bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pharmacokinetics of Extended Half-Life Factor VIII Products By myPKFiTR Is Useful for Personalized Treatment in Children with Severe Hemophilia a
Authors: Matsumura Risa , Tomioka Keita , Nishimura Shiho , Mizoguchi Yoko , Kawaguchi Hiroshi , Kobayashi Masao 
Published: November 2019

Prophylactic administration of factor VIII products is necessary to prevent bleeding and preserve normal musculoskeletal function in children with severe hemophilia A (HA). Recently, extended half-life recombinant factor VIII (EHL-rFVIII) products have been utilized in HA patients. Therefore, the pharmacokinetics (PK) of EHL-rFVIII in individuals is needed to determine the appropriate administration for personalized prophylaxis according to the age, bleeding phenotype, the presence of arthropathy, and physical activity. The myPKFiTR ver 3.0 has been developed as the device to estimate the personalized dosing with a 2 sample PK based on the population PK (Bayesian) tool. In this study we report our single-center experience to study PK profiles and to individualize dose and dosing interval based on myPKFiTR. Eight patients with severe HA aged from 10 to 20 years were enrolled in this study for personalized prophylaxis. The half-life of EHL-rFVIII was approximately 15 to 18 hours in all patients studied. The clearance of FVIII was inversely correlated with the half-life of EHL-rFVIII. The EHL-rFVIII products have been basically administered twice a week. The trough levels of FVIII were more than 3% in all patients. The prophylactic regimen in adolescents was individually determined according to the personal simulation of PK study and to patients' life style and physical activities. Adolescent patients actively participated in sports, such as track and field, basketball, and football after school. The FVIII level after school was easily estimated by the use of myPKFiTR according to the dose and duration of replacement. The doses of EHL-rFVIII products were individually determined to have more than 10 to 30% of FVIII level at the time of sports activity. During personalized prophylaxis (6 to 18 months), all of patients studied have been no bleeds during sports as well as no spontaneous bleeds. Additionally, myPKFiTR has the capability of presenting the real-time FVIII level on the screen of smartphone after the replacement of EHL-rFVIII based on the individual PK. Some patients have referred their own FVIII level before the beginning of sports through their smartphone and then have decided the necessity of the replacement. These experiences suggest the enhancement of treatment concordance in the supportive relationship between patients and medical staff. Thus, the use of myPKFiTR may be essential for the optimization of prophylactic administration of EHL-rFVIII and for the medical adherence and concordance in each individual.

Disclosures
No relevant conflicts of interest to declare.
GATA2 Zinc Finger Mutations Influence Erythroid Differentiation and Chemotherapy Response
Authors: Leubolt Georg , Redondo Monte Enric , Wilding Anja , Kerbs Paul , Cusan Monica , Bagnoli Johannes , Enard Wolfgang , Hiddemann Wolfgang , Wichmann Christian , Schotta Gunnar , Greif Philipp A. 
Published: November 2019

The transcription factor GATA2 plays an important role in cell lineage decisions during hematopoiesis. GATA2 Zinc-Finger (ZF) mutations are associated with distinct entities of myeloid malignancies. Alterations of the N-terminal ZF1 were identified in AML patients with biallelic CEBPA mutations, whereas the C-terminal ZF2 is typically affected by germline mutations predisposing to MDS and AML, or by somatic lesions in CML blast crisis. Nevertheless, the context-dependent mechanisms underlying GATA2 ZF mutations remain mostly unclear. Here, we set out to study the functional consequences of GATA2 ZF mutations.
To test the effect on differentiation, we expressed GATA2 wild-type (WT) or GATA2 ZF mutants in human CD34+ hematopoietic stem and progenitor cells, stimulated with appropriate cytokines. Differentiation was evaluated by FACS-measurements of surface marker expression (erythroid markers: CD71, CD235a; granulocytic/monocytic markers: CD15, CD14). GATA2 WT caused a block of erythroid differentiation that is overcome by the ZF1 mutants (A318T and G320D), whereas the ZF2 mutant L359V may aggravate this block. For granulocytic/monocytic differentiation an overall block was observed for GATA2 WT and all the ZF mutants tested (Figure 1 A, B).
Recently, we and others observed GATA2 mutation gain in AML relapse (Greif et al., 2018 Clin Cancer Res; Christopher et al., 2018, NEJM), pointing towards a potential role in therapy resistance. Therefore, we treated K562 cells stably expressing GATA2 WT or mutants with Daunorubicin (one of the two drugs commonly used in AML chemotherapy). Expression of GATA2 A318T in K562 cells correlated with higher sensitivity to Daunorubicin and lower expression levels of IDH2. (Figure 1 C, D). Interestingly, this particular ZF1 mutation was the only one that got lost at relapse in the study by Christopher and colleagues, consistent with therapy sensitivity, whereas most of the GATA2 mutations gained at relapse were localized in the ZF2 domain.
In summary, GATA2 ZF mutations influence hematopoietic differentiation and chemotherapy response in a position-dependent manner. In the present study, we report distinct roles for individual GATA2 mutations depending on the affected ZF domain and altered amino acid positions. Understanding the oncogenic collaboration of GATA2 mutations with other driver genes in distinct patient subgroups is a challenge ahead.
Figure.1

Disclosures
Hiddemann: Roche: Consultancy, Honoraria, Research Funding; Gilead: Consultancy, Honoraria; Bayer: Research Funding; Vector Therapeutics: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Janssen: Consultancy, Honoraria, Research Funding.
Blocking MAPK Activation and Immune Checkpoints Reverse Immune Dysfunction and Reduce Disease in a Mouse Model of LCH
Authors: Sengal Amel , Hahne Meryl , Velasquez Jessica , Abhyankar Harshal , Burke Thomas , Olea Walter , Reyes Robert , Scull Brooks , Bigenwald Camille , Bollard Catherine M. , Yu Wendong , Merad Miriam , McClain Kenneth L. , Allen Carl E. , Chakraborty Rikhia 
Published: November 2019

Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplasia characterized by granulomatous lesions containing pathological CD207+ dendritic cells (DCs) with persistent mitogen-activated protein kinase (MAPK) pathway activation. Standard of care chemotherapy strategies are inadequate or overly toxic for the majority of patients with multisystem disease. The mechanisms underlying development of inflammation in LCH lesions are poorly understood, and potential for immunotherapy has not been determined. Analysis of the LCH lesion identified the most prominent immune cells as T lymphocytes, second only to pathologic CD207+/CD1a+ DCs (LCH DC). Both CD8+ and CD4+ T cells exhibited 'exhausted' phenotypes with high expression of the immune checkpoint inhibitor receptors. LCH DC cells also showed robust expression of ligands to checkpoint inhibitor receptors. Lesion CD8+ T cells exhibited blunted expression of Th1 cytokines with impaired effector function. In contrast, regulatory T cells (Tregs) isolated from LCH lesions demonstrated intact suppressive activity. Treatment of BRAFV600ECD11c LCH mice with anti-PD-1, anti-TIM-3 and MEKi decreased the lesion size: MEKi treatment resulted in reduction of the myeloid compartment; anti-PD-1 and anti-TIM-3 were associated with reduction in the lymphoid compartment. However, combined treatment with MEKi and either anti-PD-1 or anti-TIM-3 significantly decreased both CD8+ T cell and myeloid LCH cells in a synergistic fashion. These results thus indicate MAPK hyperactivation in myeloid LCH cells drives recruitment and activation of functionally exhausted T cells within the LCH microenvironment. While the results from this study did not demonstrate a significant impact of checkpoint inhibition alone on tumor burden in experimental mice, combined MAPK and checkpoint inhibition may be a potentially effective therapeutic strategy for patients with LCH.

Disclosures
Bollard: self: Patents & Royalties: patent on HIV T cells; NexImmune: Equity Ownership; Torque: Equity Ownership; Cellectis: Membership on an entity's Board of Directors or advisory committees; Caballetta: Equity Ownership; Mana Therapeutics: Equity Ownership.
Melatonin Increases the Chemosensitivity of Diffuse Large B Cell Lymphoma Cells to Epirubicin By Inhibiting P-Glycoprotein Expression Via the NF-Κb Pathway
Authors: Sun Xiuhua , Qin Hong 
Published: November 2019

Epirubicin (EPI) is a first-line chemotherapeutic drug for the clinical treatment of diffuse large B cell lymphoma (DLBCL), but the overexpression of multidrug resistance (MDR) transporter proteins, such as P-glycoprotein (P-gp), renders EPI ineffective. On the basis of some studies, melatonin (MLT) is considered to possess the potential for chemotherapeutic synergy that can be leveraged to overcome MDR. In this study, we investigated the effect of MLT on the sensitivity of DLBCL cells to EPI. The human DLBCL cells SUDHL-6 and SUDHL-10 were incubated with MLT and/or EPI for 48 h. We found that MLT significantly enhanced the EPI-induced suppression of cell proliferation and EPI-induced apoptosis and reduced the IC50 value of the EPI-treated cells. We also found that MLT synergized with EPI to promote the activation of the mitochondria-mediated apoptosis pathway. Further mechanistic studies demonstrated that the sensitivity of DLBCL cells to EPI may be affected by the MLT influence on the expression and function of P-gp. Immunohistochemical staining studies in DLBCL cells from tumor tissues revealed that the expression of P-gp was positively correlated with NF-κB P65 expression. EPI was subsequently discovered to upregulate the expression of P-gp by activating the NF-κB pathway in the DLBCL cells. Further, we found that MLT reduced the amount of NF-κB P65 protein in the nucleus and abrogated the ability of P65 to bind to the ABCB1 promoter, decisively suppressing P-gp expression. In conclusion, our results demonstrated that MLT inactivates the NF-κB pathway and downregulates the expression of P-gp, ultimately sensitizing DLBCL cells to the EPI that suppresses their growth.

Disclosures
Qin: InnoLifes: Consultancy, Equity Ownership; Pepromene Bio: Consultancy, Equity Ownership.
Assessment of Cardiac Abnormalities in Sickle Cell Disease Patients Using Echocardiography
Authors: Mukarram Osama , Lima Gian , Crooks Samuel , Kim Min Jung , Kim Agnes S 
Published: November 2019

Background: Sickle Cell Disease (SCD) is characterized by chronic anemia and recurrent ischemia-reperfusion episodes that contribute to high output heart failure. The effects of SCD on the heart are significantly underrecognized.
Methods: SCD patients who underwent echocardiography between March 2016 and March 2018 were retrospectively analyzed. Patients with reduced Left Ventricular Ejection Fraction (LVEF) and valvular heart disease were excluded. Cardiac chamber size, systolic and diastolic function parameters, and LV and RV strain were compared between hemoglobin SS (most severe form of SCD) and SC (less severe form) subtypes and against healthy controls. Wilcoxon signed rank test was used for statistical analysis.
Results: SS patients (n = 48, mean age 31.9) had lower mean hemoglobin 8.9 g/dl vs 11.3 g/dl (p < 0.001) and hematocrit 25.8% vs 31.4% (p = 0.008) and higher LDH 437 IU/L vs 258 IU/L (p < 0.001) compared to SC patients (n = 11, mean age 34.4). Both SS and SC patients had worse diastolic function compared to healthy controls: higher E velocity 98.9 cm/s (SS), 86.4 cm/s (SC), 76.4 cm/s (control) (SS vs control, p < 0.01; SC vs control, p < 0.05) and higher E/A ratio 1.76 (SS), 1.59 (SC), 1.15 (control) (SS vs control, p < 0.001; SC vs control, p < 0.01). SS patients had larger indexed left atrial volume compared to SC patients (39.3 ml/m2 vs 28.4 ml/m2, p = 0.007). There was no significant difference in LVEF, left ventricular global longitudinal strain, or right ventricular strain between SS and SC subtypes compared to healthy controls. Furthermore, SS patients with a serum LDH > 500 IU/L had higher E/e' ratio (11.3 vs 7.2, p=0.001) and larger indexed Left Ventricular End Diastolic Volume (LVEDVi) (80.8 ml/m2 vs 53.4 ml/m2, p=0.002) compared to SS patients with LDH < 500 IU/L.
Conclusion: SCD genotype adversely determines the degree of cardiac dysfunction in patients with SCD. LVEDVi, left atrial size, E velocity, E/A ratio, and E/e' ratio may serve as useful echocardiographic parameters to follow in this patient population. Serum LDH has been associated with poor clinical outcomes in patients with SCD, and as demonstrated by our study, it also portends worsening cardiac function in this population at high risk for heart failure.
Figure

Disclosures
No relevant conflicts of interest to declare.
Impact of Primary Ambulatory Thromboprophylaxis (PATP) with Low-Molecular Weight Heparins (LMWHs) on Survival in Patients with Locally Advanced or Metastatic Lung Cancer (LC) Receiving Chemotherapy
Authors: Thein Kyaw Zin , Tijani Lukman , Oo Thein H. 
Published: November 2019

Introduction: LC is the leading cause of cancer mortality in USA. PATP was provided in experimental trials to decrease the venous thromboembolism (VTE) rates and to provide anti-tumor effect with ultimate aim to improve survival in patients with solid cancers as VTE is the second leading cause of death in cancer patients. We undertook a systematic review and meta- analysis of randomized controlled trials (RCTs) to determine the impact of PATP with LMWHs on overall survival (OS) in patients with locally advanced or metastatic LC.
Methods: We performed a comprehensive literature search using MEDLINE and EMBASE databases through July 26, 2019. The references of all potential studies were also reviewed for any additional relevant studies. RCTs utilizing PATP with LMWHs in patients with locally advanced or metastatic lung cancer were incorporated in the analysis. A generic inverse variance method was used to calculate the estimated pooled hazard ratio (HR) for progression or metastasis free survival and OS with 95% confidence interval (CI). Heterogeneity was assessed with Cochran's Q -statistic. Random effects model was applied.
Results: A total of 3,452 patients with lung cancer from six RCTs were included in our meta-analysis. The prophylactic doses of bemiparin, dalteparin, tinzaparin, nadroparin and intermediate dose of enoxaparin were used in the studies. The duration of LMWH ranged from 3 to 6 months. The randomization ratio was 1 to 1 in all studies. The I2statistic for heterogeneity was 64, suggesting moderate heterogeneity among RCTs. The pooled HR for OS was not statistically significant at 1.02 (95% CI: 0.83-1.26; P = 0.83). In a subset of small cell lung cancer (SCLC) patients, the pooled HR for OS was 1.03 (95% CI: 0.72-1.48; P = 0.85). The HR for OS was noted at 1.70 (95% CI: 0.70-4.15; P = 0.24) in patients with limited stage SCLC. In a subset of non-small cell lung cancer, the pooled HR of OS was 1.00 (95% CI: 0.79-1.26; P = 0.98). The pooled HR for progression or metastasis free survivalwas 1.03 (95% CI: 0.86-1.24; P = 0.74) according to an analysis of 5 RCTs.
Conclusions: Our meta- analysis demonstrated that no survival advantage was noted with the addition of PATP with LMWHs to routine standard chemotherapy in patients with locally advanced or metastatic LC, regardless of histology types as well as stages of SCLC.

Disclosures
Oo: Medical Education Speakers Network: Honoraria; Janssen and Janssen: Other: Research: site co-investigator .
Updated Meta-Analysis of Randomized Controlled Trials on Primary Ambulatory Thromboprophylaxis (PATP) in Patients with Advanced Pancreatic Cancer (APC) Receiving Chemotherapy
Authors: Thein Kyaw Zin , Quick Donald P. , Oo Thein H. 
Published: November 2019

Introduction: Thrombosis is the second leading cause of death in cancer patients and patients with APC are categorized as high-risk of developing venous thromboembolism (VTE). Many trials had failed to demonstrate improvement in survival with PATP. Despite decreasing VTE events, PATP in solid cancer patients is not routinely recommended. We conducted an updated meta- analysis of RCTs to determine the benefit and risk of PATP with low-molecular weight heparins (LMWH) and direct oral anticoagulants (DOAC) in patients with APC receiving chemotherapy.
Methods: We performed a comprehensive literature search using MEDLINE and EMBASE databases through June 30, 2019. The references of all potential studies were also reviewed for any additional relevant studies. The RCTs with reduction in VTE as a primary or secondary endpoint and the major bleeding (MB) as a safety outcome were incorporated in the analysis. The primary meta- analytic approach was a random effects model using the Mantel-Haenszel (MH) method. It was used to calculate the estimated pooled risk ratio (RR), and risk difference (RD) with 95% confidence interval (CI). Heterogeneity was assessed with Cochran's Q- statistic.
Results: A total of 1,013 patients with APC from two RCTs and a subgroup of another three RCTs were included in our meta-analysis. The prophylactic, intermediate and therapeutic doses of LMWH and prophylactic dose of DOAC (rivaroxaban) were used in the studies. The duration of LMWH and DOAC ranged from 3 to 6 months. The randomization ratio was 2 to 1 in PROTECHT study and 1 to 1 in all other studies. The I2statistic for heterogeneity was 60, suggesting some heterogeneity among RCTs. The VTE incidence was 28 (5.43%) in PATP group and 60 (12.07%) in control group with a RR of 0.44 (95% CI: 0.20 to 0.99, P = 0.05) and RD of -0.06 (95% CI: -0.11 to -0.01, P = 0.01). In fixed effects model, the pooled RR was 0.45 (95% CI: 0.29 to 0.70, P = 0.0003) and the absolute RD in VTE was -0.07 (95% CI: -0.10 to -0.03, P = 0.0002) with an estimate of the number needed to treat (NNT) of 15 to prevent one VTE event. MB events were reported in 9 (4.11%) patients in PATP group compared to 7 (3.27%) in control group according to an analysis of 2 RCTs. The pooled relative risk for MB was statistically non-significant at 1.25 (95% CI: 0.47 to 3.31, P = 0.65).
Conclusions: In our study, PATP in APC may statistically significantly decrease VTE events, approximately with relative risk reduction of 55% and a NNT of 15, without increasing MB events. Proper selection of patients who are high risk for VTE in outpatient setting is important. More RCTs are required to further define high risk subsets of APC patients receiving chemotherapy who may benefit from PATP.

Disclosures
Oo: Medical Education Speakers Network: Honoraria; Janssen and Janssen: Other: Research: site co-investigator .
BALB/c-hCD3E Transgenic Mice: An Ideal Animal Model for In Vivo Efficacy Study of CD3-Bispecific Antibodies
Authors: Ju Cunxiang , Zhang Mingkun , Jia Dongjing , Tang Jing , Li Shuai , Zhao Jing , Wang Demin , Gao Xiang 
Published: November 2019

T cell-mediated immunotherapy is an important strategy for treatment of a variety of tumors.T-cell bispecific antibody (TCB), which binds to a tumor associated antigen (TAA) and human CD3ε (hCD3ε) and directs specific killing of tumor cells carrying the TAA. However, there is still an unmet need for suitable animal models to evaluate the therapeutic efficacy of TCB candidates. Although immune-deficient mice transfused with human PBMCs can be used to evaluate the TCBs, these mice are not fully immune competent and have limitation in studying immune system activation by TCBs. Herewe developed a human CD3ε-transgenic mouse model with BALB/c background (BALB/c-hCD3E) to overcome the limitation.
CD3ε plays a critical role in formation and function of the TCR-CD3 complex. However, hCD3ε shares only 47% homology with mouse CD3ε (mCD3ε) in the extracellular domain, indicating that hCD3ε and mCD3ε are not functionally interchangeable. Thus, we established BALB/c-hCD3E mice that carry the human CD3E gene and its entire regulatory sequence and co-express hCD3ε and mCD3ε in over 90% of T cells. BALB/c-hCD3E mice were phenotypically normal and had normal T-, B- and NK-cell populations compared to wild-type BALB/c mice. In addition, splenic T cells from BALB/c-hCD3E mice could be activated by either anti-hCD3 or anti-mCD3 antibodies to produce IFNg, IL2 and TNFa in vitro. Moreover, we knocked out mCD3e in BALB/c-hCD3E mice to generate BALB/c-hCD3E/mCd3e-KO mice that displayed a marked reduction in the number of splenic T cells. BALB/c-hCD3E/mCd3e-KO mice also had reduced percentages and numbers of CD4+ and CD8+T cells. Importantly, anti-mCTLA4 antibodies strongly inhibited the growth of subcutaneously inoculated CT26 tumor cells in BALB/c-hCD3E but not BALB/c-hCD3E/mCd3e-KO mice.
Taken together, these findings demonstrate that mCD3ε is indispensable for T-cell development and function in BALB/c-hCD3E transgenic mice and these mice are a novel and valuable model to assessing the therapeutic efficacy of TCBs.

Disclosures
Ju: GemPharmatech Co., Ltd: Employment. Zhang:GemPharmatech Co., Ltd: Employment. Jia:GemPharmatech Co., Ltd: Employment. Tang:GemPharmatech Co., Ltd: Employment. Li:GemPharmatech Co., Ltd: Employment. Zhao:GemPharmatech Co., Ltd: Employment. Wang:GemPharmatech Co., Ltd: Employment. Gao:GemPharmatech Co., Ltd: Employment.
Delayed Time to Emergency Hip Surgery in Patients Taking Oral Anticoagulants
Authors: Cafaro Teresa , Tagalakis Vicky , Koolian Maral , Camille Simard 
Published: November 2019

Abstract
Introduction
Surgical intervention within 48 hours of admission in patients with acute hip fractures has been associated to lower morbidity and mortality. Patients anticoagulated with Vitamin K Antagonists (VKAs) have longer time to corrective surgery than those not anticoagulated. Sparse data exists on time to surgery (TTS) in patients taking direct oral anticoagulants (DOACs). The aim of this study is to establish TTS among non-anticoagulated and anticoagulated patients taking either VKA or DOAC.
Materials and Methods
We conducted a retrospective cohort study of consecutive patients admitted with acute hip fracture between July 1, 2016 and December 31, 2017. Patient-, anticoagulant- and surgery-related characteristics were collected. The primary outcome was TTS calculated from time of admission. Median TTS with interquartile range (IQR) was compared among 3 groups of patients: DOAC, VKA and No OAC.
Results
A total of 472 patients were included: 12.5% (59/472) were anticoagulated (28/472 on VKAs and 31/472 on DOACs). Median TTS was longer in the VKA group [64 hours (IQR: 50-84)] and in the DOAC group [(61 hours; IQR: 42 to 77)] versus the No OAC group [44 hours (IQR: 28-63), p =0.0006 and p=0.003 respectively]. There was no significant difference in median TTS in the VKA group versus the DOAC group (p =0.6396).
Conclusion
Patients taking either VKA or DOAC have significant delays to emergency hip fracture surgery compared to those not anticoagulated. An action plan aimed at early identification and cessation of anticoagulation is warranted in this vulnerable group of patients.
Keywords: Direct Oral anticoagulants, Vitamin K antagonists, Anticoagulation, Acute hip fracture, Emergency surgery

Disclosures
Tagalakis: BMS-Pfizer: Other: participated on ad boards; Sanofi Aventis: Other: investigator initiated grant;participated on ad boards; Pfizer: Other: participated on ad boards; Bayer: Other: participated on ad boards; Servier: Other: participated on ad boards.