bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differences in the Angiogenic Response and Subsequent Growth of Plasma Cells From Myeloma and MGUS Patients Xenografted Into Zebrafish Embryos.
Authors: Ma Dongdong , Ramachandran Aravind , Chang Yali , Lin Jianhong , Zhang Weihong , Tai Yu-Tzu , Munshi Nikhil C. , Handin Robert I 
Published: October 2019

Abstract
Abstract 2912
A monoclonal gammopathy of uncertain significance (MGUS) is an obligate precursor to each case of multiple myeloma (MM). MGUS and MM samples share many of the same gene mutations and chromosomal translocations. Despite increasing genetic and genomic information gathered from patient samples, we still do not understand which mutations are responsible for the progression from MGUS to MM. In order to learn more about this progression and the possible involvement of an angiogenic switch as seen in solid tumors, we have developed a zebrafish model to study the angiogenic potential and subsequent growth of CD138+ plasma cells derived from normal human bone marrow, MGUS and myeloma patients. Immunopurified CD138+ cells were labeled with the red fluorescent dye CM DiI, suspended in Matrigelâ and injected into the perivitelline space of Tg(fli1:GFP) zebrafish embryos that had fluorescent vasculature, 48 hpf (hour-post-fertilization). Cell growth and angiogenesis were assessed by conventional and confocal fluorescent microscopy. Inhibitors of myeloma cell growth or tumor-induced angiogenesis were added to embryo water. The injection of up to 2000 plasma cells from normal human bone marrow into zebrafish embryos did not elicit an angiogenic response and the cells did not grow after injection. In contrast, the injection of as few as 100 plasma cells purified from the marrow of patients with multiple myeloma induced a brisk angiogenic response with extension of new vessels ventrally from the sub-intestinal venous system into the site of cell injection. Angiogenesis was accompanied by the growth of the CD138+ cells and, in some cases, movement of cells out of the Matrigelâ plug to distant sites in the developing embryo. MGUS cells induced minimal or no angiogenesis and grew very slowly after injection. The growth of cells from MGUS patients and de novo myeloma patients was inhibited by the addition of the VEGFR2 inhibitor SU5416 to embryo water 24 hpi (hours-post-tumor cell-injection). If the addition of SU5416 was delayed until 48 hpi, when the angiogenic response had been established, there was no effect on cell growth. We also observed that dexamethasone, bortezimib and lenalidomide, singly and in combination, inhibited the growth of MGUS and myeloma cells. These studies document clear biologic differences between normal, MGUS and myeloma plasma cells. In addition, we provide evidence that the zebrafish embryo can be used to study tumor-induced angiogenesis, tumor cell growth and the effects of possible inhibitors. We also provide some new insights into the mechanism underlying the disappointing response of angiogenic inhibitors when given to patients with multiple myeloma.

Disclosures:
Munshi: Celgene: Consultancy; Millenium: Consultancy; Merck: Consultancy; Onyx: Consultancy.
CD82 Regulates STAT5/IL-10 and Supports Survival of Acute Myelogenous Leukemia Cells.
Authors: Nishioka Chie , Ikezoe Takayuki , Takeuchi Asako , Yang Jing , Yokoyama Akihito 
Published: October 2019

Abstract
Abstract 2981
We recently reported that adhesion molecule CD82 is aberrantly expressed in CD34+/CD38− leukemia stem cells (LSCs) (ASH 2010; Abstract 2168.). Here, we report the results of a functional analysis of CD82 in CD34+/CD38− acute myelogenous leukemia (AML) cells. Short hairpin (sh)RNA-mediated downregulation of CD82 resulted in dephosphorylation of STAT5 and a decrease in the level of IL-10. In contrast, forced expression of CD82 in CD34+/CD38+ AML cells by transduction with CD82-expressing lentiviral particles resulted in an increase in the levels of p-STAT5 and IL-10. Chromatin immunoprecipitation (ChIP) assay results indicated that STAT5A binds to the promoter region of the IL-10 gene, while reporter gene assay results indicated stimulation of IL-10 expression at the transcriptional level. These results suggest that CD82 positively regulates the STAT5/IL-10 signaling pathway. Notably, exposure of CD34+/CD38− AML cells to IL-10 stimulated clonogenic growth of these cells. Moreover, shRNA-mediated downregulation of CD82 expression in CD34+/CD38− AML cells impaired AML reconstitution in immunodeficient mice in parallel with downregulation of STAT5. Taken together, our data suggest that the CD82/STAT5/IL-10 signaling pathway is involved in the survival of CD34+/CD38− AML cells and may thus be a promising therapeutic target for eradication of AML LSCs.

Disclosures:
No relevant conflicts of interest to declare.
Outbreak of Nosocomial Respiratory Syncytial Virus Infections in a Hematology and Transplant Unit.
Authors: Lehners Nicola , Schnitzler Paul , Egerer Gerlinde , Dreger Peter , Eisenbach Christoph , Aichinger Elisabeth , Weißbrich Benedikt , Frank Uwe , Heeg Klaus , Ho Anthony D 
Published: October 2019

Abstract
Abstract 3032
Respiratory syncytial virus (RSV) is a pathogen associated with bronchiolitis in infants and small children. However, immunocompromised adults might also be at risk for severe lower respiratory tract involvement induced by RSV. We report the rapid control of an outbreak of nosocomial RSV infections in an institution with special focus on allogeneic and autologous stem cell transplantations.
Between November 2011 and March 2012 56 patients were diagnosed with RSV infection by RT-PCR at our institution. In order to identify possible risk factors for lower respiratory tract involvement in RSV infected patients, clinical course of infection as well as radiological and laboratory findings were evaluated. Characterization of RSV strains in respiratory specimens was achieved by sequence analysis of part of the RSV glycoprotein G gene and phylogenetic comparison of outbreak and community strains was performed.
Out of 56 patients with RSV infection, 39 patients showed signs of lower respiratory tract involvement. 14 fatal outcomes were observed. In most of the lethal cases coinfections with other pathogens such as Aspergillus spp. or Pseudomonas aeruginosa were present. Therefore the exact impact of RSV on the fatal outcome remains difficult, if not impossible, to assess. Age, sex, underlying disease, disease control or transplant status did not differ significantly between severe and non-severe cases. However, hypogammaglobulinemia was associated with a higher risk for fatal infection (p = 0.03) in univariate analysis, whereas therapy with oral ribavirin might show a protective effect (p = 0.02). Prolonged viral shedding was frequently observed (median 22.5 [1 – 143] days), most pronounced in patients folllowing allogeneic transplantation (p = 0.04). The major mode of RSV transmission seemed to be direct patient-to-patient contact as suggested by evaluation of patient movements on the wards. Sequence analysis on respiratory specimens obtained from 46 patients revealed the presence of a particular outbreak strain in 40 patients, characterized by an identical nucleotide sequence of RSV GA2 subtype. Phylogenetic analysis showed low prevalence of the outbreak strain in the community. By implementation of rigorous isolation measures containment of outbreak was achieved.
Highly contagious viral pathogens such as RSV are able to cause rapidly spreading outbreaks among immunocompromised patients. Hypogammaglobulinemia might be a risk factor for severe lower respiratory tract infection. Treatment with oral ribavirin might have a positive effect on outcome. Rigorous isolation measures are essential to the containment of outbreak.

Disclosures:
Off Label Use: ribavirin aerosol is FDA approved for severe cases of RSV in infants and young children.
Effective Prevention of Acute and Chronic Graft-Versus-Host Disease with Anti-Lymphocyte Globulin (ATG) without Increase of Relapse in HLA-Identical Sibling Peripheral Blood Stem Cell Transplantation.
Authors: Kröger Nicolaus , Zabelina Tatjana , Ayuk Francis , Alchalby Haefaa , Wolschke Christine , Stübig Thomas , Klyuchnikov Evgeny , Berger Jürgen , Zander Axel R. 
Published: October 2019

Abstract
Abstract 3055
Acute and chronic graft versus host disease (GvHD) are severe complications after allogeneic stem cell transplantation but occurrence of GvHD is associated with reduced risk of relapse. Therefore, strategies to prevent GvHD without increasing the risk of relapse are urgently needed.
We investigate retrospectively whether anti-lymphocyte globulin (ATG Fresenius®, Fresenius Biotech, Graefelfing, Germany) as part of the conditioning regimen may reduce the risk of GvHD without increasing the risk of relapse after peripheral blood stem cell transplantation from HLA-identical siblings or HLA-compatible relatives. Out of 462 patients who received HLA-identical stem cell transplantation between 1990 and 2011 in our institution, we selected 238 consecutive patients who received allogeneic peripheral blood stem cell grafts after the year 2000. The median age of the patients was 48 years (r.,18–73y) and diagnosis were: AML (n=93), ALL (n=24), CML (n=25), MDS (n=23) or lympho-proliferative disorders (n=73). Patients were classified as good risk (n=95) or bad risk (n=143). 79 patients did receive ATG within the conditioning regimen with a median dose of 30mg/kg (r., 20–90mg/kg) and 159 patients did not receive ATG. In the ATG group there were more HLA mismatch donors (6% vs. 1%, p=0.02), more bad risk patients (70% vs. 50%, p=0.04), more reduced intensity conditioning regimens (65% vs. 34%, p< 0.001) and older patients (median age 50 vs. 46 years, p=0.03). Acute GvHD grade I to IV was less observed in the ATG group (27% vs. 40%, p= 0.04), but the difference in severe GvHD grade III and IV did not show statistical significance (10% vs. 18%, p=0.1). Chronic GvHD was less observed in the ATG group (30% vs. 52%, p=0.002), which was most obvious for extensive chronic GvHD (14% vs. 40%, p<0.001). After a median follow up of 57 months in the non-ATG and 63 months in the ATG group the cumulative incidence of non-relapse mortality at 5 years was 20% for the ATG and 34% for the non-ATG group (p=0.06) and the cumulative incidence of relapse at 5 years was 34% for the ATG group and 29% for the non –ATG group (p=0.2), resulting in a comparable 5 year progression-free (ATG: 46% and non-ATG 37%, p=0.5) and overall survival (ATG: 54% and non-ATG 46%, p=0.4).
This retrospective study shows that ATG can prevent severe chronic GvHD without obvious risk of relapse and similar PFS and OS in HLA identical sibling transplantation. A randomized trial including QoL measurement comparing ATG vs. no ATG in HLA identical sibling transplantation is ongoing (registered NCT 00678275).

Disclosures:
Kröger: Fresenius Biotech: Honoraria, Research Funding. Off Label Use: ATG in HLA identical sibling transplantation.
Long-Term Outcomes of Autologous Transplantation in Multiple Myeloma: Prognostic Significance of Complete Response.
Authors: Krejci Marta , Hajek Roman , Adam Zdenek , Pour Ludek , Zahradova Lenka , Pelcova Jana , Mayer Jiri 
Published: October 2019

Abstract
Abstract 3126

Background:
Autologous stem cell transplantation (ASCT) after high-dose melphalan 200mg/m2 has got an important role in the treatment of symptomatic multiple myeloma (MM). The prognostic significance of achieving complete response (CR) after ASCT was cause of great debate for a long time. Some studies have shown the strong prognostic significance of achieving CR in MM, but other studies have failed to correlation between strength of the response and outcome.

Aim:
In this report we describe the long-term outcome of cohort 232 MM patients (pts) after ASCT with aim to establish the actual prognosis for the different response categories and to analyse other factors that might predict for long-term survival.

Methods:
We evaluated 232 pts with newly diagnosed symptomatic MM who received ASCT as a part of the first-line treatment between 1995 and 2005, median follow-up from ASCT was 131 months (range 61–195).

Results:
Following ASCT, overall response rate was 90% (202/232), 23% (52/232) of pts were in complete remission (CR), very good partial response (VGPR) was achieved in 45% of pts (100/232), partial response (PR) in 22% of pts (50/232), minimal response (MR) or stable disease (SD) in 10% of pts (22/232). Median progression-free survival (PFS) and overall survival (OS) from ASCT were 30.8 and 71.9 months, respectively. Progression-free survival at 12 years after ASCT in different response categories was 41% for pts with CR, 11% for pts with VGPR and 10% for pts with PR. Overall survival at 12 years after ASCT was 51% for pts with CR, 22% for pts with VGPR and 20% for pts with PR. The achievement of CR after ASCT was independent factor for long-term survival, significance differences in OS and PFS were found between CR and non-CR groups (P under 0.001 and P under 0.001, respectively).
On multivariate analysis, the other factors associated with significantly better OS were ISS stage under III (P = 0.002), no presence of renal impairment (P = 0.008), age under 60 years (P = 0.001), no presence of deletion 1q21 (P = 0.029) and lenalidomide treatment in the post-transplant relapse (P = 0.002).

Conclusion:
The achievement of complete response after ASCT in multiple myeloma is the most important prognostic factor, even after long-term follow-up. The relapse rate is low in patients who remained in CR after 12 years from ASCT. A long-term complete remission should be a goal of treatment.

Disclosures:
Hajek: Merck: Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen Cilag: Honoraria.
Functions and Regulation of Erythrocyte Equlibrative Nucleoside Transporter 1 (ENT1) in Acute Hypoxia Mediated Tissue Injury
Authors: Song Anren , Zhao Cheng , Zhang Yujin , Li Jessica , Blackburn Michael R. , Grenz Almut , Eltzschig Holger , Kellems Rodney E. , Xia Yang 
Published: October 2019

Abstract
Using a nonbiased high throughput metabolomic screen, coupled with genetic and pharmacological approaches, recent studies demonstrated that excessive adenosine signaling through the A2B adenosine receptor triggers sickling by induction of 2,3-bisphosphoglycerate (2,3-BPG), an erythroid specific metabolite that induces O2 release from hemoglobin. Adenosine is a signaling nucleoside that elicits numerous physiological and pathological effects by engaging membrane receptors. Notably, equlibrative nucleoside transporters (ENTs) on erythrocytes have been long speculated to regulate extracellular adenosine concentrations under hypoxic conditions. Thus, we hypothesize that ENT is likely a key molecule responsible for elevated circulating adenosine levels and protects tissues from hypoxia induced injury. To test this hypothesis, we first conducted in vivo Carbon-14 labeled adenosine (C14-Ado) injection and in vitro functional C14-Ado uptake assays. We found that erythrocyte plays a key role in regulation of circulating adenosine. We then conducted western blot analysis to compare expression profiles of ENTs on erythrocyte. We found that ENT1 is the major ENT expressed on both mouse and human erythrocytes. Using genetic approach, we successfully generated an erythrocyte ENT1 knockout mouse model. Using this genetic model and pharmacological approach combined with in vivo C14-Ado injection and in vitro C14-Ado uptake assay, we demonstrated that ENT1 1) is the major adenosine transporter in erythrocyte and 2) erythrocyte is the major cell type involved in regulating circulating adenosine levels through ENT1’s function. Using erythrocyte ENT1 knockout mouse model, we found that, during acute hypoxia treatment, the loss of erythrocyte ENT1 can cause faster increase in circulating adenosine level, subsequently promoting 2,3-BPG production, triggering oxygen release, and protecting acute hypoxia-mediated tissue injury. Mechanistically, we demonstrated that hypoxia regulates ENT1 activity through adenosine-ADORA2B-PKA signaling pathway. Overall, our studies demonstrate that 1) ENT1 is a major adenosine transporter expressed by erythrocytes and erythrocytes are the major cell type responsible for regulating circulating adenosine. 2) Hypoxia regulates ENT1 activity through adenosine-ADORA2B-PKA signaling pathway. 3) Inhibition or deletion of erythrocyte ENT1 results in enhanced adenosine-mediated 2,3-BPG induction and hemoglobin deoxygenation in RBCs when hypoxia is encountered. Thus, our findings suggest that erythrocyte ENT1 and ADORA2B are novel targets to prevent hypoxia-mediated tissue injury.

Disclosures
No relevant conflicts of interest to declare.
Multiple Oral Therapeutics Suppress Repressors (LSD-1, HDACs, and BCL11A) of Gamma Globin Gene Expression
Authors: Dai Yan , Faller Douglas V , Sangerman Jose I , Fucharoen Suthat , Perrine Susan P 
Published: October 2019

Abstract
Induction of gamma globin expression to certain target levels reduces clinical severity in sickle cell disease, globin imbalance, and anemia in the beta thalassemias. However, achieving high level expression in diverse patients with variable baseline HbF levels is clinically challenging. To determine whether different therapeutics have complimentary molecular actions that could be combined for higher efficacy, we evaluated a panel of oral fetal globin-inducing therapeutics in clinical use or trials for effects on transcriptional suppressors of HBG, including components of the NuRD complex, LSD1 and HDACs 1-3, and the downstream repressor BCL11A, in erythroid progenitors cultured from sickle cell and beta thalassemia patients. Using chromatin immunoprecipitation assays, immunoblot, and qRT-PCR, we investigated two pan-HDAC inhibitors, MS-275 and SB939, a short chain fatty acid derivative without global histone deacetylation effects, sodium dimethylbutyrate (ST20), and PB-04, a PK-enhancing drug identified through high throughput screening (HTS), which has a long-standing benign clinical safety profile. These therapeutics induce fetal globin expression by 1.5 to 12-fold in different individuals’ progentiors over untreated controls and are active in 60-90% of patients’ progenitors. BCL11A mRNA and protein were suppressed by 3-7 fold and 5-10 fold, respectively, over untreated cells from the same subject by all agents tested. PB-04 treatment decreased LSD1 mRNA by 5-fold. MS-275 and ST20 suppressed KLF-1 mRNA by 3- and 2.5-fold respectively. ChIP analyses demonstrated that PB-04 reduced LSD1 and HDAC-3 occupancy on the gamma globin gene promoter by 4-fold and 3-fold, respectively, coincident with gamma globin induction; ST20 decreased HDAC-2 and LSD1 binding by 6- and 2-fold respectively. Further, histone marks associated with gene activation were enriched at the gamma-globin promoter by treatment with PB-04, including acetylated Histone H3K9 (AcH3K9), the target of HDAC3, and dimethylated H3K4 (H3K4me2), a target of LSD1. Collectively, these results identify oral clinical stage therapeutics which disrupt two major mechanisms of transcriptional repression. The findings provide a basis for targeting therapies in individuals and combining pharmacologic agents which induce HbF expression through complimentary molecular mechanisms to provide high level induction.

Disclosures
Faller: Phoenicia BioSciences: Employment, Membership on an entity's Board of Directors or advisory committees. Perrine:Phoenicia BioSciences: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.
Multi-Center Study of DEP Regimen As a Salvage Therapy for Adult Refractory Hemophagocytic Lymphohistiocytosis
Authors: Wang Yini , Wang Zhao , Huang Wenqiu , Hu Liangding , Cen Xinan , Li Lihong , Wang Jijun , Shen Jianliang , Wei Na 
Published: October 2019

Abstract

Purpose: Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinflammatory syndrome. Current standard therapy increased the complete remission (CR) rate to more than 50%. Even so, about 30% patients fail the standard treatment. This study aimed to investigate the efficacy of liposomal doxorubicin together with etoposide, and high dose methylprednisolone (DEP) as a salvage therapy for adult refractory HLH.
Methods: Refractory HLH was defined as patient who did not achieve at least partial remission (PR) 2 weeks after initial standard HLH therapy. The DEP regimen included liposomal doxorubicin 25mg/m2 d1, etoposide 100mg/m2 Qw and methylprednisolone 15mg/kg d1-3, 2mg/kg d4-6, 1mg/kg d7-10,0.75mg/kg d11-14. We estimated the curative effect after 2 weeks therapy and 4 weeks therapy.
Results: We observed 63 refractory HLH patients from Jun 2013 to Jun 2014, including 42 males and 21 females. All the data were gathered from Beijing Friendship Hospital, 307 Hospital of PLA, Peking University People’s Hospital, Peking University Third Hospital, Beijing Chao-Yang Hospital, and Navy General Hospital. The median age was 34 years old (Range 18-78). Seventeen patients (27.0%) achieved CR and 31 patients (49.2%) achieved PR. The overall response reached 76.2% (48/63). The underlying disease of HLH were diagnosed within 2 to 8 weeks after HLH diagnosis, including 29 cases of lymphoma associated HLH, 22 cases of EBV associated HLH and 4 cases of familial HLH. There were still 8 cases had unknown underlying disease. The patients who had no response to DEP were died within 2 to 4 weeks after salvage therapy. Twenty-nine of the 48 patients who achieved PR or CR survived to undergo subsequent chemotherapy, allogenic hematopoietic stem cell transplantation (allo-HCT) or splenectomy. There were significant improvements on leukocytes, ferritin and soluble CD25 (sCD25) after 2 weeks therapy. Thrombocytes and glutamic-pyruvic transaminase (GPT) had improvements after 4 weeks therapy.
Conclusion: DEP regimen appears to be an effective salvage protocol for refractory HLH. Prospective randomized controlled trials of DEP regimen will further estimated the curative effect and define optimal dosing levels and schedules.

Disclosures
No relevant conflicts of interest to declare.