bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of a Functional Interferon-Gamma Response Element in the Fibrinogen FGG Gene.
Authors: Rein Chantelle M. , Farrell David H. 
Published: October 2019

Abstract
Abstract 3174
Poster Board III-114
Fibrinogen is the zymogen precursor to the main structural protein in blood coagulation, and is proteolytically activated during coagulation to form fibrin clots. Fibrinogen is an acute phase protein that is produced by the liver, and elevated fibrinogen levels are a known risk factor for cardiovascular disease, including heart attack and stroke. Fibrinogen is synthesized from three separate genes, FGA, FGB, and FGG encoding the Aαa, Bβ, and γ chains, respectively. Several inflammatory cytokines, particularly interleukin-6 (IL-6), are known to up-regulate fibrinogen synthesis, while interleukin-1β, tumor necrosis factor-αa, and transforming growth factor-β are known to antagonize IL-6 stimulation of fibrinogen synthesis. Here we report that interferon-γ (IFN-γ), a cytokine that is highly expressed in atherosclerotic lesions, decreases the production of fibrinogen by two-fold over a 24 hour period in HepG2 hepatocellular carcinoma cells, both at the protein and mRNA level. In addition, IFN-γ antagonizes IL-6 up-regulation of fibrinogen production. Since IFN-γ is known to signal via the IFN-γ receptor through a STAT1-dependent pathway, we investigated the mechanism by which IFN-γ inhibits fibrinogen production. We identify a previously unknown IFN-γ activation sequence (GAS), AGGAGCTTACATAAAGGGACAA, within the promoter of the human γ chain gene FGG. This sequence is homologous to the known GAS consensus sequence TTNCNNNAA. Nuclear extracts from IFN-γ-stimulated HepG2 cells, but not from unstimulated HepG2 cells, form a complex with oligonucleotides containing this GAS sequence, as demonstrated by electrophoretic mobility shift assays. The formation of this complex is blocked by unlabeled competing oligonucleotides and by an antibody to phosphorylated STAT1. The involvement of pSTAT1 in this complex is consistent with the known role of pSTAT1 downstream of IFN-γ receptor signaling. In addition, IFN-γ stimulates STAT1 phosphorylation in a dose-dependent manner in HepG2 cells. Furthermore, using an FGG promoter construct fused to a luciferase reporter gene in transiently transfected HepG2 cells, we show that IFN-γ inhibits luciferase expression in a dose-dependent manner. Together, these data demonstrate that IFN-γ down-regulation of fibrinogen production occurs through a pSTAT1-mediated pathway. Since IFN-γ is known to demonstrate both pro and anti-atherogenic actions, this down-regulation of fibrinogen production may serve an anti-atherogenic function in vivo.

Disclosures
No relevant conflicts of interest to declare.
Predictors of Long-Term Cytogenetic Response Following Dasatinib Therapy of Patients with Chronic-Phase Chronic Myeloid Leukemia (CML-CP).
Authors: Jabbour Elias , Bahceci Erkut , Zhu Chao , Lambert Alexandre , Cortes Jorge 
Published: October 2019

Abstract
Abstract 3296
Poster Board III-184

BACKGROUND
Achievement of major and complete cytogenetic response on imatinib in newly diagnosed CML-CP has been associated with excellent survival. Dasatinib is a potent oral tyrosine kinase inhibitor with marked activity and good tolerability in patients with any-phase CML who have failed prior therapy, including imatinib.

AIM
To investigate factors predictive of cytogenetic response and survival outcomes for patients who receive dasatinib after imatinib failure.

METHODS
Collated patient data from trials of dasatinib (START-C, START-R, and CA180-034) in patients with CML-CP were analyzed. All patients had failed prior treatment with imatinib as a result of resistance or intolerance. Patients with highly imatinib-insensitive BCR-ABL mutations (L248V, G250E, Q252H/R, Y253H/F, E255K/V, T315I/D, F317L, and H369P/R) were ineligible for START-R. Across the three trials, patients were administered dasatinib 50 (n=167) or 70 (n=655) mg twice daily, or 100 (n=165) or 140 (n=163) mg once daily. Analysis of efficacy parameters was performed on all patients receiving at least one dose of dasatinib (n=1150). A multivariate logistic regression model was applied to evaluate patient treatment characteristics as predictive factors for cytogenetic response.

RESULTS
After a median follow-up of 26 months, the rate of major cytogenetic response (MCyR) was 62% and of complete cytogenetic response (CCyR) was 51%. By multivariate analysis, the following were selected as independent favorable prognostic factors for achievement of MCyR: younger age, lower percentage of Ph+ cells, absence of the T315I mutation, prior MCyR with imatinib, imatinib intolerance (vs. resistance), no prior stem cell transplantation (SCT), shorter time from CML diagnosis to dasatinib therapy (Table 1). The same baseline factors independently predicted CCyR.

CONCLUSION
This study confirms the clinical efficacy of dasatinib among patients with CML-CP who have failed prior therapy. Furthermore, we identified baseline factors associated with improved response to dasatinib. Work to determine additional predictive factors for survival outcome with dasatinib is ongoing. The subgroups of patients who may not respond optimally to dasatinib are small, and a modification to their treatment regimen may be called for.

Disclosures
Jabbour: Bristol-Myers Squibb: Speakers Bureau; Novartis: Speakers Bureau. Bahceci:Bristol-Myers Squibb: Employment. Zhu:Bristol-Myers Squibb: Employment. Lambert:Bristol-Myers Squibb: Employment. Cortes:Bristol-Myers Squibb: Research Funding.
Prevalence of Coronary Heart Disease in Patients with Hemophilia: a Nationwide Study in Brazil.
Authors: Bley Claudia , Santos Fabio P S , Pesaro Antonio E P , Helman Ricardo , Oliveira Sergio A , Pfeferman Elcio , Serrano Carlos V , Saraiva Ana S L , Amico Elbio D , Hamerschlak Nelson 
Published: October 2019

Abstract
Abstract 3495
Poster Board III-432

Background
It is well known that patients with hemophilia (both A and B) have a lower incidence of coronary artery disease (CAD). However, patients with hemophilia may develop atherosclerosis, and with increases in life expectancy it is possible that the incidence of CAD may increase in these patients. There are no studies analyzing the prevalence of CAD in patients with hemophilia and possible risk factors.

Aims
To determine the prevalence of CAD in patients with hemophilia treated in Brazil.

Methods
All patients with hemophilia in Brazil receive therapy with Factor VIII or Factor IX at government-designed hemophilia centers. We thus contacted all centers in Brazil by e-mail and information from databases concerning health information for all patients was reviewed by the responsible physician at each center. A questionnaire about risk factors for CAD and history of coronary events was sent to patients that had a positive history of CAD.

Results
There are 6,881 patients registered with a diagnosis of hemophilia A and 1,291 patients with a diagnosis of hemophilia B in Brazil. Fifty-six percent of hemophilia centers answered the query, and information on 71.7% of patients with hemophilia A (4,934 patients) and 61.4% of patients with hemophilia B (792 patients) was reviewed. There were only 3 patients (0.05%) with a positive history of CAD. Their clinical characteristics are summarized in Table 1. During surgical treatment for CAD, replacement with Factor VIII and Factor IX was done by continuous infusion to increase Factor activity to 80-100%. Antiplatelet therapy was used in all patients at some point, and one patient (#1) developed hemarthrosis after two months of ASA. Patient #3 did not start antiplatelet therapy after CABG, and developed thrombosis of the graft. ASA and clopidogrel were started after percutaneous coronary angioplasty.

Conclusion
We found a very low incidence of CAD in hemophiliac patients, even after doing a nationwide survey that included more than 5,000 patients. No information about risk factors can be gleaned due to the small number of patients. Surgical treatment after increasing factor activity appears to be safe in such patients. Despite a theoretical risk for increased bleeding, therapy with antiplatelet agents appears to be necessary in hemophiliac patients with CAD.

Disclosures:
No relevant conflicts of interest to declare.
Long-Term Effects of In Vivo Administration of An Anti-VLA-4 Antibody (natalizumab) on Mobilization of Committed and Primitive Hematopoietic Progenitor Cells In Humans
Authors: Domenech Jorge , Goustille Julien , Pallix Maud , Bekhechi Zakia , Ducrocq Elfi , Degenne Michel , Herault Olivier , Thibault Gilles , Guennoc Anne-Marie , Binet Christian 
Published: October 2019

Abstract
Abstract 2264
The alpha4 beta1 integrin (VLA-4) exert a critical role on hematopoiesis by confining hematopoietic stem cells (HSC) and progenitor cells (HPC) within the niche. Previous preclinical studies have pointed out this role showing that HPCs can be mobilized by in vivo administration of a blocking anti-VLA-4 antibody (Ab) (Papayannopoulou et al, 1993). Very recently, two papers (Bonig et al, 2008; Zohren et al, 2008) have shown that such Ab exhibits a similar effect in humans for one month after treatment by natalizumab. In the present study, we have investigated long-term hematopoietic effects (up to 23 months) of repeated infusions of natalizumab in patients treated for multiple sclerosis (n=22). Seven patients have been explored sequentially (every month for one year) and 15 have been studied punctually (6 before and 9 after one year of treatment). We found that peripheral blood leukocytosis was consistently increased for one year in relation to lymphocytes, particularly to B lineage (by 3-fold). In parallel, an increase of circulating HPCs (CD34+ cells and total CFU) was observed and appeared more pronounced with levels above baseline values in all the patients studied (by 6-fold) while no increase of circulating mesenchymal stromal cells (CFU-F) was found. The increase was noted for both lymphoid (T and B lineages) and myeloid (granulo-monocyte, erythroid, and megakaryocyte) committed progenitor cells but also for primitive HPC (CD34+CD38- cells and CAFC). This effect was still found at long-term (up to two years of treatment) for both committed and primitive HPC. In conclusion, the HPC mobilizing effect of chronic administration of anti-VLA-4 Ab in humans involves all types of HPCs (lymphoid and myeloid, committed and primitive ones) and is not exhausted with time.

Disclosures:
No relevant conflicts of interest to declare.
Increased Red Cell Phosphatidylserine Exposure Correlates with Enhanced Thrombin Generation In Sickle Trait Patients with End Stage Renal Disease
Authors: Brittain Julia , Derebail Vimal K. , Mooberry Micah J. , Ataga Kenneth I. , Kshirsagar V. Abhijit , Key Nigel S. 
Published: October 2019

Abstract
Abstract 2665

Introduction:
Phosphatidylserine (PS) exposure by red blood cells (RBCs) is increased in sickle cell disease, where it is thought to contribute to anemia, inflammation and activation of coagulation. RBCs from subjects with sickle cell trait (SCT) may also express more PS compared to controls. We recently reported a high prevalence of SCT in African Americans with end-stage renal disease (ESRD) - a state marked by anemia, profound inflammation and coagulation activation. No study has yet examined RBC PS exposure, inflammation or coagulation activation in African Americans patients with SCT and ESRD.

Objective:
To determine whether RBC PS exposure is elevated in ESRD patients with SCT receiving hemodialysis compared to patients with normal hemoglobin genotype, and whether markers of coagulation activation and inflammation are similarly elevated. The relationship between RBC PS exposure, thrombin generation and inflammation was also examined.

Methods:
We performed a cross sectional study of 10 patients with documented SCT and 9 race-matched controls. Both groups were receiving in center hemodialysis. The percentage of RBCs demonstrating PS exposure was determined by annexin-V binding and flow cytometry detection. Plasma thrombin/anti-thrombin (TAT) complexes and C-reactive protein (CRP) levels were determined by ELISA. Mann-Whitney U tests were used to examine differences between these variables among the groups. Medians and interquartile ranges (IQR) are shown. Spearman's correlation coefficient was used to assess the relationship among the variables tested.

Results:
Consistent with previous reports in ESRD patients, we found markedly elevated RBC PS exposure among both groups; however, patients with SCT had significantly elevated PS exposure compared to those without SCT (6.65%, [IQR: 5.1, 8.8] vs. 3.0%, [IQR: 2.1,4.2], p = 0.002). Furthermore, patients with SCT had higher TAT levels compared to those without SCT (16.1 ng/L [IQR: 11.9, 28.4] vs. 5.5 ng/L [IQR: 3.8, 8.6], p<0.0001). We also noted increased plasma CRP levels in those patients with SCT compared to control (33.7 mg/L [IQR: 22.8, 62.8] vs. 11.2 mg/L [IQR: 2.2, 19.5], p = 0.016). In patients with SCT, there was a striking correlation between PS exposure and TAT levels (r = 0.897, p<0.0001), but no correlation was noted between PS exposure and levels of CRP (r=-0.17, p = 0.63).

Conclusions:
These data suggest that patients with SCT trait in ESRD may be subject to an increased risk of thrombotic complications related to increased PS exposure on RBCs.

Disclosures:
No relevant conflicts of interest to declare.
HLA Class I and EBV Positive Classical Hodgkin Lymphoma In the Chinese Population
Authors: Xin Huang , Diepstra Arjan , Gao Zifen , Visser Lydia , Vos Hans , Kooistra Wierd , Poppema Sibrand , Hepkema Bouke , Van Den Berg Anke 
Published: October 2019

Abstract
Abstract 2688
In Caucasian populations, the tumor cells of Epstein Barr virus (EBV)-positive classical Hodgkin Lymphomas (cHL) patients more frequently express HLA class I and HLA class II molecules compared to EBV-negative cHL patients. Since HRS cells also express other components of the antigen presenting pathway it can be anticipated that HRS cells in EBV-positive cHL should be able to present EBV derived antigenic peptides. In on our previous genotyping study we demonstrated that the HLA-A*02 allele was associated with a reduced and the HLA-A*01 allele with an increased risk of developing EBV-positive cHL. Intuitively, this can be explained by the generally acknowledged lack of HLA-A*01 restricted immune responses to latent EBV peptides. In this study, we analyzed HLA class I expression and the HLA association in relation to EBV in Asian cHL patients.
Formalin-fixed and paraffin embedded (FFPE) tissue blocks were available for 145 cHL patients and from 79 controls from 5 hospitals from the Northern part of China. Hematoxylin & Eosin-stained sections were used to reclassify the histological subtypes according to the WHO classification. EBV status was determined by EBER in situ hybridization. Membranous expression of HLA class I was detected by immunohistochemistry using antibodies against HLA class I (HC-10) and ß2-microglobulin. DNA was isolated from FFPE samples to detect the HLA-A*02 allele by quantitative PCR. 23 cHL cases and 5 controls were excluded due to poor DNA quality.
Positive EBV status was observed in 40% (58/145) of the Chinese cHL patients. As expected, the percentage of EBV-positive cases was much higher in the mixed cellularity subtype (71%) than in the nodular sclerosis subtype (16%; p<0.001). Expression of HLA class I was observed in 79% of the EBV-positive cHL patients and in 30% of the EBV-negative patients (p<0.001) consistent with the previous findings in the Dutch population. The HLA-A*02 allele was detected in 71% of the EBV-positive cHL and in 64% of the EBV-negative cHL patients (NS). In the controls HLA-A*02 was observed in 67% of the cases.
In this Chinese population, the tumor cells of EBV-positive cHL more frequently retained membranous HLA class I expression, similar to the Caucasian populations. The inverse correlation between presence of the HLA-A*02 allele and positive EBV status, as observed in Caucasians, is not present in these northern Chinese cHL patients. Differences in ethnic background might explain discrepancies in HLA-A association with EBV-positive cHL in different populations.

Disclosures:
No relevant conflicts of interest to declare.
Interleukin-2 Enhances Angiogenesis and Preserves Cardiac Function Following Myocardial infarction
Authors: Bouchentouf Manaf , Williams Patrick , Forner Kathy-Ann , Cuerquis Jessica , Boulassel Mohamed Rachid , Paradis Pierre , Routy Jean Pierre , Schiffrin Ernesto , Galipeau Jacques 
Published: October 2019

Abstract
Abstract 2786
Previous studies demonstrated that the addition of some cytokines improved cardiac recovery in experimental models of myocardial infarction (MI). The aim of the study is to test the hypothesis that pharmacological stimulation of lymphoid cells using recombinant human interleukin (rhIL)-2 improves angiogenesis and preserves heart function after MI. A single IV injection of rhIL-2 two days following MI improved by 27.7% the left ventricular (LV) fractional shortening of immune competent (C57Bl6) mice, but had no effect on cardiac function of immune-deficient (NOD SCID IL2Rγnull) mice. Immunohistochemical analysis of C57Bl6 cross sections of heart revealed that collagen deposition was reduced by 23.1% and that capillary density was enhanced in the scar area and the border zone of the infarct respectively by 22.4% and 33.6% following rhIL-2 injection. In addition, rhIL-2 enhanced 1.6-fold the in vivo endothelial cell proliferation index and 1.8-fold the number of NK cell infiltrating the infarcted heart, but had no effect on the number of cardiac CD4 and CD8 cells. In vitro, rhIL-2 activated NK cells enhanced cardiac endothelial cell proliferation by 17.2%. Adoptive transfer of IL-2-activated NK cells enhanced cardiac angiogenesis. Here we show that a single IV injection of rhIL-2 positively impacted cardiac function by improving angiogenesis through a process involving NK cells.

Disclosure:
No relevant conflicts of interest to declare.
The Novel JAK2 Inhibitor NVP-BSK805 Has Cytotoxic Activity on Malignant Plasma Cells
Authors: Burger Renate , Rademacher Franziska , Staudinger Matthias , Peipp Matthias , Güunther Andreas , Gramatzki Martin 
Published: October 2019

Abstract
Abstract 2993
In multiple myeloma (MM) and plasma cell leukemia, activation of the JAK/STAT pathway is induced by interleukin (IL)-6, which is produced and secreted into the tumor microenvironment primarily by stromal cells. Upon binding of IL-6 to its specific alpha-chain receptor, dimerization of the gp130 signaling subunits leads to activation of associated JAK kinases and STAT transcription factors. In particular, STAT3 has been shown to be essential for myeloma cell growth and survival. NVP-BSK805 (Novartis) is a novel substituted quinoxaline JAK2 inhibitor tool compound which displays more than 20-fold selectivity for JAK2 over the other JAK family members and more than 100-fold selectivity over a panel of additional kinases (Baffert et al., Mol Cancer Ther 9:1945, 2010). The study presented here aims at growth inhibitory effects of NVP-BSK805 in malignant plasma cells. NVP-BSK805 inhibited the growth of six human myeloma cell lines displaying dose-dependent activity with IC50 concentrations between 2.6 μ mol/L and 6.8 μ mol/L. Among the cell lines, IL-6 dependent INA-6 cells were most sensitive to the inhibitory effects of the compound: both IL-6 and bone marrow stromal cell induced proliferation as measured by [3H]-thymidine uptake was completely inhibited at 4 μ mol/L and IC50 concentrations were less than 1 μ mol/L. Viability of the stromal cells was not significantly affected. NVP-BSK805 concentrations as low as 0.5 μ mol/L were sufficient to yield a marked reduction of IL-6 induced STAT3 phosphorylation and complete abrogation at 2 μ mol/L, thereby blocking essential survival signals. Accordingly, treatment of INA-6 cells with NVP-BSK805 for 48 hours led to significant apoptosis starting at 2 μ mol/L with a 30% increase in annexin V-positive cell numbers compared to DMSO controls. Importantly, NVP-BSK805 showed potent cytotoxic activity on plasma cell-enriched primary tumor samples from patients with extramedullary plasma cell disease that are highly responsive to IL-6: in 3 out of 4 tumor samples the IC50 concentrations were between 0.5 μ mol/L and 0.6 μ mol/L. These studies are extended to combinations of NVP-BSK805 with PI3K, mToR, MAPK, HDAC, and IGF-1R inhibitors in order to optimize targeted therapy strategies facing different pathway alterations in individual myeloma patients. In INA-6 cells, synergistic activity was found combining NVP-BSK805 with rapamycin and the MEK1 inhibitor U0126. Preclinical in vivo studies are ongoing. Our results with NVP-BSK805 substantiate the use of JAK inhibitors as a therapeutic strategy for patients with MM. Since results from studies with pan-JAK inhibitors such as pyridone 6 indicate involvement of additional JAK kinases, the choice of the optimal compound will depend on its JAK family selectivity and the biology of JAK signaling in MM.

Disclosures:
No relevant conflicts of interest to declare.
Identification of a Novel T-ALL Entity with NKX2-1/NKX2-2 Rearrangements
Authors: Homminga Irene , Pieters Rob , Langerak Anton , de Rooi Johan , Stubbs Andrew , Verstegen Monique , Vuerhard Maartje , Buijs-Gladdines Jessica , Kooi Clarissa , Klous Petra , Van lierberghe Pieter , Ferrando Adolfo , Cayuela Jean-Michel , Verhaaf Brenda , Beverloo Berna , Horstmann Martin A. , de Haas Valerie , Wiekmeijer Anna-Sophia , Pike-Overzet Karin , Staal Frank , de Laat Wouter , Soulier Jean , Sigaux Francois , Meijerink Jules P.P. 
Published: October 2019

Abstract
Abstract 3139
To identify novel oncogenic pathways in T-cell acute lymphoblastic leukemia (T-ALL), we combined expression profiling of 117 pediatric patient samples and detailed molecular cytogenetic analyses. Using unsupervised and supervised analyses, we identified a novel T-ALL entity that was associated with high expression of cell cycle genes and was accordingly denoted as the proliferative cluster. Patient samples in this cluster were devoid of known oncogenic rearrangements, and clustered together with TLX1-rearranged cases in unsupervised cluster analysis suggesting pathogenic mechanisms are conserved between these cases. Alike TLX1-rearranged cases, patients within this cluster were associated with cortical thymic arrest and expressed CD1 along with CD4 and CD8. Bioinformatic analyses identified NKX2-1 as strong outlier gene that was ectopically expressed in most patients samples belonging to this cluster. One patient sample had high expression of the homologous NKX2-2 gene. Molecular-cytogenetic analyses including the Chromatine Conformation Capture on Chip (4C) method revealed various rearrangement variants of the NKX2-1 locus at 14q13 in 6 patients, including inversions to the T-cell receptor alpha/delta locus (TRAD@) at 14q11, an inversion to the IgH@ locus at 14q32 and a translocation to the TRB@ locus at 7q34. A translocation of NKX2-2 to the TRAD@ locus was identified in the case that ectopically expressed NKX2-2. All these rearrangements were novel, and have not been observed before in human cancer. NKX2-1 rearrangements could be validated in an independent T-ALL cohort. Our data strongly imply that NKX2-1/NKX2-2 represent novel oncogenes for human T-ALL.

Disclosures:
No relevant conflicts of interest to declare.