bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of New CD177 Isoform in NB1 Deficient Individuals
Authors: Bayat Behnaz , Kissel Karin , Werth Silke , Santoso Sentot 
Published: April 2020

Abstract
Human neutrophil-specific antigen CD177 (NB1; HNA-2a) belongs to the member of the cyctein-rich LY-6 superfamily. Transcriptional analysis of this family revealed intron-retaining mechanism, which produce many spliced forms of gene. In some cases, this mis-spliced isoform was the most abundant form suggesting a control mechanism of gene expression. CD177 glycoprotein carries neutrophil antigen HNA-2a, and antibodies to HNA-2a frequently causes transfusion related acute lung injury (TRALI). Recently, CD177 has been found to be associated with the expression of proteinase-3 (PR3) which plays a role in autoantibody mediated Wegener’s granulomatosis. Neutrophils from some people lack CD177 (termed HNA-2a negative), however, the mechanism responsible for this deficiency is not fully understood. In this study, we investigated whether mis-spliced form of CD177 exist in neutrophils. Neutrophil from blood donors were phenotyped for the presence or absence of CD177 by flow cytometry using two mabs 7D8, MEM166 recognising different epitopes on CD177. HNA-2a positive individuals carrying high surface CD177 density and HNA-2a negative individuals were selected. mRNA was isolated from CD177 phenotyped individuals and were analyzed by PCR using different sets of primer set. Amplification of CD177 transcript (bases 98–1401) showed the expected ~1300 bp band encoding for the entire CD177 (isoform 1), and a smaller band (~417bp) with similar intensity in HNA-2a positive as well as in HNA-2a negative individuals. Sequencing analysis of the short transcript showed the presence of intronic sequence between exon 3 and 4, which produces premature stop codon. Transfection of insect cells with related transcript resulted in production of a ~23 kDa protein which is detectable in immunoblot using rabbit polyclonal antibody against synthetic CD177 peptide. This ~23 kDa protein (isoform 4) did not react with mabs 7D8 and MEM166. Furthermore, immunoblotting analysis of other blood cells showed that isoform 4 was exclusively found on neutrophils. In contrast to the isoform 1, isoform 4 was not upregulated on neutrophils of individuals receiving GCSF. Neutrophils treated with fMLP in vitro showed down-regulation of isoform 4 transcript. This phenomenon however, has not been observed in neutrophils treated with LPS and IL-8. In summary, we characterized a new CD177 isoform in neutrophils. The presence of this isoform in both HNA-2a positive and negative CD177 phenotyped individuals and its regulation with fMLP indicate the role of this protein in inflammatory process.
Impact of Gene Expression Profiling on Diagnosis and Prognostication in Cytogenetically Normal AML.
Authors: Bullinger Lars , Hielscher Thomas , Botzenhardt Ursula , Heinrich Sabrina , Schlenk Richard , Pollack Jonathan R. , Dohner Konstanze , Benner Axel , Dohner Hartmut 
Published: April 2020

Abstract
Cytogenetically normal acute myeloid leukemia (CN-AML) comprises a biologically and clinically heterogeneous group of AML. In the past years, molecular markers like FLT3, CEBPA and NPM1 gene mutations have been identified in CN-AML, and the presence of such mutations carries important prognostic information. Furthermore, DNA microarray-based gene expression profiling (GEP) has been shown to capture the molecular heterogeneity of cancers, and has been applied to build classifiers and clinical outcome predictors in AML.
While prior studies have defined gene expression patterns associated with NPM1, CEBPA, and FLT3, we assessed the clinical relevance of gene signatures. We profiled a large set of clinically well annotated CN-AML specimens (n=296 entered on two multicenter trials for patients &lt;60 years (AMLSG HD98A and AMLSG 07-04). The 142 cases from the AMLSG HD98A trial were analyzed using a 40k cDNA microarray platform and the 154 cases from trial AMLSG 07-04 using Affymetrix microarrays (Human Genome U133 Plus 2.0 Arrays). In this data set we applied supervised analyses (LASSO penalized logistic regression) to define gene expression patterns characterizing FLT3 internal tandem duplication (ITD), CEPBA and NPM1 mutations as well as outcome signatures.
We were able to define distinct signatures associated with NPM1, CEBPA, and FLT3 consisting of 39, 27, and 47 genes, respectively. The NPM1 signature revealed a high prediction accuracy of &gt;95% in leave-one-out cross validated classification. Prediction of FLT3-ITD or CEBPA mutation performed less well with accuracies of 80% and 73%, respectively. However, for both CEBPA and FLT3-ITD the predicted mutation class labels performed slightly better than the marker itself with regard to the prognostic impact on overall survival (CEPBA: p=0.006 vs. p=0.007, FLT3-ITD p=9.57e-06 vs. p=5.11e-05; logrank test). In addition, using LASSO we also could define a signature associated with event free survival (EFS) in the cases from the AMLSG 07-04 trial. Adjusted for age, NPM1, and FLT3-ITD mutational status this signature was significantly associated with EFS (p=0.005; Wald test), and validation in our independent cDNA data set also provided significant prognostic information (p=0.02; Wald test).
Thus, GEP-based classification of CN-AML might help to identify alternative genetic changes that either phenocopy or block the effects of common molecular aberrations. Furthermore, gene expression patterns of yet unknown aberrations are reflected in prognostic signatures. Therefore, signature genes also provide a starting point to dissect “mutations” pathways, and our findings underscore the potential clinical utility of a gene expression based measure in CN-AML.
The Efficacy and Safety of Lenalidomide Oral Monotherapy in Patients with Mantle Cell Lymphoma Previously Treated with Bortezomib: Pooled Data from Two Phase II Studies (NHL-002 and NHL-003).
Authors: Reeder Craig B. , Witzig T.E. , Vose Julie M. , Zinzani Pier Luigi , Buckstein Rena , Haioun Corinne , Bouabdallah Reda , Polikoff Jonathan , Guo Pingshan , Ervin-Haynes Annette , Pietronigro Dennis , Zeldis Jerome B. , Czuczman Myron S. 
Published: April 2020

Abstract
Introduction: Mantle-cell lymphoma (MCL) is an aggressive B-cell lymphoma that if not cured with aggressive chemoimmunotherapy and stem cell transplant is usually fatal. The intravenous proteasome inhibitor bortezomib produces an overall response rate (ORR) of 33% in patients with relapsed MCL (J Clin Oncol2006;24(30):4867–74). Unfortunately, most patients eventually relapse and new agents are needed for this disease. Two recently conducted phase II trials (NHL-002 and NHL-003) tested single-agent lenalidomide for patients with relapsed MCL. Fifty-four patients with MCL were enrolled into the two studies; 26% (14/54) had received prior bortezomib and they are the subject of this report.
Methods: Patients with relapsed or refractory MCL and measurable disease 2 cm after at least one prior treatment regimen including prior bortezomib were eligible. Patients received 25 mg of lenalidomide orally once daily on days 1–21 of every 28-day cycle. Therapy was continued as tolerated or until disease progression. The 1999 IWLRC methodology was used to assess response and progression.
Results: The 14 patients in this study were heavily pretreated with a median of 4 (2–6) prior treatments (including bortezomib) and 50% were bortezomib-refractory. Median age was 66 (45–84) years and 6 (43%) patients were female. Median time from diagnosis to lenalidomide treatment was 3.3 (0.7–7.6) years. The ORR with lenalidomide was 57% (8/14), including 21% (3/14) complete response (CR)/unconfirmed CR and 36% (5/14) partial responses. One (7%) patient had stable disease. The most common grade 3 or 4 adverse events were neutropenia (50%), thrombocytopenia (43%), anemia (21%), fatigue (21%), and leukopenia (21%). Neutropenic fever occurred in 7% of patients.
Conclusion: These results demonstrate that lenalidomide oral monotherapy is very effective with manageable side effects in patients with relapsed or refractory MCL who had received prior treatment with bortezomib.
Plitidepsin Is Active in Peripheral T-Cell Lymphoma (PTCL): A Subset Analysis from An Ongoing Multicenter Phase II Trial.
Authors: Ferme Christophe , Mateos Maria Victoria , Szyldergemajn Sergio , Zucca Emanuele , Espinoza Jose , Briones Javier , Morschhauser Franck , Gisselbrecht Christian , Ribrag Vincent 
Published: April 2020

Abstract
Background: Peripheral T-cell lymphomas (PTCL) represent a small (&lt;10%) yet a particularly aggressive subset of NHL. Up to 75% of those patients (pts) eventually become relapsed/refractory, with no effective options available. Thus, a prospective open-label, multicenter phase II study to evaluate the activity of plitidepsin (Aplidin®) in adult pts with relapsed/refractory aggressive lymphomas was planned and is currently ongoing. We report the preliminary results from a cohort of non-cutaneous PTCL.
Patients and methods: As of July 2008, 14 pts were treated with plitidepsin 3.2 mg/m2 i.v. infusion over a 1-h on days 1, 8 and 15 q4wk. Eleven pts are evaluable, one is too early and two were non-evaluable as per protocol criteria: one had a hypersensitivity reaction and one had bone marrow (BM) infiltration only, but achieved a biopsy-documented BM clearance. Pts had a median of 3 (1–6) previous regimens, including 4 pts (33%) with prior autologous transplantation. Lymphoma histology: 8 PTCL-nos, 2 anaplastic large-cell, 2 angioimmunoblastic and 2 NK/T nasal type. Ten pts were male, median age was 56 y (35–74), with performance status 0 in 6 pts, 1 in 5 pts and 2 in 3 pts.
Results: One confirmed CR and 3 PR were observed for a 36% objective response rate (95% CI: 11%–70%). Median duration of response was 4 months (range: 1+ – 12+). Median overall survival was 11 months (range 1+ – 24+). Plitidepsin was tolerable in this heavily pre-treated population, particularly with low hematologic toxicity. Of 2 cases of grade (G) 4 neutropenia, 1 was already present at baseline and only 1 pt developed G3 thrombocytopenia during treatment. Transient and reversible G3 ALT/AST elevations occurred in 7 patients and 2 G3 bilirrubin increases. Clinical toxicities mainly consisted of mild to moderate muscular weakness, myalgia and cramps, plus G1-2 fatigue and nausea in 1/3 of the patients. One pt discontinued due to hypersensitivity reaction before being evaluated for response and one pt was diagnosed with Guillain-Barré syndrome while progressing after 2 cycles.
Conclusion: Plitidepsin (Aplidin®) shows promising activity and an acceptable safety profile in this difficult-to-treat subset of patients. Remarkably, no significant hematologic toxicity was seen in this heavily pretreated cohort. To confirm these preliminary data an expansion of the cohort is currently ongoing. Updated results will be presented.
Leukocytosis at Diagnosis and the Risk of Subsequent Thrombosis in Low-Risk Essential Thrombocythemia and Polycythemia Vera.
Authors: Gangat Naseema , Wolanskyj Alexandra , Schwager Susan , Hanson Curtis A. , Tefferi Ayalew 
Published: April 2020

Abstract
Background: Leukocytosis has recently been implicated as an adverse prognostic feature for thrombosis in both ET and PV. Such an association would be therapeutically most relevant in the context of “low-risk” disease. In the current study, we sought to clarify the relationship between leukocytosis at diagnosis and the subsequent occurrence of either arterial or venous thrombosis, in “low-risk” patients with ET or PV.
Methods: Data was abstracted from the medical records of a consecutive cohort of patients with WHO-defined ET or PV seen at the Mayo Clinic. Low-risk disease was defined by the absence of both thrombosis history and age 3 60 years. Cox proportional hazards model was utilized to determine the impact of clinical and laboratory variables on thrombosis-free survival (TFS). Arterial- or venous-specific TFS curves were constructed by Kaplan-Meier method.
Results: i) Patient characteristics and outcome A total of 407 “low-risk” patients were studied; 153 had PV (median age 48 years; females 43%) and 254 ET (median age 42 years; females 71%). A total of 46 thrombotic events (22 arterial and 24 venous) were recorded in 41 (27%) patients with PV during a median follow up of 130 months (range 2–562 months). The corresponding figures in ET were 54 total thrombotic events (41 arterial and 14 venous) in 47 (19%) patients at a median follow up of 104 months (range 0.25–424 months). Cytoreductive therapy was avoided in the presence of &lt; 1000 x 109/L platelet count but, at the discretion of the treating physician, some patients with higher platelet counts received prophylactic cytoreductive therapy. ii) Correlation between leukocytosis and thrombosis A leukocyte count of 3 15 x 109/L at diagnosis was documented in 42 (27%) patients with PV and 21 (8%) patients with ET; 102 patients (40%) with ET had &gt; 9.4 x 109/L leukocyte count. Leukocyte count considered as either a continuous or categorical variable (using cutoff levels of 15 x 109/L for PV and either 15 x 109/L or 9.4 x 109/L for ET) was not significantly associated with either arterial or venous thrombosis (Figure 1). iii) Correlation between other risk factors and thrombosis In univariate analysis, presence of the JAK2V617F was significantly associated with arterial thrombosis in ET (p=0.049) but significance was lost during multivariable analysis that included age as a covariate. Only advanced age was found to be significantly associated with arterial thrombosis in PV (p=0.04) and higher hemoglobin level with venous thrombosis in ET (p&lt;0.0001).
Conclusion: The current study does not identify leukocyte count at diagnosis as a marker of increased thrombosis risk in low-risk patients with either ET or PV. Instead, the study found an association between arterial thrombosis and advanced age in PV and venous thrombosis and higher hemoglobin level in ET.
Figure Figure
ADAMTS13 is a Risk Factor for MI, Stroke and Vascular Death in the Oxford Vascular Study.
Authors: Eglinton Jennifer , Harrison Paul , Segal Helen , Silver Louise , Mehta Ziyah , Rothwell Peter 
Published: April 2020

Abstract
Normal haemostasis requires a balance between ultra-high MW VWF synthesis/release and its degradation by ADAMTS13. Although a severe acquired/inherited deficiency of ADAMTS13 occurs in TTP, as VWF is an established risk factor for arterial thrombotic disease it has been recently hypothesized that intermediate to low levels of ADAMTS13 could also be an important risk indicator. A new rapid ADAMTS13 Technozym® ELISA activity assay (Technoclone, Vienna, Austria) was therefore used to measure ADAMTS13 activity within plasma samples from a cohort of 521 patients within the Oxford Vascular Study (OXVASC). OXVASC is a large population based study of all patients with acute vascular events (TIA, stroke, acute coronary syndromes and acute peripheral vascular events) and includes a substudy of various risk factors (including platelet function, aspirin responsiveness and classical coagulation markers) for thrombosis. The ADAMTS13 normal range was 68.96 – 144.36% (n = 20, mean = 106.7%, 95% CI = 95.8–115.5%). Within 521 OXVASC samples, the mean ADAMTS13 level was found to be significantly lower (p &lt; 0.0001, Mann-Whitney test) at 72.34% (95% CI = 70.33–74.35%). The minimum and maximum values obtained were 29.17% and 201.86% respectively. Although plasma VWF levels were significantly elevated, they were not inversely proportional to ADAMTS13 activity (r = 0.093). Furthermore low ADAMTS13 levels were not significantly associated with aspirin non-responsiveness (p = 0.0759) as defined by the Collagen/Epinephrine cartridge on the PFA-100, although VWF levels were significantly higher in non-responsive patients (p = &lt;0.0001). Increased VWF levels were predictive of the combined outcome of stroke, MI or vascular death (HR per decile increase 1.12, 95% CI 1.06–1.19, p = 0.0002), whereas lower ADAMTS13 levels were associated with increased risk (HR per decile decrease 0.91 95% CI, 0.85–0.97, p = 0.004). In conclusion, low ADAMTS13 activity was not associated with high VWF levels and there was no association with aspirin non-responsiveness in a high shear dependent assay (PFA-100). However, low levels of ADAMTS13 were predictive of recurrent major vascular events. ADAMTS13 may therefore have some potential as a risk factor in patients with arterial disease.
PI3 Kinase Inhibition Induces Apoptotic Cell Death in Acute Lymphoblastic Leukemia Via the Mitochondrial Pathway.
Authors: von Schwarzenberg Karin , Henkel Marco , Conzelmann Dennis , Stork Björn , Bringmann Anita , Wesselborg Sebastian , Brossart Peter 
Published: April 2020

Abstract
The phosphatidylinositol 3-kinase (PI3K) pathway regulates many cellular processes that are involved in tumor progression. Aberrant activation of the PI3K pathway due to an alteration of its elements like PTEN or Akt occurs quite frequently in malignant cells. Thus, inhibition of this pathway represents a promising option for the treatment of cancer patients. The best characterized PI3K inhibitors are LY294002 and wortmannin that were shown to disrupt downstream signaling and induce apoptotic cell death in tumor cells. Acute lymphoblastic leukemia (ALL) is a malignancy mainly found in young children and elderly with constitutive activation of PI3K pathway.
In our study we analyzed the effect of PI3K inhibition in cell lines deduced from ALL (Jurkat, BV173, SD1, T-2) and primary leukemic cells by incubating them with increasing concentrations of inhibitors of the PI3K signaling. We found that treatment of ALL cells with LY294002, the mTOR inhibitor rapamycin or Akt inhibitor SH5 induced apoptotic cell death that was accompanied by caspase-3 activation and PARP-cleavage and interfered with intracellular PI3K/Akt signaling as analyzed by phosphorylation and expression of mTOR or P70S6K. In line with these results apoptotic cell death could be inhibited by the pan-caspase inhibitor zVAD. In order to determine the pathway of apoptosis induction we took advantage of Jurkat cells (T-ALL) overexpressing or lacking molecules involved in apoptotic pathways such as FADD, an adaptor molecule recruited to the death receptor upon ligand binding, Caspase-8, Caspase-9 or Bcl-2, an anti-apoptotic protein that prevents the release of cytochrom c from mitochondria. PI3K inhibition by LY294002 induced apoptotic cell death in cells deficient of FADD or caspase-8 with no difference to wild type cells. In contrast, cells overexpressing Bcl2 or lacking caspase-9 were resistant to apoptotic death indicating that PI3-kinase inhibition is independent of the external death receptor signaling and is mediated via the mitochondrial pathway. These results were confirmed by analyzing PARP cleavage and caspase-3 activation in utilized leukemic cell lines. Furthermore, we found that the PI3K inhibitor LY294002 induced apoptosis in ALL cells that could be increased by the etoposide, a topoisomerase inhibitor, or TRAIL. In addition, in contrast to etoposide, treatment of ALL cells with TRAIL could overcome the resistance of ALL cells to PI3K inhibition even in caspase-9 deficient Jurkat cells. Our results provide an interesting approach in designing novel therapeutic strategies to target the PI3K pathway in ALL to overcome the resistance to cytotoxic agents.
Expression of Tissue Factor Is up-Regulated in U937/PR9 Cells by the PML/RARα Fusion Protein through Its Interaction with the Regulatory Region of Tissue Factor Promoter.
Authors: Yan Jingsong , Liu Hongchen , Su Xiaoyu , Wang Ping , Ruan Zhen , Wang Kankan , Chen Zhu , Chen Saijuan , Xi Xiaodong 
Published: April 2020

Abstract
Life-threatening bleeding remains a challenging complication of acute promyelocytic leukemia (APL), which is characterized by the t(15;17) translocation producing a fusion gene encoding the PML/RARα fusion protein, and is mainly ascribable to the increased levels of tissue factor (TF) in APL cells. Previous studies have shown that the TF expression levels are elevated in NB4 (APL) cells or in U937 (monoblastic) cells ectopically expressing PML/RARα indicating that the TF expression in these cells is regulated in response to PML/RARα. The molecular mechanisms by which PML/RARα regulates the TF expression are, however, still poorly understood. To examine whether the PML/RARα fusion protein regulates the TF expression through an interaction with TF promoter, a series of cell lines stably expressing luciferase reporter gene under control of different 5’ flanking regions of TF promoter were established with U937/PR9 cells, a U937 cell line expresses Zn-inducible PML/RARα. In luciferase assays a Zn-induced transcriptional activity was present in cells with truncated TF promoters (-2106~+128), (-1386~+128), (-684~+128), (-384~+128) and (-247~+128) but absent in those with a shorter version of TF promoter (-201~+128). These results suggested that the sequences within -247 and -201 of TF promoter were necessary for the regulation of TF gene expression regulated by PML/RARα in U937/PR9 cells. In ChIP experiments a markedly increased amount of PCR products in response to the expression of PML/RARα were amplified by the primers flanking the regulatory region (-247~-201) from the chromatins immunoprecipitated with an anti-RARα antibody, suggesting that the PML/RARα fusion protein interacts with TF promoter concomitant with the elevated luciferase activity. The EMSA data using probes containing distal or proximal AP-1 consensus sequences located in the regulatory region (-247~-201) showed that PML/RARα did not directly bind to this region but enhanced the binding ability of c-Jun and Jun D to the proximal AP-1 site. Collectively, our data showed that the PML/RARα fusion protein up-regulates the expression of TF through the interaction with the regulatory region of TF promoter. The more accurate localization of the regulatory region in TF promoter and the related molecular mechanisms regarding the regulatory effects of the PML/RARα fusion protein on TF expression are under investigation.