bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Genetic Alterations Associated with Mutant IL7Ralpha in T-ALL
Authors: Rodrigues Gisele O. L. , Li Wenqing , Cramer Sarah , Campos Livia W. , Zenatti Priscila P. , Laranjeira Angelo A.B.A , Hixon Julie A. , Senkevitch Emilee , Andrews Caroline , Noronha Elda P. , Pombo-de-Oliveira Maria , Brandalise Silvia , Lobo Francisco P. , Yunes Jose Andres , Durum Scott 
Published: October 2019

Abstract
The IL7/IL7R mediated signaling is essential for normal development and homeostasis of T cell precursors. Early studies have shown that around 10% of patients with Acute lymphoblastic leukemia T cell (T-ALL) have mutations in the alpha chain of the receptor for IL7 (IL-7Ralpha) driving constitutive signaling via JAK1 and independent of IL-7, gamma-chain or JAK3. Some genetic changes are important factors to initiate leukemia, but in many cases these changes are insufficient to achieve the complete leukemic phenotype, suggesting that collaborative oncogenic mutations may be present. To identify candidate mutations that work in collaboration with the oncogenic IL7R, we performed exome sequencing and SNP-CNV-Array assay on a group of eight primary T-ALL samples carrying the IL7R mutation (T-ALL-IL7Rmut). The microarray was performed using Cytoscan HD - Affymetrix and CNVs were detected by ChAs software, version 2.0.1.2. For exome sequencing we used Illumina Hiseq2000 platform and Agilent SureSelect V4 51M Capture kit (mean sequencing depths of 80X / 50X for leukemia and remission samples, respectively). Somatic Single Nucleotide Variants (SNVs) and Small Insertion/Deletion (InDels) were detected using VarScan, and mutations were functionally annotated using ANNOVAR. All somatic mutations detected were manually curated. We found 17 genes recurrently mutated (in ≥ 2 cases) and chose five of them for further analyses due to their previous involvement in ALL (PHF6, RB1, CTCF, SGK223 and DNM2). Ongoing experiments are being conducted to determine whether these recurrent mutations can collaborate functionally with mutIL7R by co-transfection into immature murine thymocytes, transplanting into mice and determining incidence of leukemia.

Disclosures
No relevant conflicts of interest to declare.
Concomitant Myeloproliferative Disorders (MPD) and Amyloidosis
Authors: Pallante Angeline J , Leung Nelson , Kyle Robert A. , Tefferi Ayalew , Russell Stephen J. , Habermann Thomas M. , Lacy Martha Q. , Hayman Suzanne R. , Al-Kali Aref , Reeder Craig , Dispenzieri Angela , Gertz Morie A. , Buadi Francis K , Hook C. Christopher , Dingli David , Pardanani Animesh 
Published: October 2019

Abstract
Background: The combination of MPD and amyloidosis is rare, and this patient population's clinical outcome is not well studied.
Methods: Pts with a MPD and amyloidosis were identified via an clinical note search engine that searches through the electronic medical records of patients seen at Mayo Clinic Rochester, Jacksonville, and Scottsdale between 1990 and 2016. Terms used included amyloid or amyloidosis, chronic myelogenous leukemia or CML, essential thrombocytopenia or ET, and polycythemia vera or PV. Demographic and clinical data were abstracted from the medical record. Pts with both disorders were analyzed and their mortality rates along with median time to death were calculated. Prevalences at the Mayo Clinic were calculated for the years 2014 and 2015.
Results: Twenty-three pts diagnosed with both a MPD and amyloidosis were identified. Thirteen (56.5%) were male, 10 (43%) were female. Eleven (47.8%) were initially diagnosed at the Mayo Clinic. Types of amyloidosis were as follows: Eleven (47%) had immunoglobulin light-chain (AL), four had localized (17%), two (8.7%) had wild-type transthyretin (ATTR), one (4.3%) had mutant ATTR, and five (21.8%) were unknown. Types of MPD were as follows: Seven (30%) had polycythemia vera (PV), seven (30%) had chronic myelogenous leukemia (CML), five (22%) had myelofibrosis, and four (17%) had essential thrombocytosis (ET). Fifteen (65%) were initially diagnosed with a MPD. Median time to last follow-up from second diagnosis was 1.7 years, and median time to death following second diagnosis was 1.4 years. The mortality rate was 87% in the total population. The median time to death for AL and PV was 2.7 years, AL and CML 1.0 years, and AL and ET 1.17 years. Myelofibrosis did not occur with AL. The most common combination was AL and PV, which accounted for five (22%) of the cases.
Treatment regimens for the patients with AL were varied. Multiple drugs were combined with dexamethasone including melphalan, velcade, pomalidomide, lenalidomide, doxorubicin, and revlimid. Cyclophosphamide, bortezomib, and dexamethasone (CyBorD) were used in two cases. The mean time to diagnosis of amyloidosis from symptom onset was 10.3 months.
Prevalences of AL and a MPD in 2014 and 2015 at the Mayo Clinic were 2.8% and 1.5%, respectively.
Conclusions: The mortality rate for the combined diagnoses is high. The most common combination of diagnoses was that of AL and PV, which was associated with a mean time to death of 2.7 years.

Disclosures
Al-Kali: Celgene: Research Funding; Onconova Therapeutics, Inc.: Research Funding.
Th17 Cells Exhibit Anti-Tumor Effects in MDS Possibly through Augmenting Functions of CD8+ T Cells
Authors: Shao Zonghong , Li Jing , Yue Lanzhu , Wang Huaquan , Liu Chunyan , Liu Hui , Jinglian Tao , Qi Weiwei , Wang Yihao , Zhang Wei , Fu Rong 
Published: October 2019

Abstract
Myelodysplastic syndromes (MDS) are a common malignant hematological disease.Th17 cells are a newly found subset of distinct CD4+ Th effector cells family and are found to play an important role in cancers. Intracellular staining and flow cytometric analysis were used to detect the percentage of Th17 cells .Here, we showed that both the percentage and the function of Th17 cells were elevated in low-risk MDS while decreased in high-risk MDS. To determine the concentration of IL-17, IL-6 and IL-23, the serum were analyzed by ELISA. Levels of upstream molecules of Th17 cells, IL - 6 and IL - 23, were higher in low-risk MDS but lower in high-risk MDS patients. The abnormal percentage of Th17 cells was closely related to clinical parameters including karyotype, morphologic blast percentage of bone marrow, peripheral absolute neutrophil count and hemoglobin concentration. Furthermore, the expression of IL-17, RORγt , STAT-3 mRNA were determinated of by qRT-PCR .Expression rates of perforin and granzyme B in BM CD3+CD8+ cells (cytotoxic T lymphocyte, CTL) positively correlated with levels of IL-17 but negatively correlated with BM blast percentage, and could be significantly increased after stimulation with human recombinant IL-17 (rhIL-17). Our results suggested Th17 cells might play an antitumor effect in the pathogenesis of MDS through IL-17/CTL pathway.

Disclosures
No relevant conflicts of interest to declare.
Autocrine and Paracrine Interactions Between Multiple Myeloma Cells and Bone Marrow Stromal Cells By Growth Arrest-Specific Gene 6 Crosstalk with Interleukin-6
Authors: Furukawa Miki , Hiroshi Ohkawara , Ogawa Kazuei , Ikeda Kazuhiko , Koki Ueda , Shichishima-Nakamura Akiko , Ito Emi , Imai Jun-ichi , Yanagisawa Yuka , Honma Reiko , Watanabe Shinya , Waguri Satoshi , Ikezoe Takayuki , Takeishi Yasuchika 
Published: October 2019

Abstract
The pathogenesis of multiple myeloma (MM) has not yet been fully elucidated. Our microarray analysis and immunohistochemistry revealed significant upregulation of growth arrest-specific gene 6 (Gas6), a vitamin K-dependent protein with a structural homology with protein S, in bone marrow (BM) cells of MM patients. ELISA showed that the serum levels of soluble Gas6 and interleukin (IL)-6were significantly increased in the MM patients when compared with healthy controls. Gas6 and TAM receptor tyrosine kinases Mer were overexpressed in human CD138-positive MM cell line RPMI-8226. Exogenous Gas6 and its signaling through Mer suppressed apoptosis induced by serum deprivation and enhanced cell proliferation of the MM cells. The conditioned medium from human BM stromal cell line HS-5 induced anti-apoptosis and cell proliferation of the MM cells with ERK, Akt and NF-kappaB phosphorylation, which were reversed by the neutralizing antibody to Gas6, IL-6 or Mer siRNA. Furthermore, the Gas6 neutralizing antibody reduced the upregulation of IL-6 and intercellular cell adhesion molecule-1 induced by a HS-5 cell-conditioned medium in MM cells. The present study provides new evidence that autocrine and paracrine stimulation of Gas6 in concert with IL-6 contributes to the pathology of MM, suggesting that Gas6/Mer signaling may be a promising novel target for treating MM.

Disclosures
No relevant conflicts of interest to declare.
Contribution of High-Density Neutrophils to Multiple Myeloma Microenvironment Dysregulation
Authors: Romano Alessandra , Giallongo Cesarina , Parrinello Nunziatina L , Tibullo Daniele , Puglisi Fabrizio , Camiolo Giusy , La Cava Piera , Parisi Marina , Consoli maria Letizia , Conticello Concetta , Di Raimondo Francesco , Luca Vanella 
Published: October 2019

Abstract
Background and objective: Multiple Myeloma (MM) is a plasma cell malignancy with a well documented immune dysfunction. Previous reports in murine models and our previous work in MM patients, showed that myeloid cells, both granulocyte-myeloid derived suppressor cells (G-MDSC) and neutrophils sedimenting on top of red cells after density gradients, defined as high-density neutrophils (HDN) are increased and exert an immunosuppressive activity. Moreover, we showed that myeloid precursors could be activate and acquire G-MDSC phenotype if cultures in presence of MM mesenchymal cells.
Thus, we characterized functionally HDN to gauge their contribution in immunesuppression, angiogenesis and bone resorption in MM microenvironment.
Methods:
In HDN freshly-isolated from a series of 60 newly-diagnosed MM, 30 smoldering MM and 30 MGUS we investigated by RT-PCR, flow-cytometry, western-blot and immunofluorescence genes and proteins related to immunesuppression (Arg-1, IDO), angiogenesis (BV8, VEGF, MMP9) and bone resorption (IL1-b, MMP9, CHI3L).
Results:
First, MM-HDN exhibited an increased expression of immunesuppressive molecules Arg-1 and IDO compared to MGUS and healthy subjects (25.5 vs 6.2 vs 1 fold changes in gene expression, p=0.003), confirmed by functional assay of enzymatic activity, immunostaining and WB, positively correlated with advanced disease.
Second, MM-HDN showed increased expression of BV8 and MMP9 (p=0.01, p=0.05) but not VEGF, as confirmed by immunofluorescence. In line with this observation, by using migration assay (CIM plate) MM-HDN induced neo-vasculogenesis in vitro by forming more and aberrant tubes than healthy or MGUS-HDN, thus to disclosure a novel proangiogenic mechanism driven by BV8.
Third, MM-HDN showed increased expression of IL1-b, MMP9, CHI3L, associated to increased bone resorption activity, evaluated by dentin-disk assay. The same findings could be obtained treating bone matrix with MM-HDN-conditioned media, suggesting the crucial role of molecules released by MM-HDN but no from MGUS-HDN.
Conclusion: taken together, our observations disclosure the contribution of aberrantly activated HDN in MM clinical features, contributing to immunosuppression, angiogenesis and bone resorption.

Disclosures
No relevant conflicts of interest to declare.
Overcoming Innate Resistance to a Beta-Catenin Inhibitor-BC2059- By Manipulating Autophagy in Multiple Myeloma
Authors: Savvidou Ioanna , Khong Tiffany T. , Horrigan Stephen K. , Spencer Andrew 
Published: October 2019

Abstract
Introduction: Chemoresistance is a major challenge in the development of new therapies in multiple myeloma [MM]. Inhibition of autophagy has been shown to restore chemosensitivity in several tumors. We have previously validated a beta-catenin inhibitor (BC2059) which targets the Wnt/beta-catenin signaling pathway.
Aim: In the present study we aim to overcome innate resistance to BC2059 by manipulating the autophagic pathway.
Methods: Autophagic flux was estimated by measurement of LC3II/LC3I in the absence and presence of hydroxychloroquine [HQ] by Western blot [WB]. Induction of autophagy was measured by the increase of LC3II/LC3I by WB, and concomitant drop of p62 expression by Flow Cytometry [FC]. Combination Indices [CI] were calculated using Calcusyn software.
Results: BC2059 induces apoptosis in a dose-dependent manner by induction of both the intrinsic and extrinsic apoptotic pathways, (increase of active -caspase-8, -caspase-9 and -caspase-3 measured by FC and cleaved PARP by WB) in all human myeloma cell lines [HMCL] tested. All HMCL tested have significant autophagic flux at baseline. Chemical inhibition of autophagy has an anti-proliferative effect, decreasing the relative cell numbers from 40% (NCI-H929) to 23% (KMS12BM) at 24hr. In parallel BC2059 is able to induce autophagy in a dose dependent manner. Induction of autophagy is BC2059 specific as treatment with melphalan or bortezomib at relative equal anti-proliferative doses did not increase LC3II/LC3I. Further autophagic inhibition by HQ was synergistic for all HMCL with CI of 0.7-0.3 (CI<1.1 indicates synergy). Interestingly, inhibition of autophagy halved the LD50 of BC2059 in a resistant HMCL (LP1). Other autophagy inhibitors (3-MA, bafilomycin A1 and NH4Cl) were also synergistic with BC2059 in LP1.
Conclusion: BC2059 exerts cytotoxicity mainly by induction of apoptosis but also induces cyto-protective autophagy. Autophagy inhibition was able to overcome innate resistance to the drug, ameliorating its cytotoxic effect. This study warrants further investigation.

Disclosures
Horrigan: betacatpharmaceuticals: Employment.
Incidence of Cytomegalovirus (CMV) Reactivation after Autologous Peripheral Blood Stem Cell Transplantation (ASCT) in 324 Patients with Multiple Myeloma and Lymphoma: A Single-Center Study at the American University of Beirut Medical Center (AUBMC)
Authors: Massoud Radwan , Assi Rita , Fares Elie , Kreidieh Nabila , Mahfouz Rami , Kanj Souha , El-Zakhem Aline , Kharfan-Dabaja Mohamed A. , Bazarbachi Ali , El Cheikh Jean 
Published: October 2019

Abstract
CONTEXT: Cytomegalovirus reactivation (R-CMV) is often diagnosed in allogeneic hematopoietic cell transplant recipients and could determine a CMV-related disease in these immunocompromised patients, involving any organ. R-CMV and end-organ disease after ASCT has not been studied thoroughly. Autograft recipients are generally considered to have low risk of R-CMV or end-organ disease.
OBJECTIVE: Evaluate the incidence, risk factors, and outcome of R-CMV in adult patients with hematologic malignancies undergoing ASCT.
DESIGN: Retrospective single center study.
SETTING: This study was approved by the institutional review board of AUBMC and conducted at our institution
PATIENTS OR OTHER PARTICIPANTS: A total of 324 consecutive ASCT were performed at AUBMC between January 2005 and March 2016. All patients and transplant-related characteristics are listed on Table 1. CMV DNA load in blood was measured by quantitative polymerase chain reaction (PCR) weekly as a routine monitoring strategy in all patients. Irrespective of PCR results, some patients suspected to have gastrointestinal involvement were biopsied. Also, in the presence of symptomatic CMV reactivation, appropriate anti-CMV therapy was instituted.
MAIN OUTCOMES MEASURES: The primary outcome is understanding the potential relationship between R-CMV and Overall Survival (OS). Secondary outcomes included the effect of CMV reactivation on transplant related mortality (TRM), and progression free survival (PFS).
RESULTS: Overall, 53 (16%) patients had R-CMV and 38 (72%) required anti-CMV treatment. Five (1,5%) had CMV disease with positive PCR on colon biopsy, yet two had PCR negative in blood. After a median follow up of 21.5 months (range: 1 to 125 months), there was no significant difference in OS or PFS between patients with or without R-CMV. TRM has increased from 1.1% in patients with no R-CMV to 13% in patients with R-CMV (P=0.003). We didn't observe any impact for age, sex, type of disease, pre-transplant treatment types/lines on the incidence R-CMV following ASCT.
CONCLUSIONS: Our data suggest that R-CMV is not uncommon in ASCT recipients and may contribute to increased TRM. Biopsy is recommended in case of high suspicion of R-CMV irrespective of PCR results.

Disclosures
No relevant conflicts of interest to declare.
Increased Mir-155 Expression Is Associated with FLT3-ITD Mutation and Poor Therapy Response in Chinese AML and Allo-HSCT Could Improve Survival of These Patients
Authors: Yang Xiaofei , Sun Aining , Hu Xingxing , Ma Wenjuan , Wu Depei 
Published: October 2019

Abstract
The aim of this study was to evaluate the diagnostic and prognostic value of miR-155 in Chinese acute myeloid leukemia (AML) patients. The miR-155 levels of 142 de novo AML patients and 28 healthy donors were detected by real-time quantitative PCR (RQ-PCR). The miR-155 expression was significantly up-regulated in AML compared with control (p<0.001). A receiver operating characteristic (ROC) curve revealed that miR-155 expression could differentiate patients with AML from control subjects (AUC=0.907, 95%CI: 0.853-0.960, P<0.001). We also monitored the miR-155 level in 22 de novo AML patients and found that the miR-155 level decreased significantly from the initial diagnosis to post-CR (P<0.001). FLT3 internal tandem duplication (FLT3-ITD) mutation was significantly more frequent in miR-155 high-expressed group versus low-expressed group (p=0.002). Considering all patients, miR-155 high-expressed cases had significantly lower complete remission (CR) and part remission (PR) rate than miR-155 low-expressed cases (p=0.04). Survival analysis was performed in non-M3-AML patients who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) as consolidation therapy. Kaplan-Meier demonstrated that there was no difference in leukemia free survival (LFS) and overall survival (OS) between miR-155 high-expressed and low-expressed patients (p>0.05). Collectively, Increased miR-155 expression is associated with FLT3-ITD mutation and poor therapy response in Chinese AML and allo-HSCT could improve survival of these patients.

Disclosures
No relevant conflicts of interest to declare.
Use of Computed Tomography Pulmonary Angiography for Suspected Pulmonary Embolism in Patients with Malignancy- a Single Center Experience
Authors: Ravulapati Sravanthi , Leung Cerena K , Mehta Mudresh R , Christopher Kara M , Woelich Susan K. , Poddar Nishant 
Published: October 2019

Abstract
Background: Pulmonary embolism (PE) is a potentially lethal condition commonly suspected in patients with malignancy. Computed Tomography Pulmonary Angiography (CTPA) is increasingly used in the diagnosis of PE, and guidelines have incorporated various screening tools including the Modified Geneva and Wells criteria to facilitate exclusion of pulmonary embolism. There is an increased risk of venous thromboembolism in patients with active malignancy and therefore an increased suspicion in patients who present to the emergency department (ED) with concerning symptoms.
Methods: This is a retrospective analysis at a single tertiary care institution. All patients initially diagnosed with an active malignancy since 2005 and underwent a CTPA between January 2010 and October 2015 were reviewed. Patients were excluded if the CTPA was performed in the setting of trauma, a history of benign malignancy, or if the diagnosis of malignancy was made subsequent to the CTPA. Data collected included patient demographics, clinical presentation, type of malignancy and treatment regimen received. The modified Geneva and Wells criteria were applied to all patients independent from the initial ED risk assessment for a PE.
Results: There were 796 patient records reviewed, of which 162 patients met inclusion criteria. Out of these 162 patients, only 8 (4.9%) were found to have a pulmonary embolism. All patients with a positive CTPA had an intermediate risk per the Geneva criteria while only 62.5% had an intermediate risk per the Wells criteria. Of the 154 patients with a negative CTPA, 71.5% and 78.7% had an intermediate risk; 22.5% and 18.7% were classified as low risk based on Wells and Geneva criteria, respectively. The median age of patients was 59 years old, and the majority were male (58%). The most common malignancies in which a CTPA was ordered were lung cancer (27.7%) followed by breast cancer (14.9%) and prostate cancer (6.8%). Despite a negative CTPA, 82 out of 154 patients (53%) were admitted to the hospital.
Conclusion: Pulmonary embolism is commonly associated with and frequently suspected in patients with active malignancy. The incidence of PE over a 5-year period in oncology patients was 5% in our emergency department. In total, 18.7% to 22.5% of patients could have avoided a CTPA if scoring was based on the Wells or Geneva criteria. Based on the review at our institution, the modified Geneva and Wells criteria are not adequate, and a new tool needs to be developed for risk stratification for the diagnosis of PE specifically in patients with active malignancy.

Disclosures
No relevant conflicts of interest to declare.
Test-Retest Reliability Analysis of the Patient Reported Outcomes Burdens and Experiences (PROBE)
Authors: Chai-Adisaksopha Chatree , Iorio Alfonso , Curtis Randall , Frick Neil , Nichol Michael , Noone Declan , O'Mahony Brian , Page David , Stonebraker Jeffrey , Skinner Mark 
Published: October 2019

Abstract
Background: Patient Reported Outcomes Burdens and Experiences (PROBE) questionnaire was developed for assessing health status in patients with bleeding disorders, specifically, hemophilia. Phase I of PROBE study confirmed robustness of methodology and feasibility of the questionnaire.
Objective: To investigate the test-retest reliability of PROBE questionnaire
Methods: Patients and non-hemophilia individuals who attended an hemophilia related workshops were invited to participate in this study. All participants were asked to fill out a PROBE questionnaire for 3 times. The paper-based questionnaires were collected immediately after being filled out on day 1 and day 2. Participants were instructed to log into the website within 2 weeks and fill out the web-based questionnaire. Test-retest reliability was analyzed.
Results: A total of 63 participants were enrolled in this study with a median age of 50 (range 14-76) years. Of these, 30 (47.6%) were hemophilia patients or carriers, 33 (52.5%) were participants with no known bleeding disorders. On general health domain, Kappa coefficients ranged from 0.69 to 0.90, indicating substantial to almost perfect agreement for all items. Reliability of web-based questionnaire showed moderate to substantial agreement for all except 1 item. For hemophilia related domain, Kappa coefficients ranged from 0.5-1.0. Of these, 5 of 11 items were in perfect agreement (Kappa=1.0). Reliability of web-based questionnaire were in substantial to almost perfect agreement. For overall health related quality of life, EQ-5D had Kappa coefficients of 0.62 to 0.89. Intraclass correlation coefficient of visual analog scale (VAS) was 0.90 (95% confidence interval; 0.83-0.94). Test-retest reliability was comparable between hemophilia patients and participants with no known bleeding disorders.
Conclusions: The results suggest that PROBE is a reliable tool to assess patients reported outcomes for hemophiliacs. Test-retest reliability analysis showed that the agreement was comparable between two participant populations. This result suggests that PROBE tool may be used to collect and compare not only patient reported outcome in hemophilia, but to generate norm values in appropriate control population. The Web-based questionnaire has an acceptable agreement with the standard paper-based version.

Disclosures
No relevant conflicts of interest to declare.