bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of Zyx Impairs Neutrophil Production in Zebrafish Embryos
Authors: Konno Katsuhiro , Kulkeaw Kasem , Nii Takenobu , Kaneyuki Ayako , Sugiyama Daisuke 
Published: November 2019

Zebrafish is a useful model to study vertebrate hematopoiesis, but lack of antibodies to zebrafish proteins has limited purification of hematopoietic cells. Here, we purified neutrophils from larval and adult zebrafish using the lectin Phaseolus vulgaris erythroagglutinin (PHA-E) and DRAQ5, a DNA-staining fluorescent dye. In adult kidney marrow, we purified neutrophil-like PHA-E4low DRAQ5low cells, which neutrophil-type granules. Specifically, at 96-hr post fertilization, we sorted large-sized cells from larvae using forward scatter and found that they consisted of PHA-Elow DRAQ5low populations. These cells had myeloperoxidase activity, were Sudan Black B-positive and expressed high levels of neutrophil-specific (csf3r and mpx) mRNAs, all neutrophil characteristics. Using this method, we conducted functional analysis suggesting that zyxin (zyx) plays a role in neutrophil generation in zebrafish larvae. Overall, PHA-E and DRAQ5-based flow cytometry may serve as a tool to purify zebrafish neutrophils.

Disclosures
No relevant conflicts of interest to declare.
Trasposon Mediated Horizontal Gene Transfer (T-HGT) of Circulating Tumor DNA Reshapes MM Clonal Architecture
Authors: Cinar Munevver , Flygare Steven , Mosunjac Marina , Nagaraju Ganji , Park Dongkyoo , Faith Sheff , Saxe Debra , Bernal-Mizrachi Leon 
Published: November 2019

Spatial genetic heterogeneity is a characteristic phenomenon that influences multiple myeloma's (MM) phenotype and drug sensitivity (Rasche L. et al and Bolli N et al.). Hence, the branch model of tumor evolution is not sufficient to explain the disorganized architecture observed in MM. In this study, we investigated whether MM ctDNA horizontal gene transfer (HGT) affect tumor genetic architecture and drug sensitivity, resembling what is seen in prokaryotes, and elucidated the mechanisms involved in the mobilization of genetic material from one cell to another. We identified that plasma from patients with MM transmits drug sensitivity or resistance to cells in culture. This transmission of drug sensitivity is mediated by ctDNA transfer of oncogenes to a host cell. Importantly, in vitro and in vivo demonstrated that ctDNA mainly targets cells resembling the cell of origin (tropism). Karyotype spreads and whole genome sequencing demonstrated that once patients ctDNA encounters host cells, it migrates into the nucleus where it ultimately integrates into the cell's genome. Integration to the genome was confirmed to be targeted to myeloma cells. Further sequencing analysis of multiple MM samples identified ctDNA tropism and integration is dependent on the 5' and 3' end presence of transposable elements (TE), particularly of the MIR and ALUsq family. These results were further validated by TE mediated delivery of GFP into MM cells in vitro and HSVTK in tumors of mouse xenografts. In conclusion, this data indicates for the first time that TE mediates MM ctDNA HGT into homologous tumor cells shaping the hierarchical architecture of tumor clones and affecting tumor response to treatment. Therapeutically, this unique quality of ctDNA can be exploited for targeted gene therapeutic approaches in MM and potentially other cancers.

Disclosures
Bernal-Mizrachi: Kodikas Therapeutic Solutions, Inc: Equity Ownership; TAKEDA: Research Funding; Winship Cancer Institute: Employment, Patents & Royalties.
The Efficacy and Safety of Cidofovir in Salvage Therapy for CMV Infection in the Patients with Haploid Hematopoietic Stem Cell Transplantation
Authors: Yu Guo-Pan , Yin Zhao , Fan Zhiping , Huang Fen , Xu Na , Jiang Ling , Wang Zhixiang , Xuan Li , Liu Qifa , Feng Ru , Sun Jing 
Published: November 2019

Cidofovir (CDV) is a nucleotide analogue with broad antiviral activity and has been approved for the treatment of cytomegalovirus (CMV) retinitis in patients with acquired immunodeficiency syndrome. Data on the use of CDV for anti-CMV treatment in the patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT) are limited. To evaluate the efficacy and safety of CDV in salvage treatment of CMV infection after allo-HSCT, we prospectively enrolled 18 patients with CMV infection after allo-HSCT in this study, of whom failed or intolerant to preemptive antiviral therapy. Dose of CDV as 5 mg/kg per week was used until CMV-DNA negative twice with quantification polymerase chain reaction (qPCR) detection or disease progression. All these 18 patients received haploid HSCT and suffered from a median of II (I~IV) ° acute graft versus host disease (aGVHD) before or at the same time of CMV infection. The patients showed as CMV DNAemia (n=7), CMV syndrome (n=5) and CMV disease (n=6) at a median time of 37 (range, 26 to 49) days post transplantation. Before CDV, all the patients received ganciclovir (n=9) or/and foscarnet (n=10) as pre-empty treatment for CMV infection, and 15 cases failed while 3 untolerated to the side effect. After a median of 2 (1-4) course of CDV therapy, 14 (77.8%) patients acquired clinical cure, 1 improved, and 3 (16.7%) failed, presenting a median response time of 2 courses. 11 (61.1%) patients had a 1-log decrease in CMV-DNA copy number after 1 cycle of CDV treatment, and 10 (55.6%) patients obtained virus clearance after 1 to 2 courses of treatment. With a median follow-up of 5 (1 to 12) months, 14 patients remained continuously CMV-DNA negative. During CDV treatment, 4 (22.2%) patients developed mild reversible renal damage and 5 (27.8%) experienced I~II° gastrointestinal reactions. Taking together, it could be effective and safe to use cidofovir in salvage treatment for CMV infection in the patients with allo-HSCT, including haplo-HSCT.

Disclosures
No relevant conflicts of interest to declare.
Preliminary Results from a Phase I Study of SHC014748M in Patients with Relapsed or Refractory Indolent B-Cell Lymphomas
Authors: Xu Wei , Jin Jie , Zhou Keshu , Wang Li , Zhu Huayuan , Qian Jiejing , Huang Jianying , Zhou Fuling , Xu Xiaohong , Wang Zhiqiang , Guo Wei , Li Jianyong 
Published: November 2019

Introduction: The phosphatidylinositol-3-kinase (PI3K) pathway plays an important role in multiple cellular functions, including proliferation, differentiation, and trafficking, and deregulation of PI3K is a hallmark of many malignancies. The preferential expression of the PI3Kδ isoform on leukocytes provides an attractive target for hematologic malignancies. SHC014748M is a novel, potent, and highly selective PI3Kδ inhibitor. This study is a phase I, first-in-human study to assess the safety, tolerability and pharmacokinetics of SHC014748M in patients with relapsed or refractory indolent B-cell lymphomas (NCT03588598).
Methods: Dose escalation was performed in a 3+3 design in sequential cohorts at doses of 50-250 mg QD, and then dose expansion was conducted at the selected cohorts to find recommended Phase II dose (RP2D). Eligible patients were those with relapsed or refractory indolent B-cell lymphomas, having at least 1 measurable lesion, and with ECOG performance status 0-2. Patients received a single dose on day 1, and then after 3 days washout multiple dosing once daily was initiated until disease progression or unacceptable toxicity. Primary endpoints included safety, tolerability and pharmacokinetics. Secondary endpoints included overall response rate (ORR), lymph node response (LNR), time to response (TTR), progression-free survival (PFS) and duration of response (DOR).
Results: As of Jul 16, 2019, 38 patients with relapsed or refractory indolent B-cell lymphomas were enrolled in the study, including 23 with follicular lymphoma (FL), 11 with chronic lymphocytic leukemia/Small lymphocytic lymphoma (CLL/SLL), 1 with marginal zone lymphoma (MZL) and 3 with Waldenstrom's macroglobulinemia (WM). 15 patients were treated in the dose escalation phase and 23 were treated in the dose expansion phase (12 in 150 mg cohort, 11 in 200 mg cohort). The average number of prior therapies was 2.2 (range 1-8), and 84 percent of patients had received rituximab before. No dose-limiting toxicities occurred. The most common adverse reactions were neutropenia (39.5%), fatigue (21.1%), thrombocytopenia (18.4%), diarrhea (13.2%), pneumonia (13.2%), cough (13.2%), lymphopenia (13.2%), anemia (13.2%) and rash (13.2%). Grade ≥ 3 adverse reactions were neutropenia (23.7%), pneumonia (13.2%), rash (7.9%) and diarrhea (5.3%). Among 14 FL patients who were available for efficacy evaluation, ORR was 57.1% (8 PR, 5 SD, 1 PD). Of 9 CLL/SLL evaluable patients, 5 (55.6%) had partial response, 3 (33.3%) had partial response with lymphocytosis, and 1 had stable disease. Of 3 WM patients, 1 had partial response and 2 had minor response.
Conclusion: SHC014748M was well tolerated and demonstrated promising clinical activity in patients with relapsed or refractory indolent B-cell lymphomas.

Disclosures
Wang: Nanjing Sanhome Pharmaceutical Co., Ltd.: Employment.
Venetoclax and DLI for Patients with Early Relapse of Acute Myeloid Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation. a Historical Prospective Multicenter Trial
Authors: Amit Odelia , Bar-On Yael , Perets Galit , Atlas Lena , Shargian Liat , Yeshurun Moshe , Yachini Bruria , Karni Chen , Ram Ron 
Published: November 2019

Introduction: Prognosis in patients with post allogeneic HCT-early relapse of AML is dismal and response to salvage treatment is less than 20%. Venetoclax has been shown to be safe and efficacious in frail/elderly patients with AML. Treatment results in an increase of pro-apoptotic function, thus the addition of alloreactive T cells may augment the anti-leukemic potency. We hypothesized that the combination of donor lymphocyte infusion may further enhance this response. Methods: Eligibility criteria for this protocol were - age>18 y, early (<6 months post allogeneic HCT) relapse and no concurrent GVHD. Venetoclax was administered at a daily dose of 400 mg for up to 6 cycles with DLI in escalating doses for up to 4 doses. Additional chemotherapy was allowed according to physician discretion. Primary endpoint of study included assessment Overall response (CR/CRi/CRp) rate at 3 months post treatment. Secondary endpoints included 1-year OS, rate of acute and chronic GVHD, documented infections, hematological complications, and other safety measures. Results: From January 2017 until May 2019, 19 patients were given the study protocol. Median age was 60 (43-75) years, Table. There were no cases of tumor lysis syndrome. Microbiology documented infections occurred in 5 patients (26%) and 2 patients were admitted during protocol (10%). No cases of significant bleeding were observed, and majority of patients were able to tolerate the protocol without any admissions. Acute GVHD was observed in 2 (10%) patients and chronic GVHD was observed in 4 (21%) patients. Overall response was observed in 8 (47%) patients (CR, n=3; CRi, n=2; CRp, n=3). Median time to response was 28 (18-67) days (median number of courses 2 (1-6)) and duration of response was 106 (31-395) days. Survival at 6 months was 3.5 months (95% CI 1.5-5.5). Median survival of patients achieving complete remission was 16.2 months (CI not det.), Figure. On univariate analysis there was no association between patients and disease's characteristics and overall survival, however any type of GVHD (entered to the model as a time dependent variable) was associated with a lower rate of mortality (HR=0.2, p= .03). Conclusions: These results may endorse the hypothesis that enhancing alloreactivity as a therapy may lead to a chemotherapy free approach to relapsed AML. A phase 2 study is under preparation with a larger sample size and a standardized approach to additional salvage chemotherapy added versus Venetoclax + DLI regimen only for a more robust analysis of this therapeutic approach.

Disclosures
No relevant conflicts of interest to declare.
Stratification of Responders and Non-Responders in the Reach-1 Trial Based on Serum Proteomic Analysis
Authors: Owens Sherry , Pratta Michael A. , Liu Hao , Arbushites Michael C. , Tian Chuan , Howell Michael D. 
Published: November 2019

Acute graft versus host disease (aGVHD) is a serious and potentially life-threatening condition that can develop in patients receiving hematopoietic stem cell transplantation (HSCT). Acute GVHD is characterized by increased levels of inflammatory mediators and signaling through the Janus Kinases (JAKs), leading to tissue and organ damage. Ruxolitinib, an oral JAK1/JAK2 inhibitor, is the first FDA-approved treatment for steroid refractory aGVHD. The current study evaluated proteomic signatures from participants in the REACH-1 clinical trial to identify those individuals which have an increased probability for success when treated with ruxolitinib in combination with corticosteroids.
Plasma was collected for 42 participants enrolled in the REACH-1 clinical trial (NCT02953678) prior to and at designated times following treatment. Broad proteomic analysis of more than 1000 proteins was conducted using the OLINK proximity extension assay. Participants were separated into responders (complete responders, very good partial responders, partial responders) (n=36) or non-responders (mixed responders, progressive disease) (n=6) based on their clinical response at Day 28 of treatment. Baseline differences in day 28 responders versus non-responders were compared using t-tests. Pharmacodynamics changes within responders and non-responders were assessed using paired t-tests. Significance of statistical tests was conferred at p<0.05.
Broad proteomic analysis of plasma identified a total of 145 differentially expressed proteins (p<0.05) between the responders and non-responders at baseline. Thirty-five proteins were elevated and 110 proteins were lower in responders compared to non-responders. At baseline, IL-8, IL-6, and IL-24 were among the most significantly down-regulated while SCF was one of the most significantly up-regulated in responders compared to non-responders. Pharmacodynamic analyses revealed 493 significantly modulated proteins in responders and 92 in non-responders. IL2-RA was among the most significantly down-regulated from baseline to day 28 in both responders and non-responders.
This study suggests that proteomic characterization has the potential to stratify responders and non-responders to ruxolitinib and corticosteroid treatment in steroid refractory aGVHD. Additionally, this study demonstrated robust pharmacodynamic and correlative changes in responders following treatment with ruxolitinib and corticosteroids suggesting that defining a responder population could enrich for patients predisposed to positive therapeutic response; however, this should be prospectively validated in additional studies.

Disclosures
Owens: Incyte Corporation: Employment, Equity Ownership. Pratta:Incyte Research Institute: Employment. Liu:Incyte: Employment, Equity Ownership. Arbushites:Incyte: Employment. Tian:Incyte: Employment, Equity Ownership. Howell:Incyte: Employment.
Study of Seroprevalence of Hepatites "B" and "C" Markers in Blood Donors from Hemonorte, Rio Grande Do Norte/ Brazil
Authors: Medeiros Ralfo Cavalcanti , Neto Manuel Josué Batista , Paiva Aldair Sousa , Sales Valéria SF , Freitas Rodrigo Villar , Paiva Hugo Diogenes De Oliveira , Araujo Rafael Santiago , Cavalcanti Geraldo Barroso 
Published: November 2019

Viral hepatitis transmissible by blood more frequent in the Brazilian population are those of type B and C. Blood banks and hemotherapy centers (Hemocenters) must carry out serological screening, in order to statistically evaluate the percentage of infected donors the virus of these hepatitis. This study aimed to characterize the blood donor population of the State of Rio Grande do Norte (RN), located in the Northeast region of Brazil, in relation to serological markers of hepatitis B and C and to compare these donors from different regions of RN. Were performed serological tests from 254.332 blood donations at the Laboratory of Serology of Hemocenters of Rio Grande do Norte (HEMONORTE) by indirect ELISA methodology. The state of RN was divided into three regions according to the presence of transfusion units or Hemocenters in: Natal city, capital of the state of RN and nearby cities and also headquarters of HEMONORTE; Western Region comprising the Hemocenter of Mossoró city and and transfusion unit of Pau dos Ferros city; and the Serido region of the hemocenter of Caicó city and the collection and transfusion unit of the Currais Novos city. The markers used for hepatitis B detection were total anti HBc (IgM and IgG) and HBsAg. The research of the virus Hepatite C virus carried out with those with third generation kits containing core proteins NS3 and NS4, in addition to NS5. The samples were considered reactive (seropositive) when optical density was obtained up to 20% below the cut-off value, which is the criterion used by most blood centers in Brazil. Among the various serological markers of hepatitis B and C, anti-HBc was the most prevalent with a percentage of 0.98%. Anti-HCV presented a prevalence of 0.77%, while HBsAg had a low prevalence (0.07%). When comparing the prevalence by region, we found that the Natal city region had the lowest rates of hepatitis B (0.93%) and hepatitis C (0.71%). The western region presented the highest percentages of reactivity for hepatitis B (1.24%) and C (1.16%). The seridó region of obtained a prevalence of hepatitis markers smaller than the western region and larger than that of the , 0.99% for hepatitis B and 0.91% for hepatitis C. This difference of percentages between regions should be investigated , to find out the reason for this prevalence variation. The strategy of clinical recruitment and screening should be reassessed and unified among hemotherapy services in Rio Grande do Norte.

Disclosures
No relevant conflicts of interest to declare.
Chronic Lymphocytic Leukemia in Kuwait
Authors: Alshemmari Salem , Pandita Ramesh , Hamadah Abdulaziz , Alhuraiji Ahmad 
Published: November 2019

Background :Chronic lymphocytic leukemia (CLL) is common malignancy in Western countries. However, little known about this disease entity in our area. This study exploring the biology in out patients' population. Method:Patients with confirmed CLL under IGHV and TP53 mutational analysis at presentation or during follow up. We also integrated other clinical and biological parameter in this study. Results: A total of 137 cases were analyzed, median age 61 years (range:34-89); 30% of the cases age was<55 years at presentation. There was 108 males vs. 29 females M:F ratio 3.7. Two patients gave a family history of CLL, while 1 patient gave a history of other lymphoproliferative disorders. Binet staging system available in 134 cases, A: 109 (81.3%), B: 12 (9%), C:13 (9.7%). B2 macroglobulin elevated in 40/112 (36%) cases and 10/103 (10%) had M-spike. CD38 positivity reported in 37/112 (33%) of cases. Cytogenetics data evaluable in 85 cases: isolated del(13q): 35%, isolated trisomy 12 (16.5%), del(11q) (4.5%), del(17p)(2.4%). IGHV mutational status mutated vs unmutated: 40% vs 60%. Cases with available treatment information on 132 cases. Fifty cases required treatment due to disease progression. First line treatment Bendamustine-Rituximab (BR) 3 cases, Fludarabine Cyclophosphamide Rituximab (FCR) 30 cases and Chlorambucil with anti-CD 20 antibody 6 cases. At the time of review, 3 cases on ibrutinib (2 in 3rdline and 1 case in the 4thline). Conclusion: This is the first study to shed light on CLL in our area. There are biological differences between our patients' population and the western countries.

Disclosures
Pandita: Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen Cilag: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees.
The Effect of Continuous Screening of Carbapenem-Resistant Enterobacteriaceae on Theprevention and Control of Bloodstream Infections in Patients with Hematopoietic Stemcell Transplantation
Authors: Yang Tingting , Luo Xueping , Yang Qing , Chen Hongchao , Luo Yi , Zhao Yanmin , Lai Xiaoyu , Yu Jian , Tan Yamin , Wei Guoqing , Huang He , Shi Jimin 
Published: November 2019

Carbapenem-resistant Enterobacteriaceae (CRE) bloodstream infections (BSIs) have become emerging cause of death in patients with hematopoietic stem cell
transplantation (HSCT). Patients underwent HSCT from September 2017 to June 2018, and from July 2018 to February 2019, were assigned as single screening group and continuous screening group; patients transplanted from January 2016 to August 2017 were assigned as control group. Continuous screening significantly improved the CRE gut detection rate compared with single screening (10% vs 1.5%, p=0.001). The CRE infection rate in the pre-intervention period, single screening period and continuous screening period were 1.6%, 2% and 0, respectively; while related mortality was were 66.7%, 50% and 0, respectively. The time from the onset of symptoms of infection to use of tigecycline in survival patients with BSIs were shorter than died patients (24 hours vs 72 hours). For 11 CRE carriers with neutropenic fever, all received tigecycline therapy-based therapy (time from detection to therapy, -3 to 17 days) and did not develop BSIs. These results suggest that continuous screening can more effectively identify high-risk groups of CRE colonization and guide targeted preemptive treatment in the presence of infection symptoms, so as to reduce the incidence of BSIs and improve the prognosis of patients.

Disclosures
No relevant conflicts of interest to declare.