bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Impact of 18f-FDG PET/CT As a Valuable Prognostic Tool for the Newly Diagnosed Multiple Myeloma with Extramedullary Disease
Authors: Baek Dong Won , Cho Hee Jeong , Sohn Sang Kyun , Jung Sung-Hoon , chae Moon Hong , Moon Joon Ho 
Published: November 2019

Purpose
18F-FDG PET/CT (PET/CT) could be a valuable tool to predict long-term survival outcomes in patients with newly diagnosed multiple myeloma (MM). It has ability to distinguish metabolically active sites such as extramedullary disease (EMD) as well as bone damage with relatively high sensitivity and specificity. In this study, we attempted to evaluate the role of PET-CT as a novel prognostic tool for patients with newly diagnosed MM who have EMD.
Patients and Methods
This study included 211 patients who were newly diagnosed with multiple myeloma from Kyunpook National University Hospital and Chonnam National University Hwasun Hospital. We retrospectively analyzed the medical records of enrolled patients. PET/CT was performed at the diagnosis and EMD was identified in 36 patients (17.1%).
Results
With a median follow-up duration of 21.5 months (range 1.4-67.7), the estimated 2-year PFS and OS rates were 46.1% and 79.6%, respectively. The presence of PET/CT positive EMD and high maximum standardized uptake value (SUVmax) on baseline PET/CT were significantly associated with inferior long-term survivals in terms of PFS (p=0.013, p=0.007) and OS (p=0.002, p=0.004). In addition, patients who underwent autologous stem cell transplantation (auto-SCT) showed superior PFS (p=0.005) and OS (p=0.022) in PET/CT positive EMD group. Meanwhile, Revised-International Staging System (R-ISS) successfully predicted the prognosis in this study. When we modified R-ISS with the presence of EMD, survival outcomes of the R-ISS stage III patients who didn't have EMD were similar to R-ISS II, while patients with PET/CT positive EMD showed even worse prognosis than the R-ISS stage III group. In the multivariate survival analysis, the presence of EMD (hazard ratio (HR), 2.397; 95% confidence internal (CI), 1.281-4.483; p=0.006) and auto-SCT (HR, 0.326; 95% CI, 0.194-0.549; p<0.001) were related to PFS, while LDH (HR, 2.56; 95% CI, 1.221-5.366; p=0.013) level and auto-SCT (HR, 0.398; 95% CI, 0.167-0.953; p=0.039) were independent prognostic factors of OS.
Conclusion
In conclusion, PET/CT positive EMD was a poor prognostic factor in patients with newly diagnosed MM. In addition, PET/CT could be a valuable tool to make better risk-adapted treatment strategies with R-ISS in EMD positive MM patients. Above all, patients with PET/CT positive EMD should be considered auto-SCT to improve long-term survivals.
Figure

Disclosures
No relevant conflicts of interest to declare.
Reduced Number and Function of Endothelial Progenitor Cells in Patients with Beta Thalassemia Major
Authors: Spectre Galia , Stark Pinhas , Steimatzky Arza , Raanani Pia , Lev Dorit , Kornowski Ran , Pazgal Idit 
Published: November 2019

There is a high incidence of venous thromboembolic complications in patients with beta thalassemia .Recently, a high incidence of asymptomatic multiple silent cerebral infarcts, presumably of arterial origin, were demonstrated in patients with beta thalassemia major (TM) and intermedia. The etiology remains unclear. Endothelial progenitor cells (EPCs) derive from the bone marrow and contribute to regeneration of endothelial damage. We hypothesized that the change in EPCs contributes to the vascular events in patients with TM and that patients have high markers for arterial thromboembolism.
Study aim: To assess the change in the number and function of EPCs in TM patients.
Methods: Blood samples were drawn from adult patients with TM before blood transfusion and from healthy volunteers. EPCs were isolated from peripheral mononuclear cells by using a Ficoll density-gradient centrifugation. EPCs' levels were detected by flow cytometry, namely by co-expression of VEGFR-2 in CD34 or CD133 positive cells. EPCs were cultured for 7 days after which functional properties were evaluated by: their capacity to form colonies using light microscopy (a colony forming unit (CFU) defined as a cluster of at least 100 flat cells surrounding a cluster of rounded cells) as well as by using a viability assay (using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay). The adhesion molecule P- selectin and the chemokine SDF1 were measured by using Elisa kits according to the manufacturer instructions.
Results: Thirty five patients (median age 35 range 27-47y) and 15 healthy volunteers (median age 34.5 range 30-50y) were included. Lower levels of CD34/VEGFR-2 cells and a trend towards CD133/ VEGFR-2 cells were observed in TM patients (Figure 1A). Viability of EPCs and their ability to perform colonies were markedly reduced in TM patients compared to controls; mean MTT 0.05± 0.05 in TM vs. 0.23±0.08 (p<0.01), median number of CFU = 0, range 0-3 in TM vs. 4, range 1-6 in controls (p<0.001), (a representative picture, Figure 1b). SDF1a and P selectin levels were significantly higher in TM compared to healthy volunteers (p=0.05 and P<0.01 respectively).
Conclusions: Our results suggest that patients with TM have reduced number and function of EPCs. This may contribute to the silent cerebral infarcts and hypercoagulable state in thalassemia. Furthermore, SDF1a and P selectin, both markers for coronary artery disease were higher in TM. Further studies are planned to establish the possible role of other cytokines in this process.

Disclosures
Spectre: Pfizer: Honoraria; Sanofi: Honoraria, Membership on an entity's Board of Directors or advisory committees; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Boeringer ingelheim: Honoraria; Bayer: Membership on an entity's Board of Directors or advisory committees.
Inhibiting the Hippo Signaling Pathway Key Molecule YAP Suppresses Mantle Cell Lymphoma Proliferation By Regulating Multiple Pathogenrelated Signaling Pathways
Authors: Xiong Wenjie , Yi Shuhua , Yan Yuting , Li Zengjun , Liu Wei , Lv Rui , Yu Zhen , Zou Dehui , Qiu Lugui 
Published: November 2019

Object: Mantle cell lymphoma (MCL) is a rare hematologic malignancy with heterogeneous course and always resistant to chemotherapy at advance or relapsed stage. Hippo signaling pathway is one of the most popular pathways in solid tumors. The major role of this pathway is regulating cell proliferation, migration, and maintaining the stemness of cells. However, the relationship between this pathway and MCL had been rarely studied. The purpose of this study is to investigate the relationship between MCL and YAP, the key molecule Hippo signaling pathway, to further clarify the mechanism of MCL resistance and to find new therapeutic targets.
Methods: In this study, expression of YAP in B cell non-hodgkin's lymphoma (B-NHL) and mantle cell lymphoma detected by real-time PCR, Western blot analysis, immunofluorescence. Knockdown of YAP by sh-RNA or inhibit the function of YAP using verteporfin (VP). Next, the effects of YAP knockdown and YAP inhibitor on MCL cells were evaluated by fluorescence detection, real-time PCR and Western blot. Cell count, CCK8, Soft Agar Assay and in vitro functional assays were performed to elucidate the function of YAP-mediated cell proliferation, the effect on signaling pathway and the relationship with chemoresistance in MCL.
Results: We detected the expression of YAP in 40 B-NHL patients and 10 healthy donor and found that YAP were overexpressed in relapsed and newly diagnosis patients and it was extremely higher in relapsed patients. Moreover, YAP is extremely high express on MCL. Knockdown YAP by shRNA or YAP inhibitor verteporfin (VP) could not only inhibit the proliferation, induce the apoptosis of MCL cell lines, but also lead to cells stopping in G1 phase. In addition, knockdown YAP could sensitize Z138 cells to the cytotoxics of BTK inhibitors ibrutinib and SYK inhibitor R788.Combined VP and ibrutinib or R788 had obvious synergy. Most importantly, knockdown YAP could obviously reduce the expression of protein involved in G1 phase, inhibit the BCR signaling pathway and PI3K AKT signaling pathway, and promote cell apoptosis and DNA damage signaling pathways. Through regulating multiple signaling pathways, YAP played an important role in cell proliferation and drug resistance in B-NHL.
Conclusion: It was the first time to demonstrate that Hippo signaling pathways may associate with the pathogenesis and drug resistance of MCL. Targeting the key molecular of Hippo signaling pathways may be potential therapeutic targets of MCL patients especially the relapse and refractory patients.

Disclosures
No relevant conflicts of interest to declare.
Maintenance Therapy in Patients with Chronic Lymphocytic Leukemia Who Achieved MRD-Positive Remissions after Induction Therapy
Authors: Kuvshinov Alexey , Voloshin Sergei , Shuvaev Vasily , Belyakova Elena , Mikhaleva Mariia , Garifullin Andrey , Rysev Georgiy , Bogdanov Alexander , Martynkevich Irina , Kleina Elizaveta , Samorodova Alena , Ruzhenkova Iuliia , Vokueva Iuliia , Chechetkin Alexander 
Published: November 2019

Introduction. There is strong evidence that the achievement of a negative status for minimal residual disease (MRD) is the single most important factor predictive of final outcome in patients with chronic lymphocytic leukemia (CLL) both in previously untreated and relapsed patients. Maintenance treatment (MT) with the CD20-directed monoclonal antibodies rituximab or ofatumumab yields better progression-free survival (PFS) compared with observation alone in individuals with CLL who have received induction therapy with chemo-immunotherapy. However maintenance therapy of anti-CD20 antibodies, although approved in some other B-cell malignancies, is not yet approved in CLL by regulators.
Aim. We wanted to assess overall and progression-free survival in MRD-positive patients who received maintenance therapy.
Methods. The study included 58 patients (median age 62 years, range 36-82). Male to female ratio - 1.7:1. We have used NCI revised guidelines (Hallek M, et al., 2008) for treatment initiation, assessment of response and MRD. Induction chemotherapy was conducted under the following programs: RB, RFC. The median line of therapy was - 1 (1-5). Evaluation of MRD was performed using 5-color flow cytometry of the bone marrow cells. The maintenance therapy was conducted MRD-positive patients (n=41): Rituximab 500 mg/m2 intravenously every 8 weeks for 2 years. The remaining MRD-negative patients (n=17) were under dynamic observation without therapy. Median observation in study was 51.5 month (15.2-134.8).
Results.The frequency of relapses in the group of patients with MT was 51.2%, in the group of patients without MT - 70.6% (p=0.18). MRD was not detected after 6-12 months of MT in 17.1% (7/41) had previously MRD-positive patients. The medians of PFS and OS were not different in the MRD-positive patients with MT versus in the MRD-negative patients without MT: PFS - 45.9 months and 57 months, respectively (p=0.83); OS - 106 and 128 months, respectively (p=0.47). Significant differences in the incidence of infectious complications between patients with MT and without of MT were not detected (p˃0.05).
Conclusions.Maintenance therapy for MRD-positive patients allows increasing of progression-free and overall survival to the level of patients with MRD-negative status. Maintenance therapy may be a means of control over the minimal residual disease and the method of its eradication. The proposed algorithm need further testing to confirm the initial results.
Figure

Disclosures
Shuvaev: Novartis: Consultancy; Pfize: Honoraria; BMS: Consultancy; Fusion Pharma: Consultancy.
Molecular Characterization and Frequencies of Different Genetic Ameliorating Factors in Transfusion Dependent Thalassemia Patients from District Peshawar
Authors: Mehmood Gulrukh , Taj Abid Sohail , Naz Arshi , Khan Tariq Masood 
Published: November 2019

Background
Thalassemia is one of the most common genetic blood disorders worldwide. The carrier rate of β thalassemia in Pakistan is found to be around 5-7%, rendering it a major health concern. Approximately 5000 children are diagnosed with thalassemia major every year in Pakistan due to consanguineous marriages. More than 200 causative molecular defects have been identified in β-globin genes. About 20 mutations account for 90% of the abnormal β-genes. Clinical severity of β-thalassemia is modified by different factors. The important ones include the type of disease causing mutation and the ability to produce α and γ globin chains. A better understanding of these ameliorating factors may have a significant impact on disease management.
Objectives
1. To re-assess diagnoses among local multi-transfused thalassemia syndrome patients.
2. To determine frequency of various genetic determinants of milder phenotypes of thalassemia among study patients.
Methods
Transfusion dependent thalassemia patients, upto 15 years age, were enrolled from Fatimid Foundation, Peshawar Pakistan. A comprehensive questionnaire encompassing demographic and clinical data was filled out for each patient. Genetic analyses for 2 alpha (α) and 13 prevalent beta (β) gene mutations and for polymorphisms at Xmn1-HBG2 and BCL11A were carried out on blood samples of the patients at National Institute of Blood Diseases, Karachi Pakistan. The data collected was analysed at Khyber Medical University (KMU) Peshawar Pakistan.
Results
A total of 54 transfusion dependent thalassemia patients were enrolled into the study. Homozygous or compound heterozygous combinations of β-globin gene mutations were identified in all the study patients. Eleven patients were found to have a co-existing heterozygous α (3.7kb) deletion, two patients had Xmn1-HBG2 polymorphism and 38 had BCL11A polymorphism. Homozygous Fr 8-9 was the most frequent mutation, found in 19 (35.2%) patients. Only 13 patients were found to have isolated β-globin gene mutations. In total, 46 (85.2%) study patients were identified to have an ameliorating genetic factor (a co-existing α-globin gene mutation, an Xmn1-HBG2 polymorphism or a BCL11A polymorphism) besides the main β-globin gene mutation.
Conclusion
It was concluded that co-existing genetic ameliorating factors are frequently found in transfusion dependent β-thalassemia patients of Peshawar District. These factors impart a milder phenotype to an otherwise severe disease. It is hence suggested that β-thalassemia patients from District Peshawar be screened for these factors and due consideration be paid by the physician in devising management plans.
Key Words
Thalassemia, Genetic modifier in thalassemia, Transfusion dependent Anaemia, Xmn1 polymorphism

Disclosures
No relevant conflicts of interest to declare.
Clinical Presentation, Management and Biomarkers of Cytokine Release Syndrome after Anti-CD19 CART-Cell Therapy for r/r ALL
Authors: Li Ping , Zhou Lili , Ye Shiguang , Li Shaoguang , Liang Aibin 
Published: November 2019

Chimeric antigen receptor T (CAR-T) cell therapy has emerged as a novel treatment modality for B-cell malignancies. CD19-specific CAR-T cells induce high rates of initial response among patients with relapsed B-cell acute lymphoblastic leukemia (ALL). However, cytokine release syndrome (CRS) is the most common and severe toxicities of CAR T-cell therapy for ALL, and clinical experience is limited. Here, we describe the clinical presentation and management of 30 patients who presented with CRS following CAR-T cell therapy for relapsed/refractory ALL at our hospital. 12 of the 30 patients (40%) developed grade 1-2 CRS, 14 patients (46.7%) presented with grade 3-4 CRS and 2 patients (6.7%) died of grade 5 CRS. Compared with grade 1-2 CRS, grade 3-4 CRS correlated negatively with overall survival and progression-free survival (P =0.02). We found that higher ferritin levels and percentages of CD19 positive cells in blood lymphocytes cells at time of CAR-T cell infusion were associated with more severe CRS. Grade 3-4 neurotoxicity was frequently present in patients with grade ≧3 CRS. We also observed that the organ disfunctions occurred in sequence after fever onset during the period of CRS. Neurotoxicity, cardiovascular disfunction and cytopenia in some patients manifest as biphasic. Compared to use of tocilizumab for CRS ≧ grade 3, early intervention of tocilizumab for hyperpyrexia duration ≧ 6h alleviates the severity of CRS, and no patients died of severe CRS since this management approach was performed. As use of novel CAR-T cell therapy expands, the data from our clinical experience may help others anticipated the clinical course of organ function and manage CRS in CAR-T therapy.
Figure

Disclosures
No relevant conflicts of interest to declare.
In Vitro and In Vivo Characterization of Marstacimab, an Anti-TFPI Antibody
Authors: Parng Chuenlei , Jin Macy , Holsti Matthew , Benard Susan , Rakhe Swapnil , Patel-Hett Sunita R. , Joyce Alison , Webster Rob , Pittman Debra D 
Published: November 2019

Hemophilia A and B are X-linked genetic disorders resulting from functional deficiencies of the intrinsic coagulation plasma proteins Factor VIII (FVIII) or Factor IX (FIX), respectively. An approach to achieving hemostatic pharmacology in hemophilia is to augment the extrinsic cascade is to neutralize Tissue Factor Pathway Inhibitor (TFPI). TFPI is a multi-Kunitz (K) domain inhibitor which binds to and inhibits Factor Xa via the K2 domain and Factor VIIa/Tissue Factor activity (K1 domain). Marstacimab is a fully human monoclonal antibody that binds to and neutralizes TPFI activity and is under development for treatment of hemophilia. To support lead identification during late discovery, five anti-tissue factor pathway inhibitor (TFPI) antibodies (EC50<5nM) were evaluated in an in vivo rabbit pharmacokinetics/pharmacodynamics (PK/PD) study and in vitro using rabbit and human hemostatic assays. Marstacimab displayed potent in vitro activities, and potent pharmacology in shortening the dilute prothrombin time (dPT) assay (1.3 nM) and in the thrombin generation assay (TGA) (2.8 nM). Following an intravenous administration in rabbits, marstacimab and the other anti-TFPI antibodies exhibited target-mediated drug disposition (TMDD) with a half-life less than 2 days, when compared to a control IgG. The pharmacokinetics of anti-TFPI antibody can be described using a Michaelis-Menten mechanistic model in which in vivo Km and Vmax were estimated for the observed nonlinear clearance. The estimated pharmacokinetic parameters for marstacimab was 45 and 38 mL/kg for the central and peripheral volume of distribution, 23 nM for Km, 131 nM/kg/hr for Vmax. In addition, the correlation between in vitro Kd, in vivo Km and in vitro/in vivo potency were assessed. Compared to other anti-TFPI antibodies, marstacimab exhibited reduced clearance and a good in vitro-in vivo relationship. Second, the binding epitopes were further characterized using competitive surface plasmon resonance (SPR) and marstacimab has distinct binding epitopes from the other antibodies. Lastly, the potential human pharmacokinetics and efficacy were evaluated using allometry scaling and in vitro potency, the results predict a minimal target occupancy of 76% following a weekly subcutaneous administration of marstacimab. These studies indicate that the differential influence of the TFPI-binding epitopes on the pharmacokinetics, but not on the pharmacodynamics. The selection strategy could be applicable to the other antibody discovery and development.

Disclosures
Parng: Pfizer: Employment. Jin:Pfizer: Employment. Holsti:Pfizer: Employment. Benard:pfizer: Employment. Rakhe:Pfizer Inc.: Employment. Patel-Hett:Pfizer: Employment. Joyce:Pfizer: Employment. Webster:pfizer: Employment. Pittman:Pfizer Inc.: Employment.
Poor Outcomes of HLA-Mismatched Allogeneic Hematopoietic Cell Transplantation and High Ferritin Levels after a Reduced-Intensity Conditioning Regimen in Patients with Advanced Myelofibrosis
Authors: Lee Han Bi , Yoon Jae-Ho , Min Gi June , Park Sung-Soo , Park Silvia , Lee Sung-Eun , Kim Yoo-Jin , Lee Seok , Kim Hee-Je , Min Chang-Ki , Cho Seok-Goo , Lee Jong Wook , Eom Ki-Seong 
Published: November 2019

Allogeneic hematopoietic cell transplantation (allo-HCT) preconditioning intensity, donor choice, and graft-versus-host disease (GVHD) prophylaxis for advanced myelofibrosis (MF) have not been fully elucidated. Thirty-five patients with advanced MF were treated with reduced-intensity conditioning (RIC) allo-HCT. We searched for matched sibling (n=16) followed by matched (n=10) or mismatched (n=5) unrelated and familial mismatched donors (n=4). Preconditioning regimen consisted of fludarabine (total 150 mg/m2) and busulfan (total 6.4 mg/kg) with total body irradiation≤ 400cGy. All showed engraftments, but four (11.4%) showed either leukemic relapse (n=3) or delayed graft failure (n=1). Two-year overall survival (OS) and non-relapse mortality (NRM) was 60.0% and 29.9%, respectively. Acute GVHD was observed in 19 patients, and grade III-IV acute GVHD was higher with HLA-mismatch (70% vs. 20%, p=0.008). Significant hepatic GVHD was observed in nine patients (5 acute, 4 chronic), and six of them died. Multivariate analysis revealed inferior OS with HLA-mismatch (HR=6.40, 95%CI 1.6-25.7, p=0.009) and in patients with high ferritin level at post-HCT D+21 (HR=7.22, 95%CI 1.9-27.5, p=0.004), which were related to hepatic GVHD and high NRM. RIC allo-HCT can be a valid choice for advanced MF. However, HLA-mismatch and high post-HCT ferritin levels related to significant hepatic GVHD should be regarded as poor-risk parameters.

Disclosures
Kim: Handok: Honoraria; Amgen: Honoraria; Celgene: Consultancy, Honoraria; Astellas: Consultancy, Honoraria; Hanmi: Consultancy, Honoraria; AGP: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; SL VaxiGen: Consultancy, Honoraria; Novartis: Consultancy; Janssen: Honoraria; Daiichi Sankyo: Honoraria, Membership on an entity's Board of Directors or advisory committees; Otsuka: Honoraria; BL & H: Research Funding; Chugai: Honoraria; Yuhan: Honoraria; Sanofi-Genzyme: Honoraria, Research Funding; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees. Lee:Alexion: Consultancy, Honoraria, Research Funding; Achillion: Research Funding.