bluuebunny/crossref_metadata_2025_split huggingface dataset:
Decisional Flow with a Score System To Start Platelet-Lowering Treatment in Patients with Essential Thrombocythemia (ET): Long-Term Results.
Authors: Latagliata Roberto , Rago Angela , Spadea Antonio , Santoro Cristina , Carmosino Ida , Breccia Massimo , Napoleone Laura , Fama Angelo , Biondo Francesca , Volpicelli Paola , Petti Maria Concetta , Alimena Giuliana , Mazzucconi Maria Gabriella 
Published: November 2019

Abstract
Treatment of ET is still a matter of debate for the difficulty in deciding between platelet(plt)-lowering agents and a “wait and see” policy with anti-aggregating agents only: a main problem is to identify what patients are “at risk” to develop severe thrombotic events during the course of the disease. Thus, we prospectively tested in ET patients at diagnosis with no clear indication to plt-lowering treatment a score system based on the following 5 variables: age, plt level, previous thrombotic events, smoking, dismetabolic diseases: the score was reassessed in each patient every 3 months during follow-up. From 04/92 to 03/98, 168 consecutive adult patients (57 males and 111 females, median age 59.5 yrs, range 20.5 – 84.8, median plt value 1,010 x 109/l, range 587 – 2,714) with ET according to PVSG criteria were diagnosed at our Institution. Among them, a plt-lowering treatment was started in 32 patients considered “symptomatic” for the presence at diagnosis or within 6 months before of severe haemorrhages, vascular accident or severe disturbances of microcirculation; plt-lowering treatment was also started in all the 33 patients aged > 70 years at diagnosis. The remaining 103 patients were “asymptomatic” and aged < 70 years and were classified according to our score system: thirty-two patients with a score ≥ 4 started plt-lowering treatment with hydroxyurea early after diagnosis. The remaining 71 patients had a score < 4 at diagnosis and received anti-aggregating agents only: of them, 24 (33.8%) started plt-lowering treatment during follow-up (18 for a score increase to ≥ 4 and 6 for the occurrence of thrombotic complications or symptoms) after a median time from diagnosis of 28 months (range 3–130) while 47 (66.2%) did not start any plt-lowering treatment. Thrombotic complications occurred in 9/103 patients (8.7%) scored at diagnosis: in particular, they occurred in 4/32 patients (12.5%) with score ≥ 4 receiving plt-lowering treatment since diagnosis and in 5/71 patients (7%) with score < 4 under anti-aggregating agents only. Overall, 6 out of 103 scored patients died and 27 were lost to follow-up, with a 15-year cumulative survival of 88%. In conclusion, a decisional flow based on present score system appears effective in patients without a clear indication for plt-lowering treatment at diagnosis to discriminate subjects at different risk of thrombotic events and could be useful to decide when a plt-lowering therapy needs to be started.
JAK2 V617F Mutation Testing in Polycythemia Vera: Use and Impact in a Single Academic Practice.
Authors: Means Robert T. 
Published: November 2019

The observation that a gain-of-function mutation in JAK2 (JAK2 V617F) is present in more than 90% of cases of polycythemia vera (PV) has substantially revised the diagnostic approach to this disease. However, most studies of the utility of JAK2 V617 derive from large PV populations: the majority of hematologists have relatively few PV patients in their individual practices. In order to assess the effect of JAK2 V617F analysis in practices with a small PV population, the utilization and impact of this test in 5 individual clinics in a single academic practice were analyzed. Sixteen patients (15 male) carrying a diagnosis of PV were identified. Five patients were followed in a clinic at a University medical center staffed by a single attending hematologist; one was followed by that attending physician in a clinic at the affiliated VA hospital; and 10 were followed by Hematology/Oncology Fellows in three continuity clinics at the affiliated VA hospital. In these latter clinics, the Fellow is primary provider, but reviews management with an attending physician. Median age of the overall population was 67.5 yrs; patients in the Fellow clinics were significantly older than those in the Attending clinics (80 yrs vs. 63 yrs; p < 0.01). Duration of disease was longer in the Fellow clinic patients than the Attending patients (8.5 yrs vs. 1.4 yrs; p=0.05). Nine patients were tested for JAK2 V617F mutation. The majority (6/9) were tested near diagnosis (median duration of erythrocytosis was 0.7 yrs for patients tested and 10 yrs for patients not tested; p = 0.02). Two of the three established patients tested did not meet PV diagnostic criteria fully. Five of the 7 not tested met WHO criteria for PV. Of the 9 patients tested, 7 had a mutation detected (5 newly diagnosed, 2 > 1 yr duration of disease). Results of mutation testing changed the diagnosis in two cases (one where it was detected and one where it was not detected). In addition, one newly diagnosed patient who met WHO criteria did not have JAK2 V617F detected and was felt to have true “JAK2 V617F-negative” PV. JAK2 exon 12 studies were not performed.
Conclusions: The use and impact of JAK2 V617F mutation testing has been reviewed in a PV population similar to what an individual or small group practice might follow. JAK2 V617F mutation testing was primarily used in the early evaluation of suspected PV, or in continuing patients whose diagnosis was not fully established. Mutation testing was infrequently used in continuing patients where the physician considered the diagnosis clearly established or where a change in diagnosis would not alter management. Negative results of mutation testing altered diagnosis if they were consistent with the patient’s clinical picture.
CD26 Expression in Mature B-Cell Neoplasia: Its Possible Role as a New Prognostic Marker in B-CLL.
Authors: Cro Lilla , Zucal Nadia , Fabris Sonia , Neri Antonino , Lionetti Marta , Morabito Fortunato , Cutrona Giovanna , Ferrarini Manlio , Molica Stefano , Deliliers Giorgio Lambertenghi , Baldini Luca 
Published: November 2019

Abstract
CD26 (dipeptidyl peptidase IV, DPP IV) is a multifunctional type II cell surface glycoprotein that is widely expressed on T and natural killer cells, as well as on epithelial, endothelial and acinar cells of different tissues; its expression on B cells is very low but it is greatly upregulated following activation. We evaluated, by means of flow cytometry, the expression of CD26 in various types of B-cell lymphoid tumors: Follicular Lymphoma (Fo-Ly, 12 cases), Mantle cell Lymphoma (MCL, 12 cases) Multiple Myeloma (MM, 20 cases), Hairy cell Leukemia (HCL, 12 cases), B-cell Chronic Lymphocytic Leukemia (B-CLL, 112 cases), CD5 negative B-cell Chronic Lymphoproliferative Diseases (CD5neg-B-CLPD, 20 cases) and Diffuse Large cell Lymphoma (DLCL, 12 cases). CD26 expression was absent or low of Fo-Ly and MCL, high on MM and HCL, variable on B-CLL, CD5neg-B-CLPD and DLCL. Fluorescence intensity of positive cells was dim in B-CLL and CD5neg-B-CLPD, heterogeneous in DLCL, and bright in HCL and MM. Interestingly, in CLL patients, CD26 expression was significantly correlated with CD49d and CD38 expressions (p<0.0001); moreover Spearman’s test revealed a significant correlation between CD26, CD38 or ZAP-70 expression and IgVH mutational status (p<0.0001). We also found a significant correlation between CD26 expression and the presence of FISH chromosomal abnormalities grouped on the basis of their prognostic relevance (p<0.0001). After a median follow-up of 36 months, 65/107 CLL pts were treated; taking 10% of positive cells as best cut-off, Kaplan-Meier curves showed that the time without therapy was significantly longer for CD26 negative cases compared with CD26 positive cases (P <0.0001). Moreover the concordant negativity of CD26 and CD38, or ZAP-70 and the combination of CD26 negativity with IgVH mutated status, identified subsets of CLL patients with a favourable outcome in terms of need of therapy.
Modulation of Gene Expression Profile and In Vivo Anti-Myeloma Activity Induced by Valproic Acid, a Histone Deacytylase Inhibitor.
Authors: Neri Paola , Calimeri Teresa , Di Martino Mariateresa , Rossi Marco , Eramo Orietta , Propato Mariagrazia , Perri Gino , Pietragalla Antonella , Ventura Monica , Bulotta Alessandra , Blotta Simona , Tagliaferri Pierosandro , Anderson Kenneth C. , Munshi Nikhil C. , Tassone Pierfrancesco 
Published: November 2019

Abstract
Valproic acid (VPA) is a well-tolerated anticonvulsant drug that has been recently recognized as powerful histone deacetylase (HDCA) inhibitor. VPA induces hyperacetylation of histone H3 and H4 and inhibits both class I and II HDCACs. Recently it has been shown that VPA exerts in vitro and in vivo anti-tumor activity against solid cancers and its in vitro anti-Multiple Myeloma (MM) activity has been previously reported. However, the molecular mechanisms are still unclear. Here we have investigated molecular changes induced by VPA as well as its in vivo activity in murine models of MM. We first studied the in vitro activity of VPA against IL-6 independent as well as IL-6 dependent MM cells. A time- and dose-dependent decrease in proliferation and survival of MM cell lines was observed (IC50 in the range of 1–3 mM). Gene expression profile following treatment with VPA at 2 and 5 mM showed down-regulation of genes involved in cell cycle regulation, DNA replication and transcription as well as up-regulation of genes implicated in apoptosis and chemokine pathways. The signaling pathway analysis performed by Ingenuity Systems Software identified the cell growth, cell cycle, cell death as well as DNA replication and repair as the most important networks modulated by VPA treatment. We next evaluated the in vivo activity of VPA using two xenograft models of human MM. A cohort of SCID mice bearing subcutaneous MM1s or OPM1 were treated i.p. daily with VPA (200 mg/kg, and 300 mg/kg, n=5 mice, respectively), or vehicle alone (n=5 mice) for 16 consecutive days. Tumors were measured every 2 days, and survival was calculated using the Kaplan Mayer method. Following VPA treatment, we found a significant (p=0.006) inhibition of tumor growth in mice bearing subcutaneous MM-1s cells treated with VPA at 200 mg/kg compared to control group, which translated into a significant (p= 0.002) survival advantage in the VPA treated animals. Similar results were obtained in animals bearing subcutaneous OPM1 cells. Flow cytometry analysis performed on retrieved tumor tissues from animals showed reduction of G2-M and S phase in tumor specimens following VPA treatment, versus untreated tumors, strongly suggesting in vivo effects of VPA on cell cycle regulation. Taken together, our data demonstrate the in vitro and in vivo anti-tumor activity of VPA, delineate potential molecular targets triggered by this agent and provide a preclinical rationale for its clinical evaluation, both as a single agent or in combination, to improve patient outcome in MM.
ITF2357, a Novel Histone Deacetylase Inhibitor, Demonstrates Significant Apoptotic Activity Against Human Multiple Myeloma Cells.
Authors: Kline Michael , Donovan Kathleen A. , Lust John A. 
Published: November 2019

Abstract
We have evaluated the efficacy of a novel hydroxamic acid-derived histone deacetylase (HDAC) inhibitor, ITF2357, to promote cell death in multiple myeloma (MM) cells. HDAC inhibitors, which promote histone hyperacetylation and increase gene expression, have been evaluated as candidate agents for combating malignancies because they impact the expression of genes related to proliferation, differentiation, and survival. Exposure of MM cell lines to 1 micromolar ITF2357 led to dramatically increased levels of histone acetylation at 4 hours and 8 hours by Western analysis. Sub-micromolar concentrations of ITF2357 promoted time- and concentration-dependent cell death in MM cell lines. Using 500 nM ITF2357, a concentration potentially achievable in vivo, viability of KAS-6/1 IL-6 dependent myeloma cells was reduced to 28% of control at 24 hrs and 2% of control at 48 hours (Figure 1). In contrast, viability of normal PBMCs was 100% at 24 hours and 80% at 48 hours (Figure 2). U266 and 8226 myeloma cells were found to be sensitive to ITF-2357 in a similar fashion with U266 being least sensitive. Cell death proceeded via apoptosis as measured using Annexin V/propidium iodide staining. ITF 2357 was superior to suberoylanilide hydroxamic acid (SAHA) at inhibition of stromal cell IL-6 production. IL-1beta (10 pg/ml) was used to stimulate bone marrow stromal cell IL-6 production (105 ng/ml) after 48 hours. Concentration of ITF2357:Stromal Cell IL-6 production after 48 hours were as follows - 10 nM: 78 ng/ml; 100 nM: 79 ng/ml; 1000 nM; 32 ng/ml. SAHA at similar concentrations showed no significant decrease in stromal cell IL-6 production compared with the no drug control. In summary, ITF2357 induces significant myeloma cell apoptosis and can inhibit stromal cell IL-6 production. It represents an attractive therapeutic candidate for MM clinical trials.
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Anti-Acute Myeloid Leukemia Activity of Chaetocin, a Novel Epigenetic Drug Inhibitor Inducing Oxidative Stress.
Authors: Chaib Hassiba , Prebet Thomas , Restouin Audrey , Castellano Remy , Opi Sandrine , Olive Daniel , Borg Jean Paul , Vey Norbert , Collette Yves 
Published: November 2019

Abstract
Recent studies have highlighted the importance of epigenetic modifications in the pathogenesis of Acute Myeloid Leukemia (AML). This results have been confirmed by the activity of new drug like DNA demethylating agents and histone deacetylase (HDAC) inhibitors in both in vivo and in vitro studies. Recently, Chaetocin, a natural fungal compound, has been identified as the first specific inhibitor of the histone methyltransferase SU(VAR)3–9 which plays a role in heterochromatin gene silencing. In this study, we decided to evaluate Chaetocin as a therapeutic agent in AML in vitro and to explore the related mechanisms. We show that Chaetocin induce dramatic cell death at nanomolar concentrations in U937 and HL60 (97.2% ± 0.4 and 91.6% ± 9 cell death at 100 nM chaetocin, respectively), and to a lesser extend in K562 (67.3% ± 1.6 cell death at 100 nM chaetocin), cell cultures. Cell death occurred at 24 h incubation time which correlated with induction of apoptosis as assessed by Annexin V/7-AAD staining and activation of downstream executioner caspase-3/7. Using transcription low-density array and quantitative RT- PCR, Chaetocin was showed to up-regulate gene transcription such as of the cell cycle inhibitor p21/WAF1 consistent with a role for the targeted SU(VAR)3–9 in heterochromatin gene silencing. In agreement with the recent report of Chaetocin being a promising new antimyeloma agent acting via imposition of oxidative stress, intracellular levels of oxidative species were increased in Chaetocin treated U937 cells in a time- and dose-dependent manner that correlated with induction of cell death. Furthermore, incubation of cells with N-acetyl cysteine, a cell-permeable precursor of intracellular glutathione reductant, prevented chaetocin-induced accumulation of oxidative species, transcription of selected genes (e.g. p21/WAF1), activation of caspase-3, and cell death. Finally, Chaetocin was found to increase the antileukemia activity of HDAC inhibitors and Aracytin, and thus appears as a promising agent for further study as a potential anti-AML therapeutic. Preliminary results obtained in vivo in xenograft models and ex vivo, using blasts of a panel of patients with AML, will be presented.
rNAPc2, a Novel Inhibitor of Tissue Factor/Factor VIIa Complex, Inhibits Tumor Growth and Metastasis in Mouse Models of Colorectal Cancer.
Authors: Zhao Jingsong , Aguilar Gerard , Imperiale Michael , Funk Walter , Abo Arie 
Published: November 2019

Abstract
Recombinant nematode anticoagulant protein c2 (rNAPc2) is a specific inhibitor of tissue factor (TF)/factor VIIa complex with novel anti-metastatic, anti-angiogenic, and anti-thrombotic activities. TF is highly expressed in human colorectal tumors and the level of TF expression positively correlates with disease stage and inversely correlates with survival. To explore the therapeutic potential of rNAPc2 during tumor growth and metastasis, we tested rNAPc2 efficacy in experimental colorectal cancers in mice. Administration of rNAPc2 inhibited pulmonary metastasis in mice systemically disseminated with CT26 murine colon carcinoma cells in a dose-dependent fashion, as measured by either number of lung surface metastases or lung mass. While rNAPc2 treatment alone moderately reduced primary tumor growth, combining rNAPc2 with the cytotoxic agent 5-fluorouracil (5-FU) resulted in synergistic growth inhibition of HCT116 human colorectal tumor xenografts in nude mice. Likewise, rNAPc2 further reduced tumor growth in HCT116 human colorectal tumor xenograft-bearing mice receiving bevacizumab (humanized anti-vascular endothelial growth factor monoclonal antibody). Using CD31 and Ki67 immunohistochemisty, we found that rNAPc2 synergized with either 5-FU or bevacizumab in inhibiting microvessel density and tumor cell proliferation in HCT116 human colorectal tumor xenografts. Furthermore, rNAPc2 synergized with CPT-11 in inhibiting hepatic metastasis in nude mice with portal vein injection of HCT116 human colorectal tumor cells. Long-term administration of rNAPc2 also significantly suppressed formation of intestinal adenomas and adenocarcinomas in ApcMin/+ mice. The dosing regimens of rNAPc2 used in these studies were well tolerated up to a three-month period by recipient mice without major hemorrhage or other adverse effects. In conclusion, the synergistic tumor inhibitory activity of rNAPc2 in pre-clinical colorectal cancer models suggests that rNAPc2 may be an effective anti-tumor agent in human colorectal cancer patients to potentiate chemo- or anti-angiogenic therapies.
High Frequencies Of Switching To 2nd TKIs and Failure To Maintain Standard Imatinib Dose In Japanese CML Patients With BIM Genetic Variants
Authors: Katagiri Seiichiro , Tauchi Tetsuzo , Umezu Tomohiro , Ohtsuki Kazuhiro , Tadokoro Kenichi , Yamamoto Yoshinori , Ohyashiki Junko H , Ohyashiki Kazuma 
Published: November 2019

Abstract
Recently, it has been demonstrated that the proapoptotic protein BIM showed a deletion polymorphism at exon 3 in eastern Asian population, and some CML patients with the BIM deletion polymorphism are resistant to imatinib treatment (Ng et al. Nature Medicine, 2012). More recently, a BIM single nucleotide polymorphism (SNP) at exon 8 (c465C>T) has also been found in French CML patients and this SNP is associated with not only imatinib resistance but also the presence of BCR-ABL mutations (Mahon et al. ASH abstract, 2012).
We aimed to investigate a possible association between such genetic variations of BIM and clinical manifestation in Japanese CML patients who experienced undetectable minimal residual disease (UMRD: so-called CMR4.5). In this study, we newly analyzed BIM SNP (c465C>T) in 47 CML-UMRD patients with known BIM deletion polymorphism status (Katagiri et al. Br J Haematol, 2013). Twenty normal subjects were used as controls.
The frequency of either BIM SNP at exon 8 or BIM deletion polymorphism did not deviate from the normal subjects in the Japanese population (P = 0.7597 and P = 0.2880, respectively). None of the subjects showed both BIM SNP at exon 8 and BIM deletion polymorphism concomitantly. We then compared the clinical features among 3 CML-UMRD groups: patients with BIM SNP, patients with BIM deletion polymorphism, and patients who showed neither BIM SNP nor BIM deletion polymorphism (no genetic variations). The frequency of CML patients who maintained 400 mg imatinib dose until stopping was significantly higher in those without genetic variations than in those with BIM SNP or BIM deletion polymorphism (P = 0.0002). Moreover, the frequency of CML patients who switched to second tyrosine kinase inhibitors (2nd TKIs) was significantly higher in those with BIM SNP or BIM deletion polymorphism than in those without such polymorphisms (P = 0.0055).Number of CML patientsMaintained IM 400 mgChange of imatinib dose2nd TKIs switchingBIM SNP (c465C>T)11/474/115/112/11BIM deletion polymorphism6/471/63/62/6BIM SNP or deletion polymorphism17/475/178/174/17No BIM genetic variationss30/4725/305/300/30
This is apparently the first study to circumstantiate the BIM genetic variants in Japanese CML patients with UMPD. Although the number of patients is small, our results suggest that CML patients without BIM deletion polymorphism/SNP could be maintained under standard imatinib dose without switching to 2nd TKIs, and thereby, have a possibility to stop TKIs therapy.

Disclosures:
Ohyashiki: Norvartis KK: Research Funding, Speakers Bureau; Bristol Meyer Squibe KK: Research Funding, Speakers Bureau.
Rapid Molecular Diagnosis Of JAK2V617F Negative MPN By Targeted Deep Sequencing Using The Ion Torrent PGM
Authors: Klose Nathan , Tallack Michael R , Magor Graham , Mollee Peter , Perkins Andrew C 
Published: November 2019

Abstract
Myeloproliferative neoplasms (MPN) are a heterogeneous group of blood disorders characterized by excess production of mature blood cells, increased risk of thrombotic complications and slow progression to myelofibrosis or, less often, leukemia. Activation of the JAK-STAT signaling pathway is a common underlying feature of these diseases and JAK kinase inhibitors are efficacious in the more advanced forms of disease. Most cases of polycythemia vera (PV) and approximately 60% of essential thrombocythemia (ET) and primary myelofibrosis (MF) harbor a point mutation in JAK2 (V617F) which leads to constitutive JAK-STAT signaling and factor independent cell growth. The remaining 40% of cases of MF and ET harbor a broad range of mutations in many genes including those involved in cytokine receptor signaling, other components or the JAK-STAT pathway or epigenetic regulators. This poses a challenge for rapid molecular diagnosis. Also, since ET is essentially a diagnosis of exclusion of reactive causes of thrombocytosis, many cases of chronic ‘ET’ may not be clonal hematological neoplasms but reactive conditions.
We have developed a rapid deep sequencing pipeline to detect mutations in 65 genes which have been implicated in MPN through previous reports of human mutations, mouse models of MPN, or other known components of hematopoietic cytokine receptor signaling. We used 10ng of DNA from blood to amplify and sequence all the exons of these 65 genes using Ampliseq and Ion Torrent PGM. Using 318 PGM chips and 8-fold multiplexing we achieved on average 200 fold coverage of the target exome. The bioinformatics of SNP validation and rapid generation of reports will be presented. From a pilot study of 30 cases referred for molecular diagnosis, we have detected the likely causative mutation in approximately 80% of ET and MF where JAK2 is wild type. Many of these mutations are known to be causative in MPN, including those in MPL, ASXL1, SET2, SH2B3 (LNK), EZH2, CBL, DNMT3A and other genes. We have identified a novel inherited mutation in a family with MPN and validated it in BAF3 factor-independency assays. We have identified further novel mutations in JAK3, EED, DNMT3A, APC and two phosphatases involved in silencing activated JAK-STAT pathway components. The biological significance of these is under investigation and progress will be reported. In many cases we find evidence for clonal evolution involving secondary mutations in epigenetic modifying proteins on top of driver mutations in the JAK-STAT pathway.
In short, targeted exome re-sequencing using Ampliseq and Ion Torrent PGM provides a rapid and relatively cheap method for molecular diagnosis and characterization of most cases of MPN.

Disclosures:
Perkins: Novartis Oncology: Consultancy, Honoraria, Membership on an entity’s Board of Directors or advisory committees.