bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene Expression Signature of Host Immune Response Is Predictive of Follicular Lymphoma Patient Survival in Independent Cohorts, and Correlates with Transformation to Diffuse Large B-Cell Lymphoma.
Authors: Alizadeh Ash A , Gentles Andrew J , Plevritis Sylvia K , Levy Ronald 
Published: October 2019

Abstract
Abstract 2951
Poster Board II-927

Background:
Expression signatures of infiltrating immune cells [1] have been shown to predict survival in follicular lymphoma (FL), but have not been cross-validated in independent patient cohorts [2,3]. These signatures may relate biologically to the frequency of infiltrating including T-cells and macrophages, or to specific transcription programs within tumor cells and/or the tumor microenvironment. We sought to evaluate the validity of this model in an independent cohort of patients with FL, assessing its relationship to outcomes including histological transformation and death.

Methods:
The immune response (IR) predictor score proposed by Dave et al. [1] was applied to gene expression data from an independent cohort of 88 FL patients [4] with known survival outcomes and history of transformation to diffuse large B-cell lymphoma (DLBCL). Genes (n=66) corresponding to IR1 and IR2 signatures were mapped from Affymetrix microarrays [1] to a custom cDNA array [4] via Entrez Gene ID, and the composite IR score was calculated per the scheme proposed by Dave et al.

Results:
The IR score was predictive of patient outcome in the 88 patient test set as a continuous variable (p=0.001, HR=2.01, 95% CI 0.50-1.30). Partitioning of patients into high and low risk groups based on the median IR score across the cohort robustly separated survival curves (Figure A). The IR score was significantly higher in FL patients known to undergo transformation to DLBCL (Figure B: mean IR score of -0.6 in non-transforming FL vs. -0.2 in transforming FL; p∼10-11, t-test).

Conclusions:
The IR score of Dave et al. was highly significant as a predictor of survival in the independent patient cohort [4]. Moreover, the score was significantly associated with propensity of FL to transform to DLBCL. To our knowledge, immune cell infiltration has not previously been implicated in transformation.
1. Dave SS et al. (2004) Prediction of survival in follicular lymphoma based on molecular features of tumor-infiltrating immune cells. N Engl J Med 351(21): 2159-2169.
2. Tibshirani R (2005) Immune signatures in follicular lymphoma. N Engl J Med 352: 1496-1497.
3. Chu G Hong WJ, Warnke R, Chu G (2005). Immune Signatures in Follicular Lymphoma (Corres). N Engl J Med. 352: 1496-1497.
4. Glas AM et al. (2005) Gene expression profiling in follicular lymphoma to assess clinical aggressiveness and to guide the choice of treatment. Blood 105(1): 301-307.

Disclosures:
No relevant conflicts of interest to declare.
Bcr/Abl Increases Bcatenin Protein and Activity in An ICSBP-Dependent Manner in Myeloid Cells.
Authors: Huang Weiqi , Horvath Elizabeth , Eklund Elizabeth A. 
Published: October 2019

Abstract
Abstract 2958
Poster Board II-934
The interferon consensus sequence binding protein (ICSBP) is an interferon regulatory transcription factor (also referred to as IRF8). Similar to other IRF proteins, ICSBP regulates transcription of genes involved in the inflammatory response. However, ICSBP also functions as a myeloid leukemia tumor suppressor. Specifically, decreased ICSBP expression in myeloid progenitor cells results in cytokine hypersensitivity and resistance to apoptosis in response to Fas-activation or IL3 withdrawal. Consistent with function as a tumor suppressor, decreased ICSBP-expression is found in the bone marrow of human subjects with chronic myeloid leukemia (CML). Expression of ICSBP increases during remission and decreasing ICSBP expression is associated with progression to CML blast crisis (BC). In murine models, ICSBP-deficiency induces a myeloproliferative disorder (MPD) which resembles CML. And, ICSBP overexpression blocks MPD in mice transplanted with Bcr/abl expressing bone marrow. Increased Bcatenin activity is also associated with CML-BC. However, the impact of ICSBP on Bcatenin expression or activity has not been previously investigated. In these studies, we hypothesized that ICSBP-deficiency in CML-BC increases Bcatenin-expression. In support of this hypothesis, we found that Bcatenin protein and activity were increased in myeloid cell lines with ICSBP-knock-down and decreased in cells with ICSBP-overexpression. Bcatenin protein and activity were also increased in primary myeloid progenitors from ICSBP-/- mice in comparison to wild type or ICSBP+/- mice, in an ICSBP-dose dependent manner. Expression of Bcr/abl decreased ICSBP-expression in myeloid cells and increased Bcatenin protein and activity. Also supporting our hypothesis, the effect of Bcr/abl on Bcatenin was abolished by re-expression of ICSBP in Bcr/abl expressing myeloid cell lines or primary murine myeloid progenitor cells. In each of these situations, the increase in Bcatenin protein was not due to increased expression of Bcatenin mRNA. These results suggest ICSBP influences expression of target genes involved in Bcatenin protein stability. Therefore, these studies identify a pathway by which Bcr/abl activity impairs ICSBP expression in immature myeloid cells, thereby increasing stability of Bcatenin protein and Bcatenin activity. Poor prognosis and BC are associated with Fas resistance and increased Bcatenin activity in CML leukemia stem cells (LSC). Bcr/abl activity is also associated with decreased expression of ICSBP. Our prior studies implicated ICSBP-deficiency in Fas-resistance in CML via the ICSBP target gene PTPN13. Our current studies implicate ICSBP-deficiency in increased Bcatenin expression in CML. These results suggest that decreased ICSBP expression drives multiple aspects of the poor prognosis LSC phenotype in CML.

Disclosures:
No relevant conflicts of interest to declare.
Role of a Conserved Helicase Motif in the Transformation of Primary Human CD34+ Cells by the NUP98-DDX10 Oncogene.
Authors: Yassin Enas R , Abdul-Nabi Anmaar M , Takeda Akiko , Yaseen Nabeel R. 
Published: October 2019

Abstract
Abstract 2966
Poster Board II-942
NUP98 gene rearrangements occur in acute myeloid leukemia and other hematopoietic malignancies, and result in the expression of fusion proteins. One of the most frequent NUP98 fusions is NUP98-DDX10 that consists of an N-terminal portion of NUP98 and a C-terminal segment of DDX10, a putative DEAD-box RNA helicase. Here we express NUP98-DDX10 in primary human CD34+ hematopoietic cells and show that it localizes within the nucleus in a punctate distribution. It dramatically increases the proliferation and self-renewal of primary human CD34+ cells and disrupts their erythroid and myeloid differentiation. Expression gene profiling shows dysregulation of many genes by NUP98-DDX10 in primary human CD34+ cells starting within 6 h. Comparison of the dysregulome of NUP98-DDX10 to that of another leukemogenic NUP98 fusion, NUP98-HOXA9, reveals a number of genes dysregulated by both oncoproteins, including HOX genes, COX-2, MYCN, angiopoietin-1, renin, HEY1, SOX4, and others, that may account for the induction of AML by these and other NUP98 fusion oncogenes. The HRAGRTAR sequence in the DDX10 portion of NUP98-DDX10 corresponds to a major motif shared by DEAD-box RNA helicases that is required for their ATP binding/hydrolysis, RNA-binding, and helicase functions. Mutating this motif diminished the transforming ability of NUP98-DDX10, indicating that it plays a role in leukemogenesis. These data demonstrate for the first time the transforming ability of NUP98-DDX10 and show that it is partially dependent on one of the consensus helicase motifs of DDX10. They also point to common pathways that may underlie leukemogenesis by different NUP98 fusions.

Disclosures:
No relevant conflicts of interest to declare.
Organization and Physiologically Mediated Differential Release of Angiogenic Regulatory Proteins From Distinct Platelet Alpha Granules.
Authors: Battinelli Elisabeth , Italiano Joseph 
Published: October 2019

Abstract 3045
Poster Board II-1021
In addition to their primary roles in hemostasis and thrombosis, platelets have been reported to participate in other physiological and pathological processes, including but not limited to inflammation, wound healing, and tumor metastasis. Although platelets are presumed to contribute to new blood vessel growth by providing numerous pro-and anti-angiogenic factors, the cellular and molecular basis by which platelets regulate angiogenesis is poorly understood. Previously we have shown that platelets differentially package angiogenic regulatory proteins in their alpha granules. More recently, we have further defined the organization of angiogenic proteins within platelet alpha granules using immunofluorescence and immunogold labeling experiments and have uncovered at least three distinct types of alpha granules. These granules were found to contain selective members of pro- and anti-angiogenic regulatory proteins, respectively. In addition, we have analyzed platelets from nine patients with metastatic renal cell carcinoma using immunofluorescence labeling for VEGF and endostatin. In comparison to volunteer control platelets without known malignancy, there is a significant increase in granules containing VEGF in patients with metastatic disease, strongly suggesting a shift of the angiogenic balance towards a more pro-angiogenic environment in the platelets from patients with malignancy. Previously we have shown that differentially packaged alpha granules are susceptible to selective release in response to proteinase-activated receptors. To explore whether selective release also occurred in physiologic conditions, we activated human platelets with known platelet agonists and looked for evidence of selective release. We have demonstrated that differential release of VEGF can be physiologically regulated by activation with ADP. Human platelets exposed to 50uM ADP result in release of VEGF and retention of endostatin, both by immunofluorecence labeling and by ELISA assay. This study provides mechanistic evidence to support an interplay between tumor cells and platelets, by illustrating a key role for platelets in tumor angiogenesis through the delivery of specific angiogenic factors. This work has implications beyond basic platelet biology as it provides a model for the platelet as a delivery system, with important clinical implications.

Disclosures
No relevant conflicts of interest to declare.
Novel SIRT1 Agonists, SRT501 and SRT2183, Induce Expression of DNA Repair Genes and Apoptosis of Ph− Acute Lymphoblastic Leukemia Cells Alone and in Combination with the HDAC Inhibitor LBH589..
Authors: Scuto Anna , Kirschbaum Mark , Cermak Jennifer M , Atadja Peter , Jove Richard 
Published: October 2019

Abstract
Abstract 3083
Poster Board III-20
Histone Deacetylase Inhibitors (HDACi) such as LBH589, which inhibit the zinc containing catalytic domain of HDAC of classes I, II, and IV, demonstrate activity against various malignancies, particularly lymphoid malignancies. SIRT1 is an NAD+ dependent class III histone deacetylase, which deacetylates histones as well as non-histone proteins and is not affected directly by HDACi such as LBH589. It remains controversial whether inhibition of SIRT1 or its activation is more efficacious in anticancer therapy. We have studied the activity of two novel SIRT1 activators, SRT501 and SRT2183, in Philadelphia chromosome negative acute lymphoblastic leukemia (ALL) cell lines. Both pre B (NALM-6, Reh) and T cell (MOLT-4) ALL lines were treated with either SRT501 or SRT2183, as well as in combination with LBH589 and evaluated for biological and gene expression responses. SRT501 induced growth arrest and apoptosis at doses ranging from 10-100 uM, with even the lowest doses inhibiting growth at 72 hours. SRT2183 is much more potent, with growth arrest and apoptosis induced at doses ranging from 1-20 uM. PCR array analysis revealed that SRT2183 treatment leads to increased mRNA levels of pro-apoptosis, growth arrest, and DNA damage response genes. We have previously demonstrated that the activity of LBH589 is mediated in part through upregulation or acetylation of proteins involved in the DNA damage response pathways. Quantitative real-time PCR confirms that the combination of LBH589 with SRT2183 leads to significantly higher expression of GADD45A and GADD45G than either agent alone. The combination of LBH589 plus SRT2183 showed enhanced inhibition of c-Myc protein levels, phosphorylation of H2A.X, and interestingly, increased acetylation of p53 (acetylation of p53 was not seen with SRT2183 alone). In summary, the novel SIRT1 activators SRT501 and SRT2183 show growth inhibitory and pro-apoptotic activity in Ph- ALL alone and enhanced activity in combination with LBH589. Clinical studies of these agents, particularly in combination with HDACi are warranted.

Disclosures
Kirschbaum: Novartis: Consultancy. Cermak:Sirtris: Employment. Atadja:Novartis: Employment.
Clinical Significance of Circulating Ki-67 Protein and Caspase-3 Activity Levels in Patients with Acute Myeloid Leukemia.
Authors: Bruey Jean-Marie , Kantarjian Hagop M. , Estrov Zeev , Ma Wanlong , Yeh Chen-Hsiung , Sferruzza Anthony , Estey Elihu , Albitar Maher 
Published: October 2019

Abstract
Abstract 3104
Poster Board III-41
Ki-67 and caspase-3 are widely accepted as proliferation and apoptosis markers in human tumors. We recently reported that Ki-67 can be detected as a circulating protein (cKi-67) in the plasma of patients with acute lymphoblastic leukemia (ALL), and that higher levels in plasma correlated with more aggressive disease. Here we investigated the levels of cKI-67 in patients with acute myeloid leukemia (AML). Since both apoptosis and proliferation are interconnected in defining the biology of a cancer, we also evaluated the level of apoptosis in AML as determined by caspase-3 activity in plasma. The study included a group of 106 newly diagnosed AML patients and 98 normal control subjects. We used the Meso Scale Discovery system for determining the levels of cKi-67. Apoptosis was determined by measuring caspase-3 activity (DEVD) in the plasma using a standard enzymatic fluorogenic assay. Median (range) levels of cKi-67 were significantly higher in patients with AML than in control subjects (1301 [0-6789] ng/mL vs 339 [35.76-2830.65] ng/mL; P<0.0001), as were caspase-3 activity levels (11.9 [5.8-218.2] pmol/min vs 8.27 [4.54-34.30] pmol/min; P<0.0001). Neither cKi-67 level nor caspase-3 activity correlated with white cell count, hemoglobin, platelet count, lactate dehydrogenase, blast count, age, cytogenetic grouping, or performance status. However, high levels (above the upper quartile of 2100 ng/mL) of cKi-67 were significantly associated with longer survival among patients >70 years of age (n=84; P=0.02) and those with unfavorable cytogenetics (n=38; P=0.006). On the other hand, high levels of caspase-3 activity (above upper quartile of 14 pmol/min) were associated with shorter survival (P=0.02) irrespective of age or cytogenetic grouping. In conclusion, proliferation and apoptosis as determined by plasma cKi-67 level and caspase-3 activity are unusual in AML. In that, higher proliferation and lower apoptosis are associated with better survival, most likely reflecting that cells progressing through the cell cycle may respond better to chemotherapy. These measurements should be considered in new therapeutic approaches that target cell cycle.

Disclosures
No relevant conflicts of interest to declare.
Hypoxia Suppresses Arsenic Trioxide-Induced Apoptosis in Human Acute Myeloid Leukemia Cells.
Authors: Yuk Yen Ju , Seo Young Jin , Kang Hyoung Jin , Jang Mi Kyoung , Lee Ji Won , Kim Hye Ry , Han Eun Jong , Shin Hee Young , Ahn Hyo Seop 
Published: October 2019

Abstract
Abstract 3109
Poster Board III-46
Acute myeloid leukemia (AML) is a class of prevalent hematopoietic malignancies, which often remains incurable because of the development of drug resistance. Leukemic cells originated from hypoxic condition in bone marrow that gives a benefit to leukemic cells by protecting them from anti-cancer drugs through physical barrier and by induction of some survival signal mediators. As the development of drug resistance is a key element in the failure of chemotherapy for leukemia, it has been studied about many factors make leukemic cells resist to chemotherapeutic drugs. However, effect of hypoxia on drug induced apoptosis is still poorly understood. Here, we investigated whether hypoxia affects the resistance to drugs in leukemic cells and whether hypoxia has some role in drug-induced apoptosis. To demonstrate whether hypoxia influences the apoptosis of human leukemic cells induced by chemotherapeutic drugs, the human AML cell lines (U937, HL-60, CCRF-CEM and K562) were treated with arsenic trioxide (ATO), actinomycin-D, 17-AAG, valproic acid and cytrabine under hypoxic condition and hypoxiamimetic agent cobalt chloride (CoCl2). Cellular proliferation was evaluated by methyl thiazolyl tetrazolium (MTT) assay. Subsequently, FACS analysis and western blot were performed to investigate the apoptosis related proteins. Among AML cell lines, HL-60 cells became resistant to apoptosis induced by chemotherapeutic drug, especially ATO, under the hypoxic condition. It was demonstrated through FACS analysis that level of Annexin-V staining in hypoxia (13.58%) was lower than in normoxia (29.07%). Also, among many apoptosis related molecules, activation of HIF-1 alpha under hypoxic condition was associated with the cell survival (HSP70) and apoptosis (BAX). Expression of HSP70 was increased and the level of BAX was dramatically down-regulated by HSP70 in ATO treated HL-60 cells. These data show that the hypoxia increases resistance to ATO induced apoptosis and the effect was mediated by HSP70/BAX dependent pathway. Collectively, this results provided several lines of direct evidence for the role of hypoxia on apoptosis of AML cells, in which HSP 70 and BAX elicit a role as an effector downstream to HIF-1alpha. These discoveries would shed new insights for understanding underlying mechanisms of hypoxia and designing new therapeutic strategy for AML.

Disclosures
No relevant conflicts of interest to declare.
The Potential Role of Galectin-1 in Hypoxia Induced AML Cell Differentiation through C/EBPαa Transcription Regulation.
Authors: Zhao Meng , Zhao Xu-Yun , Liao Shi-Hua , Zhao Ke-Wen , Wang Li-Shun , Chen Guo-Qiang 
Published: October 2019

Abstract
Abstract 3120
Poster Board III-57
Recently we reported that cobalt chloride-simulated hypoxia and mild hypoxia could induce the differentiation of human acute myeloid leukemic (AML) cells probably through hypoxia inducible factor-1 alpha (HIF-1αa) independent of its transcription activity. In this study, we used differential gel electrophoresis to compare the HIF-1αa regulated proteins between conditional induction and absence of HIF-1αa protein in U937T cells. Among all the up-regulated proteins indentified, the mRNA and protein level of galectin-1 was most remarkably changed. Knockdown of galectin-1 partially blocked the differentiation induced by HIF-1αa overexpression in U937T cells as assessed by morphological criteria and differentiation associated-antigens. Notably, reduced expression of C/EBPαa, a critical transcription factor for granulocyte differentiation, could inhibit the up-regulation of galectin-1 induced by HIF-1αa, and inducible expression of C/EBPαa in U937T cells also increased galectin-1 expression. Furthermore, by using the luciferase assay and chromatin immunoprecipitation we localized the binding site of C/EBPαa in the promoter of galectin-1. Finally the role of galectin-1 during chloride-simulated hypoxia and mild hypoxia was addressed by decreasing the expression of galectin-1 with small interfering RNA. Taken together, these results conclude that galectin-1 is required for hypoxia triggered C/EBPαa dependent AML cell differentiation.

Disclosures
No relevant conflicts of interest to declare.
Bleeding Complications of Fondaparinux Versus Nadroparine in Knee Replacement Surgery Patients.
Authors: Dijkstra Margriet , Gerritsma Carina LE , van Raay Jos JAM , van den Akker Scheek Inge , van Hulst Marinus 
Published: October 2019

Abstract
Abstract 3127
Poster Board III-64
An increase in bleeding complications was observed after the introduction of fondaparinux for prevention of deep-venous thrombosis (DVT) in knee-replacement surgery. 18 weeks after introduction fondaparinux was switched to nadroparine due to safety concerns.

Objectives
To investigate if fondaparinux (2.5mg once-daily starting 6 hours post-surgery) is associated with a higher rate of major bleeding compared to nadroparine (2,850ie once-daily starting 6 hours post-surgery) given for DVT prophylaxis after total knee-replacement in the Martini Hospital (Groningen, the Netherlands). Also, the rate of hematomas without clinical consequences was evaluated.

Methods
An observational study of all patients treated with fondaparinux after total knee replacement in 2008 and comparison with an equal number of patients treated with nadroparine after the switch. 181 Total knee replacements were included; 90 treated with fondaparinux and 91 treated with nadroparine. Major bleeding was defined as: allogenic blood transfusion during DVT prophylaxis; > 2g/dl hemoglobin drop; bleeding or hematoma requiring prolonged hospital stay, re-intervention or extra outpatient clinic visits. Hematomas not fulfilling the definition of major bleeding were regarded as hematomas without clinical consequences.

Results
Both groups were equal in gender, age and BMI. There was no significant difference in major bleeding (fondaparinux x vs nadroparine y, p=z). Also, no significant difference was found between both groups in: allogenic blood transfusion during DVT prophylaxis; number of patients with a > 2g/dl hemoglobin drop; rate of bleeding or hematoma requiring prolonged hospital stay, re-intervention or extra outpatient clinic visits. There was a significant difference in hematomas without clinical consequences (fondaparinux n=20 vs. nadroparine n=3, p=< 1*10-6 ).

Conclusions
We found no difference in major bleeding complications between fondaparinux and nadroparine. Significant more hematomas without clinical consequences were found for fondaparinux.

Disclosures
No relevant conflicts of interest to declare.