bluuebunny/crossref_metadata_2025_split huggingface dataset:
Using Auto Dendritic Cells Accompanied with CIK Plus Chemotherapy As Consolidation Treatment for 59 Patients with Acute Leukemia
Authors: Zheng Zhuanzhen , Qiao Zhenhua , Chen Wenliang , Gong Rong , Wang Yalin , Xu Lianrong , Ma Yanping , Hou Lihong , Zhang Li , Lu Yujin , Jiang Bo , Li Guoxia , Dong Chunxia 
Published: October 2019

Abstract
Abstract 4923

Objective
To explore the safety and clinic therapeutic effects of using tumor activated dendritic cells (DC) accompanied cytokine induced killer (CIK) in order to feinforce opposing anti-tumor function and to eliminate minimal residual disease.

Methods
59 patients entered into DCIK plus chemotherapy consolidation teams as achieved the 1st complete remission and after 1 or 2 cycle themotherapy. Collect 56 case's bone marrow monocyte cells at diagnosising to prepare tumor antigen, Collect bone marrow monocyte cells at complete remission to culture DC and CIK, using tumor antigen to activate DCs. After 7 days culturing separately then co-culture together for 1 day, collect these cells as DCs activated CIK, after scrubbing, infusing back to patients. Then these patients accept 10–15 day's immuno-regulation treatment, such as interleukine-2(IL-2) 500000u/d for one week or interferon -α(IFN-α)3,000,000u/d three times one week or thymopentin 1mg/d,2 times every week for two weeks. Those cases (one case AML-M3a,one case ALL-B,one case AML-M4a)only have routine culturing and infusion of DCs and CIK and the DCs are not activated by tumor antigen because of having no conservation leukemic cells.

Results
59 cases accept DCIK immuno-therapy for 6 to 16 times respectively, in the 24 to 78 months observation stage, 42 cases OS(overall survival)and 33 cases DFS(disease free survival). No one developed therapeutic-dependent auto-allergic disease, such as fever, swollen, diarrhea, rash et al. Those three cases who accepted routine culturing and infusion of CIK and DCs that had not activated by tumor antigen didn't occure any treatment-dependency side-effects and keep hematologic complete remission, minimal residual disease test outcome persist lower under 103.

Conclusion
Tumor antigen activated DCs combined with CIK can reinforce anti-tumor immune function and have no obviously side-effects,accompanying with chemotherapy,those patients maybe benefit of this kind consolidation treatment and avoid routine chemo-therapeutic side reaction, temporary curative effect are satisfaction and worth popularized.

Disclosures:
No relevant conflicts of interest to declare.
NR4A1 Mediated Apoptosis of Aggressive Lymphoma Cells Suppresses Tumor Growth in a Xenograft Model.
Authors: Deutsch Alexander JA , Rinner Beate , Pichler Martin , Reitter Sonja , Beham-Schmid Christine , Neumeister Peter 
Published: October 2019

Abstract
Abstract 2408
NR4A1 (Nur77) and NR4A3 (Nor-1) are two members of the orphan nuclear receptors (NRs). Their function as critical tumour suppressor genes (TSG) is demonstrated by the rapid development of acute myeloid leukemia (AML) of NR4A1 and NR4A3 double knock out mouse and by their reduced expression in leukemic blasts from human AML patients. The aim of our study is to comprehensively study NR4A1 and NR4A3 expression B-cell malignancies and to define and functionally characterize the nuclear orphan receptors NR4A1 as TSGs in B-cell malignancies. We found a more than 50% reduction of both, NR4A1 and NR4A3, in B-CLL (71%) and Follicular Lymphoma (70%), and in diffuse large B cell lymphoma (DLBCL) (74%) compared to normal controls. In DLBCL low NR4A1 expression was significantly associated with non-germinal center B-cell subtype and with poor overall survival (p=0.042, HR=2.2, CI=1.01–4.9). To investigate the function of NR4A1 in lymphomas, we over-expressed NR4A1 in a lymphoma cell line (Sc-1) by using an inducible lentiviral expression system and performed apoptotic assays by determing cleaved caspase 3, the sub-G1 peak and Annexin V positivity. Induction of NR4A1 expression led to apoptosis in a significantly higher proportion of induced Sc-1 cells compared to their uninduced controls in all assays analysed. Additionally, treatment of an immortalized B cell line (UH3) and three lymphoma cell lines (Karpas422, SC-1 and Ly8) with Cytosporone B (Csn-B), a NR4A1 ligand known to induce NR4A1, caused NR4A1 mediated apoptosis. To test the tumor suppressor function of NR4A1 in vivo, the stably transduced Sc-1 lymphoma cell lines were further investigated in a NOD/SCID/IL-2rγnull (NSG) mouse model. Induction of NR4A1 in Sc-1 suppressed tumor growth in the NSG mice, in contrast to vector controls and uninduced Sc-1 cells, where massive tumor formation was observed. Our data suggest that NR4A1 has pro-apoptotic functions in vitro and that Csn-B induces a NR4A1 mediated apoptosis in lymphoma cells. Our xengraft experiments define NR4A1 as novel tumor suppressor in vivo. Hence, regulation of NR4A1 is a promising new therapeutical target for future anti-lymphomatherapy.

Disclosures:
No relevant conflicts of interest to declare.
Prospective Comparison of Sperm Parameters in SCD Adult Patients Before and after a 6 Months Treatment with hydroxyurea (HU)
Authors: Bachir Dora , Lionnet Francois , Dautremer Justine Gellen , Stankovic Katia , Habibi Anoosha , Bartolucci Pablo , Chalas Céline , Berthaut Isabelle , Eustache Florence , Ravel Célia , Kuntsman Jean Marie , Girot Robert , Simon Tabassome , Mandelbaum Jacqueline , Galacteros Frederic 
Published: October 2019

Abstract
Abstract 242

Introduction
Hydroxyurea (HU) has proved its efficacy in reducing vaso-occlusive events in patients with SCD and increasing life expectancy. However, effects on fertility in adult males represent a major issue for acceptance and adherence, as HU has been reported to impair spermatogenesis through direct cytotoxic effects and hypogonadism. There are only limited data in the literature in this field and mainly retrospective.

Patients and methods
49 adult patients aged 20–52 years with homozygous SCD eligible for first line HU treatment were enrolled after informed consent between June 2010 and April 2012 in this prospective study: HYDREP.The main objective was to compare at Day 0 and 180 of HU treatment the semen parameters according to the WHO (1999) criteria(volume, sperm concentration, viability, forward motility and morphology ). Vaso-occlusive events and other complications, number of transfusions during the 6 months preceding and following HU initiation were recorded. Hematological, biochemical, hormonal parameters, HU dosage were recorded at Day 0, Day 90 and 180. HU was begun at 15mg/Kg dose (Platt *) and modulated following haematological tolerance and renal function.

Results and discussion
Preliminary results concern the 24/49 patients, who complete the 6 months protocol.
A statistically significant impairment in all sperm parameters was observed, after a 6 months HU therapy, with great variations between individuals, which are to be explained by complementary tests. These preliminary deleterious findings must be interpreted with caution as in our experience 1) spermatogenesis may recover after drug withdrawal 2) Repeated vaso-occlusive events are also potentially deleterious on fertility (Berthaut **)and 3)this study must be completed with a survey on effective fertility seen in men treated with HU. Thus, regarding to the great benefit offered by HU on prognosis allowing a real “life project” in young symptomatic adults with SCD, it is worth providing each patient of the most accurate information on fertility as an essential prerequisite for their adhesion to HU treatment.

Conclusion
Sperm cryopreservation is advised before beginning a treatment with HU. Prospective follow up studies of HU male cohort regarding fertility are warranted. Health education has a major role to play to increase HU compliance especially in view of these results.

Disclosures:
No relevant conflicts of interest to declare.
Efficacy Analysis of the Novel Dual PI3K-MTORC1/2 Inhibitors NVP-BGT226 and NVP-BEZ235 in Acute Leukemia.
Authors: Schittenhelm Marcus M , Doehner Hartmut , Doehner Konstanze , Kanz Lothar , Heinrich Michael C. , Kampa-Schittenhelm Kerstin M 
Published: October 2019

Abstract
Abstract 2474

Introduction:
Dysregulation of the PI3Kinase/AKT pathway is involved in the pathogenesis of many human malignancies. Constitutive phosphorylation of AKT is frequently found in acute leukemia but the underlying molecular mechanisms remain unclear and mutations in the PI3K/AKT pathway are uncommon in leukemia. In some cases, constitutive AKT activation can be linked to gain-of function tyrosine kinase mutations upstream of the PI3K/AKT pathway. While inhibitors of the PI3K/AKT pathway appear attractive for tumor therapy, so far response rates to PI3K inhibitor treatment of various neoplasms are moderate. Furthermore, MTORC1 inhibitors, targeting downstream of AKT, have the disadvantage of activating AKT via feed-back mechanisms. We here comparatively studied two novel dual PI3K-MTORC1/2 inhibitors, NVP-BEZ235 and NVP-BGT226, with regard to their ability to inhibit proliferation and to induce apoptosis in different leukemia cell models as well as in primary leukemia samples.

Methods:
Expression of phospho-AKT protein levels was determined in 74 leukemia patient blood and bone marrow samples by flow cytometry. Protein and functional viability assays evaluating the effects of NVP-BEZ235 and NVP-BGT226 were performed in several leukemia cell lines, mutant-tyrosine kinase cell models and with primary leukemia blasts and bone marrow cells. Antineoplastic activity was assessed in proliferation and apoptosis experiments. Immunoblots were performed to confirm consecutive suppression of AKT signaling.

Results:
AKT is generally expressed in acute leukemia at significantly higher levels as compared to healthy donor samples (p < 0.05, students t-test). Dual targeting of the PI3K-AKT-MTOR pathway profoundly inhibited AKT signaling, leading to antiproliferative effects in vitro and ex vivo. Moreover, both agents potently induced apoptosis in a subset of leukemia samples with BGT226 being the more effective agent and displaying IC50s at the low nanomolar level. Cell cycle analyses revealed that NVP-BGT226 overrides the G1-arrest observed with NVP-BEZ235-treated cells. Importantly, normal mononuclear cells revealed lower phospho-AKT expression levels and relative insensitivity towards dual PI3K-MTORC1/2 inhibition.

Conclusion:
Our data provide a strong rationale for clinical evaluation of the tested dual PI3K-MTORC1/2 inhibitors in acute leukemias.

Disclosures:
No relevant conflicts of interest to declare.
Native and Fc Enhanced Therapeutic Human Monoclonal Antibodies Targeting the Intracellular WT1 Oncogene Product in Leukemia.
Authors: Veomett Nicholas , Dao Tao , Pankov Dmitry , Yan Su , O'Reilly Richard J. , Liu Cheng , Scheinberg David A. 
Published: October 2019

Abstract
Abstract 2599
The Wilms' tumor oncogene protein (WT1) is an intracellular, oncogenic transcription factor that is over-expressed in leukemias and a wide range of cancers. WT1 may be expressed in leukemia stem cells. RMFPNAPYL, “RMF”, a WT1-derived CD8 T cell epitope presented by HLA-A0201, is a validated target for T cell-based immunotherapy, used in multiple clinical trials of vaccines and cellular therapies. Therefore, we hypothesized that a “TCR-like antibody” specific for the WT1 peptide/HLA-A0201 complex might be an effective therapeutic agent. Using phage display technology, we generated 2 lead, high avidity (Kd < 0.2nM), fully human monoclonal antibodies (mAb) specific for the WT1 RMF peptide/HLA-A0201 complex. One version, ESK1, is a native human IgG1. A second version, ESKM, with enhanced antibody dependent human effector cell cytotoxicity (ADCC) function due to altered Fc glycolsylation was also prepared. ESK mAb bind to leukemia lines and other cancer cell lines, as well as primary leukemia cells that are both WT1+ and HLA-A0201+. In vitro, both ESK mAb mediated ADCC against CML cells, at concentrations below 3 ug/ml, but ESKM was about 8–10 fold more potent and could kill targets with far fewer peptide/MHC complexes expressed on the cell surface. At therapeutic doses of ESKM, there was no difference in biodistribution between the 2 mAb in C57BL6 mice or in mice that were transgenic for HLA-A0201. Low doses of ESK (25–100ug twice) effectively treated an established disseminated, ALL or human bcr/abl + lymphoid leukemias in a NSG mouse model (T, B and NK deficient) and prolonged survival. In mice, ESKM was slightly more effective than ESK1. An F(Ab')2 version of the antibody had no anti-tumor effect, indicating that an Fc-mediated mechanism plays the major role in therapy. When combined with a single infusion of human CD34-, CD3-, human NK and monocyte effectors in the NSG mice, therapeutic effects of the mAb were more pronounced and more durable. There was no therapeutic effect of either mAb on WT1 low/A0201 negative disseminated Daudi ALL in mice. There was no observed toxicity in HLA-A0201 transgenic mice at the therapeutic mAb dose and schedule. ESK mAb are potential therapeutic agents for ALL, CML, other leukemias and cancers over-expressing the WT1 oncoprotein. Its expression in early leukemia cells may allow for elimination of the progenitors. The data also provide proof of concept for developing therapeutic mAb targeting important intracellular oncogenic proteins.

Disclosures:
Yan: Eureka: Employment, Equity Ownership. Liu:Eureka: Employment, Equity Ownership.
Divalent Cations Define the Structural Requirements for αIIbβ3 Integrin Activation
Authors: Kamata Tetsuji , Handa Makoto , Kawai Yohko , Ikeda Yasuo , Aiso Sadakazu 
Published: October 2019

Abstract
Abstract 260
Platelet αIIbβ3 integrin undergoes structural rearrangements to increase the affinity for ligands. Two major changes in integrin three-dimensional structure are critical for this activation process. One is the straightening of integrin legs (extension), and the other is the swing out movement of the β3 hybrid domain (swing-out). In this study, we have found that this structural requirement for activation differs depending upon the divalent cations present. We previously reported that the swing-out-defective mutant αIIbD319C/β3V359C did not bind fibrinogen in the presence of Ca2+/Mg2+. However, the same mutant bound fibrinogen in the presence of Mn2+, when activated by monoclonal antibody PT25-2 or by forcing the integrin legs to extend by concomitant αIIbQ595NTT mutation. We engineered another mutation that was designed specifically to prevent the swing-out. Likewise, this β3G327C/β3V419C mutation blocked fibrinogen binding in the presence of Ca2+/Mg2+, but not in the presence of Mn2+. On the other hand, when integrin extension was specifically prevented by αIIbD464C/αIIbS728C mutation, fibrinogen binding was observed neither in the presence of Ca2+/Mg2+ nor in Mn2+. Thus, swing-out is not essential for activation in Mn2+, as long as integrin legs are in extended state, while both swing-out and extension are required in Ca2+/Mg2+. To explore the underlying mechanism originating this difference, three distinct cation-binding sites in the βA domain were mutated. Among the three sites, MIDAS is occupied by Mg2+, while ADMIDAS and LIMBS/SyMBS are occupied by Ca2+ in Ca2+/Mg2+ condition. When amino acid residues composing these sites were mutated to Ala, none of them bound fibrinogen, except for D126A and D127A in the presence of Ca2+/Mg2+. Notably, these mutations restored fibrinogen binding in the swing-out-defective mutant. Amino acid residues D126 and D127 compose ADMIDAS together with S123 and M335. Swing-out of the hybrid domain disrupts M335 from ADMIDAS. This allows Ca2+ to move toward MIDAS, bringing the β1-α1 loop closer to ligand to provide contact site. However, similar rearrangement of the cation/ligand binding sites has been shown to take place in Mn2+ without the swing-out. The results suggest partial disruption of ADMIDAS is the key event for activation in Ca2+/Mg2+, while it is not so in Mn2+. Our results may reconcile the dispute over the apparently contradicting findings in the crystal structure that integrin with a closed head with Mn2+ represents an active conformation as one with an open head with Ca2+/Mg2+.

Disclosures:
Kawai: Daiichi Sankyo: Consultancy; Bayer: Consultancy; Toyama Chemical: Consultancy.
Monocytosis Has Adverse Prognostic Significance and Impacts Survival in Patients with T-Cell Lymphomas.
Authors: Sacchi Stefano , Bari Alessia , Tadmor Tamar , Marcheselli Luigi , Liardo Eliana Valentina , Pozzi Samantha , Luminari Stefano , Baldini Luca , Federico Massimo , Polliack Aaron 
Published: October 2019

Abstract
Abstract 2647
New data concerning the important role of microenvironment on lymphoma growth are emerging, and in recent years surrogate biomarkers have been identified as prognostic factors for survival in non-Hodgkin lymphoma (NHL). Unlike follicular (FL), diffuse B-cell lymphoma (DLBCL) and Hodgkin lymphoma (HL), there is still no specific internationally accepted risk stratification scoring system for peripheral T-cell lymphomas (PTCL), and the International Prognostic Index (IPI) or the Prognostic Index for T-cell lymphomas (PIT) model are been used to identify higher risk cases of PTCL. Here we retrospectively analyzed the relevance of the well recognized prognostic parameters for T-NHL in 172 patients with different types of PTCL. In 94 cases in whom peripheral blood monocyte count (PBMC) at diagnosis was available, we evaluated whether monocytosis (PBMC >800/mm3) could be used as a simple prognostic factor for overall survival (OS) and outcome in PTCL.
For the entire group with a median follow-up of 19 months (range 1–168 months), the 5-years OS was 42%, and the median OS 48 months. Monocytosis was present in 23% of the evaluable cases and patients with high PBMC (>800/mm3) at diagnosis had a worse OS (median 12 months) compared to those with PBMC < 800/mm3.This difference showed strong statistical significance (p=0.003) (Fig 1) and the Hazard ratio (HR) for PBMC >800/mm3, stratified by histopathological subtype, was 2.81. In particular 3-years OS of patients with PBMC >800/mm3 with anaplastic large cell lymphoma, angioimmunoblastic T-cell lymphoma, and peripheral T-cell lymphoma not otherwise specified was 50%, 25%, and 12%, respectively compared to 86%, 65%, and 50% for those patients with PBMC <800/mm3. In univariate analysis age >60y, advanced stage, bone marrow involvement, ECOG PS >1, LDH>UNL, PBMC >800/mm3, hemoglobin <12 gr/dL, albumin <3.5 gr/L were associated with inferior OS. In multivariate analysis, monocytosis alone retained a negative prognostic value even when adjusted for PIT and stratified by histolopathological subtype (HR 2.41, p=0.015).
In this study, as in others on B-NHL and HL, monocytosis had an independent negative impact on survival in patients with T-NHL. This data, which provide convincing support for the use of monocytosis as a simple prognostic parameter, now need to be further validated in a larger cohort of patients with T-NHL.
Figure 1: Overall survival (OS) of 94 patients with PTCL according to peripheral blood monocyte count (PBMC): monocytosis (PBMC >800/mm3) has a negative impact on OS (p=0.003) Figure 1:. Overall survival (OS) of 94 patients with PTCL according to peripheral blood monocyte count (PBMC): monocytosis (PBMC >800/mm3) has a negative impact on OS (p=0.003)

Disclosures:
No relevant conflicts of interest to declare.
“Early Lesion” of Follicular Lymphoma (FL) Precedes Clinically Overt FL: A Retrospective Study of Lymph Nodes (LNs) Obtained At Surgery with LNs Dissection for Preceding Cancer.
Authors: Ota Yasunori , Izutsu Koji , Ota Hikari , Nishida Aya , Ishiwata Kazuya , Tsuji Masanori , Yamamoto Hisashi , Yamamoto Go , Asano-Mori Yuki , Wake Atsushi , Fujii Takeshi , Taniguchi Shuichi 
Published: October 2019

Abstract
Abstract 2716
Follicular lymphoma (FL) is the most common subtype of indolent lymphoma and most patients with FL are diagnosed with advanced stage. “Early lesion” of FL including partial involvement of FL and FL in situ was proposed recently, of which clinical implication remains uncertain. One report suggested that some patients with FL in situ can develop into clinically overt FL afterward (Blood 2011; 118:2976). However, it is unclear whether clinically overt FL is usually preceded by “early lesion”. In order to answer this question, we retrospectively reviewed the archived pathological specimen of lymph nodes (LNs) that had been obtained at surgery with LNs dissection for a preceding other malignancy before the diagnosis of FL in patients with clinically overt FL.

Materials and Methods
Cases for study were diagnosed as FL at Toranomon Hospital between January 2001 and March 2012 and had undergone surgery with LNs dissection for a preceding malignancy. We reexamined the dissected LNs by morphology and immunohistochemistry. The hematoxylin and eosin stained slides were reviewed and immunohistochemical staining was performed using a streptavidin-biotin complex technique. Antibodies included the following: CD5, CD10, CD20, CD21, BCL2, BCL6, MIB1. This study was performed with the approval by the Institutional Review Board.

Results
Among the 167 patients who were diagnosed as FL at Toranomon Hospital during this period, 8 patients (4.8%) had undergone surgery with LNs dissection for a preceding malignancy (Table). Preceding malignancy was gastric carcinoma (n=3), esophageal carcinoma (n=2), and endometrial carcinoma (n=2), and colon cancer (n=1), respectively. Surgery for preceding malignancy had been performed at a median of 18 months (9–186 months) before the diagnosis of FL. No patient had lesions suggestive of lymphoma on preoperative imaging studies or endoscopic examinations. With immunostaining of the archived specimen, FL was found in all of the 8 cases, of which 6 cases were FL in situ and 2 cases were usual FL. The number of resected LN was median 38 (19–153) and the involvement of FL was detected in 60% of the resected LNs. LNs metastasis of carcinoma was found in 4 cases. Median duration from surgery for precedent malignancy to the diagnosis of overt FL was 26 mo(12–186) in patients preceded by the FL in situ and 13 mo (9,17) in patients preceded by clinically latent but pathologically usual FL.

Conclusion
We found that FL is preceded by latent FL that is only detectable by pathology. Remarkably, 6 out of 8 cases were preceded by FL in situ. This retrospective series suggests that “early lesion” of FL is a precursor lesion of clinically overt FL.

Disclosures:
No relevant conflicts of interest to declare.