bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of the Thrombin-PAR-1 Pathway in Coxsackievirus Induced Hepatitis
Authors: Tatsumi Kohei , Antoniak Silvio , Mackman Nigel 
Published: October 2019

Abstract
Objective: Coxsackievirus B3 (CVB3) can infect different tissues including the heart and liver. Recently, we found that activation of the coagulation cascade and protease-activated receptor 1 (PAR-1) enhances toll-like receptor-3 (TLR3) mediated interferon-β (IFN-β) expression and protects mice from CVB3-induced myocarditis. Here, we investigated the role of PAR-1 in early anti-viral responses in mice and isolated hepatocytes.
Methods: Wild-type (WT) and PAR-1 deficient (PAR-1-/-) mice were infected with CVB3 intraperitoneally. The innate immune response, viral load, liver enzyme plasma levels, and inflammation levels were analyzed. Bone-marrow transplantation experiments with the combination of WT mice PAR-1-/- mice were performed to identify the cellular source of PAR-1 contributing to the innate immune response to CVB3. We also analyzed the effect of the direct thrombin inhibition with dabigatran etexilate on CVB3 hepatitis. In addition, we analyzed the effect of PAR-1 activation on TLR3-dependent interferon (IFN)-β expression in primary mouse hepatocytes and the human hepatocyte cell line PH5CH8 in vitro.
Results: PAR-1-/- mice exhibited a reduced early innate immune response in the liver at day 4 after infection, which was associated at later times (day 8) to higher viral titers in the liver, increased alanine transaminase plasma levels and more remarkable inflammation compared to control WT mice. Bone marrow transplantation experiments demonstrated that PAR-1 on non-hematopoietic played the major role in the innate immune response of CVB3 hepatitis. Stimulation of PAR-1 with either thrombin or agonist peptide on primary mouse hepatocytes and human PH5CH8 cells in vitro enhanced the antiviral response to dsRNA by increasing IFN-β and C-X-C motif chemokine 10 (CXCL10) expressions, supporting the results of in vivo experiments.
Conclusion: Our results suggest that activation of PAR-1 on hepatocytes enhances the innate immune response to CVB3 in the liver.

Disclosures
No relevant conflicts of interest to declare.
C1q-Binding HLA Antibodies Do Not Predict Platelet Transfusion Failure in TRAP Study Participant
Authors: Jackman Rachael P. , Bolgiano Douglas , Lebedeva Mila , Slichter Sherrill J. , Norris Philip J. 
Published: October 2019

Abstract
Introduction: In the Trial to Reduce Alloimmunization to Platelets (TRAP) study, 101 of the 530 subjects became clinically refractory (CR) to platelet transfusions in the absence of detectable antibodies against HLA as measured by the lymphocytotoxicity assay (LCA). Using more sensitive bead-based detection methods we have previously demonstrated that while many of these LCA- patients do have anti-HLA antibodies, that these low to moderate level antibodies do not predict refractoriness. In addition to being less sensitive then bead based methods, the LCA screen only detects complement-binding antibodies. As these antibodies could be important for platelet rejection, we assessed if previously undetected complement-binding antibodies could account for some of the refractoriness seen in LCA- patients.
Methods: 169 LCA- (69 CR, 100 non-CR) and 20 LCA+ (10 CR, 10 non-CR) subjects were selected from the TRAP study. Anti-class I HLA IgG and C1q binding antibodies were measured in serum or plasma using two different multi-analyte, semi-quantitative, bead-based fluorescent antibody detection assays: the LabScreen mixed Luminex assay, and the LabScreen single antigen class I assay with or without added EDTA (One Lambda). Groups were compared using an unpaired t-test, a=0.05, and correlation between variables was also assessed.
Results: New measurements of anti-class I HLA IgG antibodies reliably reproduced earlier data with a strong correlation between the old and new measurements (R2=0.9736, p<0.0001). While some of the LCA- subjects did have detectable C1q-binding anti-class I HLA antibodies, and some LCA+ subjects did not, levels of these antibodies were significantly higher among LCA+ subjects (p<0.0001). Levels of C1q-binding anti-class I HLA antibodies were not significantly different between CR and non-CR among either the LCA- or LCA+ subjects.
Conclusions: While complement-binding anti-class I HLA antibody levels were higher in the LCA+ subjects, higher levels of these antibodies were not seen in CR LCA+ patients as compared with non-CR LCA+ patients. Complement-binding anti-class I HLA antibodies do not account for refractoriness seen among the LCA- TRAP subjects. This work confirms that low to mid level anti-class I antibodies do not drive refractoriness to platelets, and suggests that antibody-independent mechanisms cause refractoriness in patients lacking higher levels of anti-HLA antibodies.

Disclosures
No relevant conflicts of interest to declare.
Lincrna Hotset Regulates Hox Gene Transcription and Promotes Early Hematopoietic Fates
Authors: Deng Changwang , Li Ying , Tusi Besabeh , Jian Wei , Patel Bhavita , Li Yangqiu , Bungert Jorg , Zhou Lei , Qiu Yi , Huang Suming 
Published: October 2019

Abstract
Mammalian genomes encode tens of thousands of long intergenic noncoding RNAs (lincRNAs). Accumulated evidence suggests that many of lincRNAs are expressed in a development- or tissue-specific manner. However, it remains largely unknown how lincRNAs reprogram lineage-specific gene expression and regulate lineage-specific differentiation processes during early embryonic development. It is also important to identify specific lincRNA(s) that is involved in early hematopoietic lineage formation and differentiation. Here, we reported a HoxB locus-associated lincRNA HOTSET that is conservatively expressed in both mouse and human hematopoietic stem and progenitor cells (HS/PCs). During embryonic development, HOTSET RNA is dramatically induced upon embryoid body (EB) hematopoietic differentiation. Expression of HOTSET RNA is required for development of the mesoderm-derived Flk1+ bipotential precursors of blood and endothelium lineages and CD41+/c-Kit+ hematopoietic stem and progenitor cells. We demonstrated that HOTSET RNA directly interacts with the SET domains of Setd1a and MLL1 histone methyltransferases in a sense-strand dependent manner. HOTSET RNA directly binds to the HoxB1-6 gene promoters and recruits the Setd1a/MLL1 complexes to the anterior HoxB locus upon differentiation. Furthermore, HOTSET RNA mediated H3K4 methylation is essential for long-range chromatin loops that bring anterior HoxB genes into a close proximity with HOTSET RNA locus and for subsequent activation of anterior HoxB gene expression. In addition, knockdown of HOTSET RNA disrupts the recruitment of the Setd1a/MLL1 complexes, H3K4me3 levels, the three-dimensional chromatin architecture, and HoxB gene transcription at the HoxB gene cluster. Depletion of HOTSET RNA also impairs differentiation of the CD41+/c-Kit+ HS/PC population and hematopoietic transcription factor and signaling networks required for early hematopoietic differentiation. Importantly, re-expression of HoxB2-4 genes in the HOTSET-depleted embryoid bodies rescues the expression of core hematopoietic transcription factors as well as the Flk1+ mesoderm-derived precursors and CD41+/c-Kit+ HS/PCs that undergo hematopoietic differentiation. Thus, HOTSET plays an important role in early hematopoietic lineage commitment by shaping the three-dimensional chromatin landscape in the HoxB locus to activate anterior HoxB genes that subsequently modulate lineage-specific transcription networks during hematopoietic development.

Disclosures
No relevant conflicts of interest to declare.
Ki-67 Is a Strong Predictor for Central Nervous System Relapse in Patients with Mantle Cell Lymphoma (MCL)
Authors: Chihara Dai , Asano Naoko , Ken Ohmachi , Nishikori Momoko , Okamoto Masataka , Sawa Masashi , Sakai Rika , Okoshi Yasushi , Tsukamoto Norifumi , Yakushijin Yoshihiro , Nakamura Shigeo , Kinoshita Tomohiro , Ogura Michinori , Suzuki Ritsuro 
Published: October 2019

Abstract
Abstract
Background: CNS relapse is uncommon but challenging complication in patients with mantle cell lymphoma (MCL). Survival after CNS relapse is extremely poor that identification of high risk population is therefore critical in whom prophylactic chemotherapy may play a role.
Patients and Methods: A total of 405 patients (median age, 66 years; range 22-85) with MCL newly diagnosed between 1994 and 2012 at 48 institutions in Japan were analyzed.Pathological specimens were centrally reviewed in all patients. We evaluated risk factors for CNS relapse in patients with MCL using competing risk regression analysis. Patients were excluded if they had evidences of CNS involvement at initial diagnosis. Overall, 81% of the patients received rituximab containing regimen with 222 patients received CHOP, 133 patients received high-dose cytarabine containing regimen and 50 patients received other regimen such as fludarabine. At a median follow up duration of 40.4 months, 25 patients (6.2%) experienced CNS relapse. Five patients presented with brain parenchymal mass, 13 patients presented with leptomeningeal disease, and 7 patients presented with both. The median time from initial diagnosis to CNS relapse was 18.5 months. Two-year cumulative incidence of CNS relapse was 4.7% (95%CI: 2.8-7.3). No difference was seen in the incidence of CNS relapse between the treatment with or without rituximab. Univariate analysis revealed blastoid or pleomoriphic variant, Ki-67 ≥ 30, serum total protein < 6.0g/dl, leukemic presentation and high risk group in simplified-MIPI at diagnosis as significant risk factors for CNS relapse. Within these five factors, Ki-67 was the strongest predictive factor for CNS relapse with hazard ration of 8.7 (95%CI: 3.4-22.2). Two-year cumulative incidence of CNS relapse in patients with Ki-67 ≥ 30 was 25.4% (95%CI: 13.5-39.1), while that in patients with Ki-67 < 30 was 1.5% (95%CI: 0.2-2.9) (Figure). Ki-67 ≥ 30 was a significant risk factor for CNS relapse both in patients who received CHOP and hyper-CVAD/MA. Survival after CNS relapse was dismal with median survival time of 5.7 months.
Summary: We identified four risk factors for CNS relapse in patients with MCL. Within these factors, Ki-67 ≥ 30 is the strongest predictive factor with 2-year cumulative incidence of 25.4%. Evaluation for CNS involvement at the diagnosis and assessment for value of prophylactic IT chemotherapy is mandatory in patients with Ki-67 ≥ 30.
Figure 1: Cumulative incidence of CNS relapses by Ki-67 Figure 1:. Cumulative incidence of CNS relapses by Ki-67

Disclosures
No relevant conflicts of interest to declare.
Identification of a New Subclass of ALK Negative Anaplastic Large Cell Lymphoma Expressing Aberrant Levels of ERBB4 Transcripts
Authors: Scarfò Irene , Pellegrino Elisa , Kwee Ivo , Agnelli Luca , Mereu Elisabetta , Bergaggio Elisa , Abate Francesco , Machiorlatti Rodolfo , Messana Katia , Rinaldi Andrea , Tiacci Enrico , Serra Sara , Deaglio Silvia , Neri Antonino , Falini Brunangelo , Rabadan Raul , Bertoni Francesco , Inghirami Giorgio , Piva Roberto 
Published: October 2019

Abstract
Anaplastic Large Cell Lymphoma (ALCL) is a clinical and biological heterogeneous disease including the ALK+ and ALK- systemic forms. While ALK+ ALCL are molecularly characterized and can be readily diagnosed, no specific markers and molecular events leading to ALK- ALCL transformation have been identified so far.
To discover biomarkers and/or genes potentially involved in ALK- ALCL pathogenesis, we applied the Cancer Outlier Profile Analysis (COPA) algorithm to a large gene expression profiling (GEP) data set including 249 cases of T-NHLs and normal T-cells. Among the top outliers, ERBB4 and COL29A1 genes were exclusively expressed in a subset of ALK- ALCL cases, and resulted highly correlated. Differential analysis comparing the expression profiles of ERBB4/COL29A1+ to ALK+ ALCL patients identified 48 genes up- and 37 down-regulated (qval < 0.0076). The most significantly over-represented functional category included transcriptional regulators. Gene-set-enrichment-analysis (GSEA) indicated that ERBB4/COL29A1+ samples shared activation of specific pathways such as inflammatory response, TNF-NF-κB, JAK-STAT, cytokine, and angiogenesis. RNA sequencing and 5’RNA Ligase Mediated Rapid Amplification of cDNA Ends (RLM-RACE) identified two ERBB4 truncated transcripts (referred as I20ΔERBB4 and I12ΔERBB4), displaying Transcription Starting Sites (TSS) in intron 12 and 20, respectively. On the contrary, full length COL29A1 mRNA was expressed in ERBB4/COL29A1+ samples. RT-qPCR analysis performed on 170 T-NHL samples (51 PCTL-NOS, 44 ALK+ ALCL, and 75 ALK- ALCL) highlighted specific expression of ERBB4 aberrant transcripts in 25% of ALK- ALCL (18 out of 75). ERBB4+ patients showed higher expression of I20ΔERBB4 as compared to I12ΔERBB4 transcript. ERBB4 expression at protein level was confirmed by immunohistochemistry and Western Blotting on selected cases. Inspection of intronic regions spanning two hypothetical TSS revealed the presence of Human Endogenous Retrovirus (HERV) Long Terminal Repeats (LTR) displaying promoter activity, as demonstrated by luciferase assays.
In conclusion, we defined a new subclass of ALK- ALCL characterized by ectopic expression of ERBB4 and COL29A1 genes. To the best of our knowledge, ERBB4 truncated transcripts carrying intronic 5’UTR have never been described before. Further studies are required to address whether the expression of ERBB4 aberrant transcripts may contribute to the ALCL transformation and lymphoma maintenance. This information might lead to more rational therapeutic approaches for ERBB4+ ALCL patients.

Disclosures
No relevant conflicts of interest to declare.
Activation-Induced Release of the TNF-Family Member BAFF By NK Cells Facilitates Resistance of Chronic Lymphoid Leukemia Cells to Direct and Rituximab-Induced NK Lysis
Authors: Wild Julia S , Raab Stefanie , Schmiedel Benjamin J , Maurer Andreas , Schneider Pascal , Kanz Lothar , Salih Helmut R 
Published: October 2019

Abstract
NK cells are cytotoxic lymphocytes that play an important role in the immunosurveillance of leukemia and, due to their ability to mediate antibody-dependent cellular cytotoxicity (ADCC), substantially contribute to the therapeutic benefit of antitumor antibodies like Rituximab. Available data indicate that the ability of NK cells to mediate ADCC is compromised in Chronic Lymphoid Leukemia (CLL), but the underlying mechanisms are still unclear. The TNF family member B cell activating factor (BAFF) was described to be aberrantly produced in mature B cell malignancies and contributes to disease pathophysiology e.g. by acting as a growth and survival signal for CLL cells. Here we report that NK cells express and release BAFF, and NK cell activation, notably including FcγRIIIa stimulation by Rituximab, results in increased secretion of BAFF (but not its close relative APRIL). Expression on the cell surface was neither detectable in resting nor in activated state. NK cell-derived BAFF enhanced the metabolic activity of primary CLL cells and protected CLL cells from chemotherapy-induced cell death. Moreover, exposure to BAFF profoundly diminished direct and Rituximab-induced lysis of primary CLL cells by allogeneic and autologous NK cells, while NK activation and degranulation per se remained unaffected. Notably, sensitivity of CLL cells to both chemotherapeutic treatment as well as direct lysis and Rituximab-induced ADCC of NK cells could be restored by the anti-BAFF antibody Belimumab (Benlysta®), which is approved for the treatment of systemic lupus erythematosus. Thus, our data provide evidence for the involvement of BAFF in the resistance of CLL cells to chemotherapy. Moreover, our results offer a functional explanation for the reportedly compromised ability of NK cells to combat lymphoid as compared to myeloid leukemias as well as their impaired ability to mediate ADCC upon Rituximab treatment in CLL patients. Our findings point to a possible benefit of combinatory approaches employing Rituximab and Belimumab for chemo-immunochemotherapy of B cell malignancies.

Disclosures
No relevant conflicts of interest to declare.
[68Ga]Pentixafor: A Novel PET Tracer for Imaging CXCR4 Status in Patients with Multiple Myeloma
Authors: Philipp-Abbrederis Kathrin , Herrmann Ken , Schottelius Margret , Eiber Matthias , Gerngroß Carlos , Pietschmann Elke , Lückerath Katharina , Franke Katharina , Rudelius Martina , Knop Stefan , Götze Katharina , Schwamborn Kristina , Steidle Sabine , Beer Ambros , Peschel Christian , Einsele Hermann , Buck Andreas , Schwaiger Markus , Wester Hans-Jürgen , Keller Ulrich 
Published: October 2019

Abstract
Chemokine receptors form a large family of G-protein coupled receptors that mediate chemotaxis of cells towards a gradient of chemokines. The C-X-C chemokine receptor-4 (CXCR4, also known as fusin, CD184) exerts its biological effects by binding its ligand CXCL12 (or Stromal-cell derived factor-1, SDF-1) which activates downstream pathways, ultimately resulting in altered cell adhesion and cell homing. Physiologically, the CXCR4/CXCL12 interaction plays a pivotal role in the recruitment and homing of hematopoietic stem/ progenitor cells (HSPC) and immune cells. In cancer, CXCR4 expression is associated with tumor dissemination and prognosis. Using multiple myeloma (MM) as an exemplary CXCR4-expressing cancer entity, we report the evaluation of [68Ga]Pentixafor-PET as a method for in vivo mapping of CXCR4 expression density by means of Positron Emission Tomography. [68Ga]Pentixafor-PET provides images with excellent specificity and contrast when assessed in mice xenografted with human CXCR4-positive MM. We next analyzed CXCR4 expression using [68Ga]Pentixafor-PET in a cohort of fourteen patients with advanced MM. These patients also underwent standard [18F]Fluoro-Deoxyglucose-PET (FDG-PET). Nine of fourteen FDG-PET scans were rated visually positive, whereas ten of fourteen Pentixafor-PET scans revealed MM manifestations. Assessment of blood counts and standard CD34+ flow cytometry did not reveal significant blood count changes or alterations in HSPC frequency associated with tracer application. These data document the first methodology for clinical PET imaging of CXCR4 in a cohort of MM patients. [68Ga]Pentixafor-PET opens a broad field of clinical investigations on CXCR4 expression and regulation in the cancer field and beyond.

Disclosures
Wester: SCINTOMICS, > Radiopharmaceutical Technology: Employment.
Effect of Young Age on Outcomes in Pediatric Acute Promyelocytic Leukemia (APL): North American Intergroup Study CALGB 9710 (Alliance)
Authors: Kutny Matthew A. , Geyer Susan , Laumann Kristina M , Gregory John , Willman Cheryl L. , Stock Wendy , Larson Richard A. , Powell Bayard L. , Feusner James H. 
Published: October 2019

Abstract
Purpose: Clinical trials in APL have generally reported similar outcomes for children and adults. However, sub-group analysis of pediatric patients treated on 2 consecutive trials of the European APL group demonstrated that children <5 years (yr) had an increased risk of relapse (Bally et al., JCO, 2012). Here we report the effect of age < 5 yr on outcome for the North American Intergroup Study C9710.
Methods: Outcomes were compared among young children (age <5 yr, n=16) to those of older age (5-12 yr, n=25 and 13-18 yr, n=45) enrolled on C9710. Treatment on C9710 included induction with daunorubicin, cytarabine and ATRA followed by two consolidation cycles with daunorubicin and ATRA and maintenance for one year with ATRA +/- mercaptopurine and methotrexate. Patients of age ≥15 yr were randomized to receive or not receive two 5-week cycles of arsenic trioxide (ATO) before the chemotherapy consolidation. However, analysis in this study of young children compared to those of older age was restricted to patients who did not receive ATO.
Results: Clinical remission (CR) rates were similar between the 3 age groups (age <5 yr= 88%, age 5-12 yr= 80%, age 13-18 yr= 82%; P=0.93). Overall survival (OS) Kaplan-Meier estimates were not significantly different (log rank P=0.84), and 5 yr OS rates were 77% (age <5 yr), 80% (age 5-12 yr), and 84% (age 13-18 yr). Similarly, event-free survival (EFS) Kaplan-Meier estimates were not significantly different (log rank P=0.48), and 5 yr EFS rates were 56% (age <5 yr), 47% (age 5-12 yr), and 59% (age 13-18 yr). Young children did not have a higher relapse rate (5-yr cumulative incidence of relapse by age: <5 yr=29%, 5-12 yr=49%, 13-18 yr=29%).
Conclusions: Results of this intergroup trial demonstrate that ATRA administration during induction, consolidation and maintenance leads to CR rates >80% and 5 yr OS rates >75%. These results are superior to recently published results of pediatric non-APL acute myeloid leukemia and confirm results from the prior APL intergroup trial (INT0129) that demonstrated that ATRA in induction and/or maintenance significantly improved outcomes for pediatric APL (Gregory et al., Ped Blood and Cancer, 2009). Our sub-group analysis showed no association of age with outcomes in children treated with this regimen.

Disclosures
Off Label Use: Daunorubicin- labeled for use in AML for adults and for pediatric ALL (not pediatric AML) Arsenic Trioxide- labeled for use in relapsed/refractory APL (not de novo APL) Mercaptopurine and Methotrexate- labeled for use in pediatric ALL (not APL).
Molecular Mechanisms Underlying the Synergism of Erib and Hht in Treating t(8;21) AML with C-KIT mutations
Authors: Chen Bing , Chen Xin-Jie , Chen Sai-Juan 
Published: October 2019

Abstract
AML1-ETO oncoprotein (AE) resulting from t(8;21)(q22;q22) translocation is known to play a pivotal role in leukemogenesis of t(8;21) AML, yet the activating mutations or over-expression of C-KIT occur in half of the AML patients with t(8;21). Animal models have proven that activating C-KIT mutations cooperate with AE in causing overt AML. The aberrant C-KIT expression was thus thought to be the major “second hit” in stepwise leukemogenesis of t(8;21) AML.
Previous studies have demonstrated that Eriocalyxin B (EriB), a diterpenoid extracted from Isodon eriocalyx, induced apoptosis in t(8;21) leukemia cells through the NF-κB and MAPK signaling pathways and triggered degradation of AE dependent on caspase-3. By targeting AML1-ETO oncoprotein and activating apoptosis pathways simultaneously, EriB has potential to treat the t(8;21) leukemia.
In this study, we observed the synergistic effect of EriB and the chemotherapy drug Homoharringtonine (HHT) on Kasumi-1, an AML cell line harboring t(8;21) and C-KIT mutations. Compared with using EriB or HHT alone, the combination of EriB and HHT (E+H) exhibited a higher efficacy to induce apoptosis and inhibit proliferation of Kasumi-1 cells. Data further showed that HHT enhanced the AE degradation caused by EriB in a dose/time-dependent manner. In addition, HHT significantly decreased the expression of C-KIT in Kasumi-1 cells, and the combination with EriB markedly enhanced this effect. Mechanism study revealed that E+H combination down-regulated the expression of transcriptional factor Sp1 through up-regulating miR-29b, separated Sp1 protein from Sp1/NF-κB complex, and released the Sp1/NF-κB complex from the C-KIT promoter. These data point to a mechanism involving Sp1/NF-κB/miR-29b regulatory network whereby E+H combination down-regulate C-KIT mRNA expression. In addition, HHT induced a proteasome-mediated degradation of C-KIT protein, which was further enhanced by EriB.
From these observations, we conclude that the combined use of EriB and HHT might offer a potential therapeutic avenue for t(8;21) AML with C-KIT mutations.

Disclosures
No relevant conflicts of interest to declare.