bluuebunny/crossref_metadata_2025_split huggingface dataset:
PF512 NEXT‐GENERATION SEQUENCING‐BASED CLONALITY ASSESSMENT OF IMMUNOGLOBULIN GENE REARRANGEMENTS DISTINGUISHES RELAPSE FROM SECOND PRIMARY CLASSICAL HODGKIN LYMPHOMA
Authors: van Bladel D. , van den Brand M. , Rijntjes J. , Brinkman A. , Reigl T. , Darzentas N. , Hess C. , Hebeda K. , Groenen P. , van Krieken H. , Scheijen B. 
Published: June 2019

Background:Classic Hodgkin's lymphoma (cHL) has a high cure rate with current therapy, however relapse still occurs in 5%>10% of early stage and 15%>30% of advanced cHL. Interestingly, case reports and small series have shown that some of these relapses appear to be a new primary cHL. Assessment of clonal relationship between primary diagnosis and relapsed cHL samples after treatment will distinguish true relapse from second primary lymphoma. Clonality detection in cHL using conventional clonality assays has thus far been severely hampered by the low frequency of Hodgkin and Reed‐Sternberg (HRS) cells and limited DNA quality obtained from formalin‐fixed paraffin‐embedded (FFPE) material. Within the EuroClonality‐NGS Working Group, we have developed a novel approach to detect immunoglobulin (IG) heavy chain (IGH) and k light chain (IGK) gene rearrangements.Aims:The objective or our study is to determine the clonal relationship between HL diagnosis and relapse samples to assess the incidence of second primary malignancies in HL recurrences.Methods:We have collected 70 paired primary and relapse cHL cases. In 38/70 cases relapse occurred within three years after diagnosis, in 32/70 cases relapse occurred later than three years after diagnosis. EuroClonality‐NGS gene‐specific IGH‐VJ‐FR3, IGH‐DJ, IGK‐VJ and Intron‐Kde primer sets for small amplicon sizes were used to perform next‐generation sequencing (NGS)‐based clonality analysis with Ion Torrent PGM. Bioinformatic analyses with the interactive web‐based immunoprofiler ARResT/Interrogate allowed run/sample quality control followed by accurate identification of clonotypes.Results:We show that NGS‐based clonality assessment is highly suitable for detecting IG rearrangements in FFPE tissue specimens, including HL tissue specimens. Our preliminary data analysis of 7 paired diagnosis and relapse samples demonstrates the presence of identical clonotypes in 2 cases, while distinct clonotypes were observed in 3 other cases suggesting a new primary lymphoma. In the remaining 2 cases no specific clonotype could be identified in either diagnosis and/or relapse. The analysis of additional cHL recurrences by NGS‐based clonality assessment will reveal the true incidence of clonally unrelated lymphoma in relapsed cHL.Summary/Conclusion:This study is an important step towards establishment of NGS‐based clonality assessment in clinical practice for cHL, and eventually the improvement of therapeutic management of second primary cHL.
PF545 EASIX FOR PREDICTION OF SURVIVAL IN LOWER RISK MYELODYSPLASTIC SYNDROMES
Authors: Merz A.M. A. , Germing U. , Kobbe G. , Jauch A. , Radujkovic A. , Hummel M. , Benner A. , Merz M. , Müller‐Tidow C. , Dreger P. , Luft T. 
Published: June 2019

Background:Patients with myelodysplastic syndromes (MDS) are at risk of early death from cardiovascular complications due to the link between clonal hematopoiesis and endothelial dysfunction. EASIX (Endothelial Activation and Stress Index) is a novel marker of endothelial dysfunction that is calculated from serum creatinine, lactate dehydrogenase levels, and thrombocyte counts. EASIX has been established to predict endothelial complications after allogeneic transplantation.Aims:To analyze the impact of EASIX on the survival of patients with lower risk MDS and correlation with markers of endothelial activation.Methods:We investigated the impact of EASIX measured at first diagnosis on survival of lower risk MDS patients who received supportive treatment in two independent institutions: n = 100 (cohort I) and n = 338 (cohort II). Serum Angiopoietin‐2, suppressor of tumorigenicity (ST)‐2, soluble thrombomodulin (sCD141), interleukin (IL)‐18, IL37, S100A9, and HMGB1 were measured as markers of endothelial cell distress and inflammasomal activity and correlated to EASIX.Results:Univariate analysis showed that high EASIX was associated with shorter survival in both cohorts (Cohort I: hazard ratio (HR) per log2 increase: 1.50; 95% confidence interval (CI) 1.27–1.79; p‐value < 0.001 / Cohort II: HR 1.31 [1.20–1.42]; p‐value < 0.001). We visualized this continuous effect of EASIX by grouping patients into quartiles according to EASIX (Figure 1 A and B). Multivariate Cox regression analysis confirmed that EASIX remains a significant predictor of survival even after adjustment for age, sex, cytogenetic abnormalities and bone marrow blast counts (Cohort I: HR 1.35 [1.12–1.63]; p‐value = 0.002 / Cohort II HR 1.18 [1.08–1.29]; p‐value <0.001]. Models including EASIX showed lower prediction errors over time. Serum levels of Angiopoietin‐2 correlated significantly with EASIX (Spearman's rank correlation = 0.455, 95%CI 0.264–0.616, n = 74). Due to the lack of correlation between EASIX and S100A9 or IL‐1b, we performed a pathway analysis including serum HMGB1 and IL37. We observed a strong intra‐pathway correlation of S100A9 with IL1b, IL18, IL37, and HMGB1, arguing for the validity of our serum analysesSummary/Conclusion:We introduce EASIX as an easily accessible predictor for survival in patients with low‐risk MDS in best supportive care.image
PF574 TARGETED ULTRA‐DEEP NGS ASSESSMENT OF TP53 MUTATIONS IN MULTIPLE MYELOMA FROM WHOLE BONE MARROW
Authors: Petrackova A. , Sedlarikova L. , Minarik J. , Vasinek M. , Dyskova T. , Urbankova H. , Bacovsky J. , Papajik T. , Kriegova E. 
Published: June 2019

Background:Among the most important prognostic genetic aberrations in multiple myeloma (MM) are del(17p) and/or TP53 mutations. The use of the whole bone marrow (BM), when sorted MM cells are not available, may greatly simplify the NGS assessment in MM patients for clinical decisions.Aims:Therefore, the objective of our study was to assess applicability of ultra‐deep targeted NGS for analysis of TP53 mutations and other commonly mutated genes NRAS, KRAS and BRAF using the whole BM samples and compare it to results from MM sorted cells.Methods:BM samples were obtained from MM patients (n = 35, 12 M/23 F). The gDNA was isolated from whole BM (infiltration of plasma cells: median 16.4%, min‐max 1.2 ‐ 65.6%) and paired sorted MM cells. The full coding sequence and 5́and 3́UTR regions of the TP53 gene and hotspots of NRAS, KRAS and BRAF were analysed using ultra‐deep NGS with the MiSeq system (Illumina). The cut‐off for mutation detection was 1% variant allele frequency (VAF), the binomial distribution equation was used to determine a minimum number of mutant alleles.Results:Of MM patients, 22.9% (8/35) harboured mutations in TP53 gene, 20.0% (7/35) KRAS, 14.3% (5/35) NRAS and 8.6% (3/35) BRAF in the whole BM. In all patients, the VAF was in accordance with plasma cells infiltration in BM. Regarding TP53, five patients carried only TP53 mutations, two had a co‐occurrence of TP53 mutations and del(17p) and one had TP53 mutation together with 17chr trisomy as assessed by FISH. In TP53, all of the mutations except one case were observed with low frequency (VAF 2–9%). In cases when paired sorted sample was available, the mutations found in the whole BM were confirmed in sorted MM cells.Summary/Conclusion:Our results demonstrate the applicability of ultra‐deep targeted NGS in assessment of TP53 mutations in the whole BM samples in MM when sorted population is not available. Moreover, our data highlights the importance of assessment of TP53 mutations in MM patients, as they may occur regardless of del(17p). Our approach may simplify the assessment of mutational landscape in MM patients and thus impacts clinical decision making.Grant support: MZ ČR VES16–32339A, IGA UP_2019_014, MH CZ–DRO (FNOL, 00098892), NV18–03‐00500
PF598 STEM CELL YIELD AND TRANSPLANTATION IN TRANSPLANT‐ELIGIBLE NEWLY DIAGNOSED MULTIPLE MYELOMA PATIENTS RECEIVING DARATUMUMAB + BORTEZOMIB/THALIDOMIDE/DEXAMETHASONE (D‐VTD): PHASE 3 CASSIOPEIA STUDY
Authors: Hulin C. , Moreau P. , Attal M. , Belhadj K. , Benboubker L. , Caillot D. , Facon T. , Garderet L. , Kuhnowski F. , Stoppa A.‐M. , Kolb B. , Tiab M. , Sonneveld P. , Jie K.‐S. , Westerman M. , Pei L. , Kampfenkel T. , de Boer C. , Vermeulen J. , van de Donk N.W. 
Published: June 2019

Background:High‐dose therapy (HDT) followed by autologous stem‐cell transplantation (ASCT) is the standard of care in transplant‐eligible newly diagnosed multiple myeloma (NDMM). In the phase 3 CASSIOPEIA study, daratumumab + standard‐of‐care regimen VTd (D‐VTd) significantly improved stringent complete response (sCR), complete response or better (≥CR), and minimal residual disease (MRD)‐negative rates and reduced the risk of progression or death versus VTd in NDMM patients who were eligible for transplant.Aims:Here, we assessed stem cell yield and transplantation results among patients receiving D‐VTd versus VTd induction prior to HDT and ASCT in Part 1 of the CASSIOPEIA trial.Methods:In Part 1, transplant‐eligible NDMM patients ages 18–65 years were randomized 1:1 to 4 pre‐transplant induction and 2 post‐transplant consolidation cycles of D‐VTd or VTd alone. After induction, patients underwent stem cell mobilization with cyclophosphamide 3 g/m2 (recommended dose) and granulocyte colony‐stimulating factor (G‐CSF). Peripheral blood stem cells were harvested based on response to mobilization. Plerixafor was administered if stem cell collection failed at first attempt and in accordance with institutional practice. Melphalan 200 mg/m2 IV was given as HDT prior to ASCT.Results:A total of 1,085 patients were randomized to D‐VTd (n = 543) or VTd (n = 542). Among patients who completed mobilization (D‐VTd, 506; VTd, 492), more patients in the D‐VTd arm received plerixafor during mobilization than in the VTd arm (21.7% vs 7.9%). Patients underwent a median (range) of 2 (1–6) versus 1 (1–4) days of apheresis for D‐VTd versus VTd. The median number of CD34+ cells collected was lower in patients receiving D‐VTd versus VTd (6.3 × 106/kg vs 8.9 × 106/kg). Nevertheless, a similar percentage of intention‐to‐treat patients receiving D‐VTd versus VTd underwent ASCT (90.1% vs 89.3%). The median number of CD34+ cells transplanted for D‐VTd versus VTd was 3.3 × 106/kg versus 4.3 × 106/kg. Hematopoietic reconstitution rates were high and similar in transplanted patients receiving D‐VTd versus VTd (99.8% vs 99.6%). For D‐VTd versus VTd, a median (range) of 13.0 (6–54) versus 13.0 (4–43) days was required to achieve sustained absolute neutrophil counts >500 cells/mm3, and a median (range) of 14.0 (2–56) versus 12.0 (1–47) days was required to achieve sustained platelets >20,000 cells/mm3 without transfusion.Summary/Conclusion:Stem cell mobilization and collection was feasible with D‐VTd induction. Adding daratumumab to VTd allowed successful transplantation in patients with NDMM who were transplant eligible.
PF654 EPIGENETIC ALTERATION OF CSK BINDING PROTEIN(CBP) IN MYELOPROLIFERATIVE NEOPLASMS COMPLEMENTS THE JAK2V617F MUTATION
Authors: Huang J. , Ye X. 
Published: June 2019

Background:Csk binding protein (CBP), a ubiquitously expressed transmembrane protein, functions as a suppressor of Src‐mediated tumor progression by promoting the inactivation of Src. CBP is widely reported to act as a scaffold in the Csk‐mediated negative regulationof Src family kinases (SFKs). First, CBP is phosphorylated by SFKs, then it associates withC‐terminal Src kinase (Csk) through a specific site (Tyr‐317 in humans) and brings it into proximity with membrane‐associated SFKs. After that, Csk phosphorylates the C‐terminal negative regulatory tyrosine residue of SFKs, which suppresses their activation. SFKs are membrane‐associated non‐receptor protein tyrosine kinases that play pivotal roles in regulating various cellular processes including proliferation, differentiation, adhesion, migration, and survival. Thus, CBP plays the opposite role in various Csk‐mediated cellular processes and might be a significant target for Csk‐mediated tumors.Aims:To explore the role of CBP in myeloproliferative neoplasms complements the JAK2V617F mutation.Methods:Bone marrow of 78 JAK2/V617F mutation positive MPN patients were collected, including 18 of 78 idiopathic myelofibrosis, 32 of 78 polycythaemia vera and 28 of 78 essential thrombocythaemia. The mononucleated cells separated, RNA extracted and the CBP expression level detected using real‐time quantitative PCR and MSP. SPSS 23.0 used for data analysis.Results:We identified CBP as a functional tumor suppressor gene frequently methylated in myeloproliferative neoplasms complements the JAK2V617F mutation. We further uncovered CBP as one of the up regulated genes in JAK2/V617F mutation positive cell lines after pharmacologic demethylation with 5 aza 2’ deoxycytidine (Aza). Methylation of CBP was detected in several JAK2/V617F mutation positive cells. Aberrant methylation was further detected in 60% of various types of JAK2/V617F mutation positive MPN, but not in any normal PBMC sample, and is thus tumor specific. The levels of CBP mRNA were significantly lower in hypermethylated samples than unmethylated samples, suggesting CBP may be epigenetically inactivated in MPN patients. Aza treatment led to CBP promoter demethylation and transcriptional reactivation in silenced cell lines, indicating a methylation mediated silencing. Meanwhile, phosphorylated JAK2 and STAT3 were progressively reduced.Summary/Conclusion:Our results indicate that epigenetic inactivation of CBP contributes to the constitutiveactivation of JAK2/STAT signaling. CBP methylation is a frequent event in myeloproliferative neoplasms complements the JAK2V617F mutation. Restoration of CBP expression by HMA may contribute to clinical treatment for MPN patients.
PF657 USE OF A NGS MYELOID PANEL TO FIND CLONALITY IN TRIPLE NEGATIVE ESSENTIAL THROMBOCYTHEMIA. IDENTIFICATION OF NEW MUTATIONS IN DRIVER GENES
Authors: Carreno‐Tarragona G. , López N. , Gutiérrez López de Ocariz X. , Rapado I. , Martínez‐López J. , Ayala R. 
Published: June 2019

Background:Essential Thrombocythemia is a clonal haematopoietic stem cell disorder characterized by hypercellularity of the bone marrow with effective haematopoietic maturation and increased numbers of platelets in the peripheral blood. Although more than 90% of the cases present typical mutations in JAK2, MPL and CALR, the remaining cases can become a diagnostic challenge as it requires the presence of a clonal marker or the absence of evidence for reactive thrombocytosis.Aims:To evaluate the use of a Myeloid Panel for the identification of clonal markers in TN ET cases and try to identify new mutations and prognostic determinants in these patients.Methods:A cohort of 32 ET negative for JAK2 V617F mutation (RT‐PCR), CALR canonic mutations (GENESCAN) and MPL W515K and W515L mutations (RT‐PCR) was selected. Average age at diagnosis was 49,08 (range 14–88). Male:Female ratio was 11:21. Average Haemoglobin, leukocytes and platelets at diagnosis were 14.14 g/dl 8,93 x109/L and 820x109/L, respectively. We performed targeted gene sequencing by NGS (Ion Torrent S5XL System–Thermofisher Scientific) using a panel of 33 (before March 2018) or 43 genes (after March 2018) implicated in leukemia prognosis.Results:With a median follow‐up of 5 years (range 0,02–25,43) 2 patients suffered transformation to acute myeloid leukemia, 1 progression to myelofibrosis and 1 presented with suffered a thrombotic event after diagnosis. We found clonality markers in 19 (59%) out of 32 patients. These presented an average of 1,84 (range 1–3) and 1,68 (range 1–3) mutations and mutated genes, respectively. 6 patients (31,57%) presented mutations in driver genes (JAK2, MPL and CALR). Three of them, family, presented with JAK2 R683G, a mutation previously described in acute lymphoblastic leukemia but not in myeloproliferative neoplasms. Other patient had JAK2 L393 V mutation, of undetermined significance and reported in population databases. Another presented the previously described MPL S505C and W515R mutations. Finally, we identified a new deletion in CALR exon 9 (c.1153_1155delAAG, p.K385del). 17 (89,47%) mutated patients presented mutations in non driver genes. Among those the most frequently mutated ones were TET2 (29%), ETV6 (17,6%), SF3B1 and KMT2A (both 11,7%). We found one mutation in VHL, DNMT3A, WT1, TP53, EPAS, CBL, IDH1, TPHO, KIT, ASXL1, EPOR and KRAS.Summary/Conclusion:Triple Negative TE suppose a diagnostic challenge. Here we show that targeted gene sequencing by a myeloid panel is able to identify clonality in up to 59% of the cases. Also, we have described new mutations in driver genes, gaining knowledge on TE molecular basis. Finally, larger cohorts of TN ETs are needed to clearly define the determinants and prognostic factors of this disease.
PF715 THE IMPACT OF CRIZANLIZUMAB ON VOC‐RELATED MEDICAL FACILITY VISITS
Authors: Bailey M. , Thompson M. , Brown S. 
Published: June 2019

Background:A major cause of morbidity among patients with sickle cell disease (SCD) is the intense, recurrent, unpredictable attacks, known as vaso‐occlusive crises (VOCs), which are responsible for >90% of hospital admissions in SCD. An important goal of pharmacological therapy is to prevent the occurrence of these painful episodes, and in turn, reduce the clinical and economic burden of the disease.In December 2018, the United States Food and Drug Administration granted crizanlizumab (SEG101) Breakthrough Therapy designation for the prevention of VOCs in patients with SCD. This designation was granted based on positive results from the Phase II SUSTAIN trial, in which 5 mg/kg crizanlizumab was associated with a 45.3% reduction in the median annual rate of VOCs leading to healthcare visits compared with placebo.Aims:The aim of the current analysis was to investigate the impact of crizanlizumab on VOC‐related medical facility visits.Methods:sing data from the SUSTAIN trial, analyses were conducted to determine the cumulative number of VOC events across all patients, medical facility visits and by medical facility type in the 5 mg/kg crizanlizumab and placebo arms. The total cumulative number of patient days in the study was considered in determining the event rate per year, which was calculated as follows:“[Total cumulative number of VOCs across all patients]/[Total number of days observed across all patients]” ∗ 365Incidence rate ratios (IRR) were derived using a Poisson regression model. Maximum likelihood parameter estimates were generated with standard errors to produce confidence intervals and p‐values. Effect modification over time by group was assessed by maximum likelihood.Results:The analyses showed a reduction in the overall rate of VOC events requiring a medical facility visit in the 5 mg/kg crizanlizumab group compared with the placebo group. Further examination of the impact of crizanlizumab on specific sites of care demonstrated a reduction in visits to outpatient care sites, largely driven by reduced visits to emergency care units and SCD crisis centers (Table 1). A trend to reduced inpatient admissions was also observed.Summary/Conclusion:Payers aim to reimburse treatments for SCD that reduce both the clinical and economic burden of the disease. In the SUSTAIN trial, patients treated with 5 mg/kg crizanlizumab experienced a 45.3% reduction in the median annual rate of VOCs leading to healthcare visits compared with those receiving placebo. The current analysis demonstrates that this reduction in VOCs leading to a healthcare visit was seen in most medical facility visits, largely driven by reduced visits to emergency care units and specialized SCD crisis centers. These findings suggest that patients treated with crizanlizumab will experience a substantial reduction in health care resource utilization and associated costs.image
PF716 CLINICAL IMPACT OF PROGNOSTIC NUTRITIONAL INDEX IN DIFFUSE LARGE B CELL LYMPHOMA
Authors: Go S.‐I. , Lee G.‐W. 
Published: June 2019

Please indicate where the abstract has been published before: Annals of HematologyBackground:Cachexia is an indicator of tumor progression in patients with malignancy. Cachectic patients exhibit intolerance and a reduced response to antitumor therapy, and have an unfavorable prognosis. An international consensus proposed diagnostic criteria for cancer cachexia including weight loss, low body mass index (BMI), and/or presence of sarcopenia. Other potential biomarkers (e.g., albumin, C‐reactive protein, pro‐inflammatory cytokines, and microRNAs) and scoring systems (e.g., cachexia score [CASCO], Glasgow prognostic score [GPS], prognostic nutritional index [PNI]) have also been studied to diagnose cancer cachexia and assess its severity.Aims:In this study, we assessed the associations among PNI, other clinical outcomes, and survival in patients with DLBCL. Then we sought valid cachexia markers with prognostic significance and designed a novel model to improve the prognostic ability of well‐known clinical indices for patients with DLBCL.Methods:A total of 228 DLBCL patients treated with first‐line R‐CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone) were retrospectively reviewed. PNI was calculated as 10 × serum levels of albumin (g/dL) + 0.005 × absolute lymphocyte count (/mm3). Patients were categorized into low‐ and high‐PNI groups based on a cut‐off value of 40. The nomogram for predicting overall survival (OS) was constructed using a Cox regression model. PNI was positively correlated with skeletal muscle index, body mass index, and serum levels of albumin.Results:The low‐PNI group had a lower complete response rate (60.3% vs. 87.6%), increased treatment‐related toxicity, and more frequent treatment discontinuation (43.5% vs. 8.8%) than the high‐PNI group. The median OS was shorter in the low‐PNI group than the high‐PNI group (15.6 months vs. not reached; p < 0.001, Figure.1). Multivariate Cox regression analyses showed that PNI, sarcopenia, and the international prognostic index (IPI) were independent prognostic factors for OS. The nomogram developed using this regression model showed excellent discriminatory ability for predicting OS (c‐index, 0.80) compared to the IPI alone (c‐index, 0.75).Summary/Conclusion:This study supports the results of a previous study [29], whereby low PNI is an independent poor prognostic factor in patients with DLBCL treated with first‐line R‐CHOP immunochemotherapy. In addition, we newly found that treatment‐related toxicity and early discontinuation of treatment occurred more frequently in DLBCL patients with low PNI. We anticipate that the new prognostic nomogram developed using PNI, SMI, and IPI may help to realize individualized therapy that considers both the disease characteristics and the patient's tolerance to treatment in patients with DLBCL. This will be validated externally in a future study.image