bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene Expression Profiling Reveals Activation of Multiple Oncogenic Pathways and Over-Expression of Survivin in the Pathogenesis of Extranodal Nasal-Type NK/T Cell Lymphoma.
Authors: Chng Wee-Joo , Selvarajan Viknesvaran , Huang Gaofeng , Zhou Jianbiao , Law Mark , Feldman Andrew L , Manual Salto-Tellez , Ng Siok-Bian 
Published: October 2019

Abstract
Abstract 3931
Poster Board III-867
Extranodal nasal-type Natural Killer/T-cell lymphoma (NKTCL) is a distinct clinicopathologic entity most commonly affecting Asians and Central and South Americans, and characterized by a clonal proliferation of NK or T cells with a cytotoxic phenotype. There is a strong association with Epstein-Barr Virus. The tumor is aggressive with patient usually surviving short duration even with chemotherapy. We performed the first comprehensive genome-wide gene expression profiling (GEP) of extranodal nasal-type Natural Killer/T-cell lymphoma (NKTCL) using formalin-fixed paraffin embedded (FFPE) tissue (n=25) and NK cell lines (n=5) and compared the results to the GEP of normal NK cells using the Illumina DASL whole genome array, with the objective of understanding the oncogenic pathways involved in the pathogenesis of NKTCL and to identify potential therapeutic targets. Quantitative-PCR validation of the GEP findings revealed over-expression of candidate genes BIRC5 (survivin), EZH2 and STMN1 in NK cell lines compared to normal NK cells, consistent with the GEP data. We then extracted a list of genes that are differentially expressed between NKTCL and normal NK cells and tissue controls. We than subjected this list of genes to pathway and network analysis using Metacore. This revealed a significant enrichment for cell cycle related genes and pathways. Furthermore, the network analysis results demonstrated a pro-proliferative and anti-apoptotic phenotype in NKTCL characterized by activation of Myc and nuclear factor kappa B (NF-KB), and deregulation of p53. This was further corroborated using immunohistochemistry on tissue microarray of NKTCL (n=33, including all cases with GEP performed). We observed a significant percentage of NKTCL showing overexpression for c-Myc (45.4%), p53 (87.9%) and NF-KB p50 (67.7%) on immunohistochemistry. Notably, overexpression of survivin was observed in 97% of the cases. Based on our findings, we propose a model of NKTCL pathogenesis where deregulation of p53 together with activation of MYC and NF-KB, possibly driven by EBV LMP-1, result in the cumulative upregulation of survivin. When KHYG and NKYS cell lines were treated with a compound IDR E804 which inhibited survivin, there is significant inhibition of cell growth as assess by MTS assay and induction of apoptosis as measured using Annexin V staining by flow cytometry. This suggests that compounds inhibiting survivin may be a potentially useful novel therapeutic approach in NKTCL.

Disclosures:
No relevant conflicts of interest to declare.
Prognostic Impact of Mid-Treatment PET in Patients with Diffuse Large B-Cell Lymphoma Who Achieved Metabolic CR at Post-Treatment PET
Authors: Yoo Changhoon , Kim Jeong-Eun , Sohn Byeong Seok , Yoon Dok Hyun , Kim Shin , Lee Dae Ho , Kim Sang We , Lee Jung Shin , Suh Cheolwon 
Published: October 2019

Abstract
Abstract 3949
Poster Board III-885

Background
Mid-treatment positron emission tomography (PET) has been found to predict clinical outcomes in patients with aggressive non-Hodgkin's lymphoma. Currently, risk-adapted therapy based on mid-treatment PET has been widely evaluated. This study was intended to assess the prognostic value of mid-treatment PET in patients who achieved metabolic complete response (mCR) at post-treatment PET.

Methods
From February 2002 to March 2009, total 130 patients in whom post-treatment PET showed mCR were included in this study. We performed retrospective analysis of progression-free survival (PFS) and overall survival (OS) according to the results of mid-treatment PET.

Results
Median age (range) was 51 (16-85) years old, and 70 (54%) patients were male. International Prognostic Index (IPI) was low (0-2) in 91 (70%) patients and high (3-5) in 39 (30%) patients. As a front-line chemotherapy, most frequently administered regimen was R-CHOP (76%). Eighty-seven (67%) patients were mCR, and 43 (33%) patients were metabolic partial response (mPR) in mid-treatment PET. With 24 months of median follow-up, 3 year-rates of PFS and OS in overall patients were 74% and 87%, respectively. Differences of survival outcomes between patients with mCR and mPR at mid-treatment PET were not statistically significant in terms of PFS (p=0.13) and OS (p=0.76).

Conclusions
In patients with metabolic CR at post-treatment PET, survival outcome was not influenced by the results of mid-treatment PET. Therefore, risk-adapted therapy solely based on mid-treatment PET might be inappropriate in the management of newly diagnosed DLBCL.

Disclosures:
No relevant conflicts of interest to declare.
Out of Hospital Treatment of Acute Pulmonary Embolism in Patients with a Low NT-ProBNP Level.
Authors: Agterof Mariette J , Schutgens Roger E.G. , Snijder Repke J , Epping G. , Peltenburg Henny G , Posthuma Ward , Hardeman J. A. , Van der Griend Rene , Koster Ted , Prins Martin , Biesma Douwe 
Published: October 2019

Abstract
Abstract 3996
Poster Board III-932

Background
NT-proBNP is emerging to be of great importance in risk assessment of patients with acute pulmonary embolism (PE), by identifying both high and low-risk patients. The aim of the present management study was to investigate the safety of out of hospital treatment of patients with acute PE based on a NT-proBNP < 500 pg/ml.

Methods
Hemodynamically stable outpatients with objectively diagnosed acute PE and a NT-proBNP level < 500 pg/ml, were considered eligible for participation in this multicenter prospective study. Patients were discharged immediately from the emergency room or within the first 24 hours of admission. The primary objective was the 10-days mortality rate. Secondary objectives were the incidence of re-admission to hospital due to PE or its treatment, the patient's satisfaction during the first ten days of treatment and the incidence of serious adverse events in a 3-month follow-up period.

Results
Approximately 40 percent of all patients presenting at the emergency room with acute PE fulfilled the inclusion criteria. In total, 152 patients (mean age 53.4 ± 14.3 years; 51% female) with PE were treated as outpatients. No deaths, major bleedings or recurrent venous thromboembolism occurred during the first 10 days or in a follow-up period of 3 months. Seven patients required readmission. Three patients were admitted because of complaints of their PE: two patients with anxiety and pain and one patient with dyspnoea and low oxygen saturation. There were no signs of progression of PE in these 3 patients. Four patients were re-admitted because of an illness unrelated to PE. According to the PSQ18 and anxiety score, patients were satisfied with their out of hospital treatment; they had no progression of anxiety during the first ten days of treatment.

Conclusion
Out of hospital treatment is safe in a group of low risk patients with non-massive PE, based on NT-proBNP levels < 500 pg/ml. Approximately 40 percent of PE-patients can be treated in an outpatient setting. Patients consider out of hospital treatment as comfortable and have no increase in anxiety scores.

Disclosures:
No relevant conflicts of interest to declare.
Fucosyltransferase VI Induces Platelet Activation: A Novel Property of a Plasma Glycosyltransferase.
Authors: Navarro José-Tomás , Tawfiq Shwan , Wohlgemuth Roland , Hoffmeister Karin M. , Sackstein Robert 
Published: October 2019

Abstract
Abstract 4016
Poster Board III-952
A number of glycosyltransferases are present in human plasma with the α(1→3) fucosyltransferase, Fucosyltransferase VI (FTVI), having the highest plasma concentration. Notably, elevated plasma levels of FTVI are associated with a variety of cancers and correlate with tumor load/progression. The well-known association of neoplasia with thromboembolic complications prompted us to examine whether FTVI has direct effect(s) on platelet function. We obtained human platelets from blood of healthy donors and separated from platelet-rich plasma by differential centrifugation. Freshly isolated platelets (x108/ml) were stirred and exposed at 37°C to varying concentrations (20, 40, 60 and 80 mU/mL) of glycosyltransferases FTVI, β-1-4-galactosyltransferase-I (βGalT-I), or α,2-3-N-sialyltransferase (α2,3-N-ST), or to 1 U/mL thrombin. Platelet aggregation and activation was assessed by aggregometry (light transmission) or by flow cytometry of FSC/SSC characteristics and of surface expression of P-Selectin, respectively. FT-VI reproducibly induced platelet aggregation and activation, whereas other glycosyltransferases (β4GalT-I and α2,3-N-ST) had no effect on platelets. FTVI activation of platelets was concentration-dependent, and the aggregation curve for FTVI was one wave, similar to that for thrombin. FTVI-induced platelet activation was independent of catalytic conversion of surface glycans, but was inhibited by FTVI denaturation, indicating that FTVI-induced platelet activation is a lectin-mediated process. To determine the membrane target(s) mediating FTVI-induced platelet activation, biochemical studies were performed after catalytic exofucosylation of the platelet surface. Flow cytometry after platelet exofucosylation showed formation of the carbohydrate structure sLex, detected by the mAb Heca452, but no formation of Lex (CD15). Western blot showed that enforced fucosylation induced sLex on a single platelet surface protein, and further biochemical studies revealed that this protein is GPIbα. These findings unveil a previously unrecognized property of FTVI as an activator of platelets, mediated via a specific lectin/carbohydrate interaction on GP1ba, and offer novel perspectives on the pathobiology of tumor-associated thrombogenesis.

Disclosures:
No relevant conflicts of interest to declare.
Low-Dose Rituximab in Idiopathic Autoimmune Hemolytic Anemia.
Authors: Barcellini Wilma , Zaja Francesco , Zaninoni Anna , Imperiali Francesca Guia , Battista Marta , Filí Carla , Russo Domenico , Di Bona Eros , Consonni Dario , Zanella Alberto 
Published: October 2019

Abstract
Abstract 4027
Poster Board III-963

Background
Conventional therapy of warm autoimmune hemolytic anemia (WAIHA) include administration of corticosteroids and immunosuppressive agents, or splenectomy, whereas no effective treatment exists for cold hemagglutinin disease (CHD). A substantial proportion of patients with WAIHA do not respond to or relapse after corticosteroid therapy and may experience clinically relevant side effects. Favorable responses to rituximab at standard doses (375 mg/m2 weekly for 4-6 courses) have been reported in both WAIHA and CHD, idiopathic or secondary, as well as in other autoimmune diseases, such as rheumatoid arthritis and primary immune thrombocytopenia. Recently, low dose (LD) rituximab (100 mg fixed dose weekly for 4 courses) has been proven effective in patients with autoimmune cytopenias, particularly immune thrombocytopenia.

Aims
To evaluate the safety, activity and the duration of the response of LD rituximab associated with standard oral prednisone (PDN) as first line therapy in newly diagnosed WAIHA and CHD, and as second line therapy in WAIHA relapsed after standard oral PDN.

Methods
In this single-arm prospective pilot study, LD-rituximab was administered at 100 mg fixed dose weekly on days +7, +14, +21, +28 along with standard oral PDN (1 mg/kg/die p.o. from day +1 to + 30, followed by quick tapering: 10 mg/week until 0.5/mg/kg/die, then 5 mg/week until stop). Complete and partial initial responses (iCR and iPR)) were defined as Hb ≥ 12 g/dL and ≥ 10 g/dL at month +2 from the beginning of therapy, respectively; sustained response (SR) was defined as Hb ≥ 10 g/dL at month +6 and +12, in the absence of any treatment.

Results
Twelve patients (6 female, 6 male; median age 49 yrs, range 28-65) were enrolled. A iCR and iPR were observed in 6 and 4 out of 12 patients, respectively; the median Hb level increased from 9.9 g/dL (range 6.1-12.3) at enrolment to 12.3 g/dL (range 9.5-14.9) at month +2. A SR at month +6 was observed in 7 out of 7 evaluable patients, and in the first 2 patients enrolled at month +12. No side effects or serious adverse events were observed. For 9 relapsed AIHA patients laboratory data (Hb, LDH, reticulocytes) and steroid administration were available before LD rituximab treatment: a lower cumulative dose (roughly 50%) of steroid was administered to patients during LD rituximab study compared with previous therapy, without significant differences in the general trend of Hb, LDH, and reticulocytes.

Conclusions
These preliminary results seem to indicate that the addition of LD rituximab to standard corticosteroid therapy is a feasible and active treatment in AIHA. Data on SR are intriguing, particularly regarding the possible steroid sparing effect of LD rituximab, but need to be confirmed in larger survey and after longer follow up.

Disclosures:
No relevant conflicts of interest to declare.
Knockdown of Matriptase-2 Increases Hepcidin Expression in a BMP-Dependent Manner in Zebrafish Embryos.
Authors: Gibert Yann , Lattanzi Victoria J. , Fraenkel Paula G. 
Published: October 2019

Abstract
Abstract 4050
Poster Board III-985
Hepatocytes produce hepcidin, a transcriptionally regulated peptide hormone, in response to iron overload, resulting in decreased intestinal iron absorption. Hepcidin levels are inappropriately low in patients with thalassemia major and hemochromatosis. We have been developing the zebrafish embryo as a model to identify hepcidin regulators, which may be exploited to prevent and treat iron overload syndromes. Recently, it has been shown in mammalian models that the membrane-bound serine protease, matriptase-2 (also known as TMPRSS-6), inhibits hepcidin expression by cleaving membrane-bound hemojuvelin (hjv), a bone morphogenic protein (BMP) co-receptor, which is mutated in patients with juvenile hemochromatosis. Previously, we demonstrated that hepcidin expression increases in response to iron loading or activation of the BMP signaling pathway, but that hjv is not required for hepcidin expression in zebrafish embryos. We hypothesized that hepcidin expression in zebrafish embryos is normally hjv-independent, either because of low levels of hjv expression in the liver or because of the presence of matriptase-2. We used whole mount in situ hybridization, quantitative realtime RT-PCR, morpholino knockdowns and overexpression experiments to address these questions. As hjv was undetectable in the zebrafish embryonic liver by in situ hybridization, we used flow cytometry to sort zebrafish embryonic hepatocytes and assess hjv expression by RT-PCR. We found that hjv was weakly expressed in the embryonic liver, but strongly expressed in zebrafish adult liver. We overexpressed zebrafish hjv in human hepatocytes and found that it potentiated the effects of BMP6 to activate a human hepcidin promoter and a BMP response element. In contrast, overexpression of zebrafish hjv in zebrafish embryos failed to increase hepcidin expression. Knockdown of matriptase-2 in zebrafish embryos caused developmental delay and a smaller liver, but increased hepcidin expression, relative to liver size. Furthermore, treatment with the BMP inhibitor dorsomorphin abrogated the increased hepcidin expression associated with matriptase-2 knockdown. Experiments are underway to determine whether zebrafish hjv has little effect on hepcidin expression during embryonic development because of the activity of matriptase-2.

Disclosures:
No relevant conflicts of interest to declare.
Does Transcription of Remote α-Globin Regulatory Elements Affect Their Function?.
Authors: Kowalczyk Monika S , Hughes Jim R , Sharpe Jacqueline A , Brown Jill M , Buckle Veronica J , Wood William G , Higgs Douglas R 
Published: October 2019

Abstract
Abstract 4060
Poster Board III-995
Expression of many tissue and developmental stage specific genes is controlled by remote regulatory elements. The mammalian a-globin gene cluster includes four previously characterised remote regulatory elements (MCS-R1 to R4) upstream of the a-globin genes. Naturally occurring deletions and experiments in transgenic mice, have shown that one or more of these elements are necessary for fully regulated expression of the a-like globin genes. At present it is unclear exactly how these elements interact with the a-globin genes to enhance their expression, although ultimately we know that they physically interact with the promoters forming a chromatin loop. Of interest, three of the MCS-R sequences are located within introns of a widely expressed gene C16orf35, which lies adjacent to the a-globin locus and we now know that such complex, intermingled arrangements are common in the mammalian genome. We have asked two questions: can the MCS-R elements be transcribed while activating the globin genes, and does transcription of the MCS-R elements play any role in their activation?
We have analysed nascent transcription of C16orf35 and a-globin, and shown that both genes are simultaneously transcribed in a high proportion of erythroid cells. Thus it appears that the MCS-R elements can interact with the globin genes while being transcribed. We next asked whether transcription of the C16orf35 gene (through the upstream regulatory elements) is necessary for their activation. The promoter region and a putative erythroid-specific promoter of the C16orf35 gene were deleted (singly and in combination) by homologous recombination in mice. Provisional analysis has shown no evidence of a-thalassaemia in these mice, suggesting that activation of the MCS-R elements occurs independently of their transcription. In addition to their importance in globin gene regulation, these findings have general implications for the relationship between structure and function in the mammalian genome and the mechanisms by which long-range elements interact with their cognate promoters.

Disclosures:
No relevant conflicts of interest to declare.
In Vitro Generation of Cytotoxic T Lymphocytes against MAGE A1 and MAGE A3 Derived From Healthy Donors.
Authors: Bao Lei , Stamer Mindy M , Dunham Kimberly , Krishnadas Deepa Kolaseri , Lucas Kenneth G 
Published: October 2019

Abstract
Abstract 4079
Poster Board III-1014
MAGE A1 and MAGE A3 are cancer testis antigens that are expressed on a number of malignant tumor cells, but not by normal cells, except for male germ cells which lack HLA expression. Therefore, MAGE cytotoxic T lymphocytes are strictly tumor-specific. Adoptive transfer of antigen specific cytotoxic T lymphocytes (CTL) provides immediate graft-versus tumor effects while minimizing risk for graft-versus-host disease. The aim of the current study was to find ideal conditions for expansion of CTL targeting tumor-associated antigens from peripheral blood mononuclear cells (PBMCs) of healthy donors to be used in allogenic cell therapy. In this study we investigated the ability to generate MAGE A1 and MAGE A3 specific cytotoxic T cells using autologous dendritic cells (DC) loaded with MAGE A1 and MAGE A3 overlapping peptides. CTL lines specific for MAGE A1 and MAGE A3 were established by stimulating CD8 T cells from healthy donors with autologous dendritic cells loaded with MAGE A1 or MAGE A3 overlapping pooled peptides in round-bottomed, 96-well plates. CD8+ T cells were restimulated with the same ratio of peptide pulsed DC on days 7 and 14 in the presence of IL-2 (50 U/ml), IL-7 and IL-15 (5 ng/ml). These microcultures were screened 10 days after the third stimulation for their capacity to produce interferon-gamma (IFN-gamma) when stimulated with autologous EBV-transformed B lymphocytes (BLCL) transduced with lentivirus(LV) encoding MAGE A1 or MAGE A3 and autologous BLCL transduced with LV encoding GFP. MAGE A1 and MAGE-A3 specific IFN-gamma producing cells were rapidly expanded in OKT3 and IL2. The specificity of the rapidly expanded MAGE A1 and MAGE A3 specific T cells was confirmed by IFN-gamma production as measured by intracellular cytokine staining and ELISA as well as antigen specific cytotoxicity by a standard 51chromium (51Cr) release assay. We successfully generated MAGE A1 and MAGE A3 specific CTL lines from healthy donors using this method. Specific CTL lines showed cytotoxicity in vitro not only to target cells pulsed with MAGE A1 or MAGE A3 peptides but also to target cells transduced with LV-MAGE A1 or LV-MAGE A3. Specific cytolytic activity was accompanied by IFN-gamma secretion. These data indicate that tumor antigen specific CTL can be expanded using overlapping peptides regardless of an individual's HLA specificity. The ability to generate tumor specific CTL from donors of various HLA backgrounds provide a rationale for utilizing MAGE A1 and MAGE A3 overlapping peptides for expansion of antigen specific T cells for adoptive T-cell therapy against MAGE A1 or MAGE A3 expressing tumors.

Disclosures:
No relevant conflicts of interest to declare.
Adult Philadelphia-Positive Acute Lymphoblastic Leukemia (Ph + ALL) Treated at Diagnosis with Imatinib Followed by 2nd Line TKI (Imatinib or Dasatinib) and Stem Cell Transplantation.
Authors: Carella A. M. , Catania G. , Pica G. , Beltrami G. , Nati S. , Vimercati R. , Congiu A. , Rossi E. , Spriano M. 
Published: October 2019

Abstract
Abstract 4109
Imatinib has become an integral part of front-line therapy for Ph+ ALL, with remission rate exceeding 90% irrespective of whether Imatinib is given alone or combined with chemotherapy. Treatment outcome with Imatinib-based regimens has improved survival compared with historic controls, but most patients who do not undergo allogeneic stem cell transplantation (SCT) eventually relapse. Second generation TKI, e.g. Nilotinib and Dasatinib, show activity against most of the BCR-ABL tyrosine kinase domain mutations involved in acquired Imatinib resistance, but clinical benefit is generally short lived. In the attempt to improve the prognosis of these patients, we started a new intensive protocol consisting of induction phase with Imatinib (600 mg po, daily) in combination with conventional chemotherapy; moreover, two intrathecal methotrexate will be administered at the day +8 and +22. As known, this approach will determine an high number of complete hematologycal (CHR) and cytogenetic remissions (CcyR). Soon after, Imatinib will be discontinued and a 2nd TKI (Nilotinib or Dasatinib) will be offered to the patients. The idea is to reduce/eliminate as soon as possible the mutations generating under Imatinib. The primary objective of the study will be to assess the activity of sequential TKI to induce molecular remission; secondary objectives will be DFS, relapse rate, OS. This protocol will be designed for patients >18 years. Recruitment started in June 2008. Minimal residual disease will be investigate by RQ-PCR at day +30, +60, +90, +120, +150 and so on. The protocol has been designed for 20 patients. Three evaluable patients entered until now this trial; the median age was 47 years (range, 39-64 years). All 3 patients achieved CHR and CCyR at a median of 26 days (range, 18-31 days) and 46 days (range, 39-59 days), respectively. Monitoring RQ-PCR, a marked clearance of leukemic cells was found at d+60; when CCyR was achieve, Imatinib was discontinued and 2nd TKI was started: 2 patients received Dasatinib 100 mg po daily and 1 patient Nilotinib 800 mg po twice daily. Between d+100 and d+120, RQ-PCR became negative in the peripheral blood of all patients. One patient with an HLA+ sibling donor is waiting to receive an allograft; the other 2 patients, who did not find an HLA+ donor, received an autograft with molecular negative PBPC. The conditioning regimen consisted of the BU-Cy regimen. Both patients are alive and in CMR at 3 and 10 months after autografting. More patients and more time will confirm the efficacy of this new approach.

Disclosures:
No relevant conflicts of interest to declare.