bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mlkl Pores Release Neutrophil Extracellular Traps in Necroptotic Neutrophils
Authors: D'Cruz Akshay A , Bliss-Moreau Meghan , Ericcson Maria , Croker Ben A 
Published: October 2019

Abstract
Neutrophil extracellular traps (NETs) are networks of extracellular nuclear DNA and microbicidal proteins released from neutrophils in response to tissue damage and infection. Despite evidence of pathogenic roles for NETs in systemic lupus erythematosus, rheumatoid arthritis, diabetes, artherosclerosis and Alzheimer's disease, the major biochemical pathways controlling their formation remains poorly understood. Apoptosis does not contribute to NET formation but the role of regulated non-apoptotic cell death pathways such as necroptosis is not known. We have investigated the role of positive and negative regulators of necroptosis including receptor-interacting protein kinase-3 (RIPK3), mixed lineage kinase domain-like (MLKL), receptor-interacting protein kinase-1 (RIPK1) and Caspase-8. Using immunogold electron microscopy, flow cytometry, imaging flow cytometry and fluorescence microscopy, we demonstrate that necroptosis can drive NET formation via MLKL pore formation at the cell surface. This process is caspase-independent but reactive oxygen species-dependent. Genetically-modified mouse peripheral blood and bone marrow neutrophils were used to show that Caspase-8 and RIPK1 negatively regulate NET formation driven by RIPK3 and MLKL. Mice that lack MLKL are deficient in necroptosis and NET formation, and were sensitive to methicillin-resistant Staphylococcus aureus (MRSA). Neutrophil-specific Caspase-8-deficiency also leads to increased susceptibility to MRSA due to increased rates of necroptotic neutrophil death. Killing of MRSA by necroptotic neutrophils is sensitive to DNase, and is dependent on MLKL, suggesting that necroptosis-driven NET formation contributes to the bactericidal activity of neutrophils. Human peripheral blood neutrophils also generate NETs that are sensitive to pharmacological inhibitors of necroptosis, suggesting that targeting necroptosis in general may help combat autoimmune responses to DNA. This study provides a framework to investigate the role of extracellular DNA release and cell death in the setting of infection, autoimmunity and autoinflammatory disease.

Disclosures
No relevant conflicts of interest to declare.
Genetic Deletion of CCRL2 Impairs Macrophage Accumulation in Arterial Intima and Attenuates Atherosclerotic Plaque Development
Authors: Cao Yiren , Yang Fei , Tang Chaojun , Hu Shuhong , Zabel Brian Anthony , Zhu Li 
Published: October 2019

Abstract
Atherosclerosis is a chronic inflammatory disease of the arterial wall elicited by accumulation of LDL and leucocytes in the subendothelium at predilection sites with disturbed laminar flow. Chemokines and their receptors appear to act as critical players in atherosclerosis as they not only direct atherogenic recruitment of leucocytes but also exert cell hemostatic functions by chemokine ligand-receptor axes and their specific or combined contributions. Chemokine (C-C motif) receptor-like 2 (CCRL2) is an atypical chemokine receptor that cooperates with its ligand chemerin to play a role in cell trafficking and inflammatory response, the processes usually occur in atherosclerosis, but its role in atherosclerosis is not clear. Here we investigated the potential role of CCRL2 in atherogenesis using the classic apolipoprotein E-deficient (ApoE-/-) mouse model of atherosclerosis. Atherosclerosis-prone ApoE-/- mice were crossed with CCRL2-/- mice to obtain ApoE-/-CCRL2+/+ and ApoE-/-CCRL2-/- mice. Male mice of both genotypes were fed a standard chow diet until 8 weeks of age and then switched to a high fat diet for 16 weeks. Aortas were dissected and assessed by en face staining. Although CCRL2 deletion did not change mouse blood lipid profile and body weight, the atherosclerotic plaque area of the total aorta was reduced by 27.2% (P=0.0379) in ApoE-/-CCRL2-/- mice compared with ApoE-/-CCRL2+/+ mice with striking difference occurring in aortic arch. A reduction of lipid deposition by 32.6% (P=0.0089) was also observed in aortic root sections when CCRL2 was deleted. Further studies showed that deletion of CCRL2 reduced macrophage accumulation and polarization during the development of atherosclerosis. CCRL2 not only expresses in high levels in the plaques of ApoE-/- mice on a high fat diet but also co-localizes with macrophages and chemerin. Interestingly, using CMKLR1-/- chimeric mice, we showed that chemerin/CMKLR1/CCRL2 axis is involved in leucocyte infiltration and possibly affects plaque formation. More importantly, partial carotid artery ligation (PCL) model confirmed that disturbed blood flow-induced endothelial expression of CCRL2 modulates plaque formation. In conclusion, our results demonstrate that CCRL2 deficiency delays macrophage accumulation and the formation of atherosclerotic plaques potentially via the disruption of chemerin/CMKLR1/CCRL2 axis and the loss of CCRL2 response to the disturbed flow, which may represent a novel mechanism of atherosclerosis. This work was supported by Natural Science Foundation of China (grant 81370373 to L.Z. and 31300781 to C.T.)

Disclosures
No relevant conflicts of interest to declare.
Flow Cytometry-Based Platelet Function Testing Is Predictive of Symptom Burden in a Cohort of Bleeders
Authors: Boknäs Niklas , Ramström Sofia , Faxälv Lars , Lindahl Tomas L 
Published: October 2019

Abstract

Platelet function disorders (PFDs) are common in patients with mild bleeding disorders (MBDs), yet the clinical significance of laboratory findings suggestive of a PFD remain unclear due to the lack of evidence for a clear link between test results and patient phenotype. Herein, we present results from a study evaluating the potential utility of platelet function testing using whole-blood flow cytometry in a cohort of 105 patients undergoing investigation for MBD. Subjects were evaluated with a test panel comprising two different activation markers (fibrinogen binding and p-selectin exposure) and four physiologically relevant platelet agonists (ADP, PAR1-AP, PAR4-AP and CRP-XL). Abnormal test results were identified by comparison with reference ranges constructed from 24 healthy controls or the fifth percentile of the entire patient sample. We found that abnormal test results are predictive of bleeding symptom severity, and that the greatest predictive strength was achieved using a subset of the panel, comparing measurements of fibrinogen binding after activation with all four agonists with the fifth percentile of the patient sample (P = 0.00008, hazard ratio 8.7; 95 % CI 2.5-40). Our results suggest that whole-blood flow cytometry-based platelet function testing is a feasible alternative for the investigation of MBDs. We also show that platelet function testing using whole-blood flow cytometry could provide a clinically relevant quantitative assessment of platelet-related primary hemostasis.
Figure 1. Test results for each patient in comparison with reference range (A) and the fifth percentile of the entire patient cohort (B). Normal test results are colored grey, abnormal test results are colored with a continuous color gradient using the deviation from the mean divided by the standard deviation as a measure of degree of abnormality. Grey horizontal bars illustrate the number of abnormal test results for each patient. Figure 1. Test results for each patient in comparison with reference range (A) and the fifth percentile of the entire patient cohort (B). Normal test results are colored grey, abnormal test results are colored with a continuous color gradient using the deviation from the mean divided by the standard deviation as a measure of degree of abnormality. Grey horizontal bars illustrate the number of abnormal test results for each patient.

Disclosures
Lindahl: Diapensia: Equity Ownership.
Addition of Intermediate-Dose Methotrexate Is Beneficial in Terms of Both Progression-Free Survival and Overall Survival for Patients with Diffuse Large B Cell Lymphoma Receiving Either R-CHOP or CHOP
Authors: Dann Eldad J. , Goldschmidt Neta , Heffes Vered , Mashiach Tanya , Gatt Moshe E. , Horowitz Netanel A 
Published: October 2019

Abstract
Introduction: Patients with diffuse large B cell lymphoma (DLBCL) and high International Prognostic Index (IPI) or extra-nodal disease are at an increased risk of systemic and central nervous system (CNS) relapse. Presently, the management of DLBCL patients at the Rambam Health Care Campus (Haifa, Israel) and Hadassah Medical Center (Jerusalem, Israel) includes 6 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) along with 4 doses of intrathecal (IT) methotrexate (MTX), which may be followed by 2 additional cycles of an intermediate dose (3 g/m2) of intravenousMTX (ID-MTX). The current study compared progression-free survival (PFS), overall survival (OS) and CNS relapse rate in a cohort of DLBCL patients with risk factors for CNS relapse who did or did not undergo prophylactic therapy with ID-MTX.
Methods: Four hundred and eighty DLBCL patients treated at two tertiary care centers (Rambam and Hadassah) between the years 1991 and 2013 were retrospectively evaluated. The cohort included 224 females and 256 males at a median age of 58 years (range18-76). OS, PFS and CNS involvement at relapse were analyzed. The study incorporated all patients with the primary diagnosis of DLBCL and risk factors for CNS relapse, i.e., Waldeyer ring (2%), breast (1%), testicular (3%) or orbita (1%) involvement, bone marrow involvement (21%), stage IV disease (64%), LDH higher than normal (70%), ≥1 extra-nodal site (28%), IPI ≥3 (36%). Eight percent of patients had stage I-IE disease, 17% - stage II-IIE, 11% - stage III, 64% - stage IV, 29% had B symptoms. Patients with CNS involvement at diagnosis were excluded from the analysis.
Results: Four hundred and seventy three patients (99%) were treated with either CHOP or CHOP-like regimen, and 384 (80%) patients received rituximab (R). In the R-CHOP cohort, 23% of patients with IPI 0/1, 28% with IPI 2 and 42% with IPI ≥3 were given prophylaxis. One hundred and thirty patients received ID-MTX (27%), 35 patients (7%) received IT-MTX only, while no prophylaxis was administered to 315 patients (66%). The cumulative incidence of CNS relapse at 5 years was 6%, with no difference in the incidence between the patients who received prophylaxis with ID-MTX, IT-MTX or were given no prophylaxis at all. Patients with IPI ≥3 treated with ID-MTX had a decreased overall relapse rate and significantly superior OS and PFS (Table 1; Figure 1).
Conclusion: In the current study, the addition of two cycles of ID-MTX to the standard R-CHOP regimen resulted in improved 5-year PFS and OS of DLBCL patients with IPI ≥3. These data need to be further confirmed in a randomized controlled study.

Disclosures
No relevant conflicts of interest to declare.
Patterns of Microrna in Plasma Cells: From Normal Differentiation to Multiple Myeloma
Authors: Kassambara Alboukadel , Bruyer Angelique , Jourdan Michel , Robert Nicolas , Pantesco Véronique , Elemento Olivier , Klein Bernard , Moreaux Jerome 
Published: October 2019

Abstract
Background: MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression either via the degradation of target mRNAs or the inhibition of protein translation. miRNAs have been involved in fine-tuning critical cellular processes and play critical roles in cell differentiation and tumor development. Furthermore, there is an increasing recognition of the role of miRNAs in multiple myeloma, a plasma cell (PC) malignancy characterized by an accumulation of malignant PCs within the bone marrow. However, little is known about miRNA expression during human PCD as well as about the full extent to which individual miRNAs regulate fundamental processes during PCD. A complete delineation of miRNA and their target expression during normal PCD is essential to understand the role of miRNAs in plasma cell malignancies.
Methods: We analyzed the expression profile of miRNAs and mRNAs during human plasma cell differentiation (PCD) to infer miRNA-target relationships, as well as in multiple myeloma tumor plasma cells. We developed a method and an R package, that uses miRNA and mRNA expression profiles across PCD cell subpopulations to infer candidate miRNA-target interactions that could be active and functional in PCD. We inferred miRNA-target relationships from sequence-based prediction, experimentally validated target interactions curated from the literature, published data from miRNA perturbation experiments and inverse expression relationships between miRNAs and their target mRNAs.
Results: Our results reveal 63 miRNAs with significant temporal changes in their expression during normal PCD. We derived a high-confidence network of 295 target relationships comprising 47 miRNAs and 147 targets. These relationships include new examples of miRNAs that are likely to coordinately regulate multiple members of critical pathways associated with PCD. Notably, we identify new miRNAs that coordinately regulate important pathways in PCD, including members of miR-30 and miR-29 families, miR-106b and miR-16, which regulates IRF4/PRDM1 axis, active DNA methylation pathway, TGF-b signaling pathway, autophagy, ZBTB4/EZH2 axis and cell cycle progression. Furthermore, our work demonstrates that 28 PCD stage-specific miRNAs are aberrantly overexpressed in multiple myeloma cells (MMCs) compared to their normal counterpart and/or are associated with high risk myeloma, suggesting that MMCs frequently acquired expression changes in miRNAs already undergoing dynamic expression modulation during normal PCD. Finally, expression of some targets of these PCD - MM miRNAs is correlated with clinical outcome of uniformly treated MM patients.
Conclusions: Altogether, our results demonstrate that miRNAs may be important in controlling PCD and malignant plasma cell biology.

Disclosures
No relevant conflicts of interest to declare.
Interference of HMGB1 Lead to Increased Vulnerability of MM Cells to Dexamethasone and Doxorubicin
Authors: Guo Xing , He Donghua , Chen Jing , Lin Xuanru , Chen Qingxiao , Yan Haimeng , Huang Xi , Li Yi , Yang Li , Zhang Enfan , Yang Yang , Wu Wenjun , He Jingsong , Huang He , Cai Zhen 
Published: October 2019

Abstract
Background: Multiple myeloma (MM) is the second mostly diagnosed disease among hematological malignancies after lymphoma. With the novel agents, the survival of MM patients has been improved significantly but still remains incurable because of drug resistance. Studies have found that high-mobility group box 1 protein (HMGB1) was involved in inflammation, angiogenesis, and cancer invasion progression, metastasis, and drug resistance. Our research was aimed at exploring the role of HMGB1 in MM cell proliferation and drug resistance.
Methods: First, semi-quantitative real time-polymerase chain (qRT-PCR) and western blot was used to determine the levels of HMGB1 mRNA and protein expression in MM cell lines (RPMI8226, CAG, and MM.1S) and primary MM samples. Second, MM cells were transfected with HMGB1-knockdown lentivirus and the Cell Counting Kit 8 (CCK8) assay was used to determine the proliferation of MM cells with or without chemotherapeutic drugs dexamethasone (Dex) and doxorubicin (ADM). Then cell apoptosis was detected by flow cytometry. Third, Affymetrix HTA 2.0 Array was used to compare changes in gene expression levels between HMGB1-knockdown cells and the control cells and qRT-PCR was used to verify the array results. Last, Western bolt was performed to analyze changes in signaling pathways after HMGB1 knockdown.
Results: MM cell lines and primary MM samples expressed high levels of HMGB1 mRNA and protein. Although there was no difference in MM cell proliferation between HMGB1-knockdown group and the control group (P>0.05), HMGB1-knockdown significantly enhanced inhibitory effect of chemotherapy with Dex and ADM in comparison with the wildtype HMGB1 control (P<0.05). Flow cytometry results showed that apoptosis of MM cells induced by Dex and ADM was increased when HMGB1 expression was suppressed (P<0.05). Furthermore, gene array analysis on RPMI8226 and CAG cell lines showed that anti-apoptotic genes (bcl-2, HIAP1) and MM survival related genes (DEPTOR, CXCL12) were decreased and pro-apoptotic genes (TNFRSF1B, TRAIL, CXCL10) were increased in knockdown cells compared to the controls. In addition, expression levels of genes that play important roles in signaling pathways such as mTOR, NF-κB, PI3K-AKt, and p38-MAPK were also significantly changed. The gene expression microarray results were verified by qRT-PCR and Western blot demonstrated that phosphorylation of p70S6K (substrate of mTORC1 complex) and AKT-ser473 (substrate of mTORC2 complex) were both elevated in HMGB1-knockdown MM cells compared to that in the control cells.
Conclusions: Our research showed that downregulation of HMGB1 increased sensitivity of MM cells to Dex and ADM through increasing apoptosis and regulating mTOR, NF-κB, PI3K-AKt, and p38-MAPK pathways.

Disclosures
No relevant conflicts of interest to declare.
Standard of Care of Patients with CML Treated in Community Based Oncology Group Practices in Rhineland-Palatinate (Germany)
Authors: Weide Rudolf , Rendenbach Bernhard , Grundheber Monika , Burkhard Oswald , Behringer Joachim , Maasberg Michael , Ehscheidt Peter , Strehl John Werner , Hansen Richard , Feiten Stefan 
Published: October 2019

Abstract
Introduction: Significant progress has been made in CML-therapy since the introduction of imatinib and other tyrosine kinase inhibitors (TKI) into clinical care. The aim of this study was to assess diagnosis, treatment and outcome of CML-patients who received their treatment in community based oncology practices in Rhineland-Palatinate and whether European LeukemiaNET-guidelines were followed.
Methods: All Ph-/BCR-ABL-positive CML-patients who were treated between 12/2001-12/2015 in 9 oncology group practices were analyzed retrospectively concerning diagnosis, treatment and outcome according to European LeukemiaNET-guidelines. Data were collected from patient files into a central data base and analyzed statistically with SPSS.
Results: 264 patients (pts) with a median age of 60 (18-90) were analyzed. 126 (48%) were female, 138 (52%) were male. At initial diagnosis bone marrow biopsy was performed in 213 pts (81%). Cytogenetics was applied in 204 pts (77%) (38% in blood, 56% in bone marrow). FISH-analysis was used in 155 pts (59%) (33% in blood, 36% in bone marrow). PCR-testing to detect a BCR-ABL1-rearrangement was applied in 200 pts (76%) (52% blood, 37% bone marrow). 258 pts (98%) were in chronic phase, 5 (2%) in accelerated phase and 1 (0.4%) in blast crisis at diagnosis. EUTOS score could be calculated in 131 pts (50%). 20% were high risk, 80% low risk. 252 pts (95%) received some form of TKI-therapy. Out of 416 TKI-therapies 308 (74%) were PCR-based monitored, 148 (36%) were monitored by cytogenetics. First line treatment was imatinib in 201 pts (80%), 51 pts (20%) received a second generation TKI. Second line treatment consisted of dasatinib in 59%, nilotinib in 32%, imatinib in 6% and bosutinib in 3%. Third line treatment was nilotinib in 56%, dasatinib in 35%, ponatinib in 6% and imatinib in 3%. 62 pts (23%) were treated within a study protocol. 13 pts (5%) received an allogeneic transplantation. Overall survival probability was 88% after 5 years and 72% after 10 years. Disease specific survival was 95% after 5 years and 86% after 10 years.
Conclusion: The overwhelming majority of CML-patients treated in oncology group practices receive standard of care as suggested by European LeukemiaNET-guidelines. Overall survival in routine care is comparable to international studies.

Disclosures
No relevant conflicts of interest to declare.
Most up-to-Date Long Term Survival Estimates for Common Hematologic Malignancies Using the Boomerang Method
Authors: Pulte Dianne , Jansen Lina , Brenner Hermann 
Published: October 2019

Abstract
Background: Five year survival has increased dramatically for most of the common hematologic malignancies in the early part of the 21st century, probably due to new therapeutic options. However, less is known about the effects recent changes in therapy have had on longer term survival. Here, we examine 10- and 20-year survival for patients with common hematologic malignancies including myeloma, acute lymphoblastic leukemia (ALL), acute myeloblastic leukemia (AML), chronic lymphoid leukemia (CLL), chronic myeloid leukemia (CML), non-Hodgkin lymphoma (NHL), and Hodgkin lymphoma (HL).
Methods: Data were extracted from the Surveillance, Epidemiology, and End Results (SEER9) database. Patients age 15+ with appropriate ICD-10 codes for the above malignancies were included, with exclusion of those patients diagnosed by death certificate only. The boomerang method1 was used to estimate 10 and 20-year relative survival for patients in two calendar periods, 1999-2003 and 2009-13. This recently described method has been demonstrated to more closely approximate actual observed survival for a given time period compared to other methods of estimation, including period analysis, by minimizing the contribution of earlier time periods to the estimation of survival.
Results: Ten and 20-year survival increased for each malignancy examined, with increases ranging from +4.3 percent units for 20-year survival for myeloma to +28.4 percent units for 10-year survival for CML (see Table). Ten year relative survival was greater than 50% in 2009-13 for patients with CML, CLL, NHL, and HL, at 69.9%, 59.1%, 63.4%, and 63.1%, respectively. Major decreases in survival were observed between 10- and 20-year survival estimates for all malignancies except for AML, where 10- and 20-year relative survival estimates were virtually identical in 2009-13.
Discussion: Long term survival is increasing for common hematologic malignancies, with particularly strong increases being observed for CLL, CML, and NHL. Major decreases in survival were observed between 10 and 20 years for most malignancies, including NHL and ALL, which are generally considered curable. This may be partly due to the effect of recent changes in therapy leading to underestimation of 20-year survival, even using the boomerang method, but further study of long-term outcomes in curable malignancies to determine whether there is an increased risk of specific issues later in life is indicated.
Reference:
1. Brenner H, Jansen L. Timely disclosure of progress in long-term cancer survival: the boomerang method substantially improved estimates in a comparative study. J Clin Epidemiol. 2016;70:224-32.

Disclosures
Pulte: Selexys Pharmaceuticals: Research Funding; EBSCO: Other: Review of content for Dynamed medical reference product; ApoPharma: Research Funding.