bluuebunny/crossref_metadata_2025_split huggingface dataset:
Edx-17: A Novel, Safe and Efficacious Treatment for Sickle Cell Disease
Authors: Broyles Robert H. , Joshi Sunil K. , Curtis Carol D. , Roth Austin C. , Floyd Patrick A. , Trudel Marie , Belegu Visar , Baker Ashley A. , Floyd Robert A.. 
Published: October 2019

Abstract

Gene regulation of developmental hemoglobin switching holds the potential for therapeutic relief from all symptoms associated with Sickle Cell Disease (SCD). Reactivation of fetal gamma-globin expression (HbF) can replace mutant betaS-globin (HbS) to produce functional hemoglobin tetramers and eliminate the hemoglobin polymerization that is characteristic of sickled red blood cells. We have discovered a protein that regulates this developmental switch, and have identified a compound that stimulates expression of this protein. EdX-17 promotes expression of the anti-stress factor ferritin heavy chain (FtH), which enters the nucleus of erythroid precursor cells and activates expression of fetal gamma-globin, producing HbF (PNAS 98:9145-50, 2001; Blood 108:790a, 2006). Mononuclear cells were isolated from SCD patient blood and matured in vitro to the advanced erythroblast stage using a 28-day, 2-phase culture system (Methods in Molecular Biology 482:127-40, 2009; Blood 119:6296-306, 2012). Treatment with EdX-17 for 24h resulted in a dose-responsive induction of gamma-globin gene expression and a concomitant dose-responsive increase in HbF was observed after 28 days in culture. These studies demonstrate that EdX-17 doses in the picomolar range are sufficient to significantly enhance HbF. Furthermore, EdX-17 treatment reconstitutes fetal hemoglobin (HbF) in transgenic betaYAC mice to levels above 25-30% - the range thought to be sufficient to ameliorate symptoms of SCD – with no detectable ill effects. In fact, mice treated with EdX-17 tend to have shinier coats, are more alert and stronger than age-matched, untreated mice. Development of this novel therapeutic is expected to ameliorate SCD symptoms, decrease pain and morbidity, increase life-span, greatly improve patient quality of life, and significantly reduce treatment costs. Supported in part by The Sickle Cell Cure Foundation, Inc., the Bill & Melinda Gates Foundation, and EpimedX, LLC.

Disclosures
Broyles: EpimedX, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding. Curtis:EpimedX, LLC: Employment, Equity Ownership, Research Funding. Roth:EpimedX, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding. Floyd:EpimedX, LLC: Employment, Equity Ownership, Research Funding. Belegu:EpimedX, LLC: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding. Floyd:EpimedX, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding.
Inorganic Polyphosphate Activates mTOR Complexes 1 and 2 in Vascular Endothelial Cells
Authors: Hassanian Mehr Seyed Mahdi , Dinarvand Peyman , Rezaie Alireza R. 
Published: October 2019

Abstract
Inorganic polyphosphate (polyP) elicits proinflammatory signaling responses in endothelial cells through interaction with two receptors, RAGE and P2Y1. We demonstrate here that polyP elicits proinflammatory signaling responses through activation of mTOR complexes 1 (mTORC1) and 2 (mTORC2) in endothelial cells. Thus, polyP effectively induced the phosphorylation of the mTORC1 substrate, p70S6K, in endothelial cells by an AKT-dependent but ERK-independent mechanism. Furthermore, it was found that activation of the PI3K/PLC/PKC/Ca2+ signaling axis is required for the polyP-mediated phosphorylation of p70S6K. The inhibition of mTORC1 by either rapamycin or by the siRNA knockdown of raptor (mTORC1-specific component) but not rictor (mTORC2-specific component) abrogated polyP-mediated phosphorylation of p70S6K. By contrast, the siRNA knockdown of rictor but not raptor eliminated the barrier disruptive effect of polyP, suggesting that the barrier permeability regulating function of polyP is mediated through activation of mTORC2. Specific NF-kB inhibitors abrogated polyP-mediated phosphorylation of p70S6K and rapamycin suppressed polyP-induced activation of NF-kB, suggesting both proinflammatory and mTOR signaling functions of polyP are linked. Finally, inhibitors of mTOR or its upstream signaling network eliminated polyP-mediated vascular leakage and leukocyte recruitment in animal models, suggesting that findings in cellular models are physiologically relevant.

Disclosures
No relevant conflicts of interest to declare.
Usefulness of Dilute Russell’s Viper Venom Clotting Time (DRVVT) for the Routine Monitoring of New Oral Anticoagulants
Authors: Hoppensteadt Debra , Kola Jake , Brayne Mark , Iqbal Omer , Fareed Jawed 
Published: October 2019

Abstract
The new oral anticoagulants such as rivaroxaban (Bayer Healthcare) (R) apixaban (BMS/Pfizer) (A) and dabigatran (Boehringer Ingleheim) (D) have been approved to manage thrombotic and cardiovascular disorders in the US and Europe. Initially it was thought that these agents would not require monitoring at the approved dosages for specific indications. However, there have been reported bleeding complications with some of these agents suggesting that monitoring of these drugs to optimize therapy in some patient populations may be needed. The purpose of this study was to determine which of the available clotting assays could be used to determine the circulating concentration of these new oral anticoagulants in plasma.
Materials: Citrated blood was drawn from ten donors and spun at 3000 rpm to obtain platelet poor plasma (PPP). The PPP was supplemented with A, R and D in a concentration range of 0-1.0 ug/ml. In addition, five plasma samples, for each agent, containing varying concentrations of either A, R or D were also blindly analyzed. The plasma samples were analyzed using PT/INR (Innovin, Seimens, Deerfield,IL), APTT reagent (Platelin, TCoag, Ireland) dilute Russell’s viper venom time (DRVVT) and DRVVT confirm (HemosIL, Instrumentation Laboratories (Bedford, MA).
Results: In the PT and APTT assays a concentration dependent increase in clotting time was observed, however the slope of the line was flatter than with the slope observed in the DRVVT and DRVVT confirm assays. In both the DRVVT and DRVVT confirm assays the concentration-response curves for A,D and R were straight lines with a steep slope. Therefore, it was possible to extrapolate the concentrations of the unknown samples using both of these assays. When using the DRVVT and DRVVT confirm assays , there was a good correlation between the calculated concentration and the actual concentration in the range of 50-1000 ng/ml (r2=0.82). At concentrations < 50 ng/ml, the calculated concentration showed a poor correlation with the actual concentration (r2=0.26).
Conclusions: These results suggest that the PT and APTT assays are not sensitive to the new oral anticoagulants and therefore cannot be used to determine the circulating concentrations of these agents. However, the dRVVT and DRVVT confirm assays are sensitive to A, D and R at concentrations > 50 ng/ml, therefore, A,D and R can be monitored by these assays. Both of these assays are commercially available and can be readily adapted to any automated instrument. These studies warrant clinical validation of these tests in patients populations treated with the newer oral anticoagulant drugs.

Disclosures
No relevant conflicts of interest to declare.
Comprehensive Kinase Profile of Pacritinib, a Non-Myelosuppressive JAK2 Kinase Inhibitor in Phase 3 Development in Primary and Post ET/PV Myelofibrosis
Authors: Singer Jack , Al-Fayoumi Suliman , Ma Haiching , Komrokji Rami S , Mesa Ruben , Verstovsek Srdan 
Published: October 2019

Abstract
BackgroundPacritinib, a JAK2, JAK2 V617F and FLT3 kinase inhibitor is in Phase III development in myelofibrosis. In preclinical models and in Phase 1 studies in both non-Hodgkin lymphoma and myelofibrosis, it appeared unique among this class of agents in its lack of myelosuppression at doses that inhibited the JAK2/STAT3 pathway and resulted in significant clinical efficacy. This potentially important clinical observation was verified in two Phase II studies in myelofibrosis that had no exclusion restriction for low platelets. Pacritinib was effective at reducing spleen size and symptoms regardless of starting platelet counts and there was no apparent decline in either platelet count or hemoglobin levels, or an increase in transfusion requirements.
Aims and Methods: To help elucidate the mechanisms for pacritinib’s lack of myelosuppression, a kinome screening analysis against 429 recombinant kinases at 100 nM pacritinib concentration was performed, followed by titration from 1-100nM against those that were >50% inhibited at 100nM (Anastassiadis et al.; Nature Biotechnology; 2011). In humans, pacritinib achieves steady state free drug levels of approximately 200nM at the standard 400 mg daily dose.
Results:All kinases with IC50 values of <50nM are included in the table.
Table Kinase IC50 (nM) JAK2; JAK2(V617F) 6.0; 9.4 FLT3; FLT3-ITD; FLT3 (D835Y) 6.4; 12.0; 8.3 TYK2 27.0 JAK3 18.3 IRAK1 (IL-1 receptor kinase) 13.6 HIPK4 (PT kinase for P53 ser-9) 14.5 FMS 39.5 c-KIT and most common mutations except (V560G), (V559D/V654A) 43.8 c-Src; (c-Src (T341M)) 47.0 (55.5)
Of note, pacritinib did not affect JAK1 (82% control at 100nM). As the concentrations of pacritinib achieved clinically are likely inhibit signaling the kinases listed in the table, these data suggest that the lack of myelosuppression with pacritinib may be either due to inhibition of JAK2 without affecting JAK1 or to suppression of IRAK1, an IL-1 receptor kinase, downstream from Toll-Like Receptors, and associated with the inflammatory response and suppression of normal hematopoiesis in myelodysplasia models (Rhyasen et al.; Cancer Cell; 24:90, 2013).
Conclusions: The kinase profile for pacritinib is unique among agents in development for myelofibrosis combining lack of suppression of JAK1 with suppression of inflammatory signaling through IRAK1. Moreover, the kinase inhibition profile of pacritinib suggests its potential therapeutic utility in AML, MDS, and particularly in CMML due to CMML’s upregulation of JAK2, c-FMS, and IRAK1.

Disclosures
Singer: CTI BioPharma, Corp: Employment, Equity Ownership. Mesa:Incyte Corporation, CTI, NS Pharma, Inc., Gilead Science Inc., Celgene: Research Funding. Verstovsek:CTI BioPharma, Corp: Research Funding.
The Psychosocial Transplant Evaluation Rating Scale (TERS) Prospectively Predicts Inferior Overall Survival Outcome for High Risk Scoring Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Speckhart Dawn , Solomon Scott R. , Zhang Xu , Morris Lawrence E. , Bashey Asad , Holland H. Kent 
Published: October 2019

Abstract
Several studies have begun to examine the relationship between psychosocial variables and outcome in hematopoietic stem cell transplantation (HSCT). Previous studies have also looked at the relationship between the Hematopoietic Cell Transplantation Comorbidity Index (HCT-CI) and disease risk on overall survival in HSCT. Research has shown that disease risk and HCT-CI may have a negative relationship with outcome in HSCT. In this study, psychosocial data was collected prospectively on 437 patients undergoing related and unrelated allogeneic HSCT. All patients were assessed using the Transplant Evaluation Rating Scale (TERS). Based on the patient’s TERS score, each patient was stratified into one of two groups (low/moderate risk (n=371) vs. high risk (n=66)) based on their predicted psychosocial risk for problems during transplant. The 1-year overall survival (OS) for the low/mod risk TERS group versus the high TERS risk group was 78% and 61%, respectively (p=.005). Stratification by CIBMTR disease risk identified 261 patients with low and intermediate risk disease of which 228 had low/mod risk TERS score and 33 had high risk TERS score. The is a significant survival advantage for Low/Mod Risk TERS patients vs. High Risk TERS patients at 100 day, 1 year and overall survival being 97% vs 79% (P=.01), 85% vs 58% (P=.002) and 68% vs 42% (P=.002) with a median follow up of 48 months (graph attached). An adjusted logistic regression analysis showed that psychosocial factors have a significant impact on overall survival despite all patients being required to have 24 hour caregiver support, a pre-transplant psychosocial evaluation and post-transplant psychosocial intervention. The analysis controlled for other variables such as age, gender, performance status, disease risk and transplant type. These results highlight the impact of psychosocial factors on HSCT outcome, as those patients who are expected to do well based on their low/intermediate disease risk may not perform as expected due to psychosocial risk factors. These results suggest the need to perform careful psychosocial assessments on all HSCT patients in order to predict mortality risks for the individual patient and to help the transplant team strategize how to better manage the high risk psychosocial patient.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Loss of IRF8 Inhibits the Growth of Diffuse Large B-Cell Lymphoma
Authors: Xu Yulian , Jiang Lei , Fang Rachel R. , Zhou Jeff Xiwu , Morse Herbert 
Published: October 2019

Abstract
IRF8 is a transcription factor with a critical role in B lymphocyte development and biological functions. Although it has been reported that IRF8 is highly expressed in human diffuse large B-cell lymphoma (DLBCL) and the translocation of IRF8-IgH loci occurs in DLBCL, little information is available regarding the function and mechanisms for the role of IRF8 in DLBCL. In this study, by using several human DLBCL cell lines with shRNA-mediated decrease in IRF8 expression levels, we found that the loss of IRF8 significantly reduced the proliferation of lymphoma cells (Figure 1). Mechanistically, decreasing the levels of IRF8 led to a decrease in p38 and ERK phosphorylation (Figure 2), molecular events critical for B cell proliferation. Furthermore, using a xenograft lymphoma mice model, we found that the loss of IRF8 significantly inhibited the growth of lymphomas in vivo (n=5 for each group) (Figure 3). Analysis of public available data also suggested that the expression levels of IRF8 mRNA in human DLBCL tissues were inversely correlated patientsÕ overall survival time. Taken together, this study showed that IRF8 may play an oncogenic role in human DLBCL by promoting cell proliferation.
Figure 1. Loss of IRF8 decreased the proliferation of DLBCL cells in vitro. Figure 1. Loss of IRF8 decreased the proliferation of DLBCL cells in vitro.
Figure 2. Loss of IRF8 decreased the phosphorylation of p38 and ERK in DLBCL cells. Figure 2. Loss of IRF8 decreased the phosphorylation of p38 and ERK in DLBCL cells.
Figure 3. Loss of IRF8 decreased the growth of DLBCL in vivo. Figure 3. Loss of IRF8 decreased the growth of DLBCL in vivo.

Disclosures
No relevant conflicts of interest to declare.
Constitutive Activation of Shp2 Cooperates with MLL1 Fusion Proteins to Increase Expression of the Homeodomain Transcription Factors Cdx4, HoxA9 and HoxA10
Authors: Bei Ling , Shah Chirag , Wang Hao , Eklund Elizabeth 
Published: October 2019

Abstract
Leukemias with chromosomal translocation or partial tandem duplications involving the MLL (mixed lineage leukemia) gene have exceptionally poor prognosis (referred to as 11q23-leukemias). At the molecular level, 11q23-leukemias are characterized by aberrant expression of a set of homeodomain transcription factors in hematopoietic stem cells (HSC) and differentiating myeloid progenitor cells. This transcription factor set includes HoxB3, B4, A7-11, Cdx2-4 and Meis1. Cdx and Hox proteins are involved in regulating hematopoiesis. Transcription of HOX and CDX genes decreases normal myelopoiesis, but is aberrantly sustained in 11q23-leukemias. Cdx4 activates transcription of the HOXA9 and HOXA10 genes, and HoxA10 activates CDX4 transcription. The events that break this feedback loop, permitting a decrease in Cdx4-expression during normal myelopoiesis, were previously undefined. In the current study, we find that HoxA9 represses CDX4 transcription in differentiating myeloid cells; antagonizing activation by HoxA10. We determine that tyrosine phosphorylation of HoxA10 impairs transcriptional activation of CDX4, but tyrosine phosphorylation of HoxA9 facilitates repression of this gene. Since HoxA9 and HoxA10 are phosphorylated during myelopoiesis, this provides a mechanism for differentiation-stage-specific Cdx4 expression. HoxA9 and HoxA10 are increased in cells expressing Mll-Ell, a leukemia associated MLL1 fusion protein. We find that Mll-Ell induces a HoxA10-dependent increase in Cdx4-expression in myeloid progenitor cells. However, expression of Cdx4 decreases in a HoxA9-dependent manner upon exposure of Mll-Ell-expressing cells to differentiating cytokines. Leukemia associated, constitutively active mutants of Shp2 block cytokine-induced tyrosine phosphorylation of HoxA9 and HoxA10. In comparison to cells expressing Mll-Ell alone, we find that co-expression of Mll-Ell plus constitutively active Shp2 increases CDX4 transcription and Cdx4 expression in myeloid progenitor cells. And, increased Cdx4 expression is sustained upon exposure of these cells to differentiating cytokines. Our results identify a mechanism for increased and sustained CDX4 transcription in leukemias co-overexpressing HoxA9 and HoxA10 in combination with constitutive activation of Shp2. We also demonstrate that inhibition of Shp2-PTP activity decreases Cdx4 expression in Hox-overexpressing human myeloid leukemias.

Disclosures
No relevant conflicts of interest to declare.
Four-Factor Prothrombin Complex Concentrate (4F-PCC) Effectively Reverses Rivaroxaban Induced Bleeding in a Rabbit Model of Acute Bleeding
Authors: Herzog Eva , Mueller-Cohrs Jochen , Kaspereit Franz , Krege Wilfried , Niebl Peter , Doerr Baerbel , Schulte Stefan , Dickneite Gerhard 
Published: October 2019

Abstract
Rivaroxaban is an oral, selective direct factor Xa inhibitor approved for several indications in patients at risk of thrombotic events. One limitation of this agent is the lack of data pertaining to its reversal in situations where urgent response is critical (e.g. acute bleeding events or emergency surgery).
This study therefore evaluated the effectiveness of a four-factor prothrombin complex concentrate (4F-PCC; Beriplex® P/N, Kcentra®, CSL Behring), for the reversal of rivaroxaban-associated bleeding in an in vivo rabbit model of acute bleeding. In addition, the study evaluated the correlation between in vitro coagulation parameters and haemostasis in vivo in order to gain more information on the predictivity of in vitro markers for effective bleeding reversal.
Administration of single intravenous doses of rivaroxaban (150–450 μg/kg) resulted in increased and prolonged bleeding following standardised kidney injury compared to a vehicle treated control group as determined by measurements of total blood loss and time required to achieve full hemostasis. Subsequent treatment with 4F-PCC (25−100 IU/kg) resulted in a dose-dependent reversal of rivaroxaban-associated bleeding signals. Of the in vitro coagulation markers tested, thrombin generation (TGA) and whole blood clotting time (WBCT) correlated well with in vivo measures of PCC-mediated effects. Thrombin generation was highly reagent-dependent, with the assay initiated using tissue factor reagent being most sensitive to the anticoagulant effects induced by Rivaroxaban, while the phospholipid-only reagent being the most predictive of 4F-PCC mediated effective haemostasis in vivo.
In summary, in a rabbit model of acute bleeding, 4F-PCC was able to effectively reduce bleeding to control levels following rivaroxaban 150 μg/kg and 300 μg/kg administrations. The 4F-PCC mediated bleeding reversal correlated best with the in vitro endpoints thrombin generation and whole blood clotting time.

Disclosures
Herzog: CSL Behring GmbH: Employment. Off Label Use: The use of Beriplex P/N for reversal of Rivaroxaban anticoagulation represents off-label use. Mueller-Cohrs:CSL Behring GmbH: Employment. Kaspereit:CSL Behring GmbH: Employment. Krege:CSL Behring GmbH: Employment. Niebl:CSL Behring GmbH: Employment. Doerr:CSL Behring GmbH: Employment. Schulte:CSL Behring GmbH: Employment. Dickneite:CSL Behring GmbH: Employment.
The Use of Recombinant FXIII in a Major Acute Bleeding Episode of a Patient with Congenital FXIII Deficiency – the First Experience
Authors: Boda Zoltan , Arokszallasi Anita , Kerenyi Adrienne , Bereczky Zsuzsanna , Muszbek Laszlo , Pfliegler Gyorgy , Schlammadinger Agota 
Published: October 2019

Abstract
Introduction: congenital FXIII deficiency is a rare bleeding disorder with a high risk of spontaneous intracranial haemorrhage. Life-long preventive supplementation is mandatory if FXIII activity is <1%. A novel recombinant FXIII concentrate (NovoThirteen inj.) was licensed for prophylaxis in congenital FXIII-A subunit deficiency. The efficacy and safety of rFXIII in acute bleeding have not been tested.
Aim: to report the effective use of rFXIII for the treatment of a major bleeding in a patient with severe inherited FXIII-A deficiency.
Patient and methods: a 19-year-old man exhibiting severe bleeding phenotype without clear diagnosis was referred to our haemostasis centre. After presenting negative haemostasis screening tests he was evaluated for FXIII deficiency by the determination of FXIII activity, FXIII-A2B2, FXIII-A and FXIII-B concentrations and by mutation analysis.
Results: severe FXIII-A deficiency was established and prophylaxis with rFXIII was planned. Prior to the start of the prophylaxis he was hospitalized due to a large intramuscular hematoma (19.0x8.3x7.7 cm) in the right thigh and rFXIII, in the prophylactic dose (35 IU/kg) was administered for therapy. Following the administration of rFXIII, the pain disappeared and the swelling of the thigh considerable decreased. Complete resolution of the hematoma occurred without any additional FXIII supplementation. Results of the pharmacokinetic assessment were comparable with those described in the literature.
Conclusion: rFXIII can be successfully used in acute bleeding episode of severe congenital FXIII deficiency.

Disclosures
No relevant conflicts of interest to declare.