bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sphingosine-1-Phosphate (S1P) Signaling through S1P Receptor 1 (S1PR1) in Macrophage Contributes to Pain in Sickle Cell Disease
Authors: Adebiyi Morayo G. , Zhang Yujin , Sun Kaiqi , Hu Xia , Zhao Shusuan , Wu Hongyu , Couturier Jacob , Lewis Dorothy L. , Walters Edgar T. , Xia Yang 
Published: October 2019

Abstract
Sickle cell disease (SCD) is the most common autosomal recessive hemolytic disorder affecting more than 100,000 African-Americans and millions around the world. The severe health complication of patients with SCD is chronic pain, which is the leading cause of SCD patient morbidity and hospitalization. Our proposed research builds on the results from our nonbiased high-throughput metabolomics screen, which revealed erythrocyte sphingosine-1-phosphate (S1P), a bioactive lipid, is significantly elevated in the circulation of humans and mice with SCD. We identified that erythrocyte-sphingosine kinase 1 (Sphk1), the enzyme that generates S1P, promote erythrocyte sickling and the genetic deletion of Sphk1 significantly reduce pain symptoms in SCD mice. S1P is a pleiotropic molecule that signals via five g-protein coupled receptors (GPCRs) known as S1P receptors (S1PR1-5). S1PR1 is widely expressed in many cell types, including neurons and immune cells. We treated SCD mice with FDA approved drug, FTY720, to specifically target S1PR1. We demonstrated that mice had improved mechanical and thermal pain sensitivity. Notably, using flow cytometry, we found that the percentage of macrophages infiltrated to skin and percentage of S1PR1 expression in macrophages in the skin were significantly induced in SCD mice compared to the controls. This implies that elevated S1PR1 in macrophage contributes to chronic pain in SCD mice. To test this possibility, we treated SCD mice with clodronate liposomes to deplete tissue macrophages. We identified that mice had reduced sensitivity to mechanical and thermal stimuli assessments. Altogether, we provide both genetic and pharmacological evidence that elevated plasma S1P signaling via S1PR1 likely on macrophages contributes to chronic pain in SCD. Additionally, S1PR1 is known to mediate pro-inflammatory cytokine secretion in immune cells. We identified that SCD mice have elevated levels of interleukin-6 (IL-6), a pro-inflammatory cytokine, and is likely involved in SCD pain. To study the role of IL-6 in SCD pain, we generated SCD-IL-6-/- mice and observed improved mechanical and thermal pain sensitivity in these mice. From these evidences, we hypothesize that Sphk1 mediate S1P-S1PR1 activation in macrophages and IL-6 induction to promote SCD pain. Our work addresses a knowledge gap regarding an inflammatory mediated mechanism involved in SCD pain and novel therapies to counteract pain in SCD.

Disclosures
No relevant conflicts of interest to declare.
Leukemia-Associated Macrophages Educated By Organ-Specific Leukemic Microenvironment Acquire Diverse Functions and Gene-Expression Signatures
Authors: Yang Xiao , Feng Wenli , Wang Rong , Yang Feifei , Chen Chong , Wang Lina , Zheng Guoguang 
Published: October 2019

Abstract
Macrophages are important components of microenvironments and participate diverse physiological and pathological processes. Though CD68+macrophages have been demonstrated a potentially clinical marker to predict adverse prognosis in lymphoma, the role of macrophages in leukemia have not been fully elucidated. Most recently, we reported leukemia-associated macrophages (LAMs), which showed leukemia-promoting effects, in Notch1-induced mouse T-ALL leukemic microenvironment. In the present study, we studied the characteristics of LAMs from BM and spleen in MLL-AF9 induced mouse AML model, and compared the gene expression diversities among LAMs from BM and SP of AML and T-ALL models.
LAMs were sorted from leukemic BM and spleen in the middle stage of AML, control macrophages were sorted from normal mouse. Then LAMs were cocultured with AML cells for 24, 48 and 72hrs. Cell count, BrdU incorporation assay and Annexin V assay were used to detect the proliferation and apoptosis of leukemia cells, respectively. Significant increase of BrdU positive and decrease of apoptotic AML cells were detected in leukemia cells cocultured with LAMs than control Mφ. Latex-beads uptaken assay and transwell assay were used to detect the phagocytosis and migration of LAMs. Significant decrease of phagocytosis and increase of migration ability were detected in LAMs than control Mφ. Furthermore, T cell coculture assay revealed that spleen LAMs suppress the CD25 and CD69 expression in both CD4+ and CD8+T cells but increase induction of Treg cells than BM LAMs. Moreover SP LAMs had a significantly higher proliferation promoting effect on leukemia cells than BM LAMs. To explore the diverse effect among LAMs from BM and spleen, we compared the transcriptional data of LAMs from BM and spleen from AML and T-ALL. The ordered list algorithm revealed the similarity score in LAM samples with same microenvironment (991.8) is higher than those with same type of leukemia (709.1), also the expression pattern analysis revealed that LAMs from leukemic BM and spleen are much more different, the phenotypic gene validated by qRT-PCR revealed that the BM LAMs are more M1-like while the spleen LAMs are more M2-like.
Our results suggested that the diverse response of LAMs to leukemia are largely determined by organ-specific leukemic microenvironment.
The work was supported by the Grants 81370634, 81570153, 81300376 from the National Natural Science Foundation of China (NSFC).

Disclosures
No relevant conflicts of interest to declare.
Peptide Vaccination Against Cytomegalovirus (CMV) Elicits Immunological and Clinical Responses after Allogeneic Stem Cell Transplantation Even from a CMV Seronegative Donor
Authors: Schmitt Michael , Schmitt Anita , Wiesneth Markus , Hückelhoven Angela , Wu Zeguang , Kuball Juergen , Wang Lei , Schauwecker Peter , Hofmann Susanne , Götz Marlies , Michels Birgit , Maccari Birgit , Wuchter Patrick , Mertens Thomas , Schnitzler Paul , Döhner Hartmut , Ho Anthony D. , Bunjes Donald , Dreger Peter , Schrezenmeier Hubert , Greiner Jochen 
Published: October 2019

Abstract
Introduction: Patients receiving an allogeneic stem cell graft from a cytomegalovirus (CMV) seronegative donor are prone to CMV reactivation with high risk of disease and mortality. Therefore we initiated a clinical phase I trial with a novel vaccine designed by our group: a CMV phosphoprotein 65 (CMVpp65)-derived peptide in water-in-oil emulsion (Montanide™) plus administration of granulocyte-macrophage colony stimulating factor.
Material and Methods: 10 patients after allogeneic stem cell transplantation received four vaccines s.c. at a biweekly interval. Patients were monitored for clinical course and CMVpp65 antigenemia. CMV-specific CD8+ and gamma/delta T cells were analyzed by multi-color flow cytometry. Neutralizing anti-CMV antibody assays were established and correlated to clinical parameters.
Results: Peptide vaccination was well tolerated, no drug-related serious adverse events were detected. 7 of 9 patients with CMVpp65 antigenemia cleared the CMV with enduring response > 1.5 years after 4 vaccinations and are still free from antigenemia until present. 2 patients with CMV reactivation showed persisting CMV antigenemia. One patient received prophylactic vaccination and did not develop antigenemia. An up to 6-fold increase in frequency of both CMV-specific CD8+ T cells or Vdelta2- gamma/delta T cells with a maximum of 1.32% CMV-specific CD8+ T cells ex vivo without priming and 13.7% Vdelta2- gamma/delta T cells of all CD8+ T cells was detected in five patients. Also titers of neutralizing antibodies increased in four patients up to 10-fold over the time of vaccination. Humoral and cellular immune responses correlated with clearance of the CMV load.
Conclusion: Administration of CMVpp65 peptide vaccination for patients after allogeneic stem cell transplantation at high risk for CMV reactivation was safe, well tolerated and clinically encouraging. A study in solid-organ transplant patients is ongoing.

Disclosures
Kuball: Gadeta B.V.: Membership on an entity's Board of Directors or advisory committees. Wuchter:Sanofi-Aventis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Hexal: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Dreger:Gilead: Consultancy; Janssen: Consultancy; Novartis: Consultancy; Gilead: Speakers Bureau; Novartis: Speakers Bureau; Roche: Consultancy. Greiner:BMS: Research Funding.
Target Joint Status in Patients with Hemophilia a during 18 Consecutive Months of Prophylaxis with a Pegylated Full-Length Recombinant Factor VIII with Extended Half-Life
Authors: Engl Werner , Patrone Lisa , Abbuehl Brigitt E. 
Published: October 2019

Abstract
Introduction and Objective: Patients with severe hemophilia A repeatedly bleed into joints and subsequently develop target joints and arthropathy. BAX 855, a polyethylene glycol pegylated, full-length, recombinant factor VIII built on ADVATE, demonstrates an extended half-life, efficacy, and safety for prophylaxis and the treatment of bleeding in patients with severe hemophilia A. Target joint status was evaluated in an ad-hocanalysis of integrated efficacy data from previously treated adolescent and adult patients who participated in the pivotal (completed) and continuation (ongoing) studies.
Methods: The number of target joints, defined as a single joint with ≥3 spontaneous bleeding episodes in any consecutive 6-month period, were analyzed over 3 consecutive 6-month periods in patients who received twice weekly BAX 855 prophylaxis at 40-50 IU/kg.
Results: After approximately 6 months of twice weekly prophylaxis during the pivotal study, 101 adolescent and adult patients continued treatment in the continuation study, 51 of whom were treated with twice weekly prophylaxis for 18 consecutive months. At screening 29.4% (15/51) of these patients had no target joints, 19.6% (10) had 1 target joint, 21.6% (11) had 2, and the remaining 29.4% (15) had 3 or more. After the first 6-month treatment period, the percent of patients with no target joints increased to 66.7% (34 patients), including 19 patients in whom 1 or more target joints had resolved and 15 patients who remained target joint-free from screening. Of note, for the 15 patients with 3 or more target joints at screening, all of their target joints resolved after the first 6 months of treatment with BAX 855. This trend was maintained after the second 6-month treatment period. After the third 6-month period of twice weekly prophylaxis, the percent of patients with no target joints further increased to 82.4% (42), which included the 15 patients who remained target joint-free from screening. Nine patients had 1 or more unresolved target joint after 18 months of twice weekly prophylaxis, and in 6 of these patients, 1 to 3 other target joints had resolved. Overall, there were 89 target joints at screening which reduced to 14 after 18 months of twice weekly prophylaxis. Of those, 10 target joints changed status (eg, were present, resolved, and then re-appeared) and only 4 target joints (in 4 patients) persisted as unresolved through each of the 3 consecutive 6-month treatment periods.
Conclusions: These results demonstrate the efficacy of continuous twice weekly prophylaxis with BAX 855 for preventing and resolving the majority of target joints.

Disclosures
Engl: Shire, formerly Baxalta and Baxter: Employment, Equity Ownership. Patrone:Shire, formerly Baxalta and Baxter: Employment, Equity Ownership. Abbuehl:Shire, formerly Baxalta and Baxter: Employment, Equity Ownership.
Risk Factors for Venous Thromboembolism in Metastatic Colon Cancer Patients in the Contemporary Treatment Era: A SEER-Medicare Data Analysis
Authors: Pulluri Bhargavi , Littenberg Benjamin , Lal Inder , HOlmes Chris E. , Ades Steven 
Published: October 2019

Abstract
Background:The relationship between metastatic colorectal cancer (CRC) and venous thromboembolism (VTE) is not well defined in the modern treatment era. Previous population-based studies date back to a period marked by inpatient intravenous heparin therapy and inferior survival due to paucity of therapeutic anti-cancer options, and before the advent of newer therapies including oxaliplatin, irinotecan, and anti-angiogenic treatment with bevacizumab. The objectives of this retrospective study were to examine the impact of multiple putative risk factors on VTE incidence in a large representative modern cohort of older patients with metastatic CRC.
Methods:We performed a retrospective analysis of SEER-Medicare data on elderly patients with metastatic CRC diagnosed in 2004-2011. VTE and associated risk factors were analyzed using multivariate Cox proportional hazards models adjusted for sex, age at diagnosis, race, ethnicity, tumor anatomy (left/right/unknown), calendar year of diagnosis, Charlson comorbidity score, location of SEER registry and urban residence, with time-varying covariates for use of cancer therapies.
Results:Of 339,778 records, 11,086 metastatic colon cancer cases were identified. 1,338 cases had VTE with a cumulative incidence of 13% at 1 year and 19% at 3 years. The mean age was 77.9 years (range 65-106). 49.7% were women and 83.5% white. 60.5% had a Charlson comorbidity score of zero at diagnosis; 6% had scores of 6-18.
Significant predictors of VTE included female sex (Hazard Ratio (HR) 1.22; 95% Confidence Interval (CI) 1.10, 1.36; P<0.001), younger age at diagnosis (HR 1.25 per decade of age; CI 1.15, 1.37; P<0.001), urban residence (HR 1.20; CI 1.03, 1.40; P=0.02), right sided colon cancers (HR 1.19; CI, 1.06, 1.33 P=0.003), and current use of 5-fluorouracil (HR 1.33; CI 1.04, 1.70; P=0.02). The risk of VTE was significantly reduced when on bevacizumab (HR 0.77; CI 0.63, 0.93; P=.006), or irinotecan therapy (HR 0.59; CI 0.47, 0.75; P<0.001) and in those with more comorbid conditions (HR 0.97 per point of Charlson score; CI 0.935, 0.998; P=0.04).
Conclusion: The higher incidence of VTE seen in right sided colon cancers may be related to biologically aggressive tumors associated with poor survival as reported in recent studies. The lower risk of VTE in older, sicker patients was unexpected and may reflect the effect of competing mortality. In this large contemporary cohort, anti-angiogenic therapy was not associated with a higher risk of VTE; the apparent protective effects of bevacizumab and irinotecan may represent treatment assignment bias.

Disclosures
No relevant conflicts of interest to declare.
Systematic Review of Treatment for Massive and Submassive Pulmonary Embolism
Authors: Gupta Shipa , Gandara Esteban , Carrier Marc , Castellucci Lana A 
Published: October 2019

Abstract

Introduction: Anticoagulation is the standard treatment for pulmonary embolism (PE). In patients presenting with massive or submassive PE, the addition of systemic thrombolytics (STL) and catheter directed therapies (CDT) may also be considered. However, there is uncertainty about the benefits and risks of these treatments, and no randomized trials have compared STL to CDT.
Aim: We reviewed the risks and benefits of STL and CDT in the management of patients with massive or submassive PE.
Methods: A systematic literature search was conducted using MEDLINE, Embase, Cochrane Central Register of Controlled Trials, and bibliographies of relevant studies and reviews. Randomized trials and cohort studies were eligible. Registries and studies with fewer than 10 patients were ineligible. Pooled proportions and their associated 95% confidence intervals (CI) for mortality, major bleeding, intracranial hemorrhage (ICH), recurrent PE, and fatal recurrent PE were calculated.
Results: A total of 52 studies were included in the analyses: 19 studies evaluating STL; 18 studies using CDT; 5 studies using the AngioJet rheolytic thrombectomy system; and 10 studies using ultrasound-assisted CDT. The pooled proportion of outcomes for patients receiving STL were: 4.46% (95% CI: 3.04-6.13%) for mortality; 11.70% (95% CI: 7.78-16.90%) for major bleeding; 1.99% (95% CI: 1.06-3.20%) for ICH; 3.92% (95% CI: 2.56-5.55%) for recurrent PE; and 1.04% (95% CI: 0.42-1.94%) for fatal recurrent PE. For patients receiving CDT, the pooled proportion of outcomes were: 9.64% (95% CI: 5.93-14.13%) for mortality; 5.17% (95% CI: 2.18-9.33%) for major bleeding; 1.29% (95% CI: 0.48-2.48%) for ICH; 2.76% (95% CI: 1.36-4.63%) for recurrent PE; and 1.56% (95% CI: 0.53-3.12%) for fatal recurrent PE. For patients receiving AngioJet CDT, the pooled proportion of outcomes were: 10.18% (95% CI: 3.68-19.42%) for mortality; 9.14% (95% CI: 0.88-24.70%) for major bleeding; 2.30% (95% CI: 0.34-5.96%) for ICH; 4.84% (95% CI: 1.62-9.66%) for recurrent PE; and 4.84% (95% CI: 1.62-9.66%) for fatal recurrent PE. In patients receiving ultrasound-assisted CDT, the pooled proportion of outcomes were: 4.17% (95% CI: 1.64-7.79%) for mortality; 4.88% (95% CI: 2.48-8.05%) for major bleeding; 0.63% (95% CI: 0.10-1.59%) for ICH; 1.39% (95% CI: 0.48-2.77%) for recurrent PE; and 1.21% (95% CI: 0.35-2.56%) for fatal recurrent PE.
Conclusions: Our results suggest that all-cause mortality rates in patients with massive or submassive PE are similar between STL and ultrasound-assisted CDT and are lower than other types of CDT. In general, major bleeding rates were lower in CDT based groups than in STL. Importantly, ICH rates in CDT based treatment strategies were also lower than STL therapy. Randomized trials comparing STL and CDT would help solidify the role of these treatment strategies in patients with massive and submassive PE.

Disclosures
Carrier: BMS: Research Funding; Leo Pharma: Research Funding.
Staring inside of Idiopathic Hypereosinophilia: Identification of Clonality Using Targeted Exome Sequencing
Authors: Lee Jee-Soo , Seo Heewon , Im Kyongok , Park Si Nae , Kim Jung-Ah , Kim Seon Young , Lee Joon-hee , Kwon Sunghoon , Koh Insong , Hwang Seungwoo , Park Heung-Woo , Kang Hye-Ryun , Kim Ju Han , Kim Miyoung , Lee Dong Soon 
Published: October 2019

Abstract
Idiopathic hypereosinophilia (IHE) has been a disorder of uncertain etiology, characterized by persistent elevation of blood eosinophil count exceeding 1.5x103/μL without evidence of reactive and clonal causes. When end-organ damage due to eosinophilic infiltration is present, idiopathic hypereosinophilic syndrome (IHES) is diagnosed. As the name 'idiopathic' implies the uncertain etiology, the nature of eosinophilic proliferation is unclear whether benign or clonal. While T cell clonality assessment has been recommended for unexplained hypereosinophilia, this approach is not often applied to the routine practice in clinic. We hypothesized that the clonality would exist in patients classified into IHE/IHES. The bone marrow cells of 30 IHE patients were retrospectively evaluated.The T cell receptor (TCR) assay and the targeted sequencing for 88 genes related to hematologic malignancies were performed. After being analyzed, the candidate mutations were subjected to pathway analysis.Through the targeted sequencing, 140 variants in 59 genes were identified. Sixteen out of 30 patients (53.3%) harbored at least 1 candidate mutation. The most frequently affected genes were NOTCH (n=8); SCRIB and STAG2 (n=5); SH2B3 (n=4). Remarkably, we identified recurrent somatic mutations in 4 genes with 5 patients (16.7%); MPLY521X (n=2), SCRIBE795X (n=2), ASXL1G646f (n=3) and STAG2E249X (n=2). A known mutation TP53V216M was confirmed in an IHE patient. Interestingly, the TCR assay revealed that 4 of 30 patients (13.3%) had a clonal TCR rearrangement. Skin involvement was described in patients with clonal T cell population. Through the pathway analysis, the role of these genes in eosinophilia was identified. These results highly suggest that these genes are likely related to clonality of hypereosinophilia.This study revealed that somatic mutations affecting hematopoiesis and monoclonal T cell clones underlie idiopathic hypereosinophilia. It would be valuable to extend our findings on the molecular profiling of hypereosinophilia to the further study discovering the clonality of IHE.

Disclosures
No relevant conflicts of interest to declare.
Determining the Signal Source in Magnetic Resonance Imaging of Myelofibrotic Bone Marrow
Authors: Matsuura Shinobu , Patterson Shenia , Lucero Hector , Grant Aaron , Herrera Victoria L. M. , Ravid Katya 
Published: October 2019

Abstract
Magnetic resonance imaging (MRI) is a promising diagnostic method for evaluation of bone marrow myelofibrosis for offering non-invasive, wider-area coverage over conventional histology. Although the most striking feature in histology of myelofibrotic bone marrow is the proliferation of reticulin fibers, the spectrum of findings involves abnormalities in cell number and composition, vessel proliferation, as well as possible pre-fibrotic abnormalities in matrix such as inflammation/edema. Correlation of MRI observations with such histological findings to accurately define the pathogenic phases of myelofibrosis has been challenging. Here, we present the first MRI analysis of myelofibrosis in a mouse model (Gata-1low mice) in an attempt to correlate MRI signal observations with histological findings. T2-weighted MRI images of femur bone marrow of Gata-1low animals at different ages (12 to 46 weeks) showed high MR signal intensity, absent in age-matched wild-type controls, that is detected as early as 12 weeks of age, before onset of histologically-defined fibrosis in this mouse model. MRI acquisitions employing fat suppression, a technique that suppresses signals from lipids, together with the absence of chemical shift artifacts in the images, excluded the possibility of lipids as the source of the prominent MR signal. To determine the source of this prominent non-lipid signal in Gata-1low bone marrow, cellular composition, vascularization and vascular permeability were tested. Our findings demonstrate a multi-source nature of MRI signals in bone marrow, which evolves from young (12 weeks) to old (40 weeks) Gata-1low bone marrow consistent with the pathologic progression of myelofibrosis. Data support the importance of an integrative approach to further the development of this non-invasive modality for following the dynamics and progression of this multifactorial pathology in humans.

Disclosures
No relevant conflicts of interest to declare.