bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Genes with Prognostic and Therapeutic Relevance in Multiple Myeloma Using Serial Analysis of Gene Expression Technique.
Authors: Felix Roberta S. , Colleoni Gisele W.B. , Vettore Andre L. , Yamamoto Mihoko , Almeida Manuella S.S. , Andrade Valeria C.C. , Chauffaille Maria de Lourdes L.F. , da Silva Wilson A. , Caballero Otavia L. , Simpson Andrew J. , Zago Marco Antonio 
Published: October 2019

Abstract
Serial analysis of gene expression (SAGE) allows a comprehensive profiling of gene expression within a given tissue and also an assessment of transcript abundance.
Objectives: We generated SAGE libraries from normal and neoplastic plasma cells with the aim of identifying genes differentially expressed in MM that could be useful as new potential diagnostic, prognostic markers or therapeutic targets.
Material and methods: Normal plasma cells were obtained from palatine tonsils of 20 children who underwent tonsillectomy. MM SAGE library was obtained from bone marrow plasma cells of two IgGk newly diagnosed MM patients. Purified normal and neoplastic plasma cells were isolated after magnetic sorting of CD-138-positive cells.
Results: We obtained 29,918 SAGE tags from normal and 10,340 tags from tumor libraries. Computer-generated genomic analysis identified 50 overexpressed genes in MM library. After comparison of unique tags expression levels, we selected 12 overexpressed (at least 10 times) genes in the MM library (ZFHX1B, XBP1, PIM2, LGALS1, RANBP2, P53CSV, DDX5, LSM5, JUND, MAPKAPK2, SP140 and NTN1) for further investigation. Expression of 10 out of 12 genes was validated by quantitative real-time PCR in purified plasma cells of 31 MM patients and three controls. XBP1, RANBP2, P53CSV, DDX5, MAPKAPK2 were overexpressed in at least 50% of MM cases. Comparative analyses of relative expression (2-D D CT) of the 10 genes in 31 MM cases and 3 controls showed statistically significant difference between cases and controls for all genes but PIM2 and ZHFX1B, confirming the SAGE data. Also, we could identify that RANBP2 and ZHFX1B have prognostic impact in MM OS (p = 0.040 and p = 0.0502, respectively). Multivariate analysis confirmed RANBP2 expression as a good independent prognostic marker.
Conclusions: SAGE technique allowed the identification of genes differentially expressed in normal and neoplastic plasma cells. These genes may have important role in tumorigenesis or may be possible therapeutic targets for MM control, particularly P53CSV and MAPKAPK2, or prognostic impact, as RANBP2.
Impaired S Phase Arrest in AML Cells with a FLT3-ITD Treated with Clofarabine Allows Prolonged Drug exposure to Reverse the Chemoresistant Phenotype.
Authors: Seedhouse Claire , Grundy Martin , Shang Shili , Ronan John , Pimblett Heather , Russell Nigel H. , Pallis Monica 
Published: October 2019

Abstract
Abstract 1000
Poster Board I-22
We have previously reported that AML cells with a FLT3-ITD have enhanced DNA repair mechanisms following exposure to DNA-damaging drugs which may be a mechanism of chemoresistance. Clofarabine is a novel nucleoside analogue, active in S-phase, with efficacy in AML and is incorporated into DNA as clofarabine triphosphate. Here we show that in FLT3-ITD cells enhanced repair, and therefore resistance to clofarabine-induced DNA damage and toxicity, can be reversed by prolonged drug incubation. When treated with clofarabine, FLT3-ITD-harbouring MOLM13 and MV4.11 cells undergo similar levels of DNA damage (γH2A.X foci) to FLT3 wildtype (WT) cells (HL60 and KG1). After a short pulse of drug the FLT3-ITD cells have a superior repair capability than WT cells; following a 2 hour washout period γH2A.X positivity found immediately after treatment had almost completely disappeared in the FLT3-ITD cells (<10% γH2A.X remaining), whereas in the FLT3-WT cells significant damage (γH2A.X) remained (>40%). Furthermore, after a 1 hour pulse of clofarabine, whereas the FLT3-WT cells under go rapid S phase arrest the S-phase checkpoint fails in the FLT3-ITD cells: reduction in the proportion of cells synthesising DNA is >80% in FLT3-WT cells and <10% in FLT3-ITD cells. Cell cycle arrest in response to DNA damage in S phase is affected via loss of the transcriptional regulator cdc25A. This loss of expression of cdc25A fails to take place in clofarabine-treated FLT3 mutant cells compared to WT cells. In addition, cdc25A mRNA levels are maintained by the FLT3-ITD as demonstrated by siRNA to FLT3 which reduced cdc25A mRNA levels in MV4.11 cells by 87.5%. Primary FLT3 mutant samples from AML patients(n=3) also display impaired cell cycle arrest upon treatment with clofarabine and show enhanced sensitivity on prolonged treatment (24 hours) compared to wildtype samples (n=2). We conclude that there is a reversal of phenotype in mutant FLT3 cells dependant on the length of exposure to clofarabine. Efficient DNA repair renders the cells resistant to a short pulse of the drug, but a failure of cell cycle checkpoint(s) in S phase, mediated by cdc25A, renders the cells sensitive to prolonged exposure. These results may have implications for the scheduling of clofarabine in clinical studies.

Disclosures:
No relevant conflicts of interest to declare.
Prognostic Value of Monosomal Karyotype in Patients with Primary Acute Myeloid Leukemia On Behalf of Spanish CETLAM Group.
Authors: Granada Isabel , Brunet Salut , Hoyos Montserrat , Costa Dolors , Aventín Anna , Marugán Isabel , Vallespí Teresa , Barnues Marta , Duarte JJ , Solé Francesc , Teixidó Montserrat , Mascaró M. , Llorente Andreu , Guardia Ramon , Tormo Mar , Duarte Rafael F. , Millá Fuensanta , Esteve Jordi , Ribera Josep-Maria , Sierra Jordi 
Published: October 2019

Abstract
Abstract 1003
Poster Board I-25

Introduction:
Recently, the cooperative group HOVON-SAKK has refined the prognostic impact of cytogenetic abnormalities in acute myeloid leukemia (AML) by introducing the concept of monosomal karyotype (MK). This consists of ≥ 2 autosomal monosomies or one autosomal monosomy in addition to a structural alteration. In their experience, MK would explain the poor prognosis of AML with a complex karyotype.

Objective:
To investigate the prognostic impact of MK in patients with primary (de novo) AML enrolled in the Spanish CETLAM group protocols (AML 94/99/03). Also, to determine whether considering MK added predictive value to the cytogenetic classification of the Medical Research Council (MRC).

Methods:
Retrospective analysis of data from 1149 AML patients. Chromosomal formula was centrally reviewed with karyotypes being classified by the presence of MK and allocated into the MRC risk categories. Complete remission (CR) rate, disease-free survival (DFS) and overall survival (OS) were calculated.

Results:
The karyotype was assessable in 904 (79%) of the 1149 cases. In 145 of the 904 cases (16%), abnormalities involving CBF gene were detected and in 437 (48%) the karyotype was normal (NK). In 253 (28%) additional patients the karyotype was not monosomal; of them, 61 (24%) belonged to the unfavorable MRC with 17 cases harboring a complex karyotype ≥ 5 abnormalities, 7 cases with rearrangements 3q, 13 cases with -7, 9 cases with 5q abnormalities and 16 cases with t(6;9)). The remaining 69 (7.7%) patients had a MK; of them, 59 (85.5%) were from the unfavorable MRC category and included 43 cases with complex karyotype ≥ 5 abnormalities, 6 cases with rearrangements 3q, 5 cases with -7, 5 cases with alterations of 5q). The following table summarizes the results in terms of CR rate, DFS and OS:

Conclusions:
The addition of MK to the MRC cytogenetic classification refines the prognostic prediction. In our series, the dismal outcome of patients with MK is confirmed; these patients had worse prognosis than those with adverse cytogenetics without MK. Alternative treatment strategies are mandatory for MK+ patients.
Supported in part by grants: GR1-01075, ECO07/90065, PI080672 and RD06/0020/0101.

Disclosures:
No relevant conflicts of interest to declare.
Arsenic Trioxide as First Line Therapy for New Case of APL, Results and Long Term Follow up.
Authors: Ghavamzadeh Ardeshir , Alimoghaddam Kamran , Ghaffari Seyed Hamidolah , Rostami Shahrbano , Jahani Mohamad , Mossavi Seyed Asadollah , Iravani Massoud , Khodabandeh Ali , Baybordi Eissa , Bahar Babak , Jalili Mahdi 
Published: October 2019

Abstract
Abstract 1023
Poster Board I-45
Arsenic Trioxide approved for treatment of APL patients who relapsed after first line treatment failure by ATRA and chemotherapy. Its efficacy as first line treatment studied in at least three trials (including our center reports) previously. Here we are studying its efficacy, safety and long term follow up of new cases of APLwho treated by Arsenic trioxide without ATRA and/or Chemotherapy.

Material and methods:
between 1999 and 2009 we treated 190 new cases of APL by Arsenic Trioxide ( 0.15 mg/kg/day till CR ). Presence of t(15,17) confirmed in all cases by cytogenetic study and/or RT-PCR. In patients who achieved to CR we applied 4 more courses of daily ATO with the same dose for 28 days. Patients followed every 3 months by physical exam, hemogram and RT-PCR on peripheral blood for detection of relapse.

Results:
Median age of our patients was 29 years. Remission rate was 84.7% and median time to achieve to CR was 30 days. Median follow up was 37 months (maximum105 months) and for patients in remission, DFS was 73.4%. Also chance of 3 and 5 years DFS and OS were 71.3%, 83% and 67.6%, 72% respectively. Also Although early mortality was higher in patients with WBC more than 10000 at presentation, we couldn't find any significant correlation between DFS or OS and at presentation WBC count.

Conclusion:
Arsenic trioxide is effective as first line treatment of new cases of APL. Its long term results is comparable with conventional treatment of APL. optimization of treatment needs better supportive care to reduce early mortality due to APL differentiation syndrome and hemorrhagic episodes.

Disclosures:
No relevant conflicts of interest to declare.
Population Pharmacokinetics of Enoxaparin in Children at Risk for Symptomatic Thromboembolism.
Authors: Trame Mirjam Nadine , Mitchell Lesley , Male Christoph , Barrett Jeffrey S. , Hempel Georg , Nowak-Gottl Ulrike 
Published: October 2019

Abstract
Abstract 1071
Poster Board I-93

Introduction:
Enoxaparin, a low-molecular-weight-heparin (LMWH), is used off-label in children to prevent symptomatic thromboembolism when acute anticoagulation or secondary prevention is required due to venous thrombosis or stroke. This investigation was conducted because of concerns of altered pharmacokinetics and a lack of safety and efficacy data when used in children.

Patients and Methods:
Data of 126 children and adolescents with a median age of 5.9 years receiving enoxaparin either as a once or twice daily dosing regimen were analyzed. Children < 12 months of age received a starting dose of 1.5 mg/kg followed by a maintenance dose of 1.3 mg/kg. Children > 12 months of age were started on 1 mg/kg followed by a maintenance dose of 1 mg/kg. Blood samples were drawn after patients reached steady-state on their maintenance dose at baseline prior to the next dose, and at 2, 4, 8 and 12 hours after administration. The median enoxaparin concentration in our population resulted in a median anti-factor Xa activity of 0.4 U/ml (range 0 – 1 U/ml anti-factor Xa). By means of population pharmacokinetics using nonlinear mixed-effects modelling (NONMEM) plasma concentration-time data were analyzed. Several covariates such as age, body weight and body surface area were tested on their effects on the pharmacokinetic parameters.

Results:
Using a two-compartment model the enoxaparin kinetics were described sufficiently. By using body weight and age as covariates for clearance (CL) and central volume of distribution (V1) the best results were obtained. The final population estimates of enoxaparin resulted to be: CL 7.11 ml h-1 kg-1 ± 14.3%, V1 7.31 ml kg-1 ± 33.5%, intercompartimental clearance (Q) 194 ml h-1 ± 24.7%, peripheral volume of distribution (V2) 45.1 l ± 52.5% and absorption rate (ka) 0.0799 h-1 ± 21.7% (estimates ± standard errors). Interindividual variability (IIV) was found to be 75.6% for CL and 78.4% for Q, respectively. Figure 1 shows the predicted activity time-course versus the measured activities for a representative patient. The model is capable of describing all aging and dosing groups of our childhood population (neonates, infants to adolescents).

Conclusion:
The high IIV in CL and Q in our population underlines the need for monitoring the activity and individualizing the dose. Further population pharmacokinetic/-dynamic investigations should be conducted to predict target enoxaparin levels or other new antithrombotic drugs for more safety and efficacy during antithrombotic therapy when used in children.

Disclosures:
Off Label Use: Enoxaparin (LMWH) is used off-label in children to prevent symptomatic thromboembolism..
The Use of Angiotensin Converting Enzyme Inhibitors and Angiotensin Receptor Blockers and the Risk of Developing Venous Thromboembolism in Patients with Atherosclerotic Disease.
Authors: Chae Young Kwang , Khemasuwan Danai , Chebrolu Lakshmi , Gupta Shikha , Neagu Stefan , Carpio Alejandra , Kim Jongoh , Friedman Alan L. 
Published: October 2019

Abstract
Abstract 1078
Poster Board I-100

Objectives:
Arterial and venous thrombosis may share common pathophysiology involving the activation of platelets and inflammatory mediators. A growing body of evidence suggests prothrombotic effect of renin angiotensin system (RAS) including vascular inflammation and platelet activation. We hypothesized that the use of angiotensin converting enzyme inhibitors (ACEIs) or angiotensin receptor blockers (ARBs), therefore, plays a role in protecting against venous thromboembolism (VTE) in patients with history of atherosclerosis. Whether ACEIs and ARBs actually prevents VTE has never been studied in a clinical setting.

Methods:
We conducted a retrospective study, reviewing 596 consecutive patients admitted to Albert Einstein Medical Center (AEMC), Philadelphia with a diagnosis of either myocardial infarction or ischemic stroke during September 2007 to January 2009. Patients were followed up to maximum of 30 months. Patients who had been treated with anticoagulation therapy before or after the first visit at AEMC were excluded. The occurrence of VTE during the follow up period, risk factors for VTE on admission, and use of ACEIs and ARBs during the follow up period were recorded.

Results:
The mean age of the entire study population was 68.1 years. 52.0% of the patients were female and 76.5% were African American. The overall incidence of VTE was 13.4% (n=80); and 68.8% (n= 410) were on RAS inhibitors [either ACEIs only (n=348, 58.4%) or ARBs only (n=89, 14.9%) or both (n=27, 4.5%)]. Among patients on RAS inhibitors, 11.0 % (45/410) developed a VTE, compared with 18.8% (35/186) in the nonuser group [OR (Odd ratio), 0.53; 95% CI (confidence interval), 0.33 – 0.86; P=0.01]. Even after controlling for factors related to VTE (smoking, history of cancer, and immobilization, hormone use) and diabetes, the use of RAS inhibitors was still associated with lower risk of developing VTE [OR, 0.55; 95% CI, 0.34 – 0.90; P=0.02]. Although statitistically not significant due to small sample size, the OR of VTE for ACEI only users were 0.66 [CI, 0.37-1.16, p=0.15], whereas the OR for ARB only users was 0.75 [CI, 0.30-1.85, p=0.53]. Interestingly, Among patients using both ACEI and ARB, no one developed VTE (0/27), compared with 10.9% (38/348) in ACEI only users and 7.9% (7/89) in ARB only users.

Conclusions:
The use of RAS inhibitors appears to be associated with a reduction in the risk of VTE. This possible antithrombotic effect of antagonizing RAS warrants further prospective clinical investigation.

Disclosures:
No relevant conflicts of interest to declare.
Three Novel Alternative Splicing Mutations in BCR-ABL1 detected in CML Patients with Resistance to Kinase Inhibitors.
Authors: Ma Wanlong , Kantarjian Hagop , Cortes Jorge , Zhang XI , Wang Xiuqiang , Zhang Zhong , Yeh Chen-Hsiung , Sferruzza Anthony , O'Brien Susan , Albitar Maher 
Published: October 2019

Abstract
Abstract 1107
Poster Board I-129
The BCR-ABL1 tyrosine kinase is the target for kinase inhibitors in patients with chronic myeloid leukemia (CML). Multiple types of mutations in the BCR-ABL1 kinase domain have been reported. We previously reported a common alternatively spliced BCR-ABL mRNA with a 35-nucleotide insertion (35INS) between ABL1 kinase domain exons 8 and 9 that is expressed at various levels in CML patients with resistance to kinase inhibitors. Here we report 3 novel alternative splicing mutants expressed as the dominant transcripts in patient with CML and resistance to kinase inhibitors. We initially screened for ABL1 kinase domain mutations in approximately 200 patients with resistance to more than one of the three kinase inhibitors (imatinib, nilotinib, or dasatinib). Screening was conducted by first amplifying the fusion BCR-ABL1 transcript, to ensure that normal ABL1 was not amplified, then sequencing the ABL1 kinase domain coding region. We discovered 3 not previously described splice mutants, present in one patient each. All 3 showed >90% mutant transcript. The first resulted from the insertion of 79 nucleotides into the ABL1 exon 8-9 junction. The inserted sequence in fact contained a sequence from regions of intron 8, located 120 bp apart: the 35-nucleotide sequence previously described, and an additional 44-nucleotide segment downstream from 35INS. The combined 79-nucleotide insertion splice mutant showed the same protein change as 35INS (p C475YfsX11). The second splice mutation comprised an 84-nucleotide sequence from intron 7 inserted into the ABL1 exon7-8 junction, also causing a frameshift and protein truncation (p A424EfsX18). The third splice derived from a 231–nucleotide sequence from intron 4 retained in the ABL1 exon 4-5 junction. This insertion added 40 intron-encoded amino acids after K274, leading to a frameshift and early termination of BCR-ABL1 in the p-loop (p E275LfsX41). Exposure to kinase inhibitors may lead to the expression of alternatively spliced mRNA that is significantly elongated but causes a truncated fusion BCR-ABL1 that confers resistance to kinase inhibitors. These findings, when combined with the data on 35INS, support the concept that loss of the C-terminus of BCR-ABL1 is associated with significant resistance to kinase inhibitors; this mechanism appears to be a major source of resistance to kinase inhibitors.

Disclosures
No relevant conflicts of interest to declare.
Old Age Affects Survival but Not Response in Philadelphia Positive (Ph+) Chronic Myeloid Leukemia (CML) Patients Treated with Imatinib (IM): A Study of the GIMEMA CML WORKING PARTY.
Authors: Gugliotta Gabriele , Castagnetti Fausto , Palandri Francesca , Breccia Massimo , Amabile Marilina , Soverini Simona , Testoni Nicoletta , Marzocchi Giulia , Baldazzi Carmen , Annunziata Mario , Usala Emilio , Sica Simona , Tiribelli Mario , Bocchia Monica , Gozzini Antonella , Russo Sabina , Palmieri Fausto , Meneghini Vittorio , D'Emilio Anna , Alimena Giuliana , Martinelli Giovanni , Pane Fabrizio , Saglio Giuseppe , Baccarani Michele , Rosti Gianantonio 
Published: October 2019

Abstract
Abstract 1118
Poster Board I-140

Background
The median age of an unselected population of Ph+ CML patients is close to 60 years. In the prognostic classifications (Sokal, Blood 1984; Hasford, JNCI 1998) that were elaborated before the introduction of IM, age was a significant and important prognostic factor. The most recent IM studies have not clarified the prognostic importance of age and IM therapy is still denied to several elderly patients.

Aim
to asses the relationship between age (less and more than 65 years) and outcome, in CML patients treated front-line in early chronic phase (ECP).

Methods
We analyzed the data of 559 previously untreated ECP patients who were assigned to receive IM 400 mg daily (76%) or 800 mg daily (24%) in three controlled, prospective studies of GIMEMA (Clin Trials Gov. NCT00514488 and NCT00510926; and an observational study of IM 400 mg). The median follow-up is currently 42 (extremes 1 – 64) months. There were 115 patients more than 65 years old (median age 71 years), while 444 (79%) were less than 65 years (median age 46 years). The proportion of patients who were treated with IM 800 mg daily was the same in both age groups.

Results
The cumulative complete cytogenetic and major molecular response rates were identical in the two age groups (88% vs 88% and 82% vs 83%, respectively). However, overall survival (86% vs 93%, p = 0.01), failure-free survival (72% vs 81%, p=0.03) and particularly event-free survival (calculated based on the intention-to-treat principle, where events were any failure [according to the European LeukemiaNet criteria – Baccarani, Blood 2006] and treatment discontinuation for any cause) (60% vs 71%, p=0.006) were significantly inferior in the older age group. All these difference were mainly due to comorbidities leading to more deaths in CP (table).

Conclusions/Methods
These data show that response to IM was not affected by old age. Survival curves were affected because of age-related complications and comorbidities. Age should never be a contraindication to IM treatment. Acknowledgments: European LeukemiaNet, COFIN, University of Bologna and BolognAIL.

Disclosures
Saglio: Novartis: Honoraria. Baccarani:Novartis Pharma: Consultancy, Honoraria, Research Funding, Speakers Bureau; Bristol-Mayer Squibb: Consultancy, Honoraria, Research Funding, Speakers Bureau.