bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Strong and the Weak - the Dual Role of JAK1 in Inflammatory Cytokine Production,
Authors: Luft Thomas , Wagner Andreas , Conzelmann Michael , Dietrich Sascha , Krämer Oliver , Schäfer Claudia , Giese Thomas , Falk Christine , Hecker Markus 
Published: October 2019

Abstract
Abstract 3218
Inhibition of JAK1 is an emerging clinical concept that has promise for a variety of autoimmune diseases, myeloproliferative diseases and post-transplant immunosuppression, but similarly raises concerns regarding immunosuppressive side effects.
At the example of IL-12p70 production in human monocyte-derived dendritic cells we demonstrate that JAK1 has a dual role in differentially regulating effects of weak and strong activation stimuli. We have demonstrated recently that weak NF-kB-activating stimuli (e.g. CD40L or LPS) require complementary JAK1-targeting cytokines such as IFN-g to induce IL-12p70. This pathway involves RELA, CREL, JAK1 and/or JAK2, STAT1, IRF1 and IRF8 and is inhibited by RELB and TYK2 (Conzelmann et al. Biochem Pharm. 2010, 80(12):2074–86).
Here we provide evidence for an alternative IL-12 stimulating pathway depending on strong NF-kB activating stimuli (e.g. intact E. coli or LPS plus IL-1b). siRNA silencing demonstrated that this pathway is specifically inhibited by JAK1 and the transcription factor STAT3, but is not influenced by any of the other JAK/STAT family members. Both IL-12p35 and p40 mRNA expression is directly inhibited by STAT3. Furthermore, ChIP-assays revealed that STAT3 binds directly to a combined STAT/NF-kB site at the IL-12p35 promoter without altering access of RELA and CREL. Extending the cytokine panel we found that E.coli-induced IL-6 and TNF-a production is similarly inhibited by the JAK1/STAT3 pathway whereas IL-10 expression is not affected. The observed dual effects of JAK1 are clearly confirmed by the JAK1/2 inhibitor INCB018424 (Ruxolitinib) which enhances E.coli-induced cytokines whilst strongly inhibiting cytokine production stimulated by CD40L/IFN-g.
In summary, our study suggests that blockade of JAK1 specifically inhibits pro-inflammatory effects of weak, IFN-g dependent, NF-kB activating stimuli while enhancing inflammatory cytokine expression induced by strong activation stimuli. Inhibition of JAK1/2 by INCB018424 (Ruxolitinib) would therefore represent a novel immunosuppressive approach that may spare the immune defence against invading pathogens.

Disclosures:
No relevant conflicts of interest to declare.
PGE2-Independent Expression of Indoleamine 2,3-Dioxygenase-2 (IDO2) Supports the Tolerogenic Function of Monocyte-Derived Dendritic Cells,
Authors: Trabanelli Sara , Curti Antonio , Ocadlikova Darina , Evangelisti Cecilia , Salvestrini Valentina , Ciciarello Marilena , Aluigi Michela , Rossi Lara , Metz Richard , Laury-Kleintop Lisa , Prendergast George , Lemoli Roberto M. 
Published: October 2019

Abstract
Abstract 3231
The indoleamine 2,3-dioxygenases IDO1 and IDO2 participate in tryptophan catabolism along the kynurenine pathway and are specifically inhibited by 1-methyltryptophan (1-MT). IDO1 expression in dendritic cells (DCs) is upregulated by various maturation stimuli. In particular, PGE2 plays a crucial role in inducing IDO1-expressing DCs. While it is well established that IDO1-expressing DCs contribute to immunological tolerance by a number of mechanisms, little is known about the expression, function and regulation of IDO2 in DCs.
Here we show that immature monocyte-derived DCs (Mo-DCs) express IDO2 which, like IDO1, is also upregulated upon maturation with a prostaglandin-E2 (PGE2)-containing cytokine cocktail. However, while IDO1 upregulation is strictly dependent on PGE2 during maturation, IDO2 expression level is not affected by PGE2. We show that IDO2 expressed by Mo-DCs is enzymatically active, producing L-kynurenine, and that this activity affects T-cell functions by inhibiting allogeneic and autologous T-cell proliferation and by inducing formation of a population of T regulatory cells (Tregs). Taken together, our findings establish different regulatory pathways for IDO2 and IDO1 in Mo-DCs, where IDO2 is functionally active, and they demonstrate that the tolerogenic function of Mo-DCs relies in part on PGE2-independent expression of IDO2.

Disclosures:
Metz: New Link Genetics Corporation: Employment. Prendergast:New Link Genetics Corporation: Employment.
Toll-Like Receptor 3 Activation Is Involved in Venous Thrombosis Development,
Authors: Lemarie Catherine A , Laurance Sandrine , Blostein Mark 
Published: October 2019

Abstract
Abstract 3277
Venous thromboembolism (VTE) afflicts 117 people per 100,000 each year and is an important cause of morbidity and mortality. Inflammation may have a role in both the genesis and resolution of venous thrombi. Although septic thrombophlebitis occurs, most DVT are sterile. Recently, the role of toll-like receptor (TLR) signaling in modulating sterile inflammation has become better defined in experimental injury models. TLR3, for example, is activated by viral long double stranded RNA and endogenous RNA released by cellular apoptosis and necrosis, and induce a proinflammatory cellular response. Short single-stranded RNA, such as those used in RNA inhibition, stimulate TLR3 in endothelial cells and inhibit neoangiogenesis. Thus, we hypothesize that TLR3 might be involved in the inflammatory development of venous thrombosis. We first investigated whether stimulation of TLR3 influences venous thrombosis in mice. Intravenous injection of polyinosine polycytidylic acid (polyI:C), a synthetic double-stranded RNA analog and TLR3 ligand, increase the size and the cellular density of thrombi after FeCl3-induced inferior vena cava injury. TLR3 stimulation with polyI:C is also associated with an increased reactive oxygen species production, as well as increased neutrophil and macrophage content. Using a specific fluorescent probe for RNA, we found that FeCl3 induces RNA release and thus increases the RNA content in the thrombus. We next investigated the effect of RNase on thrombus development. Intravenous injection of RNase decreased thrombus size, as well as reactive oxygen species production and neutrophil and macrophage content in the thrombus. In vitro incubation of endothelial cells with polyI:C induces production of pro-inflammatory cytokines GM-CSF, G-CSF, IL-6, IL-8 and interferon-γ-induced protein 10 involved in the recruitment of neutrophils and macrophages. Taken together, these results strongly suggest that RNA release and TLR3 stimulation after endothelial cell injury participate in thrombus formation by inducing a pro-inflammatory response leading to the recruitment of neutrophils and macrophages.

Disclosures:
No relevant conflicts of interest to declare.
Coagulation Factor Activity Level and Clinical Bleeding Severity in Rare Bleeding Disorders: Results From the European Network of Rare Bleeding Disorders (EN-RBD),
Authors: Peyvandi Flora , Palla Roberta , Menegatti Marzia , Siboni Simona M , Halimeh Susan , Faeser Britta , Pergantou Helen , Platokouki Heleni , Giangrande Paul , Peerlink Kathelijne , Celkan Tiraje , Ozdemir Nihal , Bidlingmaier Christoph , Ingerslev Jorgen , Giansily-Blaizot Muriel , Schved Jean-Francois , Gilmore Ruth , Gadisseur Alain P. , Benedik-Dolnicar Majda , Kitanovski Lidija , Mikovic Danijela , Musallam Khaled M , Rosendaal Frits R. 
Published: October 2019

Abstract
Abstract 3312
The European Network of Rare Bleeding Disorders (EN-RBD) was established to bridge the gap between knowledge and practice in the care of patients with RBDs (rare bleeding disorders frequency <1/0.5-2M). The aim of this first report was to explore the relationship between the coagulation factor activity levels and clinical bleeding severity in patients with RBDs. A total of 592 records on patients with RBDs were cross-sectionally collected over a period of three years. Data on clinical bleeding episodes were available for 495 patients and were classified into four categories according to severity. The mean age of patients was 31 years (range, 7 months-95 years), with 51% being females. On linear regression analysis, there was a strong association between coagulation factor activity level and clinical bleeding severity for fibrinogen, factor (F)FX, FXIII, and combined FV and FVIII deficiencies. A weaker association was present for FV and FVII deficiencies. There was no association between coagulation factor activity level and clinical bleeding severity for FXI. It is of clinical relevance to identify levels that are not associated with spontaneous major bleeding, which was done by constructing receiver operating characteristic curves. These levels were highest for FXIII deficiency (25 U/dl), followed by FV+VII (15 U/dl), FVII (15 U/dl), FX (5 U/dl), and FV (1 U/dl) deficiencies. For fibrinogen deficiency, a factor activity level of 20 mg/dl is needed to ensure absence of major spontaneous bleeding. There is a clear relation between coagulation factor activity level and clinical bleeding severity in RBDs, which is different for the missing clotting factor.

Disclosures:
Peyvandi: NovoNordisk, CSL Behring: Honoraria. Bidlingmaier:CSL Behring, Bayer Schering: Research Funding; CSL Behring, NovoNordisk, Pfizer, Bayer Schering: Membership on an entity's Board of Directors or advisory committees; Baxter, Biotest, CSL Behring, NovoNordisk, Pfizer, Bayer Schering: Honoraria. Schved:LFB, CSL Behring, Novonordisk, Bayer Schering: Research Funding.
Critical Role of Factors II and X During Coumarin Anticoagulation and Their Combined Measurement with a New Prothrombin Time Variant (Fiix-PT),
Authors: Onundarson Pall T. , Gudmundsdottir Brynja R. , Francis Charles W. 
Published: October 2019

Abstract
Abstract 3354

Introduction:
Vitamin K antagonists (VKA) are monitored with prothrombin time (PT) based assays that are equally sensitive to reductions in factors II, VII or X. However, previous studies suggest that the anticoagulant effect of VKA depends primarily on reductions in factors II and X and not VII.

Aim and methods:
We compared the effect of vitamin K dependent (VKD) coagulation factors on PT (Quick and Owren methods) and also on rotational thromboelastometric (ROTEM) parameters. The experiments used normal platelet poor plasma (PPP) and PPP selectively immunodepleted of individual VKD factors, with and without added platelet phospholipid or washed platelets.

Results:
The PT was equally sensitive to reductions in factors II, VII or X. However, ROTEM parameters correlated poorly with the PT in anticoagulated patients` plasmas. ROTEM experiments showed that the clotting time, maximum velocity of clot formation and the maximum clot firmness were more affected by reductions in FII or FX than by FVII or FIX concentrations which had little influence except at very low concentrations. We developed a modified PT that was sensitive only to reductions in factors II and X by using factor II and X (Fiix) depleted plasma in the PT system. The Fiix-PT (Fiix-INR) correlated well with PT (INR) but the Fiix-INR fluctuated less than the INR in anticoagulated patients reflecting its insensitivity to FVII.

Conclusion:
The ROTEM results suggest that mild to moderate reductions in factors II or X are more important in clot formation than factors VII or IX at therapeutically relevant factor concentrations. Reductions in FII and X may therefore better reflect anticoagulation with VKA than FVII or IX. FVII may be a confounding source of unwanted variation in PT-INR. The new Fiix-PT that is sensitive only to FII and FX may more accurately reflect the degree of therapeutic anticoagulation in patients treated with VKA than do the current PT assays which may overestimate the fluctuation in anticoagulation.

Disclosures:
Onundarson: See i. other: Patent application for Fiix method in process. Gudmundsdottir:Other, see i: patent applicaiton filed for Fiix method.
Fludarabine, Cytarabine, and Attenuated-Dose Idarubicin (m-FLAI) Induction for Elderly Patients with Acute Myeloid Leukemia: Results of a Multicenter Phase II Study,
Authors: Lim Yoojoo , Koh Youngil , Yoon Sung-Soo , Park Seonyang , Kim Byoung Kook , Kim Dae-Young , Lee Jung-Hee , Lee Kyoo-Hyung , Cheong June-Won , Kim Hyuk , Joo Young Don , Won Jong-Ho , Park Sung-Kyu , Kim Se Hyung , Hong Dae Sik , Park Eunkyung , Kim Min Kyoung , Sohn Sang Kyun , Kim Sung Hyun , Lee Hong-Kee , Kim Chul Soo , Estey Elihu H. , Lee Je-Hwan , Min Yoo-Hong , Kim Inho 
Published: October 2019

Abstract
Abstract 3626

Introduction:
Although there have been remarkable improvements in treatment of acute myeloid leukemia (AML), the prognosis of AML in elderly patients remains poor, and the best induction chemotherapy for these patients remains yet unknown. To devise an effective induction regimen for elderly patients with AML, we conducted a phase II trial to evaluate the efficacy and safety of the modified fludarabine, cytarabine, and attenuated-dose idarubicin (m-FLAI) regimen in these patients.

Patients and Methods:
Elderly (60 years or older) AML patients who did not receive previous chemotherapy were enrolled. Patients received two consecutive cycles of m-FLAI chemotherapy as an induction. The m-FLAI regimen was comprised of fludarabine (25mg/m2, days 1–4), cytarabine (1000mg/m2, days 1–4), and attenuated-dose idarubicin (5mg/m2, days 1–3). The primary end point was complete remission (CR) rate.

Results:
A total of 108 patients (median age 68.4 years, M:F=64:44) were enrolled. CR was achieved in 62.9% of patients, and treatment-related mortality rate (TRM) was 25.8%. Median overall survival (OS) was 9.3 months, and median event-free survival (EFS) was 6.6 months. The mortality at 30 and 60 days was 18% and 24%, respectively. Performance status and comorbidity did not have prognostic value in these patients. Bone marrow expression of CD117 was related to long EFS and OS.

Conclusion:
In conclusion, m-FLAI is a safe and effective induction regimen for previously untreated AML in elderly patients. Bone marrow CD117 expression is an independent good prognostic factor in these patients. (ClinicalTrials.gov number, NCT01247493)

Disclosures:
No relevant conflicts of interest to declare.
Meta-Analysis of Diffuse Large B-Cell Lymphoma Gene Expression Identifies Novel and Recurrent Biological Connections,
Authors: Care Matthew , Barrans Sharon , Worrillow Lisa , Jack Andrew , Westhead David , Tooze Reuben 
Published: October 2019

Abstract
Abstract 3682
Cell of origin classification of diffuse large B-cell lymphoma (DLBCL) identifies biologically and clinically meaningful subsets. In future these classes may inform distinct treatment. However a meta-analysis to assess consistency of class and gene signature associations across multiple DLBCL gene expression datasets is lacking. Furthermore diagnostic material is routinely available in formalin-fixed paraffin embedded (FFPE) form, and there is limited evidence that for DLBCL FFPE derived RNA can produce gene expression datasets comparable to freshly extracted RNA. Using an FFPE derived data-set we develop a platform robust implementation of the cell of origin classifier. With this classifier we observe comparable separation of survival in distinct data-sets derived from fresh and FFPE material. Classification of six data-sets encompassing a total of >900 DLBCL samples with this implementation establishes very similar patterns of gene expression for each DLBCL class. When tested against 12,000 signatures derived from LLMPP, MSigDB, GeneSigDB and genome wide transcription factor motif analysis of all TRANSFAC and JASPAR matrices, this communality translates into reproducible association of ABC and GCB-DLBCL with distinct molecular pathways, chromosomal regions and transcription factor motifs. This meta-analysis thus confirms the validity of FFPE derived data, identifies the most robust DLBCL class and signature associations and provides evidence for specific transcription factor signatures in both ABC and GCB-DLBCL.

Disclosures:
No relevant conflicts of interest to declare.
Inhibition of Caspase-8 Activity Caused by Overexpression of BCL10 Contributes to the Pathogenesis of High-Grade MALT Lymphoma,
Authors: Chen Yan , Yang Yishu , Sun Min , Yan Zhuohong , Cui Xiaoli , Zhang Ge , Morris Stephan W. , Zhang Quangeng 
Published: October 2019

Abstract
Abstract 3694
Mucosa-associated lymphoid tissue (MALT) lymphoma comprises approximately 8% of all non-Hodgkin lymphomas and is the most common lymphoma in the gastrointestinal tract. B-cell lymphoma/leukemia 10 (BCL10) overexpression and nuclear expression has been associated with high-grade MALT lymphomas unresponsive to H. pylori eradication treatment. To explore the molecular mechanism of BCL10 overexpression on the pathogenesis and malignant phenotype of MALT lymphoma, we generated EμSR-BCL10 transgenic mice. Results in our previous report showed that predisposition to marginal zone B-cell lymphoma (MZL) development was enhanced in these transgenic mice. MALT lymphoma precursor cells - marginal zone (MZ) B cells - are expanded in BCL10 transgenic mice. Constitutive activation of both canonical and noncanonical NF-κB pathways was identified in these MZ B cells. Elevated expression of BAFF, a NF-κB downstream gene, may be in part responsible for increased tumor cell survival in MALT lymphomas. In this continuing study, quantitative analysis of BCL10 protein levels and the extent of MZ B cell expansion in heterozygous (Tg/+) and homozygous (Tg/Tg) EμSR-BCL10 transgenic mice demonstrated a direct pathogenic effect of BCL10 protein on MALT lymphomagenesis. The activities of caspase-8 and caspase-3, but not caspase-9, were inhibited with increasing BCL10 protein levels in a dose-dependent manner. Apoptosis induced by anti-IgM was selectively inhibited in MZ B cells from transgenic mice, but no differences were observed following treatment with dexamethasone, γ-irradiation or anti-CD95, implying that overexpressed BCL10 exerts anti-apoptotic effects through the B-cell antigen receptor pathway. Expression of API2, another NF-κB downstream gene, was also elevated in a dose-dependent manner with BCL10 protein level in the MZ B cells. Overexpressed BCL10 protein co-immunoprecipitated with caspase-8 and API2, suggesting an in vivo interaction leading to inhibition of caspase-8 activity. Our current data suggests a novel effect of overexpressed BCL10 in the pathogenesis of high-grade MALT lymphoma by increasing expression of API2 and recruitment of API2 and caspase-8 to form a protein complex leading to the suppression of caspase-8 activity in MZ B cells.

Disclosures:
No relevant conflicts of interest to declare.
Efficacy of 90yttrium-Ibritumomab Tiuxetan in Extranodal Marginal-Zone Lymphoma,
Authors: Vanazzi Anna , Grana Chiara , Crosta Cristiano , Pruneri Giancarlo , Rizzo Stefania , Radice Davide , Pinto Antonello , Calabrese Liliana , Negri Mara , Paganelli Giovanni , Martinelli Giovanni 
Published: October 2019

Abstract
Abstract 3713

BACKGROUND:
To evaluate clinical activity of 90Y-Ibritumomab Tiuxetan in Extranodal Marginal-Zone Lymphoma.
PATIENTS AND METHODS: From May 2004 to April 2011 thirty patients affected by relapsed/refractory Marginal-Zone Lymphoma - arisen at any extranodal site - were enrolled to receive 90Y-Ibritumomab Tiuxetan at the activity of 0,4 mCi/kg. Median age was 57 years (range 36–83 ys); 16 female and 14 male. Thirtheen patients had Helicobacter Pylori-negative gastric Mucosa-Associated Lymphoid tissue Non Hodgkin Lymphoma, while 17 patients had extra-gastric Marginal-Zone Lymphoma. At time of treatment 13 out of 30 patients had disseminated disease (stage III/IV; Ann-Arbor Staging System); 4 patients bone marrow involvement. All patients had received a previous treatment with a maximum of 7. The median number of previous treatments administered - including antibiotics, CT, RT or surgery - was 1 with a maximum of 7; 14/30 had received at least 2 previous treatments, 5 of them more than 3.

RESULTS:
Toxicities were primarily hematological, as expected, and reversible; infection or febrile neutropenia was observed in 3 patients. Up to now no myelodisplastic syndrome occurred. Overall Response Rate was 90%: twenty-three patients achieved a complete response (77%); partial response occurred in 4 patients (13%), stable disease in 2 patients (7%) and 1 progression (3%). With a median follow-up of 47 months, 2 patients relapsed after complete response; 8 out of 20 complete responses are still responder after > 3 years and 5 patients after > 5 years.

CONCLUSIONS:
90Y-Ibritumomab Tiuxetan seems to be active in patients with Extranodal Marginal-Zone Lymphoma resistant or refractory to conventional systemic treatment. These results suggest as a single shot of radioimmunotherapy could represent a possible option for the treatment of relapsed Marginal Zone Lymphoma.

Disclosures:
No relevant conflicts of interest to declare.
The Allosteric AKT Inhibitor MK-2206 Demonstrates Potent Antiproliferative Activity in Lymphoma Cells and Synergizes with the HDAC Inhibitor Vorinostat,
Authors: Buglio Daniela , Lemoine Manuela , Estrella Jaymie , Neelapu Sattva S. , Davis Richard Eric , Doyle Laurence , Little Richard F , Berry Donald , Younes Anas 
Published: October 2019

Abstract
Abstract 3729
The serine/threonine kinase Akt plays a critical signaling role downstream of phosphatidylinositol-3-kinase (PI3K) and is important in promoting cell survival and inhibiting apoptosis. Indeed, Akt activation and overexpression is often associated with resistance to chemotherapy or radiotherapy. Previous studies demonstrated the potential therapeutic value of targeting the PI3K pathway in lymphoma, as both the selective PI3Kδ inhibitor CAL-101, and everolimus and temsirolimus, which target PI3K and mTOR, produce clinical responses in a variety of lymphomas. We evaluated the effect of the novel allosteric Akt inhibitor, MK-2206, in a panel of lymphoma cell lines and primary lymphoma cells. We found that Akt, and activated pAkt, are highly expressed in lymphoma cells. After 72 hours of incubation, the Akt inhibitor MK-2206 demonstrated antiproliferative activity in a variety of lymphoma cell lines, with an IC50 ranging between 0.1 and 5μM. There was no correlation between pre-treatment levels of pAKT, PI3K isoforms, or PTEN protein expression and sensitivity to MK-2206. Within the diffuse large B cell lymphoma cell lines, those of GCB cell of origin were more sensitive to MK-2206, compared with the ABC-derived cell lines. Resistant cell lines tended to had weak or absent expression of p-GSK3 and p-4EBPI. Mechanistically, MK-2206 treatment decreased the level of p-Akt (Ser473), and p-Akt (Thr308), irrespective of drug sensitivity. Furthermore, MK-2206 decreased the phosphorylation level of Akt downstream targets, including p-GSK3 beta and p-PRAS40, upregulated p27. and dephosphorylated p70S6K. Moreover, MK-2206 treatment decreased HIF-1 alpha and VEGF expression. Depending on the cell of origin, the antiproliferative effect resulted from cycle arrest at the G0/G1 phase, autophagy, orapoptosis. MK-2206 showed synergistic effect in combination with the HDAC inhibitor, Vorinostat. Using pathway-specific protein arrays focusing on apoptosis, kinases, and transcription factors, the combination of MK-2206 and Vorinostat effectively altered p53 and p27 levels, which were associated with increased PARP cleavage and induction of apoptosis. Our data demonstrate that AKT is a promising target for the treatment of lymphoma, and provide a rationale for an ongoing trial, evaluating MK-2206 for the treatment of patients with relapsed lymphoma.

Disclosures:
No relevant conflicts of interest to declare.