bluuebunny/crossref_metadata_2025_split huggingface dataset:
Decreased NO Bioavailability Leading to Endothelial Dysfunction in Children with ALL Results From Increased eNOS Inhibiting by Asymmetrical Dimethylarginine and May Be Associated with Increased Mortality
Authors: Porwolik Julita , Niedzielska Ewa , Doroszko Adrian , Piasecka-Grzeszek Alicja , Solska Urszula , Chybicka Alicja , Szuba Andrzej 
Published: October 2019

Abstract
Abstract 5318

Background
Endothelial dysfunction worsens the prognosis in numerous severe diseases. Impaired vascular reactivity, increased procoagulative and proinflammatory action are commonly observed when the nitric oxide bioavailability is limited. Asymmetrical dimethylarginine (ADMA) is a competive inhibitor of the endothelial nitric oxide synthase (eNOS). ADMA is produced during proteolysis of methylated proteins, especially histones. The aim this study was to determine if ADMA may play important role in pathogenesis of endothelial dysfunction in children with ALL.

Material and Methods
N=14 children at age of 4–18 years with ALL treated with the ALLIC protocol were investigated. Plasma levels of the NO pathway metabolites (L-Arginine, ADMA) were analyzed at baseline, then during the 33rd and 78th day of protocol. The control group constituted of N=14, age-matched healthy children.

Results
Plasma ADMA levels were significantly higher in children with ALL at baseline as compared to the control group (1.92±0.42 vs. 0.56±0.1 ng/ml, p<0.05), and were significantly decreasing to 0.63±0.15ng/ml, p<0.05 following the steroid therapy. During the 78th day the ADMA levels were maintained at similar levels as compared to the 33rd day. Opposite trends were observed with the L-Arginine levels and the L-Arg/ADMA ratio reflecting the NO synthesis. L-Arg and L-Arg/ADMA ratio were significantly lower in children with ALL at baseline vs. control group (41.65±4.12vs.52.4±2.31pg/ml and 28.7±6.61vs.83.41±14.13, respectively, p<0.05). Moreover, the L-Arg/ADMA ratio at the 78th day was significantly higer in cases treated unsuccesfully in comparison with those who survived (88.1±21.3vs.43.0±10.2, p<0.05).

Conclusions
Decreased NO bioavailability in children with ALL at baseline results from the eNOS inhibiting by ADMA and may be associated with increased mortality.

Disclosures:
No relevant conflicts of interest to declare.
Effects of Low ADAMTS 13 Levels and the Role of Von Willebrand Factor in Cardiovascular Disease
Authors: Corinaldesi Giorgio 
Published: October 2019

Abstract 5320
Von Willebrand factor (vWF) is a large multimeric glycoprotein that plays an important role as a co-factor in platelet adhesion and aggregation and as the circulating carrier for factor VIII; increased vWF levels, and the persistence of unprocessed ultra-large vWF multimers (ULVWF) that are hyperactive to binding GPIb-IX-V complex with high-strength bonds, precipitate platelet clumping in arterioles and capillaries, causing platelet aggregation and thrombotic microangiopathy resulting in tissue ischemia with an increased risk of CAD and stroke. vWF is the major ligand binding the subunit of trombospondin, P-selectin, collagen and SLAM family (signaling lymphocytic activation molecule), mediates platelet translocation, tethering and rolling under high shear stress and adhesion.
The activity of VWF is modulated by ADAMTS-13 a Zn2+/Ca2+ dependent metalloprotease that cleaves vWF to smaller and less active forms at the tyr842/met843 within the A2 domain, and cleaves the tyr 1605/met 1606; this mechanism reduces the molecular weight and platelet-tethering function of vWF to the sub endothelium at high shear stress. ADAMTS-13 levels also correlate positively with cholesterol, triglycerides and body mass index (BMI). In addition, we have observed a negative association with fibrinogen, C-reactive protein, and ion trasport K+dependent Na+/Ca2+ exchange (SLC24A3); the phosphorylation of tyrosine 1605 and threonine/serine kinase inhibitors have negative effects on integrin GPIIb-IIIa that showed decrease expression.
In patients with a level of ADAMTS-13 below 38%, the presence of ultra large vWF multimers is high and the relative risk of ischemic heart disease was 18.2% (95% CI, 0.8 to 4.2), and the relative risk of stroke was 11.5 % (95% CI, 0.6 to 3.9).
We investigated the relation between the low activity of ADAMTS-13 and AMI or stroke; we measured VWF and ADAMTS-13 antigen levels in 16 patients with AMI, in 10 patients with stroke, and in 16 control subjects.
We demonstrated that vWF levels are significantly higher in patients with AMI or stroke, while a severe decrease of ADAMTS-13 activity augments the risk of cardiovascular disease, and cerebral ischemia; measurement of both VWF and ADAMTS-13 may provide a good indicator for the likelihood of AMI and stroke.

Disclosures:
No relevant conflicts of interest to declare.
Sox4 Is Essential for Initiation and Progression of Acute Lymphoblastic Leukemia Induced by P190-BCR-ABL Transformation
Authors: Ma Haiqing , Mallampati Saradhi , Sun Baohua , Gong Yun , Lefebvre Véronique , Sun Xiaoping 
Published: October 2019

Abstract
Abstract 564
Sox4 is an Sry-related HMG box transcription factor that plays critical roles in development. Sox4-null mice die at embryonic day 14 due to cardiac malformation. Recent studies have suggested that SOX4 may contribute to tumorigenesis, based on the fact that SOX4 is upregulated in many human tumor types, including brain, bladder, prostate, colon, and lung cancers. In addition, increased expression of Sox4 induced by retroviral insertional mutagenesis in mice has been shown to be associated with leukemia and lymphoma. We also found that SOX4 is highly expressed in human acute lymphoblastic leukemia (ALL) cells. We therefore set out in this study to determine the function of Sox4 and its underlying mechanisms in ALL. To overcome embryonic lethality caused by conventional Sox4 inactivation, we used Sox4 conditional null mice (Sox4fl/fl) to establish Sox4-null ALL cell lines. First, we obtained pure B220+ pro-B cells by co-culturing Sox4fl/fl fetal liver cells and OP-9 bone marrow stromal cells in the presence of IL-7. Subsequently, pro-B cells were transformed with p190-BCR-ABL and Sox4 inactivation was induced with Cre recombinase. We found that Sox4−/− pro-B transformed cells exhibited a significantly slower proliferation rate than Sox4+/− pro-B cells. Notably, cell cycle analysis indicated a lower percentage of S phase cells in Sox4−/− pro-B transformed cells and these cells yielded much fewer colonies than transformed Sox4+/− cells in colony formation assay. We further labeled the transformed cells with a luciferase reporter and injected them intravenously into sublethally irradiated NOD/SCID mice. Bioluminescence imaging showed that mice injected with Sox4−/− cells developed leukemia significantly more slowly and survive longer than mice injected with Sox4+/− cells. In conclusion, our data show that Sox4 significantly contributes to initiation and progression of leukemia from p190-BCR-ABL-transformed ALL cells in the mouse and thereby strongly support the notion that SOX4 overexpression critically contributes to acute lymphoblastic leukemia development in humans.

Disclosures:
No relevant conflicts of interest to declare.
MYC
+ Diffuse Large B Cell Lymphomas (DLBCL) Treated in Randomized Prospective Salvage Therapy, RICE or RDHAP Followed by BEAM Plus Autologous Stem Cell Transplantation (ASCT). A BioCORAL Report
Authors: Cuccuini Wendy , Briere Josette , Mounier Nicolas , Voelker Hans-Ullrich , Rosenwald Andreas , Sundstrom Christer , Cogliatti Sergio , Bron Dominique , Hirchaud Edouard , Soulier Jean , Gaulard Philippe , Houlgatte Remi , Gisselbrecht C. , Thieblemont Catherine 
Published: October 2019

Abstract
Abstract 594FN2

Background
. Approximately 5% to 10% of DLBCLs harbor a MYC/8q24 oncogene rearrangement. We determined the prognostic significance of MYC+ DLBCL in a selected population of patients with relapsed/refractory DLBCL included in the CORAL and prospectively treated by RICE or RDHAP followed by BEAM and ASCT1. METHODS. Among the 396 patients included, 150 patients were analysed with break-apart FISH probes for MYC/8q24, BCL2/18q21, and BCL6/3q27. Correlations with clinical characteristics at diagnosis and at relapse, immunohistochemistry (IHC) for CD10, BCL6, MUM1, FOXP1, and BCL2 expression, cell of origin determined according to different IHC algorithms2-5 and by transcriptomic analysis, response to treatment and survivals were analysed considering presence or absence of MYC rearrangement. RESULTS. Twenty-four (16%) cases exhibited a MYC rearrangement, associated in 87.5% (n=21) of the cases with a BCL2/18q21 rearrangement (n=16), with a BCL6/3q27 rearrangement (n=4), and with both BCL2/18q21 and BCL6/3q27 rearrangements (n=4). Among the matched cases analysed (n=57), biological characteristics including tumor immunophenotype and breakpoints occurrence, were identical between primary and secondary biopsies. Presence of MYC+ rearrangement was significantly associated with CD10 expression (p= 0.0067) and a GC phenotype, based either on Hans algorithm (p<104) or on Alizadeh GC signature (p=0.0015). Patients with MYC+ DLBCL presented significantly with more advanced aaIPI (p =.03). Sixty-three percent of them presented a time to relapse < 12 months, compared to 51% in patients with MYC− DLBCL (p=0.2925). Before HDT+ASCT, CR and ORR (PR+CR) were observed in 56% and 63% in patients with MYC+ DLBCL, compared to 75% and 87% in patients with MYC− DLBCL (p=0.3442). Outcome of patients with MYC+ DLBCL was significantly worse than MYC− DLBCL, with a 4-y PFS at 22% vs 45% (p =.0105) and a 4-y OS at 33% vs 62% (p =.0192). Type of treatment, RDHAP or RICE did not have any impact on survival with a 4-y PFS at 23% vs 22% (p=NS) and a 4-y OS at 44% vs 28% (p=NS), respectively. No difference of outcome was observed between patients with simple hit and with complex hits MYC+ DLBCL. CONCLUSION. MYC rearrangement is an early event in DLBCL and is significantly associated with a GC phenotype. Patients with MYC+ DLBCL presented with a more advanced disease, and have a significant inferior prognosis than patients with MYC− DLBCLs. Their outcome seems not influenced by the proposed salvage therapy.

Disclosures:
No relevant conflicts of interest to declare.
Development of a Novel Small Molecule CRM-1 Inhibitor for Non Hodgkin's Lymphoma
Authors: Azmi Asfar Sohail , Al-Katib Ayad , Aboukameel Amro , Kauffman Michael , McCauley Dilara , Mohammad Ramzi , Shacham Sharon 
Published: October 2019

Abstract
Abstract 598
Recent evidence demonstrates that Non Hodgkin's Lymphoma (NHL) tumors have elevated expression of the XPO1 gene that codes for the nuclear exporter protein CRM-1 controlling the localization of critical tumor suppressors including p53 family members. For effective apoptosis and cell cycle regulation, p53/p73 nuclear localization and DNA binding are highly critical making CRM-1 an attractive therapeutic target for NHL that carries >90% wild type/functional p53 and only rare mutations in p73. We have identified novel small molecule inhibitors of CRM-1 (KPTs) that bind irreversibly and lock target proteins (including p53 and p73) in the nucleus leading to apoptosis of tumor cells. We demonstrate for the first time that KPTs can induce apoptosis in resistant NHL cell lines and corresponding xenograft models. The most potent CRM-1 inhibitor (KPT-185) induced growth inhibition and apoptosis in a panel of NHL cell lines with a median IC50 ∼25 nM. Western blot and confocal microscopy analyses demonstrated that KPT-185 treatment resulted in nuclear localization of p53 in wt-p53 and p73 in mut-p53 cell lines. Additionally, we observed KPT-185 mediated activation of p21 and Bax, known downstream executioners of p53/p73 cell cycle control and apoptosis, respectively. Most significantly, siRNA knockdown of p53 in WSU-FSCCL and p73 in WSU-DLCL2 abrogated the apoptotic potential of KPT-185, confirming that these were indeed p53/p73 dependent apoptotic events. KPT-185 showed a substantial enhancement in apoptosis when combined with genotoxic p53/p73 re-activating regimen CHOP. The KPTs selectively kill cancer cells with minimal toxicity to normal tissues, and possess clinically acceptable pharmacokinetic parameters. Using WSU-DLCL2 SCID models, we further show that oral administration of related CRM-1 inhibitor KPT-276 (75 and 150 mg/Kg p.o) resulted in 65 and 70% tumor reduction, respectively and subcutaneous injections of KPT-251 (25 and 75 mg/Kg) resulted in 70 and 74% suppression of tumor growth with no observed toxicity to the host. Remnant tumor tissue analysis (histology and protein markers) fell in line with our in vitro results with clear activation of p73 pathway. Our study verifies CRM-1 as a potential therapeutic target in NHL irrespective of the functional status of p53. These results build a strong case for the clinical use of our novel CRM-1 inhibitors either as single agents or in combination with CHOP.

Disclosures:
Kauffman: Karyopharm: Equity Ownership. McCauley:Karyopharm: Employment. Shacham:Karyopharm: Equity Ownership.
Mini-Hepcidins Prevent Iron Overload In A Mouse Model of Hereditary Hemochromatosis
Authors: Nemeth Elizabeta , Ramos Emilio , Ruchala Peter , Preza Gloria , Ganz Tomas 
Published: October 2019

Abstract
Abstract 689
Mini-hepcidins are synthetic peptide analogues of the hepcidin N-terminus which is crucial for hepcidin interaction with ferroportin. Due to their small size and relative ease of synthesis, mini-hepcidins are better candidates than native hepcidin as therapeutics for the prevention and treatment of iron overload. In our previous studies, the first 9 amino acids of hepcidin (DTHFPICIF) were sufficient for in vitro activity (measured as ferroportin-GFP degradation). We modified the amino acid sequence of hepcidin-9 to improve resistance to proteolysis and to enhance the biophysical interactions with ferroportin. From >70 modified mini-hepcidins, we selected several that were more potent than native hepcidin in vitro, and tested them in mouse models. Bioactivity was assessed by measuring the hypoferremic effect of minihepcidins in C57BL/6 mice 4h after administration. Several minihepcidins administered by intraperitoneal or subcutaneous injection were more potent than the native hepcidin in reducing serum iron. Remarkably, retroinverted mini-hepcidins modified by palmitoylation or bile acid conjugation were active also by gavage. None of the tested mini-hepcidins altered endogenous hepcidin production. We also examined the ability of parenteral minihepcidins to prevent tissue iron loading in hepcidin knockout mice. Daily injections for 12 days completely prevented iron loading of the liver, decreased serum iron and increased splenic iron content. At high doses, minihepcidins were sufficiently potent that they caused iron-restricted microcytic anemia in hepcidin knockout mice. Minihepcidins may be useful for the treatment of human iron overload conditions caused by hepcidin deficiency.

Disclosures:
No relevant conflicts of interest to declare.
Thrombotic and Inflammatory Mediators in Cancer Patients Are Downregulated by Enoxaparin. Biochip and Protein Array Analysis
Authors: Hoppensteadt Debra , Litinas Evangelos , Khan Hussein , Cunanan Josephine , Thethi Indermohan , Iqbal Omer , Fareed Jawed 
Published: October 2019

Abstract
Abstract 713

Introduction:
The pathogenesis of cancer is known to upregulate inflammatory and thrombotic processes which contribute to the increased mortality in these patients. We hypothesized that the baseline inflammatory and thrombotic mediators are upregulated in cancer and treatment with LMWHs such as enoxaparin may downregulate these mediators. Methods: To test this hypothesis plasma samples were retrospectively analyzed from an open label multi dose active comparator parallel design study in which all patients (n=110) were initially treated with enoxaparin (1-1.5 mg/kg sc) for 5 days. These patients were subdivided into two groups. Group A continued to receive enoxaparin whereas Group B received warfarin. Baseline blood samples, 5 days and 12 weeks post treatment samples were analyzed using bio chip arrays (Randox analyzer) and protein chip array using surface enhanced laser desorption ionization (SELDI) technique.Results: In the cerebral biochip array analysis, levels of CRP, TNFRI, D DIMER, NGAL and TM were elevated at baseline which reduced after treatment with enoxaparin at three months except for NSE and TNFRI. In the cytokine biochip array, IL2, IL4, IL6, IL8, IL10, VEGF, IFNG, TNFA, IL1A, IL1B, MCP1 and EGF showed marked upregulation at baseline with enoxaparin treatment resulting in a decrease of IL6 alone. In contrast the warfarin treated group, both the cerebral and cytokine biochip array showed only marginal decreases.The baseline plasma samples from the patients recruited with multiple cancers in the Oncenox study showed a greater prevalence of the 11.6kDa biomarker (76%) with average amplitude of 23.6. The samples collected after 3 Months of enoxaparin treatment revealed markedly reduced prevalence (38%) and average amplitude of 5.4, whereas the warfarin treated group showed a modest decline in the 11-6kD a biomarker with a 52% prevalence and an average amplitude of 18.2. Conclusions: These results confirm that inflammatory and thrombotic mediators are differentially downregulated by treatment with enoxaparin in comparison to warfarin. The biochip and protein arrays provide unique tools to profile the known mediators and identify newer biomarkers. which may be useful in differentiating target sites for the newer drugs in cancer management.

Disclosures:
No relevant conflicts of interest to declare.
A “canonical” Npm1 mutation Knock-in Mouse Model Revealed Subtle but Definitive Myeloid Expansion with Poor HSC Niche Interaction
Authors: Chou Wen-Chien , Chou Shiu-Huey , Chiou Ji-Shain , Ko Bor-Sheng , Lin Shu-Wha , Hsu Yueh-Chwen , Tien Hwei-Fang 
Published: October 2019

Abstract
Abstract 762
Nucleophosmin (NPM1) is a ubiquitous multifunctional phosphoprotein, which has nucleocytoplasmic shuttling activity. Somatic mutations in NPM1 gene result in cytoplasmic dislocation of NPM1 (NPM1c) and are frequently associated with acute myeloid leukemia (AML). The pathogenetic effects of mutated NPM1 protein have been explored by animal models including transgenic or “humanized” knock-in mouse models. Here, we demonstrate the first “canonical” mouse Npm1 mutant knock-in model. Different from the previously report of humanized NPM1 mutant knock-in model, we inserted TCTG after nucleotide c.857 of murine Npm1 coding sequence (c.854–857dupTCTG), a pattern identical to human NPM1 mutation, without any “humanized” sequence. This mutation caused a shift of peptide sequence from WQWRKSL* (amino acid 286–292) to LCLAVEEISLRKGFKQFEIFCLHFCNS* (amino acid 285 to 311), a pattern mildly different from the change in human NPM1 mutation, but is still predicted to generate a nuclear export signal. NPM1c+ genotype and protein accumulation in cytoplasm were confirmed with PCR and immunocytochemistry, respectively. We found that the homozygous NPM mutant (NPMc+/c+) mice were embryonic lethal before E10.5 day, while hetrozygote (NPMwt/c+) mice survived and were fertile, and born with Mendelian ratio. Most NPMwt/c+ mice had normal hematologic parameters and remained disease-free, however, these mice developed a delayed-onset aberration on the distribution of granulocyte-monocye progenitors (GMP), monocytes, and B lymphocytes in blood, spleen, and bone marrow. Colony forming unit assay showed normal hematopoietic development of marrow hematopoietic stem cells (HSC), but poor cobblestone formation while HSCs contacted with stroma microenvironment in NPMwt/c+ mice, suggesting the NPMc mutant may affect the ability of HSCs on contact signal expression. Three (12.5%) of 24 NPMwt/c+ mice developed leukocytosis and splenomegaly mimicking myeloproliferative neoplasm of human. Microscopic examination showed panmyelosis of the bone marrow and existence of hematopoiesis in the spleen. In addition, the immune activities of NPMwt/c+ mice's splenocytes and thymocytes with mitogen stimulation were decreased. In summary, our “canonical” NPMwt/c+ mouse model demonstrated subtle but definitive phenotypes in hematopoietic cells and provided insight into the pathogenesis of NPM1 mutation in human acute myeloid leukemia. A second hit may be necessary for the development of AML in NPMwt/c+ mice since no AML was detected in these mice till 20 months of age.

Disclosures:
No relevant conflicts of interest to declare.
Role of Microparticles in Hemostasis and Thrombosis
Authors: Mackman Nigel 
Published: October 2019

Abstract SCI-10
Microparticles (MPs) (also known as or microvesicles) are submicron plasma membrane vesicles released from activated or apoptotic cells. Current estimates indicate that MPs are ∼200 nm in diameter. They are formed as membrane blebs and have phosphatidylserine exposed on the surface due to a loss of membrane asymmetry. MPs containing tissue factor (TF) are highly procoagulant. We recently developed an assay to measure TF activity of MPs isolated from plasma. We found that healthy individuals contain low levels of TF+ MPs, but levels are increased in many pathological conditions associated with thrombosis, such as sepsis, cardiovascular disease, sickle cell disease, diabetes mellitus, and cancer. We have analyzed the role of TF+ MPs in various murine models of thrombosis and in different patient populations. Work in collaboration with Bruce and Barbara Furie demonstrated that leukocyte-derived TF+ MPs contributed to microvascular thrombosis in a model of laser injured cremaster arterioles using healthy mice. Further studies have been performed using different murine models of disease. Arterial thrombosis is mostly caused by rupture of atherosclerotic plaques and leads to myocardial infarction and stroke. In hypercholesterolemic LDLR−/− mice we observed increased levels of hematopoietic cell-derived TF+ MPs that were associated with a prothrombotic state. These TF+ MPs may enhance thrombosis after plaque rupture. Indeed, we observed more fibrin deposition in the laser injury model of thrombosis in hypercholesterolemic mice than controls. Importantly, we found that simvastatin reduced TF expression in mononuclear cells and levels of TF+ MPs in the plasma in these hypercholesterolemic mice. Venous thrombosis occurs without significant damage to the vessel wall and is associated with embolization of clot fragments that can block pulmonary arteries. Venous clots appear to form due to changes in the vessel wall, stasis, and/or increased coagulability of the blood. We have analyzed levels of TF+ MPs in murine cancer models and cancer patient samples. We and other have found that tumors release TF+ MPs into the circulation. In one study, we found that pancreatic patients with the highest levels of MP TF activity develop thrombosis. These results suggest that TF+ MPs may enhance venous thrombosis in cancer patients by binding to activated endothelium. Further studies are needed to determine if levels of TF+ MPs in the circulation can be used to identify patients at risk for thrombosis.

Disclosures:
Mackman: Merck: Consultancy; Daiichi: Consultancy; Daiichi: Research Funding.