bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Role of Cbx Proteins in Human Benign and Malignant Hematopoiesis
Authors: Jung Johannes , Schepers Hein , Lazare Seka , Buisman Sonja , Weersing Ellen , Dethmers Bertien , Klauke Karin , Bystrykh Lenja , De Haan Gerald 
Published: October 2019

Abstract
Introduction:
Hematopoietic stem cells (HSCs) need to properly balance self-renewal and differentiation to prevent malignant transformation or HSC exhaustion. We recently showed that the presence of specific Cbx proteins in the Polycomb Repressive Complex 1 (PRC1) plays a crucial role in balancing self-renewal and differentiation of murine HSCs. Whereas Cbx7 induces self-renewal, Cbx4 and Cbx8 instead induce differentiation. In this project, we investigate the role of various CBX Polycomb proteins in human normal and malignant hematopoiesis. We aim to identify genes which are controlled by CBX7 in normal and malignant human hematopoiesis.
Methods:
We overexpressed CBX2, -4, -6, -7 and -8 in human CD34+HSPCs. We assessed functional consequences by measuring cobblestone area-forming cell (CAFC), colony-forming unit (CFU) frequencies, and performed stem cell xenotransplantation studies in NSG mice. To identify genes which are controlled by CBX7 or CBX8, we performed RNA- and Chip-Seq in CD34+ HSPCs. To explore the role of CBX7 in leukemic cells, we performed short hairpin RNA-mediated knockdown of protein expression in leukemic cell lines.
Results:
Overexpression of CBX7 and CBX8 in human CD34+ HSPCs led to a significant 5-10-fold increase of week 5 CAFC (fold increase: 9,62 for CBX7; 4,92 for CBX8) and 1,5-fold increase CFU-frequencies (fold increase: 1,32 for CBX7; 1,41 for CBX8) after 14 days. In contrast, overexpression of CBX4 and CBX2 led to equal or decreased CAFC- and CFU-frequencies. Transplantation of CBX7 overexpressing human CD34+ HSPCs in NSG mice led to higher long-term engraftment in comparison to the empty vector control (p<0,05 after 18 weeks). Even after one week of in vitro culture of CBX7-overexpressing CD34+cells significant engraftment levels were obtained. Conversely, downregulation of CBX7 in various leukemic cell lines resulted in markedly decreased proliferation and strongly induced differentiation. Expression analysis showed that genes that are upregulated upon CBX7 overexpression are preferentially expressed in primitive stem cells, and are repressed in more differentiated cell types.
Conclusions:
Our study indicates that CBX7 and CBX8 regulate self-renewal of human HSPCs. Furthermore, our data show that repression of CBX7 markedly inhibits proliferation of leukemic cells and can release the differentiation block of leukemic cells in vitro. Collectively our results indicate that targeting CBX7 may be a viable strategy to induce terminal differentiation of leukemic cells.

Disclosures
No relevant conflicts of interest to declare.
A Novel Humanised ROR1 Bi-Specific T-Cell Engager Molecule for the Treatment of Chronic Lymphocytic Leukaemia
Authors: Gohil Satyen Harish , Harrasser Micaela , Paredes-Moscosso Solange R , Nathwani Amit C , Della Peruta Marco 
Published: October 2019

Abstract
Bispecific T-cell Engagers (BiTEs) are a promising immunotherapeutic approach, usually consisting of two single chain variable fragments (scFv) linked in tandem. One arm binds to tumour-associated antigens, whilst the other binds CD3 on T-cells, facilitating the formation of cytotoxic synapses leading to killing of tumour targets in a MHC and co-stimulatory molecule independent manner. Blinatumomab, the most advanced BiTE in clinic targets CD19 and has shown impressive results in patients with relapsed/refractory B cell acute lymphoid leukaemia but results in elimination of normal and malignant B lymphocytes.
Receptor Tyrosine Kinase Like Orphan receptor is a surface antigen expressed on a range of haematological and solid malignancies, including Chronic Lymphocytic Leukaemia (CLL), Acute Lymphoblastic Leukaemia (ALL), and cancer stem cell-like cells (CSC). It is however expressed at low or undetectable levels in normal tissues making it an attractive target for immunotherapy approaches. We generated a panel of novel fully humanised ROR1 antibodies and selected an optimal scFv based on ROR1 binding to use in our BiTE format.
Our ROR1 BiTE (ROR1xCD3), at low concentrations, was efficient inducing T-cell mediated cytotoxicity and consequent inflammatory cytokine secretion against a panel of haematopoietic cell lines with constitutive ROR1 expression in vitro. Importantly cytotoxicity was comparable to that seen with a control CD19 BITE. Moreover, ROR1xCD3 led to significant and selective cytotoxicity of primary CLL samples. We transitioned our in vitro studies to animal models and showed that our ROR1xCD3 is able to retard growth of established SKW cells engrafted in immunocompromised mice, as detected by bioluminescence imaging. Additionally ROR1xCD3 improved survival, with the ROR1xCD3 treated cohort showing improved median survival of 50 days compared to control mice of 18 days despite only receiving a single infusion of T-cells.
In summary, we have developed a novel ROR1 BiTE able to target an array of haematopoietic malignancies. Our ROR1 BiTE induces T-cell cytotoxicity against ROR1 positive cell lines in vitro and in vivo as well as primary CLL cells and has the potential to be a beneficial therapeutic for CLL.

Disclosures
Gohil: UCL Business: Patents & Royalties: ROR1 based immunotherapies. Paredes-Moscosso:UCL Business: Patents & Royalties: ROR1 based immunotherapies. Nathwani:UCL Business: Patents & Royalties: ROR1 based Immunotherapies. Della Peruta:UCL Business: Patents & Royalties: ROR1 based immunotherapies.
Economic Evaluation of the Direct Oral Anticoagulants Compared to Vitamin K Antagonists in the Treatment of Deep Venous Thrombosis and Pulmonary Embolism in the Outpatient Setting: Analysis from the Queen Elizabeth II Health Sciences Centre in Halifax, Nova Scotia
Authors: Al Saleh Abdullah S. , Berrigan Patrick , Anderson David , Shivakumar Sudeep 
Published: October 2019

Abstract
Background: To date, there have been few economic evaluations of the direct oral anticoagulants in the treatment of venous thromboembolism (VTE) conducted from a Canadian perspective. As a result, there is a lack of consensus within existing literature regarding which treatment strategy should be adopted into the clinical setting. To our knowledge, this is the first Canadian economic evaluation in patients with VTE who are treated solely on an outpatient basis.
Objectives: We conducted a cost-minimization analysis to determine the least costly intervention for the prevention of recurrent venous thrombosis in patients with acute unprovoked VTE including apixaban, rivaroxaban, low molecular weight heparin (LMWH) in combination with dabigatran, and LMWH overlapped with vitamin K antagonists (VKA). We also conducted a cost-effectiveness analysis to assess value for money in terms of cost per quality adjusted life year (QALY) gained.
Methods: We used Microsoft Excel to develop a decision model. Decision model parameters were determined using published literature, local hospital data, expert opinion, and chart review. Our cost-minimization analysis aggregated costs related to pharmaceuticals, laboratory testing, and hematologist fees. Our cost-effectiveness analysis also included costs related to health outcomes.
Results: Our cost-minimization analysis found apixaban to be the least costly intervention at three months ($609.28). At six months and one year LMWH overlapped with VKA was the least costly intervention ($896.08 and $1,193.88, respectively). With respect to cost-effectiveness, at three months apixaban was the dominant strategy. At six months, apixaban was cost-effective given a willingness-to-pay per QALY as low as $4,986.81. At one year, LMWH overlapped with VKA was cost-effective given a willingness-to-pay per QALY below $135,670.28. Probabilistic sensitivity analysis of our cost-effectiveness analysis found apixaban to be the dominant strategy compared to LMWH overlapped with VKA in 42.6% of iterations and 70.6% of iterations resulted in incremental cost-effectiveness ratio below $100,000 per QALY gained.
Conclusions: Our findings suggest apixaban is the least costly and most cost-effective strategy for short-term anticoagulant treatment for VTE but favor LMWH overlapped with VKA for long-term treatment.

Disclosures
No relevant conflicts of interest to declare.
Global Prospective Cohort Study of Factors Associated with Venous Thromboembolism in Patients with Lung Cancer Receiving Cancer Therapy
Authors: Kuderer Nicole M. , Poniewierski Marek S. , Culakova Eva , Lyman Gary H. , Khorana Alok A. , Pabinger Ingrid , Agnelli Giancarlo , Liebman Howard A. , Vicaut Eric , Meyer Guy , Shepherd Frances A. 
Published: October 2019

Abstract
BACKGROUND: Patients with lung cancer are known to be at increased risk for venous thromboembolism (VTE). However, there have been few studies of risk factors for VTE in lung cancer patients undergoing systemic chemotherapy.
METHODS: CANTARISK was a prospective, non-interventional, global cohort study including patients with lung cancer initiating a new chemotherapy regimen. Clinical data were collected at baseline and at 2, 4 and 6 months follow-up. The impact of patient-, disease- and treatment-related factors on the occurrence of VTE in the first 6 months was evaluated in univariable and multivariable Cox regression analyses.
RESULTS: A total of 1,980 patients with lung cancer were enrolled from 2011-12 of which 84% were diagnosed with non-small cell lung cancer (NSCLC). Median age was 63 years (range, 25-91) and 63% were male while 82% were active or former smokers. Race was white (70%), Asian (22%), or black (4%) with similar numbers from North America, Europe, and other regions including Asia. Metastatic disease was reported in 70% and ECOG PS was ≥2 in 13%. During the first six months, 121 patients developed a VTE (6.1%), of which 47.1% had pulmonary embolism (PE), 45.5% deep venous thrombosis (DVT), 3.3% catheter-associated thrombosis, and 4.1% visceral thrombosis. Among significant factors in univariable analysis, independent predictors for VTE in multivariable Cox regression analysis included female gender, US geographic region, leg immobilization, and presence of a central venous catheter (Table) with a trend toward greater risk for higher grade histology. Although predictive of early all-cause mortality in this study population (Kuderer et al ASCO 2016), the previously validated risk score for VTE in ambulatory cancer patients (Khorana et al: Blood 2008) was not significantly associated with VTE in either univariable or multivariable analysis.
CONCLUSIONS: Several demographic, geographic, and clinical factors are significantly associated with an increased risk of VTE in patients with lung cancer receiving systemic chemotherapy. Future analysis will attempt to assess how novel targeted treatment options might impact the Khorana score's predictive ability across all lung cancer patients.

Disclosures
Kuderer: Janssen Scientific Affairs, LLC: Consultancy, Honoraria. Lyman:Amgen: Research Funding. Khorana:Bayer: Consultancy, Honoraria; Leo: Consultancy, Honoraria, Research Funding; Halozyme: Consultancy, Honoraria; Sanofi: Consultancy, Honoraria; Amgen: Consultancy, Honoraria, Research Funding; Roche: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria; Janssen Scientific Affairs, LLC: Consultancy, Honoraria, Research Funding.
Knock-Down of the NFAT2 Long and Intermediate Isoforms Leads to CLL Acceleration
Authors: Heitmann Jonas S , Märklin Melanie , Anastasia Posdorowkin , Fuchs Alexander R , Truckenmüller Felicia M , Kopp Hans-Georg , Kanz Lothar , Rao Anjana , Wirths Stefan , Mueller Martin R 
Published: October 2019

Abstract
NFAT2 is a highly phosphorylated transcription factor which regulates developmental and activation programs in diverse cell types. We and others have previously described a significant overexpression of NFAT2 in CLL cells as compared to physiological B cells. Three major isoforms of NFAT2 with different regulatory properties have been described (700aa short isoform, 800aa intermediate isoform, 900 aa long isoform). Here, we analyzed the role of different NFAT2 transcripts in CLL with respect to disease phenotype and cell proliferation.
We investigated primary samples from CLL patients (n=30) for their expression profile of different NFAT2 isoforms using RT-PCR. Applying an shRNA approach, we generated stable knock-down cells of the CLL cell line MEC-1 for the long and intermediate isoforms and for the entire NFAT2 gene resulting in the complete ablation of all isoforms. The proliferation properties of the different MEC-1 cell lines was subsequently assessed in xeno-transplant experiments into NSG mice.
While physiological B cells express comparable levels of the short and intermediate/long isoforms, we could detect a five fold overexpression of the intermediate/long isoforms in primary CLL samples. To further analyze the differential regulation of the different NFAT2 transcripts on tumor cell proliferation and cell cycle regulation, we injected NSG mice with MEC-1 cells with intact NFAT2 (n=6), MEC-1 cells with a knock-down of the intermediate and long isoforms (n=6) and MEC-1 cells with a complete NFAT2 knock-down (n=6). MEC-1 cells with selective ablation of the intermediate and long NFAT2 isoforms grew significantly faster in NSG mice than MEC-1 cells with intact NFAT2 expression or MEC-1 cells with a complete NFAT2 knock-down. MEC-1 cells selectively lacking the intermediate and long isoforms led to accelerated tumor proliferation upon subcutaneous injection. Cell cycle analysis as assessed by flow cytometry showed a significantly increased number of cells in the G1/S-Phase for the group without expression of the short isoform, while the group with complete NFAT knock-down exhibited a compromised growth pattern as compared to wild-type MEC-1 cells.
In summary, our data demonstrate that genetic loss of the intermediate and long isoforms of NFAT2 leads to CLL acceleration

Disclosures
No relevant conflicts of interest to declare.
Rate of Rise of EBV Viral Load By Quantitative PCR after Allogeneic Transplantation Correlates with PTLD Facilitates Timely Institution of Rituximab
Authors: Muffly Lori S. , Shiraz Parveen , Nguyen Isabelle T , Brown Janice (Wes) M. 
Published: October 2019

Abstract
A growing body of data supports the benefit of first-line rituximab therapy for EBV- associated post-transplant lymphoproliferative disorder (PTLD) the potential for adoptive cellular therapy, and the benefit of early diagnosis. Quantitative PCR has proven to be an important, albeit not infallible, diagnostic test; however, data regarding the utility We reviewed the records of 731 adult patients who underwent allogeneic hematopoietic cell transplantation (HCT) at Stanford University Medical Center from January 2010 through Feb 2016.
(Table 1. Demographics) With respect to EBV screening, all patients received 400 mg po acyclovir tid for one year, preemptive strategies based on plasma PCR for CMV were used with either IV ganciclovir (MA) or valganciclovir (NMA) administered. Quantitative EBV PCR was performed on whole blood (Jan 2010 through mid-2011) or plasma (mid-2011 to present) every other week in asymptomatic patients or at any time a patient developed symptoms that may be attributed to EBV.
Observations/Conclusions:
· Both the initial measureable and the rate of increase in viral load were significantly greater in patients with PTLD than those with isolated viremia.
· Promptly administered rituximab was effective in improving resolution of PTLD when compared to historically reported data.
· Conversely, patients with low grade viremia without increase in viral load may be watched expectantly.
· Although not standard practice, importance of EBV serology of donor has been demonstrated in SOT and pediatric HCT and would be anticipated to be of equal importance in adults, especially
· Early biopsy or flow cytometric analysis of EBV+ lymphocyte to determine polymorphic versus monomorphic disease can further refine our understanding the pathophysiology of disease
· The crucial limitation of our study is that determination of EBV serology was not routine; however, by limiting our analysis to patients with EBV reactivation and the use of a standardized protocol to respond to viremia has permitted the current observations to be made.
Table 2 Table 2.

Disclosures
No relevant conflicts of interest to declare.
RAD51 Modulators Induce Mitotic Catastrophe in Aid Expressing Cells through Multiple Pathways
Authors: Mills Kevin , Hasham Muneer , Lamont Kristin , Philip Vivek , Branca Jane 
Published: October 2019

Abstract
The B-cell specific DNA mutase/recombinase Activation Induced Cytidine Deaminase (AID) initiates DNA double strand breaks (DSB) necessary for immunoglobulin class switch recombination in activated B-cells. However, AID-induced DSBs are not restricted to immunoglobulin loci, and we previously demonstrated that the homologous recombination (HR) factor RAD51 is critical to repair widespread AID-initiated DSBs. While it is known that failure to properly repair AID-induced genomic DSBs is cytotoxic to B-cells, the pathways culminating in cell death were not fully known. Here, we demonstrate that AID-initiated DSBs can trigger mitotic catastrophe, a non-apoptotic cell death mechanism. Using genetic and chemical biology approaches we further show that AID-induced mitotic catastrophe involves multiple pathways, including mTOR. We also demonstrate that AID-induced cytotoxicity can be used to effectively and preferentially target AID-expressing lymphoid malignancies, in xenograft cancer model. This study reveals a cell death response to genomic damage by AID, providing new insight into the mechanisms that ensure cellular survival during B-cell maturation. Our findings support the idea that RAD51 modulation may be leveraged as a novel "synthetic lethal" therapeutic approach for treating AID-expressing malignancies, via the induction of mitotic catastrophe.

Disclosures
Mills: Cyteir Therapeutics, inc: Employment, Equity Ownership, Patents & Royalties. Hasham:The Jackson Laboratory: Patents & Royalties.
Anemia in Diabetic Patients without Underlying Nephropathy. a Retrospective Cohort Study
Authors: Mahjoub Ali R. , Patel Eshan , Ali Sami , Webb Kendal , Astrow Alan , Kalavar Madhumati 
Published: October 2019

Background: Anemia (defined by the World Health Organization as a hemoglobin level of less than 13 mg/dl in men and less than 12 mg/dl in women) is common in diabetic patients, particularly in those with reduced renal function. Most studies attribute anemia in diabetic patients to kidney function impairment. There are no controlled systematic studies of the prevalence and predictors of anemia in patients with diabetes in the absence of overt nephropathy.
Objective: This retrospective cohort study was designed to measure the prevalence of anemia in diabetic patients with normal kidney function (estimated glomerular filtration rate (EGFR) greater than 90 ml/min/1.73 m2 and negative micro albuminuria) and compare it to a control group of non-diabetic patients. The study was designed to investigate diabetes as an independent risk for causing anemia.
Methods: We undertook a retrospective review of medical records of 400 patients (older than 18 years old) visiting the outpatient clinic in our institution between January and June 2015. 200 patients with diabetes (glycosylated hemoglobin (HgA1C) greater than 7) were compared to 200 non-diabetic patients (HgA1C level below 5.6), to identify the prevalence of anemia, with and without kidney disease and any other associated factors.
Results:The prevalence of anemia in all diabetic patients was 22% vs. 9% in non-diabetic group (OR: 2.69, 95% CI; 1.49 to 4.86, P = 0.001). Out of the 22% (n = 44) anemic patients, 18 patients had anemia with normal kidney function, out of those; 5 had iron deficiency anemia and one had autoimmune disease, while the remaining 12 patients (6%) did not have any obvious cause of anemia other than diabetes vs. 2 patients (1%) in non-diabetic patients who we did not find any explanation for the their anemia (OR: 6.6, 95% CI; 1.45 to 30, P < 0.015).
Conclusion: Most studies have highlighted an association between anemia and nephropathy in diabetic patients. This retrospective cohort study suggests a high incidence of anemia with unidentified pathology in the diabetic population. The mechanism of this anemia is not well understood, although direct glucose toxicity to erythrocyte precursors in the bone marrow or oxidative stress to mature erythrocytes are both possibilities. Prospective study of hemoglobin levels in patients with diabetes may help elucidate the precise mechanism of anemia in this group.

Disclosures
No relevant conflicts of interest to declare.
Observational Study of Magnetic Resonance Imaging Monitoring in Patients with Aplastic Anemia (AA) and Low or Int-1 Risk of Myelodysplastic Syndromes(MDS) Complicated with Iron Overload
Authors: Du Yali , Han Bing , Long Zhangbiao , Chen Miao , Hou Bo , Feng Feng , Wang Zhao 
Published: October 2019

Abstract

Objective: To observe the clinical characters and magnetic resonance imaging (MRI) T2* change in patients with aplastic anemia (AA) and low or int-1 risk of myelodysplastic syndromes(MDS) complicated with iron overload.
Methods:This is a prospective study. Patients with AA and low and intermediate-1 risk factors of MDS complicated with iron overload were enrolled. Clinical parameters and T2* of liver, heart and pancreas were collected regularly.
Results: Of all the 92 recruited patients (54 males and 38 females), the median age was 47.5 (15~18) year-old, median disease duration was 26 (3~407) months. There were 54 AA patients and 38 MDS patients. SF was related to liver T2* and pancreatic T2*(AA group r=-0.476,-0.509, respectively, P<0.05; MDS group r=-0.401, -0.565, respectively, P<0.05) in both groups, but not related to cardiac T2* in either group. SF level was correlated with transfusion amount in AA patients (r=0.403, P<0.05), but not in MDS patients (r=0.234, P>0.05. Of the 20 patients who received iron-chelation therapy, 3 (15%) had diarrheas, 2 (10%) decreased in hemoglobin, 2 (10%) decreased in platelet, 2 (10%) had increased creatinine, 1 (5%) had allergic dermatitis, but the symptoms relieved after drug withdrawal. 2 patients died of bone marrow failure during the follow-up period. 38 patients finished the half-year evaluation. Those (18 cases) who chelated adequately had significant decrease in SF (3354.9±3287.3 vs 2119.3±1742.0 ng/ml, P=0.044). Meanwhile an increase in hemoglobin level (83.5±30.0 vs 96.7±31.6 g/L, P=0.01) was noticed in those with decreased SF. 26 patients finished one year follow-up, those (15 cases) who chelated adequately had significant decrease in SF (3583.9±3543.8 vs 1693.9±1532.5 ng/ml, P=0.006), increase in cardiac T2* (25.0±13.4 vs 32.3±15.7 ms, P=0.015) and left ventricular ejection fraction (LVEF) (63.6±8.6 vs 67.1±5.2 %, P=0.028) , whereas no such changes were found in those did not chelate adequately. No change was found either in chelated or non-chelated groups in hemoglobin, WBC count, platelet count, SGPT, TBil, creatinine level, liver or pancreas T2* value. For those with decreased SF, even without chelation, increase in hemoglobin level (91.1 ±29.9 vs 108.4±31.4 g/L, P=0.046),cardiac T2* (25.5±13.7 vs 33.1±15.9 ms, P=0.014)) and LVEF level (63.6±8.6 vs 67.2±5.4 %, P=0.027) were also documented.
Conclusion: SF was related to liver T2* and pancreatic T2* in both AA and MDS groups. SF was relevant to transfusion amount in patients with AA but not MDS. Adequate iron chelation can decrease the SF level, and might bring improvement in cardiac T2* and LVEF level. Decrease of SF even without chelation might also benefit patients.

Disclosures
No relevant conflicts of interest to declare.