bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Study of Proliferation Relative Long Non-Coding RNA in CD59- Cell from Paroxysmalnocturnal Hemoglobinuria Patients
Authors: Fu Rong , Wang Honglei , Liu Hui , Shao Zonghong , Li Liyan 
Published: November 2019

Background: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonogenic disease of hematopoietic stem cells. LncRNA has a wide range of biological functions, including regulation of gene expression, cell differentiation, cell proliferation and substance metabolism.LncRNA maybe contribute to the proliferation of PNH clone.
Methods:
PNH clone (CD59-cells) and (CD59+cells) from PNH patients were sorted and analyzed by RNA sequencing in 5 PNH patients. The results were analyzed by KEGG, we focus on the proliferation relative pathway-NF-kB pathway. The mRNA(TAB2, TLR4, LYN,CFLAR, TNFAIP3, PTGS2, TRIM25, CXCL8) which FPKM>10 and over 3 pathients were chosen to search out the upstream regulation LncRNA. MALAT1 and LINC01002 were screened by Co-expression. Then the expression of MALAT1 and LINC01002 in 30 PNH patients were detected by qRT-PCR to vertify the LncRNA sequencing results.
Results: Transcription analysis revealed that 742 upregulation LncRNA and 3276 upregulation mRNA were identified in CD59- cells. The highly expressed NF-kB pathway mRNA (TAB2, TLR4, LYN,CFLAR, TNFAIP3, PTGS2, TRIM25, CXCL8) were analysed by LncRNA co-expression, after that MALAT1and LINC01002 were concerned with the 8 mRNA. The results of PNH primary cells (CD59-cells) showed that the level of MALAT1and LINC01002 expression was significantly higher than that of the CD59+ cell in 30 PNH patients (p<0.05).
Conclusion: MALAT1and LINC01002 seems to be involved in the proliferation of PNH clones. Its mechanism of action in patients with PNH needs further study.

Disclosures
No relevant conflicts of interest to declare.
Faster Diagnosis of TTP Using New Technoscreen Adamts-13 Activity Assay
Authors: Braf Annica , von Horn Henrik , Bark Niklas , Norberg Eva-Marie , Truedsson Asa , Binder Nikolaus B , Antovic Jovan P 
Published: November 2019

Background
The thrombocytopenic purpura (TTP) is a life-threatening condition characterized by microvascular thrombosis where it is of great importance to shorten the time to the treatment. TTP can be caused by congenital or acquired deficiency of ADAMTS13 (an enzyme that cleaves the von Willebrand factor). Deficiency leads to the accumulation of ultra-large von Willebrand multimers, which causes platelet aggregation and the formation of microthrombi. TTP laboratory findings include thrombocytopenia, hemolytic anemia, the presence of schistocytes and a markedly reduced activity of ADAMTS13. ADAMTS13 activity can be analyzed using an ELISA method. We have evaluated whether a new semi-quantitative method, TECHNOSCREEN from Technoclone, can be used as a screening method to shorten the time for patient diagnosis.
Method
We analyzed 43 unidentified samples with known ADAMTS 13 activity, which consisted of 20 samples with an ADAMTS13 activity <10%. The new screening method (TECHNOSCREEN ADAMTS13 Activity) was compared to the existing ELISA method (TECHNOZYM ADAMTS13 Activity ELISA). Both methods were from Technoclone, Vienna, Austria. The methods are based on incubating patient plasma with von Willebrand fragments and specific antibodies to cleavage fragment. Color intensity correlates with the amount of cleavage and thus ADAMTS13 activity of the sample. In the screening method, color development takes place in a flow through test device and is semi-quantitatively graded by visual comparison against a color chart graded as 0.0, 0.1, 0.4 and 0.8 IU /mL.
Results and conclusion
The results showed good agreement between the existing ELISA and the newTECHNOSCREEN method. Sensitivity was 100% and specificity 71% if the ELISA is used as a gold standard. In May 2019, the TECHNOSCREEN method was introduced as a first-line analysis. This has resulted in faster patient diagnosis as well as saving laboratory staff time. The TECHNOSCREEN assay takes about 1 hour to perform compared to about 5 hours for the ELISA. To exactly quantify ADAMTS13 activity or detect the presence of antibodies, the ELISAs are performed approximately once a week.

Disclosures
Binder: Technoclone: Employment. Antovic:Shire: Other: Grants and Personal Fees; Roche: Other: Personal Fees; Stago: Other: Personal Fees; Werfen: Other: Personal Fees; Siemens: Other: Personal Fees; Sobi: Other: Personal Fees; Novo Nordisk: Membership on an entity's Board of Directors or advisory committees, Other: Personal fees; CSL Behring: Other: Personal Fees.
Prevalence of Bronchiolitis Obliterans Syndrome (BOS) Following Allogeneic Hematopoietic Stem Cell Transplant (alloHSCT) in the USA, Europe and Japan
Authors: Hofstetter Emilie , Henig Noreen Roth , Kappeler Dominik 
Published: November 2019

Background:
Bronchiolitis obliterans syndrome (BOS), also known as respiratory chronic Graft versus Host Disease (cGvHD) is an obstructive airway disease of the lungs mainly associated with alloHSCT or lung transplantation. BOS is characterized by T-cell mediated inflammation and fibrosis of bronchiolar walls that reduces the diameter of the bronchioles resulting in progressive and irreversible airflow obstruction.
BOS is a well described complication following lung transplant with a 5-year prevalence rate of 50%. It can also be a complication following alloHSCT with similar histopathology and clinical symptoms. Currently, there are no approved treatments for BOS. The aim of this study is to describe the prevalence of BOS following alloHSCT.
A review and analysis of the following data sources was performed: AlloHSCT activity reports from The European Society for Blood and Marrow Transplant 2016 the Center for International Blood and Marrow Transplant Research 2016, and the Japanese Data Center Hematopoeitic Stem Cell Transplantation 2015 in order to identify the number of procedures performed. Further a PubMED literature search was also conducted and identified publications with > 100 patients using the key terms "bronchiolitis obliterans and haematopoietic stem cell", and "bronchiolitis obliterans after stem cell transplant" from 2011 to 2017. Case reports and redundant publications were excluded from the analysis.
Results
Approximately 30,000 alloHSCTs were performed each year in the USA (8766 in 2016), Europe (17,617 in 2016) and Japan (3700 in 2015). Based on seven eligible studies published between 2011 and 2016, the reported prevalence of BOS in patients who underwent alloHSCT ranged from 3.4% to 10%. Median time to diagnosis of BOS following alloHSCT was 273 to 547 days post-transplant. Those with routine Forced Expiratory Volume at 1 second (FEV1) monitoring had a higher likelihood of being diagnosed with BOS and a shorter time to diagnosis.
Conclusion
Based on estimated prevalence, up to 3000 patients following alloHSCT per year could be diagnosed with BOS. Routine monitoring may positively impact early and shorten time to diagnosis and allow treating such a population.
Figure

Disclosures
Hofstetter: Breath Therapeutics GmbH: Consultancy. Henig:Breath Therapeutics Inc.: Employment. Kappeler:Breath Therapeutics GmbH: Consultancy.
Free Fatty Acid Uptake By Hematopoietic Stem and Progenitor Cells Drives Immune Cell Expansion in Response to Salmonella Typhimurium infection
Authors: Mistry Jayna J , Moore Jamie A , Hellmich Charlotte , Marlein Christopher R , Beraza Naiara , Di Palma Federica , Bowles Kristian M , Rushworth Stuart A 
Published: November 2019

Haematopoietic stem and progenitor cells (HSPC) are tightly regulated in a balance between self-renewal and differentiation and undergo rapid expansion in response to infection. In the context of acute bacterial infection, the hematopoietic system needs energy to drive the immune response necessary for host survival. In an adult human, bone marrow adipose tissue accounts for up to 70% of bone marrow and we have previously shown that free fatty acids are transferred from adipocytes to AML blasts providing the AML blasts more energy for proliferation and survival (Shafat, Blood 2017). Therefore, we hypothesis that during an acute bacterial infection, the HSPC population uptakes free fatty acids (FFA) to support and sustain the rapid response to bacterial infection. To determine if HSPC acquired FFA we infected C57BL/6 with Salmonella Typhimurium (S.typhimurium) or treated C57BL/6 with lipopolysaccharide (LPS) for 2 and 16 hours. Using both models we show that the HSPC populations (HSC (haematopoietic stem cell), MPP (multipotent progenitor) and LSK (LN-, CD117+, Sca1+)) all accumulate FFA as measured by staining with neutral lipid-specific BODIPY 493/503 dye. To determine if this was uptake or accumulation of fatty acid within the HSPC of animals infected with S.typhimurium we used BODIPY®-dodecanoic acid fluorescent fatty acid to trace FFA uptake. HSPC populations from animals treated with LPS had a higher uptake of BODIPY®-dodecanoic acid when compared to the control treated mice. To assess metabolic changes within HSC population in response to infection, cells were sorted (LN-, CD117+, Sca1+, CD48-, CD150+ and CD34+) and analysed for gene expression changes. In HSC, increases were observed in several metabolic genes including FABP4, CD36 and CPT1 in response to S.typhimurium as well as LPS at 16 hours post stimulation. Moreover, following cell sorting, the LSK (LN-, CD117+, Sca1+) population were assessed for metabolic activity for different substrates. The LSK population from LPS treated animals had an increased dependency on beta oxidation when compared to the control. Moreover, etoximir (a beta-oxidation inhibitor) inhibited LSK basal metabolism and reduced LSK expansion (as measured by colony forming cell assay and Ki-67 staining) in animals treated with LPS. Our data indicates that infection drives uptake of FFA in different HSPC populations. The FFA are metabolised by beta oxidation which provides the energy for the expansion of leukocytes.

Disclosures
Bowles: Abbvie: Research Funding; Janssen: Research Funding. Rushworth:Abbvie: Research Funding; Janssen: Research Funding.
Novel Immune Checkpoint Deficiency and Susceptibility to EBV-Associated Lymphoma
Authors: Somekh Ido , Magg Thomas , Duque Alejandro Gallón , Stauber Tali , Lev Atar , Unal Ekrem , Simon Amos J. , Rohlfs Meino , Conca Raffaele , Tatematsu Megumi , Hauck Fabian , Somech Raz , Klein Christoph 
Published: November 2019

Background: Understanding genetic predisposition to cancer is of paramount importance to tailor therapeutic strategies. Several defects in immune checkpoint regulators have been discovered in patients with EBV-induced lymphoma (e.g. CD27, PRKCD, RASGRP1, MAGT1, SH2D1A, ITK). Here, we describe the clinical and immune phenotype of 2 unrelated patients from consanguineous families presenting with a primary immune deficiency (PID) and EBV-associated lymphoproliferation due to biallelic mutations in CD137 (TNFRSF9/4-1BB).
Methods: One patient of Turkish origin (P1) and Palestinian origin (P2), respectively were evaluated. Genetic analysis using whole exome sequencing was conducted. Immunological and biochemical assays were performed on primary patient material.
Results: Clinical findings included recurrent sinopulmonary and herpes virus infections from childhood on. One patient suffered from auto-immunity (AIHA and auto-immune thrombocytopenia). Abnormal immunoglobulin levels were documented (IgG 413-1670, IgM 105-714, IgA 49-68 mg/dL). Both patients developed EBV-associated lymphoproliferative disorders: P1 developed Burkitt lymphoma and was treated according to NHL-BFM2000 regimen in combination with rituximab. He is currently in remission. P2 had monoclonal EBV-positive lymphoproliferation and was successfully treated with immunosuppressive therapy (e.g. cellcept, glucocorticoids). To shed light on the underlying genetic etiology, we performed whole exome sequencing. A large homozygous deletion in CD137 (c.1_545+1716del) was identified for P1, while P2 harbored a homozygous missense mutation (c.C452T, p.Thr151Met). Impaired anti-CD3 T cell lymphocyte activation and proliferation were observed, amenable to correction upon addition of anti-CD28 monoclonal antibodies. In an attempt to provide definitive proof that the CD137 gene variant causes the activation defect of T-cells, we designed a genetic rescue experiment in patient T cells. Upon retrovirus-mediated recombinant expression of WT CD137, proliferation and activation defects in T cells were restored.
Conclusions: In sum, we here show that a genetic defect in the immune checkpoint molecule CD137 causes a new primary immunodeficiency disorder with susceptibility to EBV-induced lymphomagenesis.

Disclosures
No relevant conflicts of interest to declare.
Improved Outcomes of All-Trans-Retinoic Acid and Arsenic Trioxide Plus Idarubicin As a Frontline Treatment in Adult Patients with Acute Promyelocytic Leukemia
Authors: Yang Shiwei , Zhu Zunmin , Zhu Xiaojian , Zhang Yicheng 
Published: November 2019

Objective To analyze the outcomes of all-trans-retinoic acid (ATRA) and IV arsenic trioxide (ATO) plus idarubicin (IDA) as a frontline induction therapy in 118 acute promyelocytic leukemia (APL) patients with high risk (HR) and standard risk (SR) from January 2008 to December 2017. Methods The medical records of 118 APL patients (HR, n=45; SR, n=73) who received the frontline triple combination regimen at Henan Provincial People's Hospital and Tongji Hospital were retrospectively reviewed. Consolidation therapy comprised 6 cycles of ATO and ATRA plus IDA, and IDA was administered in 1~4 cycles of consolidation therapy based on the comparable clinical efficacy compared with 6 cycles and fewer side effects to myocardium without subsequent maintenance therapy. Results Of 118 patients, there were 3 (2.5%) early deaths and 115 (97.5%) hematologic complete remissions (hCR), and 102 (88.7%) of 115 patients achieved molecular complete remissions (mCR) following induction therapy, and all patients were PCR-negative for PML-RARa after the first cycle of consolidation therapy. The 5-year overall survival (OS) and event-free survival (EFS) were 93.0±2.9% and 92.4±3.0%, respectively. Early death, hCR, mCR and toxicities were comparable between the HR and SR groups. The cumulative incidence of relapse (CIR) in the HR group was 4.7% (n=2), and the CIR in the SR group was 0. The OS and EFS of the SR and HR groups were comparable (92.3±4.5 vs. 92.8±4.0%, X2=0.263, P=0.608; 92.3±4.5 vs. 91.1±4.2%, X2=0.917, P=0.338). The total dosage of IDA was approximately 181~258 mg, and no patient experienced cardiotoxicity. OS and EFS were not influenced by fms-related tyrosine kinase 3 internal tandem duplication (FLT3-ITD) mutation status (P=0.405, 0.528, respectively). Conclusion The triple combination of ATRA and ATO plus IDA as both an induction and consolidation therapy for the HR and SR groups attained excellent outcomes, and this regimen was effective, safe and easy, without maintenance therapy. The triple combination treatment might be a preferred frontline therapy for APL patients, especially for HR patients or patients with the APL FLT3-ITD mutation.

Disclosures
No relevant conflicts of interest to declare.
This Is a Title in Title Case: A Combination of Low-Dose Decitabine and Chidamide Resulted in Synergistic Effects on the Proliferation and Apoptosis of Human Myeloid Leukemia Cell Lines
Authors: Xu Fangfang , Guo Honggang , Shi Mingyue , Liu Siwei , Wei Min , Sun Kai , Chen Yuqing 
Published: November 2019

Introduction: Myeloid leukemia is a malignant disease caused by both genetic and epigenetic changes. Aberrant DNA methylation and histone modifications are prevalent in cancers including leukemia and the process of epigenetic modifications is reversible, which makes them potential treatment targets using specific inhibitors. In this study, we sought to determine the antileukemic effects of low-dose decitabine (one of the most widely used DNA methyltransferase inhibitor) combined with chidamide (a novel orally active histone deacetylase inhibitor) on myeloid leukemia cells by detecting cell proliferation, cell cycle distribution and cell apoptosis to provide a promising regimen for clinical application.
Methods: We conducted CCK-8 assay to determine the effect of low-dose decitabine combinated with chidamide on cell viability, cell cycle distribution analysis by use of flow cytometry accompanied with RT-qPCR and western blotting analysis to reveal its inhibitory mechanism of cell growth, and cell apoptotic analysis also by use of flow cytometry accompanied with RT-qPCR, western blotting analysis, intracellular ROS production and mitochondrial transmembrane potential assay to determine the apoptotic mechanism of combination of low-dose decitabine and chidamide in K562 and THP-1 cells.
Result: Low-dose decitabine and chidamide alone played an inhibitory role in the proliferation of K562 and THP-1 cells in a dose- and time-dependent manner, and the combination of the two enhanced this effect, accompanied by induction of p21 expression and cell cycle arrest at G0/G1 phase. Chidamide instead of decitabine had pronounced effects on the histone H3 acetylation levels of leukemia cells. Meanwhile, combination of the two significantly induced cell apoptosis via mitochondrial transmembrane potential loss, resulting in upregulation of the cell apoptosis-related gene Bax and the protein cleaved PARP-1 and downregulation of antiapoptosis gene Bcl-2, and proteins, including pro-caspase 9, pro-caspase 3 and PARP-1.
Conclusion: Our results provided evidence for the antileukemia effects of low-dose DNA methyltransferase inhibitor decitabine combinated with histone deacetylase inhibitor chidamide in preclinical myeloid leukemia models and suggested that combination of the two showed therapeutic potential for myeloid leukemia treatment.

Disclosures
No relevant conflicts of interest to declare.
Obinutuzumab-Related Adverse Events: A Systematic Review and Meta-Analysis
Authors: Amitai Irina , Gafter-Gvili Anat , Raanani Pia , Gurion Ronit 
Published: November 2019

Background:
Rituximab, the first FDA-approved anti-CD20 monoclonal antibody has dramatically improved outcomes for patients with CD20 positive lymphoproliferative disorders for 2 decades. Obinutuzumab was developed to potentiate activity and overcome resistance to rituximab. Clinical data suggest obinutuzumab is superior to rituximab in specific lymphoproliferative disorder, at the potential cost of increased toxicity, especially infections.
In order to better define the toxicity profile of obinutuzumab as opposed to rituximab, we conducted a systematic review and meta‐analysis of all randomized controlled trials comparing obinutuzumab with rituximab, either alone or combined with chemotherapy for any CD20 positive lymphoproliferative disorder, in order to assess safety.
Methods:
A comprehensive search was conducted until June 2019. Two reviewers appraised the quality of trials and extracted data. The Primary outcome was grade 3 to 4 infections; secondary outcomes included any adverse events, serious adverse events, and other grade 3 to 4 hematologic toxicity. Relative risks (RRs) with 95% confidence intervals (CIs) were estimated and pooled. A fixed‐effect model was used to pool data unless there was significant heterogeneity, in which case a random‐effects model was used.
Results:
Our search yielded 4 randomized controlled trials conducted between the years 2009 and 2014, including 3429 patients. Median age of patients in the trials was 58 to 62 years. The trials included patients with follicular lymphoma (FL) (2 trials), chronic lymphocytic leukemia, (1 trial), and diffuse large B cell lymphoma (1 trial). All trials compared between obinutuzumab and rituximab therapy: in 3 trials the antibody was combined with chemotherapy and in 1 trial it was given alone. In 2 trials, which included FL patients, the antibody was given also as maintenance therapy for up to 2 years.
There was a statistically marginally increased rate of grade 3-4 infections (RR 1.17 [95% CI, 1.0‐1.36]) and febrile neutropenia (RR 1.23 [95% CI, 1.0‐1.5]) and a statistically significant increased rate of grade 3-4 adverse events (RR 1.15 [95% CI, 1.09‐1.2]) in the obinutuzumab arm vs rituximab. Also, the obinutuzumab arm showed increased risk of grade 3-4 thrombocytopenia (RR 2.8 [95% CI, 1.92‐4.06]), infusion related reactions (RR 2.8 [95% CI, 2.16‐3.64]) and cardiac events (RR 1.65 [95% CI, 1.11‐2.46]). There was no significant difference in grade 3-4 anemia and neutropenia rates, nor in the mortality rate at 36 months (RR 0.93 [95% CI, 0.78‐1.11]).
Conclusions:
Our systematic review and meta-analysis demonstrates that obinutuzumab has a more severe toxicity profile as compared to rituximab. As better clinical outcomes were reported with obinutuzumab in several CD20 positive lymphoproliferative disorders, but with more toxicity, clinicians are faced with a challenge upon deciding which therapy to choose for their patients.

Disclosures
Gurion: Roche: Consultancy.
Unveiling a Critical Role of Thrombin in Enhanced CD8+ T Cell Response to Cancer
Authors: Cantrell Rachel , Rosenfeldt Leah A. , Mureb Duaa , Sharma Balkrishan , Revenko Alexey , Monia Brett P , Palumbo Joseph S. 
Published: November 2019

A serious and life-threatening cancer-associated sequelae is venous thromboembolism (VTE). Indeed, VTE is the second leading cause of death in cancer patients, second only to the malignancy itself. Cancer patients with VTE or at high risk for VTE are generally treated with anticoagulants, which limit thrombin generation. While it's been widely accepted that thrombin plays a role in cancer progression, the effects thrombin has on other cell types within the tumor microenvironment (TME) have not been thoroughly studied.
An understudied role of thrombin may be found in its ability to drive T cell functions. Recently, we have identified thrombin as a potential enhancer of CD8+ T cell effector functions by signaling through the protease activated receptor 1 (PAR-1). Our preliminary data shows that thrombin increases CD8+ T cell survival in a PAR-1 dependent fashion. CD8+ tumor infiltrating lymphocytes (TILs) play a critical role in tumor clearance through their cytolytic and anti-tumor cytokine producing capacity. However, the hostile tumor microenvironment (TME) promotes a gradual reduction in CD8+ TIL capacity to produce cytokines and kill targets. Specific components of the TME, including PDL1 expression, are associated with loss of T cell functionality. A promising strategy to block the interaction of CD8+ TILs and the inhibitory TME components is immune checkpoint inhibition (ICI) therapy, as shown by effective blockade of PD1 signaling by anti-PD1 antibodies. However, these ICI therapies leave many patients unresponsive, highlighting the necessity to uncover additional underlying mechanisms involved in modulating CD8+ T cell responses against cancer. Our preliminary findings lead us to hypothesize that thrombin, in conjunction with PD1 blockade, may work in synergy to promote CD8+ T cell killing of tumors.
Consistent with our hypothesis, preliminary results suggest that thrombin is necessary for a robust anti-tumor immune response following ICI in vivo. Here, cohorts of C57BL/6 mice with low or normal circulating prothrombin levels bearing B16 tumors were treated with an anti-PD1 antibody or control IgG. Anti-PD1 therapy significantly limited tumor growth in mice with normal prothrombin levels, but had no impact on tumor growth in mice with low prothrombin levels.
A major implication of our findings is that limiting thrombin generation with anticoagulants may be detrimental in the context of immune checkpoint inhibition treatment. Better defining the potential risk of reducing ICI efficacy by concurrent treatment with anticoagulants will require a detailed understanding of the role thrombin plays in cancer immunobiology.

Disclosures
Revenko: Ionis Pharmaceuticals: Employment. Monia:Ionis Pharmaceuticals: Employment. Palumbo:Ionis Pharmaceuticals: Research Funding.