bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nodular Lymphocyte Predominant Hodgkin Lymphoma. Experience in a Single Institution in Argentina
Authors: Alfaro Elizabeth M , Sanchez La Rosa Cristian G , Guitter Myriam , Millan Natalia , Montanari Andrea , Galluzzo Laura , Felice Maria Sara , Zubizarreta Pedro 
Published: October 2019

Abstract
Background: Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL) is a rare subtype of Hodgkin Lymphoma (HL) that represents approximately 5% of HL and discloses a distinct clinical-pathological features with an overall good prognosis. Treatment strategies have been based on protocols for HL. Selected patients can be treated with surgery alone.
Objective: Our aim was to describe the clinical characteristics, management and outcome of patients with NLPH.
Methods: Between March 2000 and June 2016, 16 patients (14 M/2 F), median age: 9.8 (range 4.9-14.5) years, were consecutively diagnosed in our institution. Initial chemotherapy schedules were ABVD (Doxorubicin, Bleomycin, Vinblastine and Dacarbazine), COPP/ABVD or AV-PC (Adriamycin/Vincristine/Prednisone/Cyclophosphamide) were administered to 14 patients. When response was partial or null, additional chemotherapy was administered, IF-RT, Anti CD20 and/or HDC/Autologous SCT. A watch-and-wait strategy after lymph node complete surgery was used in two cases when this strategy became trustworthy.
Results: Fourteen patients (87.5%) showed nodal compromise being cervical localization the most frequent localization. Thirteen patients (81.2%) presented localized disease (stage IA: 8, stage IB: 1, stage IIA:4) and 3 (18.7%) advanced disease (stage IIIA:1, stage IVA:1, IVB:1). Two patients had B symptoms. Two IA stage cases (12.5%) had complete node resection. Five patients (31.2%) received 4 cycles of ABVD. One patient (6.2%) received COPP/ABVD. Five patients (31,2%) received 3 cycles of AV-PC. One patient (6.2%) received 6 cycles of AV-PC plus 4 doses of Rituximab. One patient (6.2%) presented progressive disease after 3 cycles of ABVD, additional chemotherapy was administered and HDC/Autologous SCT. Three (18%) patients received IF-RT plus chemotherapy. One patient is still on treatment. Response to treatment was: CR in 12 pts (75%), Partial-response: 2 pts (12.5%), NR: 1 pts (6.2%). Median follow up: 58.4 (range 2-89) months. All patients remain in CR. Conclusion: Although the management of NLPHL was not uniform in our retrospective analysis, the results were promising. Watch and wait strategy plays an important role to avoid adverse effects associated with therapy. Tailored therapy according to staging and disease response, seemed to be a good strategy in our setting.

Disclosures
No relevant conflicts of interest to declare.
Pharmacogenetic Markers of Treatment Response of Imatinib Mesylate in Chronic Myeloid Leukemia Brazilian Patients
Authors: Vieira Natália M , Leite Renata , Reginato Fabíola , Zandoná Marília , Vanelli Costa Tito , Fogliatto Laura , Linden Rafael , Venzon Antunes Marina , Capron Arnaud , Wallemacq Pierre , Wagner Sandrine , Mattevi Vanessa , Capra Marcelo 
Published: October 2019

Abstract
The treatment of chronic myeloid leukemia (CML) was revolutionized by the introduction of imatinib mesylate (IM). However, approximately 20% of patients are non-responsive and interpatient variability in response to IM is still a phenomenon lacking explanation. Single nucleotide polymorphisms (SNPs) located in genes encoding proteins involved in IM pharmacokinetics are potentially involved in the causes of this variation. In this study, we investigated the association of SNPs in the genes encoding IM metabolizing enzymes CYP3A4 (rs35599367 and rs2740574) and CYP3A5 (rs776746) and efflux transporter proteins ABCB1 (rs3213619, rs1128503, rs2032582 and rs1045642), ABCG2 (rs2231142) and ABCC4 (rs9561765) with response to treatment and with IM plasma through levels and hair concentrations. The analyzed sample was constituted of 182 CML patients on IM treatment. DNA samples were genotyped for the nine SNPs using real-time polymerase chain reaction. Hair and plasma trough IM concentrations were measured through liquid chromatography coupled to mass spectrometry methods. Clinical response was defined according to the European Leukemia Net guidelines. The number of responders to standard IM therapy was 104. A trend to a higher frequency of CYP3A4 *22 (rs35599367) allele carriers was observed among responders to IM (12.5 vs. 3.4% of non-responders, P = 0.087), although no significant differences in plasma or hair concentrations between genotypes were found. Pharmacogenetics may become a valuable approach to optimize therapy with IM in CML, but many factors still need to be clarified to make possible its application in clinical practice.

Disclosures
No relevant conflicts of interest to declare.
Germline Calr Mutation and Thrombocytosis Presenting with Concomitant BCR-ABL1+ CML
Authors: Baughn Linda B. , Gilles Scott , Courville Elizabeth L. , Nelson Andrew C. , Sachs Zohar 
Published: October 2019

Abstract
CALR mutations are present in 70-84% of JAK2 wild-type myeloproliferative neoplasms (MPN) and 67% and 88% of essential thrombocytopenia (ET) and primary myelofibrosis (PMF) respectively. Most cases of MPN are apparently sporadic, but 7-11% have evidence of familial predisposition. While germline mutations in ET-associated genes, MPL and JAK2, have been described in hereditary thrombocytosis, germline mutations in CALR have not been described in any setting. Two types of CALR mutations are common in MPN: a 52-base pair deletion (bp) and a 5 bp insertion, both in exon 9. With rare exceptions, CALR mutations are generally mutually exclusive with JAK2 or MPL mutations and have very rarely been reported in conjunction with the BCR-ABL1translocation.
Here, we report a patient with a germline CALR mutation, thrombocytosis, and subsequent development of BCR-ABL+ CML. A 67-year-old female with no significant medical history presented with severe abdominal pain and nausea. Peripheral blood analysis revealed a marked leukocytosis composed of 66% neutrophils, 16% myelocytes, 6.5% monocytes, 3.5% basophils, 2.5% promyelocytes, 2.5% metamyelocytes, 1.5% lymphocytes, 1.5% blasts, and no eosinophils. The patient was non-anemic and had a normal platelet count (340,000/mm3). Bone marrow biopsy revealed a hypercellular marrow with myeloid predominant trilineage hematopoiesis and 1-2% blasts with morphology consistent with chronic myelogenous leukemia (CML). Fluorescence in-situ hybridization analysis of peripheral blood identified a BCR-ABL1fusion in 98.5% of interphase cells.
After 3 months of standard imatinib therapy, quantitative RT-PCR showed a reduction of BCR-ABL1/ABL1 in the peripheral blood, however platelet count was elevated at 539,000/mm3. Thrombocytosis persisted over 2 years with a maximal platelet count of 584,000/mm3. Given the patient's thrombocytosis, her peripheral blood was subjected to a next generation sequencing of JAK2, MPL, and CALR genes. A 52-bp out-of-frame deletion in exon 9 of the CALR gene was detected (52% allele frequency) in peripheral blood. In addition, the same 52-bp CALR deletion (63% allele frequency) was present at the time of diagnosis and within a buccal specimen (47% allele frequency) when the BCR-ABL1 transcript was 1% in the peripheral blood. Immunostain of the buccal sample was strongly positive for cytokeratin (CK) AE1/AE3 but CD45 was not detected indicating no leukocyte contamination. This case reports the first instance of a germline CALR mutation associated with thrombocytosis and is the fourth report of the co-occurrence of BCR-ABL1 and CALR mutation in a single patient. Evolution to BCR-ABL1+ CML suppressed the CALR-mutant thrombocytosis phenotype, emphasizing the effect of these genes on lineage determination in abnormal myeloid proliferation.

Disclosures
No relevant conflicts of interest to declare.
Luspatercept Response in ESA-NaïVe/RS+ Patients and RS- Patients with Low-Intermediate Risk Myelodysplastic Syndromes (MDS)
Authors: Platzbecker Uwe , Germing Ulrich , Götze Katharina , Kiewe Philipp , Wolff Thomas , Mayer Karin , Chromik Joerg , Radsak Markus , Donovan Eileen , Wilson Dawn M. , Zhang Xiaosha , Laadem Abderrahmane , Sherman Matthew L. , Attie Kenneth M. , Giagounidis Aristoteles 
Published: October 2019

Abstract
Background: Management of anemia is a common therapeutic challenge in patients with myelodysplastic syndromes (MDS). Luspatercept (ACE-536), a fusion protein containing modified activin receptor type IIB, is being developed for treatment of anemia in lower-risk MDS. Luspatercept binds GDF11 and other TGF-β superfamily ligands to promote late-stage erythroid differentiation and increase hemoglobin (Hgb) levels (Suragani R, Nat Med, 2014 and Attie K, Am J Hematol, 2014).
Aims: This is an ongoing, phase 2, multicenter, open-label study to evaluate the effects of luspatercept in patient (pts) with low-intermediate risk MDS. Endpoints included erythroid response (IWG HI-E), RBC transfusion independence (RBC-TI, ≥ 8 weeks), duration of HI-E, pharmacodynamic and iron metabolism biomarkers, safety, and pt-reported QoL.
Methods: Inclusion criteria included age ≥ 18 yr, Hgb < 10 g/dL (if < 4U RBC/8 weeks), no prior HMA, and no current lenalidomide or erythropoiesis-stimulating agent (ESA). An expansion cohort of up to 56 patients was added to this phase 2 study to evaluate response to luspatercept in pts who do not qualify for the phase 3 MEDALIST trial (for RS+ positive patients with baseline EPO ≥ 200 U/L and ≥ 2U RBC/8 weeks). These include pts with low transfusion burden (< 4U RBC/8 weeks) who are either 1) ring sideroblast (RS)+ (≥ 15% RS in bone marrow) with baseline EPO ≤ 200 U/L and no prior ESA use, or 2) RS- with any baseline EPO level and any prior ESA use. Patients are treated with 1.0 mg/kg of luspatercept every 3 weeks for up to 5 doses, with titration up to 1.75 mg/kg. Patients may rollover to an open-label extension study for up to an additional 2 years of treatment.
Results: Results for the initial patient cohorts have demonstrated a high proportion of HI-E and RBC-TI responses in RS+ patients. Data for the additional ESA-naïve RS+ patients with low EPO levels and RS- patients with 3 months of treatment will be presented at the meeting.
Conclusions: Erythroid response to luspatercept has been demonstrated in RS+ patients with lower-risk MDS and is being explored in ESA-naïve RS+ patients with low EPO levels and RS- patients. A Phase 3 study of luspatercept in regularly-transfused RS+ patients with lower-risk MDS according to IPSS-R is ongoing (MEDALIST study; NCT02631070).

Disclosures
Donovan: Acceleron Pharma: Employment. Wilson:Acceleron Pharma: Employment, Equity Ownership. Zhang:Acceleron Pharma: Employment. Laadem:Celgene Corporation: Employment, Equity Ownership. Sherman:Acceleron Pharma: Employment, Equity Ownership, Patents & Royalties. Attie:Acceleron Pharma: Employment, Equity Ownership. Giagounidis:Celgene Corporation: Consultancy.
Long-Term Use of Azacitidine in Private Practice in Brazil: An Analysis of Patients Profiles
Authors: Centrone Renato , Garcia Laura Yolanda Chialanza , Santucci Rodrigo , Bellesso Marcelo 
Published: October 2019

Abstract
The AZA-001 study showed that azacitidine has a significant role in prolonging overall survival (OS) in High-Risk Myelodysplastic Syndromes (HR-MDS), particularly in patients with unfavorable cytogenetics, even in the subset of patients without any recorded response except progression. Otherwise, the benefit of azacitidine treatment in OS of HRMDS shows a lack of improvement in survival over the years in most of the patients in a Multicentric Spanish Study Group.
Since in our institution we had 34 patients who received Azacitidine for HR-MDS and 20-30% blast Acute Myelogenous Leukemia (AML) during last 36 months, we performed a retrospective analysis of our current long-term treatment patients currently taking Azacitidine, to understand their characteristics. We used WHO classification, IPSS- R score, bone marrow (aspirate) blast count, conventional cytogenetics-based risk, performance status (ECOG) and transfusional dependence.
We studied 11 patients with MDS who received at least 06 cycles of Azacitidine,(75 mg/ m2 D1 to D7) until data cut on 1st August 2016. The median age was 71 (range 46-84 years) and 72 % of the patients are male. WHO 2008 classification before beginning of treatment was: 3 AML (2 former RAEB-2 with recent progression), 03 Refractory Anemia with Excess Blasts-2 (RAEB-2), 02 Refractory Anemia with Excess Blasts-1 (RAEB-1), one Refractory Cytopenia with Multilineage Dysplasia (RCMLD) and 02 Refractory Anemia (RA). All but one patient had "good" cytogenetic risk and IPSS-R ranged from 2 to 8,5 (low to high risk), median 4,7 points.
Regarding treatment, patients received from 6 to 31 cycles of Azacitidine, with responses according to IWG including one complete response (CR), 03 partial responses (PR), 06 stable diseases (SD) and one Hematological improvement (HI). Transfusion dependence was present at the beginning of treatment in 07 cases, and no change in this parameter happened until those patient's last visit. Performance Status was very good (ECOG=0) in 06 patients before the first dose of azacitidine and good (ECOG-1) in the remaining subjects. At data cut, we had 09 patients with ECOG=0 in the group.
This analysis showed that patients had no real improvement of disease parameters since 07 of 11 did not achieve even a partial response, and the same number of patients stay as transfusion-dependents. However, even the worse IPSS-R classified patients had at least a better quality of life and they survived at least one year with treatment. The main finding of this evaluation is that patients who achieve a response other than progression or refractoriness have a good cytogenetic risk, most of them with normal Karyotype, confirming literature.
Table Table.

Disclosures
No relevant conflicts of interest to declare.
Functional Consequences of Spliceosome Mutations
Authors: Halene Stephanie 
Published: October 2019

Abstract
Mutations in key factors of the spliceosome occur in over 50% of patients with myelodysplastic syndromes (MDS) and thus splicing represents the most frequently affected pathway in MDS. The vast majority of mutations cluster in four genes: SF3B1, SRSF2, U2AF1, and ZRSR2. Mutations occur in a mutually exclusive manner suggesting a common mechanism by which they cause disease, and yet they segregate to specific subtypes of myeloid malignancies suggesting unique, mutation-specific functions. With the exclusion of ZRSR2, mutations occur at highly restricted residues and are strictly heterozygous, suggestive of gain of function mutations. We and others have shown that mutations affect RNA binding affinity and specificity, and can lead to a variety of outcomes including exon inclusion or skipping, alternative splice site selection, intron retention, and most recently alternative polyadenylation. Overall, these splice changes are subtle: only a small subset of all splice events is affected by these mutations and changes in isoform usage are < 10% for most alternative splice events identified. Furthermore, a direct connection between alternatively spliced transcripts and differentially expressed genes is often not seen. However, these alternative splice events affect genes critical to cellular function and hematopoiesis. Examples include EZH2, ABCB7, macroH2A1, and ATG7. Mutations in SRSF2 result in inclusion of a poison exon in EZH2 resulting in reduced protein levels of this critical epigenetic regulator. Mutations in SF3B1 cause alternative splicing of ABCB7 slating its RNA for non-sense mediated decay. Intriguingly, mutations in U2AF1 promote usage of a distal polyadenylation site in an essential autophagy factor, ATG7, resulting in inefficient translation and decreased protein levels. As a consequence, autophagy is impaired, which can promote cellular transformation, providing a possible explanation for the clonal advantage of cells with splicing factor mutations. Additional research is critical to further determine how alternative splicing contributes to the MDS phenotype with impaired hematopoietic differentiation and cellular function and eventual malignant transformation. Latest findings will be presented here with a focus on mechanistic, functional, and biologic effects of splicing factor mutations.

Disclosures
No relevant conflicts of interest to declare.
Ferroportin Mediated Control of Iron Metabolism and Disease
Authors: Lakhal-Littleton Samira 
Published: October 2019

Abstract
Ferroportin, the only known mammalian iron export protein, releases iron from the duodenum, reticuloendothelial system and liver, the sites of iron absorption, recycling and storage respectively. By downregulating ferroportin, the liver-derived hormone hepcidin controls systemic iron availability in response to erythroid demand and inflammation. This ferroportin/hepcidin axis has long been recognized as essential for systemic iron homeostasis. However, both ferroportin and hepcidin are found in tissues not recognized for their role in systemic iron control, such as the heart, the kidney, the brain and the placenta. Co-existence within the same tissue suggests a possible function for hepcidin and ferroportin in local iron homeostasis. However, this hypothesis has not been formally explored. Using mouse models with cardiac-specific manipulation of hepcidin and ferroportin, we have uncovered a role for the cardiac hepcidin/ferroportin axis in cell-autonomous iron homeostasis within cardiomyocytes. Disruption of this cardiac pathway leads to fatal cardiac dysfunction, even against a background of normal systemic iron homeostasis. One the one hand, loss of cardiac ferroportin causes by fatal cardiac iron overload that is preventable by dietary iron restriction 1. On the other hand, loss of cardiac hepcidin or of cardiac hepcidin responsiveness causes fatal cardiomyocyte iron deficiency that is preventable by intravenous iron administration. Comparative study of cardiac iron homeostasis and function in cardiac versus systemic models of ferroportin/hepcidin disruption provides insight into the interplay between systemic and cellular iron homeostasis. A role for the hepcidin/ferroportin axis in cell-autonomous iron control, demonstrated here in the context of the heart, has not previously been described in any other tissue. A pertinent question is whether our findings in the heart extend to other tissues that express both hepcidin and ferroportin, such as the kidney, brain and placenta. Disturbances in iron homeostasis are of clinical importance in cardiovascular disease, renal failure, neurodegeneration and developmental defects. Our findings have two clinically relevant implications: a) that disruption of the local hepcidin/ferroportin axis may in itself have a disease-modifying effect, and b) that therapeutic strategies developed to target the systemic hepcidin/ferroportin axis may have off-target effects relating to local iron control within some tissues.
Reference
1.Lakhal-Littleton S, Wolna M, Carr C, et al. Cardiac ferroportin regulates cellular iron homeostasis and is important for cardiac function. PNAS. 2015; 10;112(10):3164-3169.

Disclosures
No relevant conflicts of interest to declare.
Long-term Live Single Cell Quantification of Transcription Factor Dynamics
Authors: Schroeder Timm 
Published: October 2019

Abstract
Hematopoiesis is highly complex and dynamic, and consist of large numbers of different cells expressing many molecules. Despite intensive research, many long-standing questions in hematopoiesis research remain disputed. One major reason is the fact that we usually only analyze populations of cells - rather than individual cells - at very few time points of an experiment. Tracking of individual cells would be an extremely powerful approach to improve our understanding of molecular cell fate control. We are therefore developing imaging systems to follow the fate of single cells over many generations. We program new software to help recording and displaying the divisional history, position, properties, interaction, etc. of all individual cells over many generations. In addition, novel microfluidics devices are designed and produced to allow improved observation and manipulation of cells. Our technologies allow continuous long-term quantification of protein expression or activity in living cells. Among other approaches, we generate knock in models expressing transcription factor to fluorescent protein fusions from endogenous gene loci. This enables non-invasive long-term live quantification of transcription factor protein dynamics in single stem and progenitor cells throughout their differentiation. The resulting novel kind of continuous quantitative single cell data is used for the generation and falsification of models describing the molecular control of hematopoietic cell fates.

Disclosures
No relevant conflicts of interest to declare.
Toll-Like Receptors: Mechanism and Relation to Human Populations
Authors: Freedman Jane 
Published: October 2019

Abstract
Inflammation and infection are known to alter platelet count, production, and function and major studies have demonstrated that acute infection is associated with a transient 5-fold increased risk of thrombotic vascular syndromes including stroke, acute myocardial infarction, and peripheral vascular occlusion. Platelets play an intricate role in thrombosis, myocardial infarction, and thrombotic stroke and are now being appreciated for their functional innate immune receptors. More recently, platelets have been connected to the host's immune system via their ability to trap bacteria and present them to leukocytes for destruction. This interaction appears crucial to the host's immune defense. Platelets are known to express functional interleukin 1 receptor (IL1R), toll like receptor (TLR)2, TLR3, TLR4, TLR7, and TLR9 with other immune receptors under investigation. It is known that activation of TLR2, TLR7, or TLR4 leads to platelet-neutrophil interaction.Platelet-leukocyte communication has also been observed during different bacterial or viral infections.
We have used animal models and large-scale human clinical data to examine if platelets demonstrate specific immunity or broadly express TLRs and characterize how this expression is causative in immune responses. Our studies have included thousands of participants of the Framingham Heart Study in addition to investigations of active infection and inflammatory diseases. We have also examined if presence of immune receptors in platelets is associated with vascular inflammation and cardiovascular disease or risk factors. We have found widespread and varied immune and inflammatory platelet receptor and gene expression that varies remarkably between individuals and diseases.
Additionally, anucleate platelets contain transcripts that might relate to other physiological or pathophysiological conditions and are known to be released into the circulation, participate in protein formation, and engage in horizontal RNA transfer to other vascular cells. These platelet transcripts include microRNAs (miRNAs), which are small noncoding RNAs involved in many molecular processes, most notably regulation of gene expression and release of these noncoding RNAs also may be regulated by innate immune receptors. In platelets and their released exosomes, these noncoding RNAs seem to participate in vascular homeostasis, inflammation, and platelet immune function. In addition, they are associated with the presence or extent of cardiovascular diseases, such as atrial fibrillation and peripheral vascular disease. Thus, platelets are known to contain a myriad of receptors that regulate hemostasis and thrombosis; however, there is growing appreciation of the diverse expression of receptors traditionally associated with immune and inflammatory cells.

Disclosures
No relevant conflicts of interest to declare.