bluuebunny/crossref_metadata_2025_split huggingface dataset:
PS1120 IRON MOBILIZATION IN A REAL LIFE COHORT OF APLASTIC ANEMIA PATIENTS TREATED WITH ELTROMBOPAG
Authors: Cavallaro F. , Fattizzo B. , Levati G. , Milesi G. , Barcellini W. 
Published: June 2019

Background:Most aplastic anemia (AA) patients are heavily transfusion dependent of both platelets and red blood cells (RBC), and iron overload may become an issue. Excess iron not only hampers the function of several systems (liver, heart, kidney, endocrine system), but has also a negative effect on normal hematopoiesis, which is already particularly poor in AA patients. TPO mimetic eltrombopag (EPAG) improves hematopoiesis in about 40% of patients with both relapsed/refractory and newly‐diagnosed AA, and recent reports demonstrated iron‐mobilizing activity in vitro; furthermore, harmonic oscillation of serum ferritin concomitant to EPAG initiation and discontinuation have been also observed in vivo, indicating an iron mobilization effectAims:To evaluate iron mobilization mediated by EPAG in a real life cohort of AA patientsMethods:Clinical and hematologic parameters, including iron panel, of 10 AA patients treated with EPAG at our Institution in the last 18 months were collected. Changes in iron status and their relationship with EPAG treatment were systematically recordedResults:Table 1 shows hematologic parameters and iron status at baseline and after EPAG treatment. Median age was 43 years (23–91), and male to female ratio 1:1. At the time of EPAG initiation 6 cases presented with severe AA and 60% were RBC transfusion dependent. Three patients were therapy naïve and received EPAG together with frontline IST, starting at 150 mg/day since day+14; seven cases were relapsed/refractory ones, previously treated with ATG and cyclosporine (2), cyclosporine alone (2), or steroids only (2). As regards iron status, ferritin levels and plasma iron increased from EPAG start to month +1 and +3 (p = 0.04), and transferrin saturation was persistently augmented versus baseline at month +1,+3, and +6 (p = 0.01, p = 0.03, P = 0.04, respectively). Considering non transfused patients, we observed a similar increase of these parameters after EPAG start, although baseline values were lower than in transfusion‐dependent cases. At month +12 an amelioration of iron status was observed, both in transfusion dependent and not, particularly iron and transferring saturation that did not further increase. Notably, iron chelation was performed in 2 transfused cases only, possibly accounting for this improvement. Considering hematologic parameters, Hb, ANC, and PLT values significantly ameliorated from baseline to month +3, and along the follow up. Mean Hb increase from baseline was 0.7 ± 1.6 g/dL, mean ANC increase 0.3 ± 0.48x109/L, and mean PLT 27 ± 38x109/L. Transfusion requirement was progressively lost, and ORR increased from 56% at month+3 to 100% at month +6 and thereafter (although CR was observed in 1 patient only)Summary/Conclusion:In our real life cohort of AA patients we observed an increase of iron and transferrin saturation levels soon after EPAG initiation, irrespective of the transfusion status. This data support the iron‐mobilizing effect of EPAGimage
PS1132 INHERITED DNA REPAIR AND CELL CYCLE GENE DEFECTS IN CHRONIC LYMPHOCYTIC LEUKEMIA
Authors: Moore N.S. , Aldubayan S.H. , Taylor‐Weiner A. , Stilgenbauer S. , Getz G. , Wu C.J. , Van Allen E.M. , Brown J.R. 
Published: June 2019

Background:Chronic lymphocytic leukemia (CLL) is among the most heritable cancers, with 60% of disease risk genetically determined. However, most of the genetic heritability of CLL remains unexplained. Previously, we identified ATM as the first CLL risk gene.Aims:Here we leverage a deep‐learning‐based germline variant calling algorithm to explore germline mutational enrichment in DNA repair and cell cycle genes in CLL.Methods:A two‐stage case‐control analysis was conducted using gene‐based mutational enrichment analysis of 50 established cancer predisposition DNA repair and cell cycle genes. In the discovery phase, a total of 285 Spanish patients and 5,608 ancestry‐matched controls were evaluated. In the validation stage, an independent cohort of 514 European patients and 27,173 ancestry‐matched controls were analyzed. An FDR correction was applied to both datasets and genes with a q‐value <0.2 in both cohorts were considered significant.Results:Our joint analysis of 799 CLL patients from 2 genetically distinct cohorts and 32,781 ancestry‐matched cancer‐free controls identified ATM and CHEK2 as significantly enriched in both CLL datasets. First, our analysis recaptured the previously reported finding of ATM variant enrichment in CLL patients. Carriers of pathogenic ATM mutations in our cohorts (n = 9 patients, discovery: 1.05%, validation: 1.17%) were 2.8–3.7 times more likely to develop CLL compared to cancer‐free individuals (discovery: OR = 2.8, 95%CI = 0.7–9.0, q‐value = 0.181; validation: OR = 3.7, 95%CI = 1.6–8.3, q‐value = 0.0454). In addition, our analysis identified 21 CLL patients carrying pathogenic CHEK2 alterations (discovery: 1.40%, validation: 3.31%), making CLL patients 4.4–8.0 times more likely to carry such alterations compared to controls (discovery: OR = 8.0, 95%CI = 2.3–27.0, q‐value = 0.026; validation: OR = 4.4, 95%CI = 2.5–7.3, q‐value <0.001).Summary/Conclusion:Our analysis of genetically distinct CLL cohorts, using a high‐sensitivity variant calling algorithm, supports CHEK2 as a potentially novel CLL predisposition gene that may explain a portion of the missing monogenic heritability of CLL. In addition, this study highlights the DNA repair and cell cycle regulation pathways as potential drivers of CLL susceptibility.
PS1195 TIME TO HEALTH‐RELATED QUALITY OF LIFE IMPROVEMENT IN THE PHASE 3 BFORE TRIAL: FIRST‐LINE BOSUTINIB VS IMATINIB IN CHRONIC PHASE CHRONIC MYELOID LEUKEMIA
Authors: Brümmendorf T.H. , Bushmakin A.G. , Cappelleri J.C. , Reisman A. , Viqueira A. , Mamolo C. , Isfort S. 
Published: June 2019

Background:Bosutinib showed improved efficacy vs imatinib for newly diagnosed chronic phase chronic myeloid leukemia (CP CML) in the ongoing BFORE trial (NCT02130557). Gastrointestinal events and transaminase increases were more frequent with bosutinib; musculoskeletal events and edema were more common with imatinib. Improvement or preservation of health‐related quality of life (HRQoL) was seen in both treatment arms after 12 months.Aims:In post hoc analyses, we examined time to improvement of HRQoL with bosutinib or imatinib.Methods:HRQoL was assessed at study visits with the patient‐reported Functional Assessment of Cancer Therapy–Leukemia (FACT‐Leu) questionnaire. Nonparametric models (with SAS Proc Lifetest) were used to estimate median times to transient improvement (any increase in a FACT‐Leu domain score from baseline) and sustained improvement (any increase for 2 consecutive visits) of HRQoL.Results:Median times to transient improvement of FACT‐Leu scores were similar with bosutinib and imatinib (Table). Median time to sustained improvement was numerically shorter with bosutinib for FACT‐Leu total, FACT‐General (FACT‐G) total, emotional well‐being and leukemia subscale scores. Differences between treatment arms for both models (transient or sustained events) were not statistically significant (P>0.05).Summary/Conclusion:Median time to HRQoL improvement was comparable with bosutinib and imatinib, suggesting differences in rates of specific side effects of individual tyrosine kinase inhibitors for CP CML do not necessarily translate into measurable differences in patient‐reported HRQoL.image
PS1200 IDENTIFICATION OF MUTATIONS IN RED CELL MEMBRANOPATHIES USING NGS‐TARGETED SEQUENCING PANEL
Authors: More T. , Kedar P. , Warang P. , Devendra R. , Madkaikar M. 
Published: June 2019

Background:Red cell membranopathies refers to genetically and phenotypically heterogenous disorders due to defects in red cell membrane and skeletal proteins. Phenotypes vary from severe transfusion‐dependent hemolytic anemia to fully compensated hemolysis. Causative genes implicated in RBC membranopathies are numerous, making a gene‐by‐gene approach time consuming, expensive and labor intensive. Use of Next Generation Sequencing (NGS) targeted sequencing panel can expedite the molecular diagnosis after routine laboratory tests.Aims:This study aimed at defining a strategy for effective diagnosis of RBC membrane defects and identification of pathogenic mutations associated with the disease.Methods:Patients with clinical symptoms of hemolytic anemia, jaundice, hepatosplenomegaly and some requiring frequent blood transfusions were recruited in this study. Blood was collected from the patients after informed consent. Peripheral blood smear examination and flow cytometric based EMA binding test were performed and identification of mutations in red cell membrane proteins was done using NGS‐targeted sequencing panel. The relationship between gene mutations and clinical phenotypes was analyzed by In silico prediction tools such as SIFT, Polyphen2.Results:We identified mutations in 21 cases of Hemolytic Anemia (HA) in membrane protein‐coding genes using NGS‐targeted sequencing panel. Five Hereditary Spherocytosis (HS) patients were diagnosed with mutations in ankyrin (ANK1) gene, 3 patients with alpha spectrin (SPTA1) gene and 4 patients with beta spectrin (SPTB) gene. Band 3 (SLC4A1) gene mutations was identified in two HS cases and in 2 with dRTA cases. Five patients showed mutation in PIEZO1 gene causing Hereditary xerocytosis (HX). Twelve novel mutations were found as shown in Table 1.Summary/Conclusion:RBC membranopathies showed highly varied etiology. Our experience demonstrates the high diagnostic yield of NGS for molecular diagnosis of RBC membrane disorders associated with defects in large genes. Timely detection of the disorder not only offers therapeutic benefits for our patients but is likely to help in genetic counseling and future antenatal diagnosis, if required.image
PS1202 DIFFERENTIAL DIAGNOSIS OF CONGENITAL HEMOLYTIC ANEMIA OF NEONATES AND INFANTS IN JAPAN
Authors: Ogura H. , Utsugisawa T. , Aoki T. , Kinoshita A. , Okamoto Y. , Kawakami T. , Yamamoto T. , Kanno H. 
Published: June 2019

Background:As a reference center, we analyze erythrocyte membranes, enzymes, and hemoglobin for congenital hemolytic anemia (CHA) in cases that are not diagnosed using ordinary laboratory examinations, and we introduce genetic testing using a hemolytic anemia‐related gene panel. We analyzed 136 cases from 2016 to 2017, including 36 patients under 1 year of age and 100 patients aged 1 year and over. Consequently, a definitive diagnosis was obtained for 84% of patients aged 1 and over but for only 72% of patients under 1 year of age.Aims:We aimed to summarize the final diagnosis for hemolytic anemia in patients less than 1 year of age analyzed over 3 years from 2016 to 2018 and discuss the common clinical features and laboratory data of undiagnosed cases.Methods:We analyzed 58 infantile CHA casesusing the following laboratory tests: flow‐cytometric osmotic fragility (FCM‐OF) test, eosin 5’‐maleimide binding test, assay of 15 red blood cell (RBC) enzymes, reduced glutathione content, and isopropanol stability test for unstable hemoglobinopathy. For the cases with a difficult diagnosis, genetic analysis with target‐captured sequencing (TCS) using a gene panel of 74 genes related to CHA was carried out.Results:Among 58 infants with CHA, we diagnosed 30 cases (52%); there were 18 cases of hereditary spherocytosis (HS), 2 cases of hereditary elliptocytosis (HE), 1 case of hereditary pyropoikilocytosis, 4 cases of erythro‐enzymopathies, and 2 cases of unstable hemoglobinopathies. Among the 28 undiagnosed cases, 14 showed an elevated percentage of residual RBCs in the FCM‐OF test. These cases also showed abnormal RBC morphologies such as target cells, stomatocytes, and/or pyknocytes. We performed TCS for 4 of the 14 cases, but none harbored mutations such as PIEZO1 or KCNN4. Of the 7 cases, which could be followed up to at the age of one year, all recovered from CHA within a year.Summary/Conclusion:Mild hemolytic anemia accompanied by stomatocytes and/or target cells is a typical clinical feature of dehydrated hereditary stomatocytosis (DHSt) in adults. We previously showed that FCM‐OF clearly discriminated DHSt from HS or HE (Utsugisawa et al., Blood 2017;130:929). In this study, approximately half of the infants with undiagnosed CHA showed similar RBC morphology and higher osmotic resistance with no causative gene mutations. We noticed that clinical pictures of these infants resembled the description of infantile pyknocytosis (IP), which is a transient neonatal hemolytic anemia with the characteristic RBC morphology. As IP cases follow a relatively benign clinical course, we propose that FCM‐OF may be used as a biomarker for the tentative diagnosis of IP.
PS1221 LONG‐TERM EFFICACY AND SAFETY OF INOTERSEN FOR HEREDITARY TRANSTHYRETIN AMYLOIDOSIS: NEURO‐TTR OPEN‐LABEL EXTENSION 2‐YEAR UPDATE
Authors: Brannagan T. , Cruz M. Waddington , Wang A.K. , Polydefkis M.J. , Dyck P.J. , Khella S. , Plante‐Bordeneuve V. , Burk J.L. , Barroso F. , Merlini G. , Conceição I. , Hughes S.G. , Kwoh J. , Jung S.W. , Guthrie S. , Pollock M.P. , Benson M.D. , Gertz M. , Coelho T. 
Published: June 2019

Background:Hereditary transthyretin amyloidosis (hATTR) is a rare protein misfolding disorder that causes progressive and debilitating polyneuropathy. A randomized, controlled phase 3 trial (NEURO‐TTR; NCT01737398) demonstrated efficacy and safety of inotersen in patients with hATTR polyneuropathy (Benson 2018 NEJM).Aims:To provide an update on the long‐term efficacy and safety of inotersen, an antisense oligonucleotide inhibitor of transthyretin protein production, in patients with hereditary transthyretin amyloidosis (hATTR) with polyneuropathy.Methods:Patients who completed NEURO‐TTR were eligible to enroll in an ongoing open‐label extension (OLE) study (NCT02175004). We report an update on the long‐term efficacy and safety of inotersen in patients with hATTR polyneuropathy after 24 months in the OLE. Assessments included modified Neuropathy Impairment Score +7 neurophysiologic tests composite score (mNIS+7), Norfolk Quality of Life–Diabetic Neuropathy questionnaire total score (Norfolk QoL‐DN), Short Form 36 Health Survey version 2 (SF‐36v2) physical component summary (PCS), and safety monitoring.Results:Of 139 patients who completed NEURO‐TTR, 135 (97.1%) enrolled in the OLE. As of 5/31/2018, some patients were ongoing but had not yet completed 24 months in the OLE, and longest inotersen exposure in both studies was 5.2 years. Patients who switched from placebo to inotersen in the OLE demonstrated slowing of neurologic disease progression by mNIS+7 and Norfolk QoL‐DN as early as 6 months after starting inotersen (mean change from OLE baseline to month 6/year 2: 6.22/5.08 for mNIS+7 and 0.54/2.26 for Norfolk QoL‐DN). Patients who received inotersen for 39 months (15 months NEURO‐TTR + 24 months OLE) continued to show benefit (mean change from OLE baseline to year 2: 11.18 for mNIS+7 and 5.22 for Norfolk QoL‐DN). Patients who continued inotersen also showed stabilization of health‐related quality of life measured by SF‐36v2 PCS. No evidence of increased risk for grade 4 thrombocytopenia or severe renal events has been observed with increased exposure duration; no new safety concerns have been identified.Summary/Conclusion:In the OLE, inotersen improved, halted, or slowed progression of hATTR polyneuropathy, with greater stabilization observed in patients who initiated inotersen earlier.
PS1233 HES1 AND HES4 HAVE SIMILAR AND DIVERSE CAPACITIES TO MEDIATE HUMAN NOTCH‐DEPENDENT HEMATOPOIETIC LINEAGE DECISIONS
Authors: De Decker M. , Lavaert M. , Roels J. , Tilleman L. , Van Nieuwerburgh F. , Taghon T. 
Published: June 2019

Background:Multipotent hematopoietic stem cells (HSCs) originate in the bone marrow and give rise to all the different cell types in the blood. This process is orchestrated by interactions between cell‐intrinsic mechanisms and the environmental niche that results in the activation of particular molecular networks that drive specific hematopoietic lineage differentiation. One important example is the development of T cells in the thymus. Following colonization by thymus seeding progenitors, Delta‐like‐ligand‐4 (DLL4) that is expressed by thymic epithelial cells triggers the Notch1 receptor that is expressed on these multipotent hematopoietic progenitor cells (HPCs). Activation of the Notch signaling pathway results in the induction of T‐lineage differentiation through the upregulation of T cell‐specific genes that also prevent differentiation towards other hematopoietic lineages.Aims:In both mouse and human, Notch1 activation is the main initial driver to induce T cell development in hematopoietic stem cells. However, although well described in mice, the role of Notch1 target genes during human hematopoietic developmental choices is still rather unclear. Previous studies in human have shown that TCF1, GATA3 and BCL11B are necessary for the initial stages of T cell differentiation by repressing other hematopoietic lineages, but other transcription factors may also function during these developmental choices. Here, we focused on the function of the Notch1 target genes HES1 and HES4.Methods:We cocultured CB CD34+ Lin− hematopoietic precursor cells with enforced HES1 and HES4 expression on OP9‐DLL4 and MS‐5 stromal cells in conditions that support T‐, B‐, NK‐ and myeloid‐lineage differentiation. In addition, we performed RNA sequencing to identify HES1‐ and HES4‐induced molecular changes during human early hematopoiesis.Results:Our data illustrate that HES1 mainly acts as a repressor of differentiation by maintaining a quiescent stem cell signature in CD34+ HPCs. In contrast, while HES4 can inhibit the NK and myeloid potential of HPCs, it does not affect B cell development. Furthermore, even though HES4 does not alter the Notch ligand requirements to induce T cell development, it promotes early T cell differentiation when Notch1 signaling is present.Summary/Conclusion:Overall, we show that even though HES1 and HES4 are both induced through Notch1 activation during normal human T cell development, they display both unique and similar features during human hematopoiesis.
PS1234 APPLICATION OF FLOW SOM TO CLASSICAL FLOW CYTOMETRY: UNSUPERVISED ANALYSIS OF HEMATOPOIESIS AND HEMATOLOGICAL MALIGNANCIES
Authors: Lacombe F. , Dupont B. , Vial J.‐P. , Lechevalier N. , Béné M.C. 
Published: June 2019

Background:Multiparameter flow cytometry (MFC) is an essential tool for the diagnosis and follow‐up of hematological malignancies. Numerous panels have been devised that best allow to achieve these goals. However, analysis of MFC graphs still heavily relies on the expertise of biologists and on specifically well adapted cascade‐gating strategies. Because of the development of mass cytometry, which generates data in a multidimensional environment of up to 50 dimensions, new strategies have been developed, that could be applied to classical MFC.Aims:In this study, the capacities of a rapid new tool for multidimensional analysis, FlowSOM (self‐organizing maps) have been combined to a classical MFC analysis tool, Kaluza®, for the identification of normal and diseased human bone marrow (BM) cell subpopulations.Methods:Normal BM samples collected during bone surgery were analyzed in MFC with four antibody panels adapted to the exploration of acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) respectively. The listmode files obtained were merged and submitted to FlowSOM analysis, using R packages, which yielded trees of 100 nodes defining normal BM cell subsets. FlowSOM parameters were then transferred to Kaluza® for a bidimensional representation of the trees within which selected navigation could be performed with Kaluza® tools. In a second part of the study, the same stainings were performed on myelodysplastic or leukemic (AML or ALL) samples (diagnosis or follow‐up), which were submitted to FlowSOM together with the reference merged BM.Results:Node‐by‐node selection allowed for a precise definition of each of the unsupervised subsets generated by the software. This disclosed a selective segregation of delicate subsets such as non‐classical monocytes, hematogones or stem cells. Direct simultaneous comparison of diseased samples and normal BM allowed for a precise definition of abnormal cells (blasts) at diagnosis, and their immediate recognition as minimal residual disease in follow‐up samples.Summary/Conclusion:Flow SOM appears as a new and efficient tool for a rapid identification of BM cell subsets, highlighting differences from normal BM in dysplastic of leukemic samples. The node‐by‐node approach facilitates the definition of abnormal cells and their quantification, supervision intervening only after the generation of subsets delineated by artificial intelligence.
PS1278 LACTATE CLEARANCE AS A SURVIVAL PREDICTOR IN PATIENTS WITH HEMATOLOGIC DISEASES WITH SEPTIC SHOCK
Authors: Osipov I. , Salogub G. , Bezrukikh V. , Ivanov V. , Mikhaylov E. , Shnalieva N. , Stepanova N. 
Published: June 2019

Background:Infectious complications are one of the main causes of mortality in patients with hematological diseases. Neutropenia is common adverse effect of antitumor treatment. These patients have high risks of development severe infectious complications (sepsis and septic shock). It is known, that poor lactate clearance is associated with increased mortality in the general population of patient's. We evaluated the prognostic value of lactate clearance in patients with neutropenia grade 4 (CTCAE v4.0)Aims:To evaluate the clinical outcomes of septic shock treatment in patients with myelotoxic agranulocytosis depending on the clearance of lactate.Methods:127 patients with septic shock were included (between July 2015 and December 2018). A prospective and retrospective diagnosis of septic shock was established on the basis of Sepsis 3 criteria: arterial hypotension in combination with an increase in lactate of more than 2.0 mmol / l. The highest incidence of septic shock (up to 35%) was registered in high‐risk group patients: during the induction remission of acute myeloid leukemia and recipients of allogeneic hematopoietic stem cells. 88 percent cases were associated with gram‐negative bacteria (Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, Serratia spp. and E. coli). The median duration of neutropenia before the development of septic shock was 15.1 (13.26–17.06) and 17.72 (16.33–19.12) days in patients who died and survived (p = 0.03, Log Rank). The levels of C‐reactive protein and procalcitonin did not differ.Results:The median lactate values at the time of diagnosis of septic shock in survivors and non‐survivors patients were 2.9 (3.071 ‐ 3.997) and 4.26 (3.665–4.867) mmol / l (p = 0.05, Log rank). 30 ‐ day overall survival (OS) in patients with a lactate level of more than 4.0 mmol/l was 34%, with a lactate level of 2.0–4.0 mmol/l was 56% (p <0.001, Log Rank). All patients received serial lactate measurements were divided into lactate clearance >10% (n = 26), >20% (n = 46), and <10% (n = 55) groups. Patients with a lactate clearance of more than 10% had higher 30‐day OS (68% vs. 29%, p < 0.01, Log Rank). The rate of lactate elimination (more than 10% versus more than 20%) did not have an additional effect on OS. In multivariate analysis with an additional assessment of the venous‐to‐arterial carbon dioxide tension, absolute lactate values, the prognostic value was shown only for the lactate clearance.Summary/Conclusion:Achieving lactate clearance during the first 6 hours treatment septic shock more than 10% is associated with improved 30‐day OS in patients with severe neutropenia. The advantage of this marker is an early assessment of the effectiveness of therapy. In the future, on the basis of lactate kinetics, it is possible to create the strategy of ”Early Goal‐Directed Therapy“.image