bluuebunny/crossref_metadata_2025 huggingface dataset:
PS1109 DENTAL HEALTH STATUS IN PATIENTS WITH HEMOPHILIA. SINGLE CENTER EXPERIENCE IN ROMANIA
Authors: Ursu C.E. , Serban M. , Clevet O. , Traila A. , Boeriu E. , Jinca C. , Boia E. , Arghirescu S.
Published: June 2019

Background:

Shortage of replacement therapy in haemophilia is responsible for the devastating consequences of uncontrolled bleeds, with unfavourable impact on the quality of life, strongly dependent on oro‐dental health.

Aims:

The objective of our study was to analyse the dental status in persons with haemophilia (PwH) in our treatment conditions, characterised by the introduction of on demand replacement in 1997 and of prophylaxis in 2014‐2015.

Methods:

This descriptive non‐interventional cross‐sectional study was conducted on 47 PwH (12 children, 35 adults), all with severe form of disease and on a control matched group of 47 persons. For each PwH were recorded laboratory data and history of replacement therapy. Oral health status evaluation was performed according to the criteria suggested by World Health Organization (WHO).

Results:

We assessed a very bad dental status in PwH with very high values for the majority of the parameters. In haemophilic children comparable to matched control group there were significant differences regarding the D index and PL index (p = 0.04, p = 0.01). Concerning the domain of decayed teeth in children we observed the existence of an age correlation (r = 0.47/0.81), unfortunately with a lowering appeal for dental care in children with haemophilia, the F index (0.2 ± 0.4) being decreased compared to the children from control group (2.2 ± 2.2), with significant difference (p = 0.003). The dental indexes in adult haemophiliacs were found high, significantly higher in the field of D and F than those belonging to the control group. F index was found significantly higher in the control group than in PwH, proving the low addressability for dental care in haemophiliacs. All these results, with the exception of Missing teeth and DMFT index (r = 0.63/r = 0.65), were not correlated with the age. Adult PwH, with mean age of 34.8 ± 9.81 years, had a dismal dental profile, only 7/35 had a complete denture, 7/35 missing 25%, and 3/35 missing more than 50% of their teeth.

Summary/Conclusion:

Lack of access to treatment or a low consumption, especially in young age, can influence and have a decisive impact on oral health status in PwH. Our results highlight the importance of an accessible dental care for this vulnerable group of patients in the frame of a comprehensive haemophilia care, including preventive dental treatment and an adequate replacement therapy.
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.

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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.

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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“.

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PS1289 THE ATYPICAL LYMPHOCYTE COUNT: A NOVEL PREDICTIVE FACTOR FOR SEVERE DENGUE INFECTION
Authors: Abeysuriya V. , Choong C. , de Mel S. , Thilakawardana B.U. , de Mel P. , Shalindi M. , de Mel C. , Chandrasena L. , Seneviratne S.L. , Yap E.‐S.
Published: June 2019

Background:

Dengue haemorrhagic fever (DHF) and Dengue Shock Syndrome (DSS) are life threatening complications of DI which occur in a minority of patients. The early identification of these individuals is critical to guide clinical management. There is currently no validated laboratory test which can predict severe complications of DI. The Atypical lymphocyte count (ALC) is a research parameter generated on Sysmex FBC analysers which has been correlated with the presence of circulating atypical lymphocytes (AL). Pilot studies have suggested that the presence of AL may be associated with severe DI.

Aims:

We evaluated the utility of the ALC as a predictive factor for severe dengue related thrombocytopaenia as well as DHF and DSS.

Methods:

We prospectively collected data on patients admitted to Nawaloka Hospital Sri Lanka (NH) with DI between December 2016 and August 2018. DI was diagnosed based on a positive Non‐structural antigen 1 or dengue IgM antibody. ALC data were extracted from the Sysmex XS500i automated FBC analyzer from day of admission to day seven. Clinical data was recorded from medical records and the computerized data base maintained by NH. Statistical analyses were performed using SPSS version 20.

Results:

201 patients were included in the study. DSS and DHF occurred in 6.5% (n = 13) and 6% (n = 12) of patients respectively. Patients with DHF and DSS as well as those with low Glasgow Coma Scale and respiratory compromise had a higher mean ALC on each day of FBC assessment compared to those without these complications. We detected a statistically significant negative correlation between the ALC on admission and platelet count on days five to seven (the period during which nadir thrombocytopaenia is expected in DI). [Spearmen's correlation; Day 5:‐0.485, Day 6:‐0.428, and Day 7:‐0.344] (P = 0.001) We performed Receiver Operator Curve (ROC) analysis to determine the predictive value of an admission ALC more than 0.5x 103/L for severe thrombocytopaenia (defined as a platelet count less than 50 x 109/l) on day five. We found that this ALC cut off had 90% sensitivity and 70% specificity for severe thrombocytopaenia on day 5. The positive and negative predictive values were 74.4% and 91.2% respectively. We also showed that patients with cardiac and abdominal ultrasonographic abnormalities had a higher mean ALC.

Summary/Conclusion:

We propose that an admission ALC of greater than 0.5 x 103/L has a strong predictive value for severe thrombocytopaenia on days five to seven of DI. Our data also suggest that a higher ALC on admission maybe predictive for severe DI. ALC on admission maybe a valuable modality for early identification of patients at risk of severe dengue related complications.

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PS1336 DISCREPANCE OF ALLELIC BURDEN CHANGES AND CLINICAL RESPONSE IN PATIENTS WITH HIGHER RISK MYELODYSPLASTIC SYNDROME TREATED WITH HYPOMETHYLATING AGENTS
Authors: Moon J.H. , Cho H.J. , Baek D.W. , Ahn J.‐S. , Ahn S.‐Y. , Choi S.H. , Kim T. , Zhang Z. , Sohn S.K. , Kim D.D. H. , Kim H.‐J.
Published: June 2019

Background:

Hypomethylating agents (HMAs) such as azacitidine (AZA) and decitabine (DEC) are used to treat higher risk myelodysplastic syndrome (HR‐MDS). However, underlying genetics and allelic burden change after HMAs needs to be clarified.

Aims:

This study investigated the effects of alleleic burden changes on the clinical outcomes after treatment with HMAs.

Methods:

This study included bone marrow samples from 64 patients (34 AZA and 30 DEC) taken at diagnosis and follow‐up after median 4 cycles. Targeted deep sequencing on a custom myeloid gene panel of 84 genes (Agilent SureSelect) was performed on trios of T‐cell, pre‐HMA, and post‐HMA samples on 64 HR‐MDS patients.

Results:

At diagnosis, we detected 135 mutations from 54 patients. Most frequently mutated genes were TP53 (n = 9), DNMT3A (n = 7), ASXL1 (n = 7), DDX41 (n = 7) and U2AF1 (n = 6). HMAs were administered median 4 cycles (range 1–14). At follow‐up, allelic burdens were decreased from median 13.7% (range 1.5–87.7) to 1.7% (range 0.0–49.4%), where from median 9.9% to 0.8% in AZA group (p < 0.001) and from median 17.8% to 6.7% in DEC group (p = 0.005). CR/mCR rate were higher in patients with allelic burden reduction <2% (n = 26/43, 60%) than those without (24%, n = 5/21), but not significantly different (p = 0.765). With median follow‐up duration of 19.3 months (range 0.2–99.6 months), 2‐year overall survival (OS) rates were 54.6 ± 8.1% and 44.8 ± 11.3% in patients with and without allelic burden reduction, respectively (p = 0.907). The presence of TP53 mutations did not affect the achievement of CR/mCR (p = 0.379). However, the presence of TP53 mutations were associated with inferior OS rates: 2‐year OS rates of 56.5 ± 7.2% and 22.2 ± 13.9% (p = 0.001). In the multivariate analysis, IPSS high (HR 5.34) and the presence of TP53 mutations (HR 3.42) affected adversely on OS, while the achievement of CR/mCR (HR 0.23) and the performance of allogeneic transplantation (HR 0.33) were favorable factors for OS.

Summary/Conclusion:

This study showed that allelic burden is decreased after treatment with both azacitidine and decitabine in HR‐MDS patients. However, the allelic burden reduction is independent to the achievement of CR and OS. The presence of TP53 mutation remained independent prognostic factor in HR‐MDS patients treated with HMAs, even with decreased allelic burden.
PS1365 NEXT‐GENERATION OPTICAL MAPPING REVEALS LARGE GENETIC HETEROGENEITY IN MULTIPLE MYELOMA
Authors: Petrackova A. , Fillerova R. , Minarik J. , Mikulkova Z. , Balcarkova J. , Krhovska P. , Pika T. , Gajdos P. , Behalek M. , Papajik T. , Kriegova E.
Published: June 2019

Background:

Genetic heterogeneity in multiple myeloma (MM) affects not only the disease development but also therapeutic response and prognosis. Nowadays, karyotype and FISH are standard diagnostic tools for the detection of genetic aberrations in MM. However, comprehensive analysis of genome architecture, particularly structural variations (SVs), still remain a challenge.

Aims:

To study the genomic architecture of MM cells using a novel nanochannel technology based on optical mapping of high‐molecular weight DNA in order to refine genomic variability in human MM.

Methods:

We analysed five samples of bone marrow aspirates from patients with newly diagnosed MM. High‐molecular weight DNA was isolated from sorted MM cells, labelled by DLS chemistry and visualised by Saphyr optical system (BioNano Genomics). Created single molecules were de novo assembled into physical maps spanning whole genome based on known labelled specific motifs. After filtering of structural variants against the database of healthy controls, only rare de novo variants were analysed. Karyotype, FISH, and arrayCGH results were available for the enrolled patients and were compared to the results of optical mapping.

Results:

For all MM samples, the optical genome maps (480 Gbp) were assembled with an average effective coverage of 124X and aligned to a human reference genome (GRCh38). In our cohort, numerous copy number variations and large somatic SVs, including deletions, insertions, inversions, duplications and translocations, were identified (Table 1) with majority of SVs being deletions (∼60%). All genetic variants detected by arrayCGH were also confirmed by mapping. Additional SVs with novel intra‐ and inter‐chromosomal translocations and copy number variants were found in all patients, not detected by FISH/arrayCGH. Regarding novel translocations, majority were intra‐chromosomal (64%) within chromosomes 3, 4, 6, 14, 16, 17, and 18. Additional translocations (36%), detected only by mapping, involved unusual chromosomes in MM: t(2;8), t(3;6), t(3;12), t(6;8), t(6;17), t(8;9), and t(8;22). All samples carried novel SVs on chromosome 17, including two intra‐ (P1) and one inter‐chromosomal translocations (P2), insertions in four (P2‐P5) and deletions in two (P4‐P5) patients. Although no IGH locus rearrangements were detected by standard methods, monosomy on chromosome 14 within IGH locus was observed in four samples (P2‐P5) by mapping. The analyses of associations of SVs and affected genes with clinical characteristics are ongoing.

Summary/Conclusion:

A large spectrum of novel genomic rearrangements was detected in MM using next‐generation mapping technology, showing a high potential of optical maps for refinement of genomic variability in human MM.

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Grant support: research grant Celgene, MZ ČR VES16–32339A, NV18–03–00500, MZ ČR – RVO (FNOl, 00098892)
PS1369 AMITRIPTYLINE STRENGTHENS THE EFFECTS OF BORTEZOMIB AND MELPHALAN TREATMENT IN MULTIPLE MYELOMA BY INHIBITING ACID SPHINGOMYELINASE
Authors: Faict S. , Oudaert I. , De Veirman K. , Maes K. , Jacobs L. , De Bruyne E. , Swinnen J. , Schots R. , Fostier K. , Vandebroek I. , Vanderkerken K. , D’Auria L. , Menu E.
Published: June 2019

Background:

Malignant cells can adapt their lipid metabolism to attain higher rates of proliferation, and resist chemotherapy‐induced apoptosis.

Aims:

In this study, we aimed to identify the changes in lipid metabolism in multiple myeloma (MM) patients, and to further investigate the role of this altered metabolism in vitro.

Methods:

We performed a lipidomics analysis to compare plasma from MM patients to healthy controls. Based on these results, lipid spike‐in assays enabled us to evaluate effects on MM cell proliferation. Sphingomyelinase (SMase) content of MM cells and their exosomes was measured, before and after treatment with standard‐of‐care drugs, using an AmplexRed assay. We inhibited both neutral and acid SMase (ASM) in MM cells to evaluate effects on drug efficacy. Finally, we investigated whether MM exosomes rich in ASM are able to induce drug resistance in MM cells low in ASM.

Results:

Lipidomics analysis revealed that several ceramide species were significantly more present in MM patient plasma, while sphingomyelin seemed to be decreased. When adding ceramide in vitro to MM cells, we observed higher viability and proliferation. The increased ceramide/sphingomyelin ratio in plasma lead us to believe that the sphingomyelinase enzyme, which converts sphingomyelin into ceramide, was upregulated in MM cells. This was confirmed in BM samples, comparing CD138+ MM cells to the CD138‐ fraction. In the MM cell lines JJN3, OPM2, LP1 and U266, we observed an increased expression of acid and total SMase when treating these cells with melphalan or bortezomib for 24 h. Furthermore, we investigated whether MM exosomes were involved in the secretion and transfer of acid SMase. Interestingly, amitriptyline, an inhibitor of acid SMase, often prescribed in MM patients for neuropathic pain, increased the efficacy of both drugs in vitro, and also induced apoptosis in primary MM cells as a single agent.

Summary/Conclusion:

SMase is increased in primary MM cells and cell lines upon treatment with melphalan and bortezomib, while inhibiting this expression using amitriptyline leads to a higher efficacy of drug therapy.
PS1379 SAFETY AND EFFICACY OF ONCE‐WEEKLY CARFILZOMIB DOSING IN FRAIL PATIENTS: A SUBGROUP ANALYSIS FROM THE PHASE 3 A.R.R.O.W. STUDY
Authors: Mateos M.‐V. , Ludwig H. , Kumar S. , Rosenbaum C. , Huang M. , Goldrick A. , Blaedel J. , Dimopoulos M.
Published: June 2019

Background:

A.R.R.O.W. demonstrated superior progression‐free survival (PFS) with once‐weekly carfilzomib (K, 70 mg/m2) and dexamethasone (Kd70) vs twice‐weekly K (27 mg/m2) and dexamethasone (Kd27) in relapsed and refractory (RRMM) patients (pts), regardless of age. For a comprehensive measure of fitness, frailty scales were developed incorporating age, comorbidities, and functional status (Palumbo Blood 2015;125:2068–74; Facon Blood 2015;126:4239).

Aims:

Here, we assessed post‐hoc pt outcomes by frailty status.

Methods:

PFS and safety were assessed by treatment arm and a frailty algorithm which incorporated age, Charlson Comorbidity Index derived from medical history, and ECOG performance status; pts with frailty scores of 0, 1, or ≥2 were classified as fit, intermediate, or frail, respectively. PFS was assessed using the Kaplan‐Meier method. Safety was assessed in pts who received ≥1 treatment dose.

Results:

The distribution of pts by frailty status was generally balanced between treatment arms (Table). Once‐weekly Kd70 vs twice‐weekly Kd27 resulted in median PFS for fit, intermediate, and frail pts of 15.7 vs 5.7 months (hazard ratio [HR] 0.53 [95% confidence interval (CI), 0.33–0.86]), 11.1 vs 7.7 months (HR 0.81 [95% CI, 0.55–1.19]), and 10.3 vs 6.6 months (HR 0.76 [95% CI, 0.49–1.16]), respectively.

Rates of grade ≥3 treatment‐emergent adverse events (TEAEs) of interest were similar between treatment arms across frailty subgroups (Table). In the once‐weekly Kd70 subgroups, there was no grade ≥3 peripheral neuropathy and grade ≥3 cardiac failure rates were ≤4%.

Summary/Conclusion:

Once‐weekly Kd70 resulted in PFS benefits vs twice‐weekly Kd27, regardless of frailty level. These results support weekly Kd70 as a treatment option for fit and frail RRMM pts.

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PS1402 OUTCOME IN TRANSPLANT ELIGIBLE PATIENTS WITH MULTIPLE MYELOMA IN LATIN AMERICA. AN INTERNATIONAL STUDY OF GELAMM
Authors: Peña C. , Schutz N.P. , Bove V. , Villano F. , Osorio R. , Chandía M. , Cardemil D. , Contreras C. , Contreras C.G. , Donoso J. , Espinoza M. , Gabriel L.R. , López‐Vidal H. , Rojas C. , Soto P. , Ochoa P. , Duarte P. , Remaggi G. , Yantorno S. , Corzo A. , Zabaljauregui S. , Shanley C. , Lopresti S. , Orlando S. , Verri V. , Quiroga L. , García C.A. , Fernández V. , Fantl D. , Riva E.
Published: June 2019

Background:

Multiple myeloma (MM) is a frequent hematologic malignancy. Despite the important advances in treatment strategies during the last decades, the gold standard remains being a proteasome inhibitor (PI)‐based induction, followed by autologous stem cell transplant (ASCT).

In Latin America (LA) there is great heterogeneity in access to new drugs and ASCT, and there is scarce data regarding patient́s outcomes in the region.

Aims:

The aim of this study was to describe clinical characteristics and outcomes of MM transplant eligible patients in LA countries.

Methods:

Retrospective international multicenter cohort study. Unselected consecutive MM transplant‐ eligible patients diagnosed between 2010 and 2018 from participating centers in Chile, Argentina and Uruguay were included. Data was collected from clinical records in a standardized report form. We analyzed clinical characteristics at diagnosis and frontline therapy outcomes, including ASCT.

Transplant‐eligible patients were defined as fit patients younger than 66 years old. Response to treatment was defined according to current IMWG criteria.

Statistical analysis was performed by using STATA 13.

Results:

Six hundred and seventy one patients were included in the study, 282 from Chile, 258 from Argentina and 131 from Uruguay. The main characteristics are shown in Table 1. Significant differences in access to PI induction and ASCT were found between private and public institutions, with patients from the latter receiving suboptimal frontline therapy. Optimal response (sCR, CR and VGPR) was achieved in 51% of the patients in the CyBorD group, in 65% in the VTD group, and in 45% in the CTD group, the 3 main induction regimens used. Only 52% of patients finally underwent ASCT. With a median follow up of 33 months (range 1–109), 5 year overall survival (OS) was 58,4%, with a median survival of 73 months in the whole cohort. When analyzed according to induction treatment, 5 year OS was 74% for PI‐based, 48% for thalidomide‐based, and 23% for those not treated with novel agents (P < 0,0001). In the multivariable analysis, not undergoing ASCT (p < 0,001), renal failure (p = 0,004), hypercalcemia (p = 0,001), and achieving less than VGPR (p = 0,001) response were independent factors associated with worse outcome.

Summary/Conclusion:

Survival in transplant‐eligible patients is lower than the “optimal” described survival in other regions. Unequal access to PI‐based regimens and therefore to ASCT between public and private centers was observed. Reasons for 48% of candidates not being transplanted merit further analysis.

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PS1406 WEEKLY CARFILZOMIB, LENALIDOMIDE AND DEXAMETHASONE UNTIL PROGRESSION IN RELAPSED REFRACTORY MULTIPLE MYELOMA
Authors: Gruchet C. , Richez V. , Guidez S. , Tomowiak C. , Fouquet G. , Machet A. , Moya N. , Sabirou F. , Levy A. , Bobin A. , Gardeney H. , Bouyer S. , Bridoux F. , Javaugue V. , Azais I. , Durand G. , Loiseau H. Avet , Leleu X.
Published: June 2019

Background:

Carfilzomib, Lenalidomide and Dexamethasone association (KRd) has led to approval in early RRMM. On ASPIRE International phase 3 study Carfilzomib was used on a twice a week basis at 27 mg/m2 and limited to 18 months exposure. We have reported already that KRd on a weekly basis at 56 mg/m2 had similar and safe properties. In this abstract we report the long‐term exposure data on KRd weekly given at 56 mg/m2 until progression.

Aims:

We aimed to evaluate the efficacy of KRd given on a prolong duration beyond 18months, and to validate the safety profile of continuous exposure to Carfilzomib.

Methods:

28 patients were prospectively recruited. Carfilzomib 20/56 mg/m2 was administered on days 1,8,15 on 28 days cycles. Lenalidomide (25 mg/day) was given 21/28 days and Dexamethasone was administered weekly. Patients were treated until progression.

Results:

With a median follow‐up at start of KRd of 30 months, 50% of patients relapsed and 39% died. 24/28 patients received 1 prior line of treatment. 8/28 patients are still on treatment with duration > 24 month and 6/28 with duration > 30 months. The median number of cycles was 15. ORR and CBR was 85.7% and 89.3%, whom 46% ≥ CR; with a median DOR of 13 months and 43% having more than 18 months. 6 patients had negative MRD at 10–6 and normalized PET CT. Median of OS is not reached, and the 30 month‐expected OS from the start of KRd was 56%. The median PFS and EFS was at 29 months, and the 30 month‐expected PFS and EFS was 45%. PFS and EFS being superimposable speaks to that there was no safety concern related to prolonged exposure to K. Only 4 patients stopped KRd for safety issues. Hematologic and non‐hematologic adverse events ≥ grade 3 were reported in 16/28 and 10/28 patients. Adverse events ≥ grade 3 seen in ≥10% of patients were neutropenia, thrombocytopenia, vomiting and pyrexia. Of note, 5 patients (18%) were 365 years old and these patients showed similar data compared to the studied cohort.

Summary/Conclusion:

KRd weekly at 20/56 mg/m2 is effective and safe to early RRMM patients, provides improved safety profile to patients allowing treating patients until progression. Further studies are warranted to confirm this data on a larger early RRMM population and validate the concept of long duration of treatment using Carfilzomib combination.
PS1412 POMALIDOMIDE 3RD LINE VERSUS 4TH LINE FOR IN EARLY RELAPSED REFRACTORY MULTIPLE MYELOMA
Authors: Gruchet C. , Guidez S. , Tomowiak C. , Fouquet G. , Machet A. , Moya N. , Sabirou F. , Levy A. , Bobin A. , Gardeney H. , Bouyer S. , Bridoux F. , Javaugue V. , Azais I. , Durand G. , Loiseau H. Avet , Leleu X.
Published: June 2019

Background:

Pomalidomide in association with dexamethasone is approved for relapsed and refractory Mutiple Myeloma (RRMM) in 3rd line and beyond based on the multicenter international phase 3MM‐003 study that demonstrated greater efficacy for Pomalidomide plus dexamethasone over high dose dexamethasone. However, MM‐003 mainly recruited RRMM with 5 prior lines +, and very few data are available regarding real life Pomalidomide‐based treatment in 3rd and 4th line RRMM.

Aims:

The aim of this study was to study efficacy and safety of Pomalidomide‐based treatment in 3rd and 4th line for RRMM.

Methods:

This study is a retrospective, multicenter study based on 108 consecutive RRMM treated with Pomalidomide‐based treatment in 3rd and 4th line. All assessment made according to IMWG.

Results:

Median age was 62.5 (range, 30–86), with 36% older than 65 years, sex ratio M/F 1.25 and ISS disease stage 2 or 3 in 58%. 100% patients received previous treatment with Bortezomib and Lenalidomide and 34,2% had previously an autologous stem cell transplantation. 52 patients (48%) had pomalidomide‐based therapy as 3rd, and 56 (52%) as 4th line. 74% of patients received a double‐based therapy (Pomalidomide plus Dexamethasone) and 26% received a triple based therapy (Pomalidomide, Cyclophosphamide and Dexamethasone). Overall ORR was 55%, with 25% more than VGPR including 3 complete response with Pomalidomide‐based as 3rd line. ORR was 53% with 16% ≥ VGPR including 1 CR for Pomalidomide‐based as 4th line. With a median follow‐up of 24 months, 58% and 48% at relapsed and died, respectively, in 3rd line, and similarly, 74% and 50% in 4th line. The median TTP was 7 (CI95% 1.4;12.6) and 9 (6.1;11.9) months in 3rd and 4th line, and the median OS 17 (7.8;26.1) and 23 (12.6;33.4) months, respectively. 22% and 30% have discontinued and reduced pomalidomide‐based treatment respectively. No patient died related to adverse events. AEs leading to pomalidomide‐based modification in schema included hematological AEs in 40% and non‐hematological AEs in 12% of patients. Overall, the most common adverse events grade 3 or 4 were neutropenia (22%), anemia (11%), thrombocytopenia (7%) and infectious disease (5%).

Summary/Conclusion:

Pomalidomide‐based therapy demonstrates efficacy in the real life with a manageable safety profile. Further study ill look into triplet versus doublet pomalidomide‐based regimen, with a benefit expected for the former. Further prospective studies are warranted to confirm this data on a larger MM population.
PS1478 EVALUATION OF COMBINED MEASUREMENT OF IMMATURE PLATELET FRACTION AND SCHISTOCYTES IN PREGNANCY‐ASSOCIATED THROMBOTIC THROMBOCYTOPENIC PURPURA AND SEVERE PREECLAMPSIA/ HELLP SYNDROME
Authors: Fayek M. , Mekawy M. , El‐Gamal R. , Abdelkader A. , Abdelbary H.
Published: June 2019

Background:

The occurrence of thrombotic microangiopathy (TMA) in pregnancy is an unfortunate emergency condition. Proper diagnosis is mandatory which requires the consideration of two overlapping diagnoses: severe preeclampsia/ haemolysis, elevated liver enzymes, and low platelet syndrome (SPE/HELLP) and thrombotic thrombocytopenic purpura (TTP). The long turn‐around times of ADAMTS13 testing precludes the timely distinction between the two conditions.

Aims:

We aimed at evaluating schistocyte counts and immature platelet fraction (IPF%), as both increase in TMAs, to discriminate between TTP and SPE/HELLP of pregnancy.

Methods:

IPF% was measured using SysmexXE‐2100 automated hematology analyzer, and schistocyte counts were estimated microscopically as per the International Council for Standardization in Hematology (ICSH)‐Schistocyte Working Group guidelines. The study included 30 pregnant patients with SPE/HELLP, 13 pregnant patients with TTP, and 30 women with normal pregnancy. The discrimination between the two patient categories was based on clinical judgment and TTP cases were identified using the PLASMIC score.

Results:

TTP patients had higher values of IPF% than SPE/HELLP 19.5 (16.9–27.1) vs 13 (9.5 – 23.25); p <0.001); similar results were revealed regarding schistocyte counts 6.5 (3.9–8.6) vs 2.1 (1.6 – 3.5); p <0.001). IPF% and schistocyte counts were able to discriminate between TMA patients and normal pregnant women, and between and SPE/HELLP and TTP patients. Moreover, the discriminatory function of each was improved when the two parameters were used in combination.

Summary/Conclusion:

IPF% analysis should be used in conjunction with manual schistocyte counting in TMA cases to distinguish TTP pregnant patients from patients having SPE/HELLP.