bluuebunny/crossref_metadata_2025_split huggingface dataset:
High Efficiency and Tolerance of Temozolomide in Relapse/Refractory Primary Intra-Ocular Lymphoma (R/R PIOL). a Retrospective Multicentric Study from the LOC Network
Authors: Baron Marine , Gyan Emmanuel , Merle-Beral Hélène , Leblond Veronique , Cassoux Nathalie , Touitou Valerie , Bodaghi Bahram , Darugar Adil , Choquet Sylvain 
Published: November 2019

Abstract
Background: PIOL, now named “primary vitreo-retinal lymphoma”, is a very rare subset of non Hodgkin lymphoma, usually arising in elderly patients and characterized by a high level of relapse, mainly in the first year, with more than 20% of cases relapsing in brain, and a short survival. There is no consensus on treatment procedures, even in first line, and prospective comparative studies do not exist. Classical attitudes are systemic chemotherapy (SC), often high dose methotrexate, radiotherapy or intraocular injection of methotrexate but publication on R/R PIOL are exceptional. New treatments are necessary, especially with a good tolerance profile. As Temozolomide (Te) hassome efficiency on primary-central-nervous-system lymphoma (PCNSL) we used this drug in R/R PIOL in our center.
Methods: Inclusion criteria were R/R PIOL and/or PIOL not eligible for IV chemotherapy. Diagnosis was established on cytological and molecular analysis after vitrectomy, and the absence of brain or meningeal localization verified by MRI and lumbar puncture. Treatment consisted in Te at 150mg/m2 orally 5 days per month, without corticosteroid use, in absence of any response, dosage was increased to 200mg/m². A complete response was defined as a normalization of eye exam and intraocular interleukins 10 and 6 levels.
Results: Sixteen patients were analyzed, 3 males and 13 females, mean age 75 years [35-90]. All but two received systemic chemotherapy with at least high dose methotrexate and high dose cytarabin before Te, 8 were in second line, 4 in third, with 1 patient who relapsed after autologous stem-cell transplantation (ASCT) conditioned by thiotepa, cyclophosphamide and busulfan, and 2 in forth. The 2 patients treated in first line where more than 80 years old. Median duration of treatment was 5 months. The median follow-up (fu) is 16 months. Overall response rate is 75%, with 10 CR (63%) and 2 PR (13%). At the last fu, 7 patients are still in CR, with a median DFS of 13 months. The patient treated after ASCT relapse is still in CR at 62 months. The two old patients treated in first line are in CR at last fu. Two patients where treated a second time by Te after relapse, obtaining 2 new CR, one of 4 months, one persistent at 14 months. Median OS is not reached. Only 3 patients experienced hematological grade 3/4 toxicity.
Conclusion: This work represents the biggest study with an homogeneous treatment in R/R PIOL. Temozolomide appears as a safe and efficient treatment of R/R PIOL or in first line in patients in bad condition, even after high dose chemotherapy and/or in elderly patients. Longer follow-up, prospective and larger studies are necessary to confirm these data

Disclosures
Leblond: Roche: Honoraria, Speakers Bureau.
JAK2 Is a Direct BCL6 Target Gene: Implications for Therapy in Diffuse Large B-Cell Lymphoma
Authors: Barber Ruth , Zobel Jenny , Beck Daniel , Evans Sian , Elliott Richard , Lord Christopher J , Ashworth Alan , Porter Andrew G.C. , Wagner Simon D 
Published: November 2019

Abstract
Increased STAT3 signalling is a factor in driving ~50% of diffuse large B-cell lymphoma. In some cases increased cytokine production by the lymphoma is responsible for activation of JAK2 and STAT3 but the regulation of signalling through this pathway is not clear. We constructed a conditional BCL6 deficient cell line through disruption of the endogenous BCL6 loci of a genetically tractable human B-cell lymphoma by homologous recombination, and insertion of a tetracycline regulatable BCL6 transgene. On induction of BCL6 deficiency growth of the cell line slowed by 3 to 4-fold. A synthetic lethal screen employing a library of small molecule inhibitors in genetically BCL6 deficient lymphoma cells showed that lestaurtinib, a JAK2 inhibitor, enhanced loss of viability. We investigated the hypothesis that JAK2 is a direct BCL6 target gene. JAK2 mRNA and protein expression were induced by BCL6 deficiency. Inspection of the JAK2 proximal promoter region demonstrated a potential BCL6 binding site. BCL6 bound to this sequence in vitro and mutagenesis of the binding site relieved BCL6 mediated transcriptional repression in luciferase reporter assays. Analysis of ChIP-seq data showed a binding peak in the JAK2 proximal promoter region confirming in vivo BCL6 binding. Data from a large and publicly available gene expression dataset confirmed an inverse correlation between BCL6 and JAK2 mRNA whilst there was a positive correlation between JAK2 and STAT3 mRNA. STAT3 is a known target of BCL6 transcriptional repression. We suggest that BCL6 represses both JAK2 and STAT3 and that relatively high BCL6 levels will tend to reduce JAK2-STAT3 signalling whereas lower BCL6 levels will amplify this pathway. Mouse xenografts utilising our conditional BCL6 deficient cell line showed that lestaurtinib alone caused minimum reduction to tumour size but the combination of BCL6 deficiency and JAK2 inhibitor caused growth reduction with central necrosis. In summary we identify JAK2 as a BCL6 target gene and propose that BCL6 has an important role in regulation of JAK2-STAT3 signalling in DLBCL. BCL6 mRNA expression may be a biomarker to enable the rational use of JAK2 inhibitors in DLBCL.

Disclosures
No relevant conflicts of interest to declare.
The Experience of Gruppo Laziale for the Study of Ph-, Chronic Myeloproliferative Syndromes in the Treatment of Myeloproliferative Neoplasms in Blastic Phase with Azacytidine
Authors: Andriani Alessandro , Montanaro Marco , Voso Maria Teresa , Villivà Nicoletta , Ciccone Fabrizio , Andrizzi Cristina , De Gregoris Cinzia , Buccisano Francesco , Alimena Giuliana , Latagliata Roberto 
Published: November 2019

Abstract
Prognosis of patients with myeloproliferative neoplasms developing blast phase (MPN-BP) is very poor with a median survival of about 2-3 months; nowadays, no treatment can induce a durable complete response (CR) in patients who are not candidate to BMT. Very recently, a possible role of azacytidine (AZA) was reported in these patients, but there is still a lack of data as to its efficacy; to address this issue, we evaluated retrospectively 19 patients (M/F 15/4 F, median age 71.3 years, interquartile range (IQR) 64.5 – 77.7] with MPN-BP treated with AZA and collected in the database of our cooperative group. The primary MPN diagnosis was Essential Thrombocythemia in 6 cases (31,6%), Policythemia Vera in 3 (15,8%), Primary Myelofibrosis in 7 (36,8%) and MDS/MPN in 3 (15,8%); the JAK-2 V617F mutation was present in 11/17 patients tested (64.7%). All but 1 patient received previous chemotherapies during chronic phase. Median time from diagnosis to BP evolution was 52.6 months (IQR 11.2 – 181.8). All patients were treated with AZA at the standard dosage of 75 mg/m2: 16 patients received a 7-day schedule and 3 a 5-day schedule every 28 days. Two patients died early after 5-AZA initiation for pulmonary fungal infection and respiratory failure respectively, 4 patients had a disease progression, 4 patients maintained stable disease, 3 patients had a hematological improvement, 1 patient achieved partial response and 5 patients (26.3%) a CR after 4, 4, 4, 5, and 12 months. The median cumulative survival from BP evolution was 9.9 months (95%CI 6.7 – 13.1).A comparison with the historical cohort of 72 patients of our database presenting MPN-BP and treated with approaches other than AZA (median cumulative survival 3.1 months, 95%CI 1.1 – 5.1) showed a significant advantage for patients treated with AZA (p= 0.02). Our data confirm the relative good efficacy and safety of AZA in this group of patients with otherwise dismal prognosis, underlining the possible achievement of long-lasting responses in a sizeable portion of them.

Disclosures
No relevant conflicts of interest to declare.
Mir-148a and Mir-25 Target TULA-2 and Regulate Human Platelet FcγRIIA Signaling
Authors: Zhou Yuhang , Abraham Shaji , Andre Pierrette , Edelstein Leonard C. , Shaw Chad , Tsygankov Alexander , Kunapuli Satya P. , Bray Paul F , McKenzie Steven E. 
Published: November 2019

Abstract
FcγRIIA-mediated platelet activation is important in the pathogenesis of heparin-induced thrombocytopenia (HIT) and other immune-mediated thrombocytopenia and thrombosis disorders. FcgRIIa is also the major transmembrane signaling adapter for the human platelet fibrinogen receptor, integrin aIIbb3. In HIT, antibody to PF4/heparin is necessary but not sufficient for the disease. There is considerable inter-individual variation in platelet FcγRIIA activation; the reasons remain unclear. To identify potential regulators in the FcγRIIA pathway, we determined the platelet RNA expression profile in 154 healthy human donors who were phenotyped for platelet aggregation via FcgRIIA. 87 mRNAs and 4 DE miRNAs were identified as differentially expressed (DE) between hyper- and hypo-responsive donors by microarray and Nanostring analysis, respectively. Among them, hyper-responders show significantly lower T-cell ubiquitin ligand-2 (TULA-2) mRNA level and higher miR-148a-3p and miR-25-3p levels than hyporesponders. TULA-2, also known as UBASH3B, is a known protein tyrosine phosphatase for Syk, and thus a biologically plausible regulator of platelet FcgRIIA.
Both miR-148a-3p and miR-25-3p are predicted to target the 3’UTR region on TULA-2 mRNA (Fig., A). Overexpression of miR-148a-3p or miR-25-3p results in down-regulation of the TULA-2 protein level in HCT116 cells and HEL cells (Fig., B). Inhibition of miR-148a-3p by anti-miR (LNA) leads to elevated TULA-2 expression. A luciferase construct with the 3’UTR miR binding sites was generated. A decrease in luciferase activity was observed in assays with co-transfection of the construct and a miR, supporting the binding between miR-148a-3p and miR-25-3p with TULA-2 3’UTR (Fig., C). Finally, we show that knockdown of TULA-2 caused increased Syk-pY (Y323) level in HEL megakaryocytic cell line in response to FcγRIIA crosslinking by IV.3 (mAb against FcγRIIA) + GAM (Goat anti-mouse F(ab)’2)(Fig., D). These results for the first time show that TULA-2 serves as a negative regulator of the FcγRIIA pathway and that miR-25 and miR-148a may contribute to HIT susceptibility. Studies are in progress for the translation of these findings in vivo.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Comparison of Safety Reports to the FDA from a Pilot Introduction of Peginesatide, a Third Generation Erythropoiesis Stimulating Agent Versus Those from the Usual Care Setting: Manufacturers of Biosimilars and the FDA Should Consider Pilot Introductions of These Agents
Authors: Bennett Charles L. , Jacob Sony , Georgantopoulos Peter , Nichols Judy , Macdougall Iain C 
Published: November 2019

Abstract
Background: Thirty eight anaphylaxis cases were reported to the FDA during the first year of peginesatide marketing in 2012. Marketing was discontinued as a result. We compare 28 reports from a pilot peginesatide introduction conducted by the largest dialysis organization (LDO) in North America versus the 10 reports of anaphylaxis from usual care settings. The pilot introduction was conducted at 10 LDO centers, a nurse was assigned to each center, and staff were educated on peginesatide dosing, safety, pharmacokinetics, and handling.
Methods: Reports in the FDA of anaphylaxis occurring within 30 minutes of peginesatide administration were reviewed for information on administration site, patient characteristics, time to report to FDA, and patient outcome.
Findings: In comparison to 28 anaphylaxis events reported to the FDA from the pilot introduction (as described in Bennett CL et al NEJM 2014), 10 anaphylaxis reports to the FDA from the usual care settings were less often reported as fatal (0% versus 22%) or grade IV severity (10% versus 22%) associated with hypotension (20% versus 57%), or cardiorespiratory arrest (0% versus 29%), and were reported to the FDA later (median of 81 days (range, 14- 172) versus 46 days (range, 4 – 136 days). They were more often associated with clinical findings of diaphoresis (40% versus 18%), syncope (30% versus 18%), or angioedema (20% versus 11%). Onset was a median of four to five minutes after peginesatide infusion in either setting. Among 25,000 peginesatide-treated patients (19,430 in the pilot introduction), anaphylaxis rates were 1.4 per 1,000 persons in either setting.
Conclusion: Clinical characteristics of anaphylaxis events were more serious and FDA reporting more timely for peginesatide-associated anaphylaxis reported to the FDA from the pilot introduction versus usual care settings. With the anticipation of biosimiars in the United States, consideration should be given to requiring pilot introductions of these agents to facilitate safety assessment.

Disclosures
No relevant conflicts of interest to declare.
Selective Inhibition of EZH2 and EZH1 Enzymatic Activity By a Small Molecule Suppresses MLL rearrangement-Induced Leukemia
Authors: Wang Gang Greg 
Published: November 2019

Enhancer of Zeste Homolog-2 (EZH2) and Related EZH1 Control Gene Expression and Promote Oncogenesis Via Methylating Histone H3 Lysine 27 (H3K27). These Methyltransferases Are Proposed to be Therapeutic Targets Due to Their Hyperactive Mutations and Overexpression Found in Cancer Including Various Hematological Malignancies. Here, We Characterized a Set of Small Molecules That Allow Pharmacological Manipulation of EZH2 and EZH1, Which Include UNC1999, a Highly Selective Inhibitor of Both Enzymes, and Its Di-Methylated Analog UNC2400, an Inactive Compound Useful to Assess Off-Target Effects. We Show That UNC1999 Suppresses H3K27 Tri- and Di-Methylation Globally and Inhibits Growth of MLL-Rearranged Leukemia Lines. Mechanistically, UNC1999 Preferentially Affects the Regulatory Elements Such As Enhancers and Distal Promoters, Leading to De-Repression of Many Known Polycomb Target Genes. Gene De-Repression Correlates with Decrease in H3K27me3 and Concurrent Gain in H3K27 Acetylation. UNC2400 Does Not Induce These Effects, Demonstrating UNC1999’s on-Target Inhibition. Oral Administration of UNC1999 Significantly Prolongs the Survival of a Well-Defined Murine Leukemia Model Generated By MLL-AF9. Thus, Our Studies Provide Detailed Profiling of a Set of Compounds for Studying EZH2 and EZH1 in Cancer and Establish Specific Inhibition of EZH2 and EZH1 Enzymatic Activities By Small Molecules As a Promising Therapeutics for MLL-Rearranged Leukemia.

Disclosures
No relevant conflicts of interest to declare.
Value of Procalcitonin Arise Markedly after Use of ATG in Conditioning of HSCT without Value in Differentiating Bacterial and Non-Bacteria Inflammation
Authors: Ge Hui , An Shengli , Li Chunfu 
Published: November 2019

Abstract
Background It is difficult how to differentiate bacterial infections from viral infections and to differentiate bacterial infections from other non-infective systemic inflammation. Especially in the patients with lack of neutrophil bacteria infection is usually life-threaten. The prognosis for those patients is high dependent on early diagnosis and treatment. We still feel confusing in spite of monitoring serum procalcitonin (PCT) and C-reactive protein (CRP) levels with temperature.
Aim To find out whether PCT and CRP are helpful to differentiate bacterial infection and non-bacterial inflammation after use of anti-human T-lymphocyte globulin (ATG) in conditioning of hematopoietic stem cell transplantation (HSCT).
Method Total 60 patients were involved in current study, who underwent HSCT and received conditioning included Cyclophosphamide, Fludarabine, ivBusufan, Thiotepa and ATG (at total dose of 5mg/kg for thymoglobulin, and 15-30mg/kg for ATG-Fresenius, respectively, on day-3 to -1). Blood samples were obtained in the morning from day-3 to 0 for testing PCT and CRP associated by daily recording temperature. Patients were grouped into Group 1 (G1, with antibiotics, n=30), Group 2 (G2, without antibiotics, n=30), Group 3 (G3, bacteria culture positive, n=7) and Group 4 (G4, bacteria culture negative, n=47)
Results The value of PCT was not significant difference when G1 vs. G2 (p=0.061), and G3 vs. G4 (p=0.891), but CRP value and temperature were of significant difference between G1 and G2 (p=0.001 and 0.000, respectively). When comparing G3 with G4, CRP was significant (p=0.021) but temperature was no (0.377). However, the deference of daily CRP value was proved not to be enough to do a differential diagnosis by ROC (AUROC=0.685, 0.665, 0.643 and 0.643 on day-3 to 0, respectively).
Conclusion The current study shows the value of PCT is insignificant to differentiate bacterial infection and non-bacterial inflammation after use of ATG in conditioning of HSCT.

Disclosures
No relevant conflicts of interest to declare.
Direct Contact with Stromal Cells in Association with SDF-1 Is Important for B-Lineage Differentiation Toward CD19+ ProB Cells from Human Hematopoietic Precursors, but Dispensable for Generation of CD7+CD45RA+ Multipotent Lymphoid Precursors
Authors: Minami Hirohito , Ohishi Kohshi , Nakamori Yoshiki , Masuya Masahiro , Katayama Naoyuki 
Published: November 2019

Abstract
The regulatory mechanism of human early B- and T- lymphoid differentiation has not been well studied. Coculture on telomerized human bone marrow stromal cells supported the generation of CD7+CD45RA+ multipotent precursors with differentiation potential for T-, B-, NK-lineage, and monocytic cells, CD10+CD19-CD45RA+ B-biased precursors, and CD10+CD19+CD45RA+ proB cells from human CD34+lin- hematopoietic progenitors. CD7+CD45RA+ and CD10+CD19- lymphoid precursors can be developed from hematopoietic progenitors by soluble factors produced from stromal cells, but the generation of CD19+ proB cells was reduced. Replating analysis showed that direct contact with stromal cells promoted B-lineage differentiation toward CD19+ proB cells from CD34+lin- cell-derived CD7+CD45RA+ and CD10+CD19- lymphoid precursors.
SDF-1 is shown to be critical for B-lineage differentiation from studies of mice. SDF-1 was produced from the human telomerized stromal cells. Inhibition of binding to SDF-1 with neutralizing antibody against CXCR4 suppressed B-lineage differentiation to CD19+ proB cells from hematopoietic precursors, while the generation of CD7+CD45RA+ cells was not significantly affected. By replating analysis, anti-CXCR4 Ab inhibited the differentiation to CD19+ proB cells from CD7+CD45RA+ and CD10+CD19- lymphoid precursors on stromal cells.
These data indicate that direct contact with stromal cells in association with SDF-1 produced from stromal cells is important for B-lineage differentiation toward CD19+ proB cells from CD7+CD45RA+ and CD10+CD19- lymphoid precursors but dispensable for differentiation toward CD7+CD45RA+ multipotent lymphoid precursors.

Disclosures
No relevant conflicts of interest to declare.
Assessing the Mechanism of Cytarabine-Induced Killing in Acute Leukemia
Authors: Vincelette Nicole Dionne , Yun Seongseok 
Published: November 2019

Abstract
The nucleoside analogue cytarabine is part of standard therapy for acute myelogenous leukemia (AML) and is also widely used in various acute lymphocytic leukemia (ALL) regimens. During a 7-day infusion at 100 mg/m2/d, which is part of the “7 + 3” AML induction regimen, steady-state cytarabine concentrations in the 100-200 nM range are achieved. Previous literature suggesting the mechanism of cytarabine-induced killing in AML examined cytarabine concentrations in the micromolar range, which might be less pertinent to routinely achieved drug exposure. To better understand the mechanism of cytarabine-induced killing, AML and ALL cell lines were exposed to varying cytarabine concentrations (25-200 nM) for 24-48 hours and assayed for cell death using a variety of assays. Dose- and time-dependent induction of apoptosis was observed in both AML and ALL cell lines, as detected by phosphatidylserine externalization (Annexin V staining), DNA fragmentation (propidium iodide staining for subdiploid cells), nuclear morphological changes, and cleavage of caspase substrates. Jurkat T cell ALL variants lacking critical components of the death receptor pathway (FADD or procaspase 8) underwent apoptosis with kinetics indistinguishable from parental cells, arguing against involvement of the death receptor pathway. Immunoblotting for Bcl-2 family members in AML and ALL cell lines revealed upregulation of the BH3-only proteins Bim, Puma and Noxa across multiple lines. In further experiments, small interfering RNA (siRNA)-mediated knockdown of Bim, Puma, and Noxa or the mitochondrial proteins Bax and Bak markedly diminished the apoptotic response to cytarabine. These results suggest that cytarabine at nanomolar concentrations kills cells through the mitochondrial apoptotic pathway but, in contrast to more targeted agents, does so by upregulating multiple BH3-only Bcl-2 family members simultaneously. Additional studies to determine the mechanisms of Bim, Puma, and Noxa upregulation are underway. (Supported in part by R01 CA166741 and predoctoral fellowships from the Mayo Foundation for Education and Research.)

Disclosures
No relevant conflicts of interest to declare.
The Clinical Significance of Minimal Residual Disease Detected By Multiparametric Flow Cytometry in Acute Leukemia
Authors: Huang Fen , Jing Hui , Yi Zhengshan , Wei Xiaolei , Zheng Zhongxin , Xie Muchen , Feng Ru 
Published: November 2019

Abstract
Purpose
Minimal residual disease (MRD) detected by multiparametric flow cytometry is an important method for predicting relapse. According to the dynamic changes of MRD, through the first clinical treatment to prevent relapse in patients with elevated MRD levels.
Method
We retrospectively analyze dynamic level of MRD of 75 patients with acute leukemia in our department of hematology from January 2011 to June 2013. According to the retrospective data, to analyze the expression of LAIP and discuss the relationship between changes of MRD and prognosis.
Result
In this study there were 75 patients, including 20 patients with acute lymphocytic leukemia (ALL) and 55 patients with acute myeloid leukemia (AML). The specific LAIP were detected in 49 patients with AML, and the cross lineage and non-synchronous expression were majority. In cross lineage expression, detected frequency of CD45/CD34/CD33/CD13/CD56 was highest, accounted for 67.3% (33/49). In non-synchronous expression, CD45/CD34/CD117/CD33/CD64 accounted for 57.1% (28/49). In 20 patients with ALL, there were 16 patients with B-ALL and 4 patients with T-ALL. The specific LAIP were detected in all of the patients. In patients with B-ALL, detected frequency of CD45/CD19/CD22/CD10/CD13 was highest, accounting for 43.8% (7/16). In abnormal quantity of antigens, CD45/CD19/CD22/CD10/CD38 was the most common type, accounting for 50%. The abnormal expression of CD7/TDT was detected in four patients with T-ALL. In monitoring period 7 patients relapsed. We analyzed the relationship between the clinical data and replase. The results showed that level of peripheral hemoglobin at diagnosis and the times of remission induction were significantly associated with replase (P=0.021 and P=0.017, respectively). To analyze the relationship between change of MRD and prognosis, the results showed that 0.05% as the threshold was significantly related with recurrence. 20 patients of persistent MRD≥0.05% had significant differences with 28 patients of persistent MRD<0.05% in replase free survival (P=0.005).
Conclusion
It has important significance to predict relapse in patients with acute leukemia by detecting minimal residual disease. Patients of MRD ≥ 0.05% should receive early intensive therapy or hematopoietic stem cell transplantation. Even patients without relapse, it should be closely monitor dynamic levels of MRD and highly vigilant extramedullary relapse.

Disclosures
No relevant conflicts of interest to declare.