bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of p53-Dependent Pathways In RPL11 Mutant Zebrafish
Authors: Danilova Nadia , Sakamoto Kathleen M. , Lin Shuo 
Published: October 2019

Abstract
Abstract 2227
Deficiency of ribosomal protein (RPs) is associated with Diamond Blackfan Anemia, a congenital syndrome with bone marrow failure and variable malformations. Recent studies have demonstrated that deficiency of several RPs leads to activation of p53 and related family members, ΔNp53 and ΔNp63. In addition, P53 inhibition rescues hematopoiesis in RP-deficient mice and zebrafish. p53 activation, however, results in the alteration of many pathways and specific mechanisms leading to bone marrow failure and other defects in DBA remain unidentified. To understand the global changes in gene expression downstream of RP insufficiency, we took a system biology approach to characterize the molecular pathways involving p53. We studied pathways downstream of p53 that contribute to the RPL mutant phenotype in zebrafish. Changes in p53-dependent pathways in RPL11 mutants included dysregulation of cell cycle and apoptosis, shift in energy production from glycolysis to aerobic respiration, suppression of biosynthesis of structurally important proteins, downregulation of detoxifying enzymes, and upregulation of factors leading to activation of adrenal-pituitary axis and inflammation. Among multiple metabolic changes were those that led to increased levels of insulin and glucose. Both differentiation and production of blood were affected. Expression of genes that regulate the development and function of neural system were notably decreased, in particular, those involved in the development of anterior brain structures and eyes. The production and migration of neural crest cells were disrupted suggesting possible explanation for craniofacial and other defects observed in patients with DBA. The RPL11 mutants also had abnormal development of hematopoietic stem cells and shortened life span of mature red blood cells. Dexamethasone treatment led to downregulation of p53 expression; suppression of p53 targets mediating cell cycle arrest and apoptosis; and rescue of expression of HSC markers such as runx1. The life span of erythroid cells was also increased in response to dexamethasone treatment. These data contribute to our understanding of the molecular pathophysiology of DBA and suggest new potential targets for therapeutic intervention.

Disclosures:
Sakamoto: Abbott Laboratories, Inc.: Research Funding; Genentech, Inc.: Research Funding.
Fecal Calprotectin In Allogeneic Stem Cell Trasplantation as Surrogate Marker of Gastrointestinal Graft Versus Host Disease
Authors: Oreiro Mariana Bastos , de la Guia Ana Lopez , Nieto Jose B , De Paz Raquel , Baltasar Patricia , Hernandez Dolores , de la Rua Ana Rodriguez , Gallardo David , Canales Miguel 
Published: October 2019

Abstract
Abstract 2327

Background:
Calprotectin is a major cytosolic protein of neutrophils that have showed to be a sensitive marker of intestinal inflammation. The aim of our study has been to evaluate fecal calprotectin (FC) as a diagnosis tool in patients with acute gastrointestinal graft versus host disease (GI GVHD).

Methods:
Since March 2009, patients with suspicion of acute GI GVHD were consecutively included. Patients were tested for FC (reference range: 0–30 mg/kg) before starting treatment. Infections by Clostridium difficile, cytomegalovirus, fecal bacteria and intestinal parasites were also excluded. Colonoscopy was performed in all patients and biopsy samples were taken for histopathological examination.

Results:
To date, eleven patients have been included. The median age was 48.2 (r: 21–67). Indications for transplantation included acute myeloid leukemia (6 patients), myelodysplastic syndrome (2 patients), acute lymphoid leukemia, severe aplastic anemia and refractory follicular lymphoma (1 patient, respectively). Ten patients received myeloablative conditioning and one received reduced intensity conditioning regimen. Five patients were histologically diagnosed with acute GI GVHD. The median for FC in this group was 510.5 (r: 107.4–629). The median of FC in patients without GI GVHD was 117 (r: 13.9–205). Patients with GI GVHD had higher values of FC than patients without GI GVHD (p:≤0,009) For a cut-off point value of 205, sensitivity for the test was 83.3% (IC 61.3–100%), specificity was 100%, positive predictive value was 100%, and negative predictive value was 83.3% (IC 61.3–100%). In 3 patients who were diagnosed with cytomegalovirus enteritis and had no criteria for GI GVHD, the FC values were 15.4, 105, and 100.7. In a patient with Candida spp. infection FC was 13.9. The FC was higher in patients with grade IV GI GVHD, with median of 590 (r: 502–629).

Conclusion:
FC appears to be a promising non-invasive biomarker of acute GI GVHD. If these findings are confirmed, it may provide a useful non-invasive test for the diagnosis of GI GVHD in patients following allogeneic transplantation.

Disclosures:
No relevant conflicts of interest to declare.
Successful Treatment of High-Risk and Refractory Acute Myeloid Leukemia with haploidentical Stem Cell Transplantation Plus NK cell therapy
Authors: Uharek Lutz , Friedrichs Birte , Nogai Axel , Gentilini Chiara , Basara Nadezda , Niederwieser Dietger , Thiel Eckhard 
Published: October 2019

Abstract
Abstract 2370
Regardless of the treatment applied, the outcome for patients with refractory hematological malignancies is extremely poor. For haploidentical stem cell transplantation (haploSCT), which has been evaluated especially in high-risk AML patients lacking an HLA-identical donor, registry data show an overall survival below 10% for AML not transplanted in remission. In order to improve immune competence and to reduce the risk of relapse, we have combined the early transfer of NK cells with the haploSCT approach. PATIENTS AND METHODS: Twenty-five patients (AML = 16, ALL = 5, CML = 2, Hodgkin = 1, MDS = 1) received a haploSCT followed by transfer of purified CD56+CD3-NK cells at day +2. Conditioning consisted of total body irradiation, thiotepa, fludarabine, and OKT3. NK cells were isolated from the CD34- fraction using an automated two-step procedure of CD3+ depletion and subsequent CD56+ selection. No other immunosuppression was routinely delivered. RESULTS: Patients received a mean of 13.3 × 106 CD34+ cells/kg (range 6.12–26.97) with 2.89 × 104/kg (0.95-7.4) contaminating CD3+ cells. A mean of 9.8 × 106 CD56+CD3- NK cells/kg (range 1.61–32.2) was adoptively transferred. Most of the patients developed early GVHD of the skin (median onset day=12), which promptly resolved after short term treatment with steroids and CSA. The main cause of death consisted of infections (n = 10), chronic GvHD (n= 2), and relapse (n = 4). After a median follow-up of 1442 days (3.9 years) (range 0.1 to 7.1 years) 9 of 25 patients are alive and in complete remission resulting in a 2-year overall survival (OS) estimate of 29%. Out of 16 high-risk patients with AML, 10 had refractory or active disease prior to transplantation. For a matched pair analysis, these patients were matched for age, disease status, conditioning regimen and year of transplantation using the EBMT database. AML patients treated with haploSCT plus NK cell transfer had a superior 2-year overall survival (40 vs 11%, 95% CI) in comparison to matched controls with haploSCT only (p=0.02) CONCLUSIONS: HaploSCT plus NK cells is feasible and shows promising survival rates for a group of patients lacking other treatment options. Best results were achieved in patients with AML not only in remission but also with refractory disease.

Disclosures:
No relevant conflicts of interest to declare.
Association of HLA-G Haplotype Risk Model with Treatment Free Survival In Chronic Lymphocytic Leukemia Patients
Authors: Kapp Johanna , Ochsenfarth Crista , Switala Magdalena , Frey Ulrich , Sellmann Ludger , Duerig Jan , Horn Peter A , Duehrsen Ulrich , Rebmann Vera , Nuckel Holger 
Published: October 2019

Abstract
Abstract 2418

Introduction:
The human leukocyte antigen G (HLA-G) molecule exhibits limited tissue distribution and exerts multiple immunoregulatory functions. Since the net result of these effects is immunosuppression, HLA-G expression in tumor cells may favor their escape from antitumor immune responses, thus allowing tumor progression. The HLA-G protein polymorphism is extremely restricted in comparison to the classical HLA antigens. However, specific HLA-G polymorphisms in exons 3, 4, 5, 7 and parts of the 3`UTR encode for different haplotypes. Demonstrating the extensive and proven prognostic role of HLA-G in various diseases and especially in B-CLL we investigated the role of the HLA-G haplotypes in B-CLL.

Methods:
Genotyping was performed on the basis of examining exons 3, 4, 5, 7 and parts of the 3`UTR by pyrosequencing in 190 patients with B-CLL. In total, we found 17 different haplotypes in the cohort.

Results:
The haplotype distributions between 190 B-CLL patients and 190 healthy controls were not different, arguing that the haplotypes of HLA-G do not increase the susceptibility for B-CLL. However, combining four different haplotypes (*01:06, *01:12; *01:12 variant and *01:13N) we evaluated a risk score (0=no risk allele; 1=one risk allele; 2=two risk alleles) for TFS. 77 patients showed no risk allele, 90 patients one risk allele and 23 patients two risk alleles. The TFS for those patients with two risk alleles were significantly shorter (median TFS 33 months) than for those with one risk allele (median TFS 49 months) and those with no risk alleles (median TFS 88 months)(log rank test: p=0.03). In multivariate analysis we could show that the stage according to Binet (HR 1.7, 95% CI 1.2–2.6, p=0.005), CD38 status (HR 1.8, 95% CI 1.0–3.2, p=0.04) and the haplotype risk model (HR 1.7, 95% CI 1.1–2.6, p=0.011) were independent predictors for first therapy.

Conclusion:
Our study is the first study demonstrating that the combination of different alleles of the HLA-G gene is associated with the risk of first therapy in B-CLL patients. This fact emphasizes the extensive role of this gene by the tumor escape mechanism and could be responsible for progress in other cancers.

Disclosures:
No relevant conflicts of interest to declare.
Transcriptional and Translational Regulation of the Cancer-Testis Antigen PRAME Mediated by Pathogen-Associated Molecular Patterns
Authors: Wadelin Frances R , Spriggs Keith A , Falcone Franco H , Heery David M 
Published: October 2019

Abstract
Abstract 2475
PRAME (preferentially expressed antigen in melanoma) is a germinal tissue-specific gene that is expressed at high levels in a number of haematological malignancies. In AML, PRAME is a positive prognostic indicator, being associated with a favourable response to chemotherapy and increased overall survival, even in the presence of unfavourable karyotypes. In contrast, over-expression of PRAME mRNA is associated with poor prognosis in CML, NHL, HD, CLL and myeloma, being associated with progressive disease and chemo-resistance. Despite proposals to implement PRAME as a tool for monitoring minimal residual disease and as an immunotherapeutic target, the physiological functions of PRAME in normal and tumour cells are unclear. A role in the regulation of retinoic acid signalling has been proposed, although interactions of PRAME with retinoic acid receptors are weak compared to other coregulator proteins. Sequence homology and structural predictions suggest that PRAME is a member of the family of leucine-rich repeat (LRR) proteins that function in cellular immunity, bearing most resemblance to the LRR domains of Toll-like receptors (in particular TLR3 and TLR4) and internalin proteins. LRR proteins can bind pathogen-associated molecular patterns (PAMPs) including proteins, nucleic acids and sugars, and consistent with this PRAME has been reported to interact with OPA proteins from Neisseriae gonorrhoeae. Here we show that expression of PRAME in HL60 leukaemic cells is strongly induced by lipopolysaccharide (LPS) or peptidoglycan in the presence of interferon (IFN)-gamma, whereas no induction was observed after exposure to other PAMPs such as zymosan A. This burst of PRAME expression peaked between 1 and 4 hrs post treatment, but was not mediated by the proximal promoter region. Moreover, we show that treatment of HL60 with LPS and IFN-gamma increases the association of PRAME transcripts with polysomes indicating PRAME is also translationally regulated, and we provide evidence that PRAME 5'UTR contains an internal ribosome entry sequence for CAP-independent translation. Finally, we show that the subcellular localisation of PRAME protein in HL60 and U937 cells is modulated by treatment with LPS and IFN-gamma. In summary, our studies indicate that the LRR protein PRAME is subject to transcriptional and translational regulation following cellular exposure to bacterial PAMPs and interferon, suggesting it may have a role in innate immunity.

Disclosures:
No relevant conflicts of interest to declare.
Gene Expression Profiling Using the Illumina ‘DASL’ Platform on RNA Extracted From Formalin Fixed Paraffin Embedded (FFPE) Tissue Identifies Distinct Prognostic Groups In CHOP-R Treated DLBCL
Authors: Barrans Sharon , Worrillow Lisa , Care Matthew , Crouch Simon , Smith Alex , Patmore Russell , Davies Andrew , Tooze Reuben , Roman Eve , Jack Andrew 
Published: October 2019

Abstract
Abstract 2485
Diffuse large B cell lymphoma (DLBCL) is a heterogenous disease, which has been subclassified into germinal centre (GCB) and activated B-cell (ABC) type using gene expression profiling. This has been shown to separate DLBCL into distinct prognostic sub-groups in patients treated with either CHOP or CHOP-R therapy. Previous studies have required the use of fresh or frozen samples for the extraction of RNA of sufficient quality to permit whole genome expression analysis. The Illumina ‘DASL' platform allows for highly reproducible gene expression data to be generated from FFPE material, which opens up large series' of retrospective data for detailed expression studies.
The aim of this study was therefore to determine whether the Illumina DASL platform could yield reproducible results on formalin fixed paraffin embedded (FFPE) biopsies from a large series of archival CHOP-R treated DLBCL samples.
RNA was extracted from paraffin sections using the Ambion Recoverall extraction kit, with 179/206 (87%) of cases yielding >200ng of RNA sufficient for DASL analysis. The DASL assay was performed according to Illumina protocols. Using stringent exclusion criteria, 157/179 (88%) cases yielding results that were considered to be of sufficiently high quality to be included in the analysis. To fully assess the reproducibility of the assay, 35 cases were analysed on 2–8 occasions across multiple experimental days.
Using Pearson's correlation, with full-linkage clustering, four discrete clusters were identified (n=28, 40, 46 and 43). Of important note, 95% of the samples were seen to cluster more tightly with their repeats than with any other sample, with all duplicated samples being called in the same cluster with 100% accuracy, suggesting that the technique is highly reproducible.
Univariate Kaplan-Meier survival analysis showed that the clusters identified patients with very different outcomes. Two of the clusters showed identical survival curves and therefore these clusters were merged to give 3 clusters with 2-year overall survivals (OS) of 51% (n=71), 65% (n=46) and 77% (n=40), log rank p=0.03, with a 3.7 year follow-up.
This data supports the use of gene expression profiling to classify DLBCL patients into clinically relevant prognostic groups. The Illumina DASL assay allows for highly reproducible gene expression data to be produced in valuable, archival data series, and also in the context of clinical trials, where the majority of the tissue available for study is FFPE. The patients identified in this study as having a sub-optimal response to CHOP-R should be considered for alternative therapies, which should be validated in the context of a clinical trial.

Disclosures:
No relevant conflicts of interest to declare.
Expression of the Neonatal Fc Receptor FcRn Is Not Required for the Amelioration of Murine ITP by IVIg or a Monoclonal Antibody
Authors: Crow Andrew R. , Suppa Sara J , Lazarus Alan H 
Published: October 2019

Abstract
Abstract 2529
There are several theories as to the mechanism of intravenous immunoglobulin (IVIg) in the treatment of autoimmune diseases such as immune thrombocytopenia (ITP). One prominent theory involves accelerated pathogenic autoantibody clearance by saturation of the neonatal Fc Receptor (FcRn). FcRn is an IgG receptor, and FcRn within endosomes binds endocytosed IgG and diverts IgG from degradation. In the treatment of ITP, it has been theorized that high concentrations of IVIg saturate FcRn, reducing the ability of the pathogenic anti-platelet antibodies to bind FcRn, increasing their catabolism, and thus rapidly decreasing their serum concentration which results in a decrease in their ability to induce platelet clearance. Mice lacking FcRn have only 20–30% of the level of endogenous IgG as compared with wild-type mice and have an accelerated clearance rate of both endogenous and injected IgG. We have utilized these mice in a murine model of ITP to help understand the role, if any, that FcRn plays in IVIg's therapeutic activity.
Both IVIg (2 g/kg body weight) and monoclonal antibodies to CD44 (2 mg/kg body weight; please see submitted abstract ID# 29504, “Amelioration of murine immune thrombocytopenia by CD44 antibodies: a potential therapy for ITP?”) successfully ameliorate murine ITP. To definitively determine if FcRn is required for the acute amelioration of ITP by these two therapeutics, we employed FcRn deficient mice in the murine ITP model. Here, we demonstrate that FcRn deficient mice (B6.129X1-Fcgrttm1Dcr/DcrJ) injected with an anti-platelet antibody exhibit a slightly more profound degree of thrombocytopenia than wild-type mice. FcRn deficient mice treated with IVIg or the anti-CD44 antibody KM114 (at a 3 log fold lower dosage than IVIg) were protected from ITP to the same extent as wild-type mice.
FcRn has an absolute requirement for the protein β2 microglobulin (β2M) to be functionally expressed. Specifically, β2M deficient mice do not possess functional FcRn and also show low endogenous IgG levels and increased clearance of IgG. To verify and substantiate the results found with FcRn deficient mice, we next employed β2M deficient mice in the murine ITP model and found that β2M deficient mice (B6.129P2-B2mtm1Unc/J) treated with IVIg or KM114 were also protected from ITP to the same extent as wild-type mice.
These data suggest that for both high dose IVIg as well as low dose monoclonal CD44 antibody treatment in an acute ITP model, FcRn expression is dispensable.

Disclosures:
No relevant conflicts of interest to declare.
Resveratrol, a Natural Antioxidant From Grapes and Red Wines, Prevents EBV-Associated Lymphoproliferation and Transformation through Inducing Apoptosis
Authors: Espinoza Luis J. , Takami Akiyoshi , Nakata Katsuya , Kato Shunichi , Kasahara Yoshihito , Nakao Shinji 
Published: October 2019

Abstract
Abstract 2543
Epstein-Barr virus (EBV) infection and reactivation is an increasing complication in immune deficient patients, particularly after allogeneic hematopoietic stem cell transplantation (HSCT) or solid organ transplantation (SOT). Whereas tapering of immunosuppression, donor lymphocyte infusion, and rituximab may be effective for this complication after HSCT, severe adverse events such as fatal GVHD and infections could develop thereafter. Resveratrol, a natural antioxidant compound found in grapes and red wines, possesses various biological activities including antitumor, anti-inflammatory, and antimicrobial efficacies “(Nature Reviews Drug Discovery 5, 493–506, 2006). We report here anti-EBV effects of resveratrol in eliminating EBV infected cells and preventing EBV-associated B-cell transformation. Resveratrol, at nontoxic concentrations to normal cells, effectively prevented the transformation of primary B cells derived from a panel of 16 healthy donors as well as cord blood mononuclear cells derived from 10 different donors. In addition, resveratrol induced dose-dependent apoptosis, assessed by FACS-based detection of Annexin V, in 7 different established EBV-transformed lymphoblastoid cell lines (LCLs) and in the Burkitt lymphoma EBV+ Raji and Daudi cell lines. These effects were consistently associated with the activation of caspase-8 and a 60 % down-regulation of the anti-apoptotic protein BCL-2. The addition of the caspase-8 inhibitor c-FLIPL to the cell culture vigorously inhibited the resveratrol induced apoptosis. Moreover, the resveratrol treatment decreased by 80% the EBV nuclear antigen 1 (EBNA1) expression, a critical protein required for the persistency of EBV infection. These results suggest that resveratrol may have potential roles in preventing EBV-associated lymphoproliferative diseases in immune compromised patients. Since resveratrol is a nontoxic, inexpensive and clinically available agent, it may be useful in preventing EBV- infection and reactivation after HSCT and SOT.

Disclosures:
No relevant conflicts of interest to declare.
New Instrument for Comprehensive Symptom Profile Assessment In Patients with Malignant Lymphomas
Authors: Novik Andrei A , Ionova Tatyana I , Kalyadina Svetlana A , Fedorenko Denis A , Mochkin Nikita E , Kurbatova Kira A , Nikitina Tatyana P , Gorodokin Gary I 
Published: October 2019

Abstract
Abstract 2561
Comprehensive symptom assessment before and during treatment in lymphoma patients is worthwhile. We aimed to develop a new symptom assessment tool — Comprehensive Symptom Profile in Lymphoma Patients (CSP-Lym) and test its applicability to this patient population.
The CSP-Lym is a self-reported tool which allows the assessment of the severity of 45 symptoms in lymphoma patients. It consists of numerical rating scales, scored from “0” (no symptom) to “10” (most expressed symptom). Thirteen symptom clusters have been identified, which were clinically relevant and increased the practicability of the tool. Utility of the CSP-Lym was demonstrated by means of interviewing both patients and physicians. Applicability of the tool with preliminary analysis of psychometric properties was tested in a pilot study. 75 patients with different types of malignant lymphomas (Stage – I-IV) were included in the study: Non-Hodgkin's lymphoma – 26; Hodgkin's lymphoma – 49. Mean age was 33.5 years old; male/female distribution –28/47. Patients filled out the CSP-Lym before and at different time-points of treatment.
The utility of the CSP-Lym was demonstrated. The patients acknowledged the comprehensiveness of the tool. All of the items were easy for the patients to read and understand. Completion of the CSP-Lym in paper and pencil format took 5–7 min. The data produced by the tool were clear for interpretation by physicians and were used by them in day-to-day decision making.
The construct validity of the CSP-Lym was proven by factor analysis and “known-group” comparison. A six factor structure was found for the core symptom items, which explained 65% of the total variance. The analysis of symptom severity in the groups with and without B-symptoms revealed differences between the groups: all of the symptoms were more severe in patients with B-symptoms. Statistically significant differences between the groups (p<0.05) were found for the majority of symptoms. Reliability of the CSP-Lym was satisfactory: Chronbach's alpha coefficient varied from 0.63 to 0.92. Sensitivity to changes was demonstrated by the comparison of symptom severity before and after treatment. The majority of symptoms after treatment were less severe than at base-line.
Thus, the CSP-Lym is an appropriate and practical tool to assess the symptom severity in lymphoma patients. The utility of the questionnaire was shown; preliminary psychometric properties appeared to be satisfactory. Further studies are needed before the wide-spread use of the CSP-Lym in clinical practice and clinical trials.

Disclosures:
No relevant conflicts of interest to declare.