion:

In conclusion, we demonstrate that both IEM and LDM‐MS are sensitive and specific methods for the identification of amyloidogenic proteins in amyloid deposits in biopsies from various organs.
PF650 AGE DOES NOT CONFER DIFFERENT FEATURES AND OUTCOMES IN PATIENTS WITH NEWLY DIAGNOSED ACTIVE MULTIPLE MYELOMA YOUNGER THAN 66 YEARS OF AGE TREATED IN A RECENT PERIOD
Authors: Bove V. , Ranero S. , Villano F. , Riva E.
Published: June 2019

Background:

Multiple myeloma (MM) is a disease of the elderly, with median age at diagnosis of 66 years. However, 10% of patients are younger than 50 years. Publications regarding age as prognostic factor for clinical characteristics at diagnosis and outcomes are heterogeneous, with some authors reporting aggressive presentation and worse outcomes in young patients and others showing better outcomes.

Aims:

To describe clinical and laboratory characteristics, response to treatment and survival in active newly‐diagnosed MM (NDMM) under 66 years. To compare outcomes between <51 and 51–65 years of age.

Methods:

Retrospective review of medical records of active MM diagnosed from jan2010 to dec2018 at Hospital de Clínicas, Montevideo, Uruguay. Smoldering MM and plasma cell leukemia were excluded. Characteristics at diagnosis were analyzed using descriptive statistics. Survival outcomes were calculated by Kaplan‐Meier method and compared using log‐rank test. In all cases, p < 0,05 was considered significant.

Results:

131 patients <66 years were included, 36 (27.5%) <51 years. Characteristics at diagnosis are shown in Table 1. Most patients were diagnosed at an advanced Durie‐Salmon and prognostic score (ISS) stages, showing no differences according to age. The mean monoclonal protein was lower in younger patients (p = 0,048). CRAB features and bone marrow infiltration were comparable. Bone plasmacytomas and extramedullary infiltration were more frequent in younger patients. Most used induction regimens were CyBorD and CTD in both groups. Rates of upfront autologous stem cell transplant (ASCT) consolidation were similar. Overall response rate (> = PR [partial remission]) was 77,4% in <51 and 79,9% in 51–65 years (p = 0,590) with comparable depth of response. Median progression free survival (PFS) was 25 months in <51 years and 26 months in 51–65 years (p = 0,683). Mortality rate was 33.3% in <51 years and 31.5% in older. The main causes of death were progression (53,2%) and infections 14,9%, similar in both groups. With a median follow up of 34 months, overall survival (OS) was 68,1%, median not reached in patients <51 years and 75 months in patients 51 – 65 years (p = 0,528).

Summary/Conclusion:

Clinical and laboratory features are similar in NDMM patients <50 and 51–65 years, although bone plasmacytoma and extramedullary disease were more frequent in the younger. However, this finding did not translate into differences in response or survival. Despite its efficacy and availability, upfront ASCT rate is low.

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Disclosures: no conflicts of interest to declare.

Funding: none.
PF651 POMALIDOMIDE, CYCLOPHOSPHAMIDE AND DEXAMETHASONE IN RELAPSED/REFRACTORY MULTIPLE MYELOMA PATIENTS IN A REAL LIFE SETTING: A SINGLE CENTER RETROSPECTIVE STUDY
Authors: Trudel S. , Tessoulin B. , Jullien M. , Blin N. , Gastinne T. , Mahé B. , Dubruille V. , Bonnet A. , Lok A. , Chevallier P. , Peterlin P. , Garnier A. , Guillaume T. , Le Bourgeois A. , Le Gouill S. , Moreau P. , Touzeau C.
Published: June 2019

Background:

Pomalidomide dexamethasone (Pd) is a standard of care for relapsed and refractory multiple myeloma (RRMM) patients who received at least two prior lines of therapy, including both lenalidomide and proteasome inhibitor (PI). In the phase 3 trial MM‐003, patients randomized in the Pd arm achieved an overall response rate (ORR) of 31% and a median progression‐free‐survival (PFS) of 4 months. Similar results were confirmed in the STRATUS trial (MM‐010). In order to improve these results, the triplet oral combination Pomalidomide Cyclophosphamide and Dexamethasone (PCD) has been investigated. To the best of our knowledge, we still need “real life” data evaluating the efficacy and safety of the triplet PCD combination in relapsed MM patients.

Aims:

To evaluate efficacy and safety of PCD in unselected RRMM patients outside clinical trials.

Methods:

All consecutive patients treated in our institution between January 2015 to July 2018 were included in this retrospective institutional review board approved analysis if they fulfilled the following criteria: (i) diagnosis of multiple myeloma according to International Myeloma Working Group (IMWG) consensus criteria (ii) relapsed and/or refractory to at least two priors therapies, including lenalidomide and PI and (iii) treated with pomalidomide, cyclophosphamide and dexamethasone. Response to treatment was determined according to IMWG criteria. Progression‐free‐survival (PFS) and OS were estimated using the Kaplan‐Meier method.

Results:

49 consecutive patients were included. The median age was 66 years, including eight patients (16%) above 75 years. Nine (18%) patients had high‐risk cytogenetic (t(4;14) or 17p deletion). The median number of prior therapies was 3 (range, 2–8) and all patients were previously exposed to proteasome inhibitors and lenalidomide. Thirty‐nine (80%) patients were refractory to lenalidomide. Twenty‐eight (57%) patients were refractory to PI. A double refractory status (lenalidomide and PI) was identified in 21 (43%) patients. The ORR was 76%, including 27% very good partial response or better. With a median follow‐up of 16 months, the median PFS was 7.3 months and the median OS was not reached. Regarding safety, most frequent toxicity was hematologic, including 37% grade 3–4 cytopenias. Deep venous thrombosis were reported in 4% of patients. Nine patients (18%) discontinued therapy due to adverse event.

Summary/Conclusion:

This real‐life study demonstrated that PCD is therefore a manageable, cost‐effective and oral triplet combination for unselected RRMM patients eligible to Pd.
PF666 LOW JAK2 V617F ALLELE BURDEN PHI‐NEGATIVE CHRONIC MYELOPROLIFERATIVE NEOPLASMS MAY ASSOCIATE WITH ADDITIONAL CALR OR MPL MUTATIONS
Authors: Makarik T. , Odilov A. , Abdullaev A. , Nikulina E. , Treglazova S. , Stepanova E. , Soubortseva I. , Kovrigina A. , Melikyan A. , Sudarikov A.
Published: June 2019

Background:

Jak2 (Janus kinase 2) V617F, CALR (Calreticulin) exon 9 and MPL (receptor for thrombopoietin) exon 10 mutations are associated with the vast majority of Phi‐negative chronic myeloproliferative neoplasms (MPNs). These mutations affect sequential stages of prolipherative signal transduction and therefore after the emergence of one type of mutation another types basically should not have any selective advantages for clonal expansion. However simultaneous findings of these mutations have been reported by different investigators in up to 10% of MPN cases.

Aims:

To analyze cases with combined mutations in Russian cohort of MPN patients

Methods:

Study includes 3267 samples, directed for genetic analysis to the National Research Center for Hematology between 2014 and 2019 from the patients with clinical evidence of MPN. DNAs and RNAs were extracted from blood using reagent kit from Interlabservice (Russia). Jak2 V617F mutation was quantitated by real‐time PCR kit from Syntol (Russia) according to manufacturers instructions. CALR exon 9 deletions/insertions were analyzed by fragment analysis (sensitivity > = 3%). MPL W515L/K mutations were assessed by in‐house allele specific PCR. All cases were tested for phi‐negativity using BCR‐ABl p210 PCR kit from Interlabservice (Russia).

Results:

In 406 of 3267 cases (12.5%) CALR mutations were detected. In 24 of these 406 cases (6%) Jak2 V617F mutation was found in addition to CALR mutation. Matched measures were obtained at least twice at different time points during the course of disease for these cases. In 19 from 24 (80%) cases with combined CALR and Jak2 mutations V617F allele burden was lower than 3%. Also combination of Jak2 V617F with MPL W515L/K was observed in 3 out of 3267 cases only. Two of them had V617F allele burden lower than 3% (Tabl.1).

Summary/Conclusion:

Additional mutations may coexist over low background of JAK2 V617F allele. Therefore cases of MPNs with low initial JAK2 V617F allele should be further tested for CALR (and possibly MPL) mutations to ensure correct disease and treatment monitoring. Further dynamic observation studies are required to assess the value of additional mutations for MPN disease progression and treatment outcome.

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PF685 THE RELATIONSHIP BETWEEN TOTAL ANTIOXIDANT CAPACITY AND JAK2V617F MUTATION IN ESSENTIAL THROMBOCYTHEMIA
Authors: Gaman M.‐A. , Moisa C. , Gaman A.M.
Published: June 2019

Background:

Essential thrombocythemia (ET) is a BCR‐ABL1‐negative myeloproliferative neoplasm associated in 50–60% of cases with the presence of the JAK2V617F mutation. Several studies have revealed that oxidative stress induces a growth advantage to JAK2V617F‐ positive clones, but little is known about the antioxidant status of ET patients.

Essential thrombocythemia (ET) is a BCR‐ABL1‐negative myeloproliferative neoplasm associated in 50–60% of cases with the presence of the JAK2V617F mutation. Several studies have revealed that oxidative stress induces a growth advantage to JAK2V617F‐ positive clones, but little is known about the antioxidant status of ET patients.

Aims:

To evaluate the total antioxidant capacity (TAC) in patients with ET and to investigate if there are any differences between JAK2V617F‐positive cases and JAK2V617F‐negative ET cases in terms of TAC.

Methods:

We evaluated patients diagnosed with ET according to the WHO criteria (2016 revised) vs. healthy subjects (control group). Patients were classified based on age, sex and JAK2V617F mutational status. Haematological and biochemical parameters, as well as bone marrow aspirate or biopsy, were analyzed. Detection of JAK2V617F mutation was performed by RT‐PCR. TAC was measured using a multi‐detection microplate reader FLUOstar Omega (reagents from Sigma‐Aldrich). Data analysis was conducted using the student T‐test and a p‐value ≤ 0.05 was considered statistically significant. The study was approved by the Ethics Committee of the University of Medicine and Pharmacy of Craiova, Romania, approval no. 79/23.02.2017. All subjects agreed to partake in the study and signed a written informed consent.

Results:

The study group involved 58 ET patients (mean age 59.72 ± 14.63 years, age range 22–82): 25 males (40.32%, mean age 55.77 ± 15.80 years, age range 22–82) and 37 females (59.68%, mean age 62.14 ± 13.30 years, age range 23–81). 36 patients (62.07%) were JAK2V617F‐positive and 22 patients (37.93%) were JAK2V617F‐negative. 7 patients (12.07%) had homozygous genotype and 29 patients (50.00%) had a heterozygous genotype. TAC was decreased in patients with ET compared with healthy subjects and in JAK2V617F‐positive vs. JAK2V617F‐negative cases (p < 0.05). We found no significant differences between sexes in terms of age (men = 55.77 ± 15.80 years vs. women = 62.14 ± 13.30 years, p = 0.1116) or TAC (men = 0.50 ± 0.04 mM/L vs. women = 0.49 ± 0.06 mM/L, p = 0.3043). Also, we did not record significant differences in terms of TAC between JAK2V617F homozygous genotype vs. JAK2V617F heterozygous genotype (0.48 ± 0.05 mM/L vs. 0.49 ± 0.06 mM/L, p = 0.1686).

Summary/Conclusion:

In patients diagnosed with ET, TAC is decreased compared to normal individuals. Also, the decrease in TAC is more expressed in JAK2V617F‐positive cases vs. JAK2V617F‐negative cases or controls. No significant differences in terms of TAC were found between sexes or JAK2V617F homozygous vs. heterozygous genotype.
PF698 LUSUTROMBOPAG RELIABLY INCREASES PLATELETS REGARDLESS OF BASELINE PLATELET COUNTS IN THROMBOCYTOPENIC CHRONIC LIVER DISEASE PATIENTS UNDERGOING PLANNED INVASIVE PROCEDURES: RESULTS OF 2 PHASE 3 TRIALS
Authors: Brown R.S. , Imawari M. , Izumi N. , Osaki Y. , Bentley R. , Baykal T. , Ochiai T. , Kano T.
Published: June 2019

Background:

Lusutrombopag is an oral, small molecule thrombopoietin receptor agonist approved in Japan and US for improvement of thrombocytopenia, and in EU for severe thrombocytonia, associated with chronic liver disease in patients undergoing planned invasive procedures. The degree of thrombocytopenia may impact the treatment response to lusutrombopag and need for platelet transfusion.

Aims:

This analysis evaluates the increase in platelet count and maximum platelet count achieved according to baseline platelet count.

Methods:

L‐PLUS 1 and L‐PLUS 2 were two similar phase 3 multicenter, randomized, double‐blind, placebo‐controlled studies; L‐PLUS 1 (JapicCTI‐132323) was conducted in Japan and L–PLUS 2 (NCT02389621) was conducted globally. Patients with thrombocytopenia due to chronic liver disease received lusutrombopag 3 mg or placebo for up to 7 days prior to an invasive procedure scheduled 9–14 days after randomization. Patients were adults with platelet count (x109/L) <50 at baseline. Platelet transfusion was mandated if the platelet count remained <50 no more than two days prior to the planned invasive procedure. The change in platelet count over time was a key efficacy endpoint. A per‐protocol population from the two trials was integrated for this analysis; post‐platelet transfusion platelet counts were excluded from the analysis when calculating the proportion of patients who met a criterion and summary statistics for maximum platelet counts.

Results:

312 patients were randomized. Of the 270 patients in the per‐protocol population, 78.1% of patients in the lusutrombopag group (n = 137) achieved a platelet count ≥50 at least once during the study versus 30.1% of patients in the placebo group (n = 133). The platelet count increased by double or more in 47.4% and 3.8% in the lusutrombopag and placebo groups, respectively, and increased by 50% or more in 77.4% and 12.8%, respectively. The average maximum platelet count in the lusutrombopag group stratified according to baseline platelet count of <20, ≥20‐<30, ≥30‐<35, ≥35‐<40, and ≥40‐<50 was 56, 45, 64, 82, and 87, respectively (Figure). The average maximum change in platelet count in these strata were +41, 21, 32, 45, and 42, respectively.

Summary/Conclusion:

Patients on lusutrombopag experienced a clinically meaningful response in platelet count regardless of whether the baseline platelet count was above or below 35x109/L, thereby avoiding the need for use of platelet transfusion.

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