bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Evidence Against the Continuum of Low-Primed Uncommitted Hematopoietic and Progenitor Cells (CLOUD-HSPC) Concept for Hematopoiesis
Authors: Mohanlal Ramon , Blayney Douglas W. , Huang Lan 
Published: November 2019

Introduction: The classical model of blood stem cell differentiation is branch-like linear HSPC progression from progenitor multi/oligo potent stem cells to differentiated mature cell types. An alternative model, referred to as CLOUD-HSPC, proposes bone marrow (BM) HSPCs continuous and direct differentiation into unipotent cells of myeloid lineage (neutrophils (N), basophils (B), eosinophils (E), monocytes (M), mast cells, megakaryocytes (platelets) and erythrocytes without linear stem cell commitment (Velten Nat Cell Biol 2017). Pegfilgrastim (Peg), and the non-colony stimulating agent Plinabulin (Plin) prevent chemotherapy (chemo) induced neutropenia (CIN) in cancer patients (pts), and both exert BM effects leading to CD34+ cell mobilization (Blayney ASH 2017, 2018). In mice, Plin enhances LSK (Lin-Sca+cKit3+) cell differentiation in BM (Ghosh, AACR 2018). Here we clinically validated the CLOUD-HSPC concept by analyzing peripheral blood mononuclear cell (PBMC) counts derived from two bifurcating progenitor cells 1. Granulocyte-Monocyte Progenitor (GMP) producing N,M,E and B and 2. Common Lymphoid Progenitor (CLP) producing lymphocytes (L), after exposure to Peg or Plin in a CIN setting. With linear stem cell commitment, increase in the differentiated mature cell counts would positively inter-correlate, and with CLOUD-HSPC, they would not.
Methods: In the Phase 3 portion of study BPI-2358-105 (NCT03102606), pts with NSCLC, BC or HRPC received pre-medication with dexamethasone (Dex) on day (D) -1,0, and 1 and docetaxel (Doc) on day (D) 1. Pts were randomized 1:1 to either Plin 40 mg (n=52), given 30 min after Doc infusion D1, or Peg 6 mg (n=53), D2. Central laboratory (Covance Laboratory) PBMC counts were obtained at screening, D 1,2, 6,7,8,9,10, 15 in Cycle 1, and correlated with each other at a pre-planned interim analysis. D1 and D2 data points were omitted due to confounding effects of Dex and demargination. Data of Plin and Peg were combined, since both drugs exert BM effects leading to increased N cell counts and showed similar correlation trends.
Results: Maximum increases in % from screening value on D5-D15 of each N,M,E,B and L counts correlated with each other. Pearson correlation coefficients (r) after linear regression and corresponding p-value are summarized below.
Conclusions: There is a linear and positive inter-correlation between differentiated mature cells derived from the GMP and CLP lineage, proving strong clinical evidence against CLOUD-HSPC, but in favor of the classical linear commitment model.
Table

Disclosures
Mohanlal: BeyondSpring Pharmaceuticals: Employment. Blayney:BeyondSpring Pharmaceuticals: Research Funding. Huang:BeyondSpring Pharmaceuticals: Employment.
Fast Onset of Action of Subcutaneously Administered Marzeptacog Alfa (Activated) Supports on-Demand Treatment in Hemophilia a Mice
Authors: Del Greco Frank , Neuman Linda , Levy Howard , Reckless Jill , Knudsen Tom , Blouse Grant E. 
Published: November 2019

Background: Patients with Hemophilia A or B with and without inhibitors lack treatment options that are fast and easy to use for treating acute bleeding. Currently, their only options are intravenous administration of replacement or bypass therapies. Marzeptacog alfa (activated) (MarzAA) is a novel increased potency recombinant FVIIa in clinical development. We hypothesized that, in addition to its clinically demonstrated efficacy in SC prophylaxis, MarzAA could potentially be used SC on-demand to control acute hemorrhage after bleeding has started.
Objective: To demonstrate the procoagulant effect of dosing MarzAA subcutaneously at timepoints both before and after injury using a tail clip bleeding model in hemophilia A mice.
Methods: Each mouse was initially weighed and briefly anesthetized with isoflurane for collection of 5 µL blood to assess baseline hemoglobin levels. Later, anaesthetized (100 mg/kg ketamine + 10 mg/kg xylazine) F8-/- (Haem A, B6;129S4-F8tm1Kaz/J) mice were submitted to a tail clip injury model completely transecting the tail at a diameter of 1.25 mm from the tip using a sharp razor blade. Following injury, blood loss was monitored with the tail submerged in warm saline for 20 minutes and quantified by hemoglobin content. Mice were dosed with MarzAA SC (1.5 mg/kg) or vehicle at 15, 30, 60, or 90 minutes before or one minute after injury (1.5 and 4.5 mg/kg). An ED50 for MarzAA was determined using various doses ranging from 0.15 mg/kg to 3.0 mg/kg. Statistical significance was set at α = 0.05.
Results: SC administration of MarzAA resulted in a dose dependent reduction in bleeding when administered to hemophilia A mice 15 minutes before injury. At the highest dose of MarzAA, full efficacy was achieved as bleeding in the treated mice was comparable to the blood loss observed in normal hemostatically competent mice. The ED50 for MarzAA was calculated to be 387 µg/kg after SC administration in this model. More importantly, when dosed as a rescue therapy one minute after injury, SC MarzAA significantly reduced bleeding to 350±46 µL from 635±50 µL (vehicle), p = 0.02.
Conclusion: MarzAA demonstrated efficacy when administered SC both before and after injury. Taken together these data suggest that SC MarzAA could potentially be used on-demand to treat patients with acute bleeding. These results provide a basis for further clinical investigation of the acute treatment of a bleed with SC MarzAA in hemophilia patients.

Disclosures
Del Greco: Catalyst Biosciences: Consultancy, Equity Ownership. Neuman:Catalyst Biosciences: Employment, Equity Ownership. Levy:Catalyst Biosciences: Employment, Equity Ownership. Reckless:RxCelerate: Employment. Knudsen:Catalyst Biosciences: Employment, Equity Ownership. Blouse:Catalyst Biosciences: Employment, Equity Ownership.
Quality of Life in Patients with Multiple Sclerosis after Autologous Hematopoietic Stem Cell Transplantation
Authors: Olivares-Gazca Juan Carlos , Murrieta-Álvarez Iván , Olivares-Gazca Jesús Mauricio , García-Navarrete Yarely Itzayana , Cantero-Fortiz Yahveth , León-Peña Andrés A. , Ruiz-Delgado Guillermo J. , Ruiz-Arguelles Guillermo J. 
Published: November 2019

Introduction
Multiple sclerosis (MS) is an inflammatory, demyelinating and neurodegenerative disease of the central nervous system (CNS) that causes a whole spectrum of neurological disorders associated with a profound decrease in the quality of life of affected patients. Currently, autologous hematopoietic cell transplantation (ASCT) is a validated therapeutic approach and has been shown to be superior to new immunomodulatory agents. However, the impact of these therapies on the quality of life of patients with MS is unknown.
Objective
Identify the impact on the quality of life in patients with multiple sclerosis after ASCT at our center.
Methods
A quasi-experimental, longitudinal, prospective and single-center study was conducted in which the quality of life was determined in patients with MS before and after ASCT. Patients who could not answer the questionnaire themselves were excluded and incomplete questionnaires were eliminated. The quality of life was determined by applying the MS-QoL 29 instrument which is validated instrument for this pathology (Cronbach 0.88-0.90 and Pearson with high correlation with MS-QoL56). The variables related to the physical and mental components of the instrument as well as demographic characteristics were studied. The statistical analysis of the data included measures of central tendency as well as inferential for the comparison of means and proportions (NC 95%, p <0.05).
Results
We included 52 patients prospectively from October 2018 to June 2019, 71% of the patients were women and the remaining 29% men. The median of age of the subset is 50 years (Interval 27-65). Of the selected patients, 45% has PPMS, 39% has SPMS and 16% has PPMS. Twenty six patients were followed at 3 months and seventeen were followed 6 months after ASCT. The statistical differences between the quality of life in the patients prior to the ASCT and the follow-up at 3 and 6 months in both the physical and mental components was analyzed. In the physical component the differences at 3 months (A) were significant (p = 0.019, 95% NC) as well as the differences at 6 months (b) after ASCT (p = 0.0024, 95% NC). In the mental component the differences were significant at 3 months (C) (p = 0.0012, NC 95%) as well as the differences at 6 months (D) after ASCT (p = 0.0007, NC 95%).
Conclusions
The study suggests that ASCT is a feasible and safe therapeutic alternative to improve the quality of life in patients with multiple sclerosis.
Figure

Disclosures
No relevant conflicts of interest to declare.
An Institutional Survey of Warfarin Reversal in the Setting of Life-Threatening Bleeds
Authors: Garrahy Ian , Pawar Tushar , Donato Anthony , Stevenson Amber , Forman Daniel 
Published: November 2019

Introduction:
The bleeding risk of warfarin is well known. Therapeutic options for warfarin reversal in life-threatening bleeds include fresh frozen plasma (FFP), recombinant factor VIIa (rFVIIa), and prothrombin complex concentrate (PCC). Despite the theoretical advantage and clinical evidence supporting PCC, it is not widely used in the US.
Methods:
An online anonymous questionnaire was sent to all providers in the Tower Health System asking them about their practice, specialty, degree, years in practice, and basic questions regarding their comfort and frequency of prescribing anticoagulants. The questionnaire also asked the providers how they would manage ten various clinical scenarios related to anticoagulation management. One question specifically addressed the management of warfarin reversal in an 85-year-old presenting with hemorrhagic shock. Analysis of variance was used to compare the scoring means between groups while linear regression and Pearson's correlation coefficient measured the relationship between years of practice and test scores.
Results:
Out of 404 responders, 232 (57.4%) selected an incorrect answer and 212 (52.5%) incorrectly selected fresh frozen plasma as the answer to the question (see uploaded image) that addressed warfarin reversal in the setting of major bleeding. Those providers who answered this question correctly and those who answered it incorrectly had mean scores of 68.26% and 52.16% respectively on the overall survey (p value 0.000).
Conclusion:
As compared to FFP, the use of PCC for warfarin reversal is associated with a significant reduction in all-cause mortality. This project demonstrates a professional practice gap and serves to highlight an area in medicine where many providers are not practicing in accordance with evidence-based practice. Furthermore, this particular question from the survey discovered that providers who performed better on the survey were more likely to answer questions regarding anticoagulation reversal correctly.
Figure

Disclosures
No relevant conflicts of interest to declare.
Molecular Profiling of Aggressive Non-Hodgkin Lymphoma - Results from a Phase 1/2 Preben Study
Authors: Leivonen Suvi-Katri , Jørgensen Judit , Larsen Thomas Stauffer , Pasanen Annika , Karjalainen-Lindsberg Marja-Liisa , Toldbod Helle M , d'Amore Francesco , Leppa Sirpa 
Published: November 2019

Background: Aggressive non-Hodgkin lymphoma (NHL) relapsing after standard first line chemotherapy represents an unmet clinical need. Currently, a phase 1/2 study with the combination of pixantrone, etoposide, bendamustine, and in CD20 positive tumors, rituximab, in patients with relapsed aggressive NHL of B- or T- cell phenotype (the PREBEN study) is ongoing. Here our aim was to molecularly characterize samples from the PREBEN trial and find clinical correlates for predicting treatment response.
Methods: The profiling cohort consisted of 21 patients with pre-treatment RNA samples and clinical data. Nanostring PanCancer Pathways and PanCancer Immune profiling panels (altogether 1348 genes) were utilized for the gene expression analyses. The findings from gene expression analyses were correlated with clinical parameters.
Results: Fourteen patients had diffuse large B-cell lymphoma (DLBCL), whereas seven had peripheral T-cell lymphoma (PTCL). In general, the expression of DNA replication genes distinguished DLBCL from PTCL. Additionally, gene expression analyses identified genes having differential expression based on the response to the treatment. Supervised hierarchical clustering of the ten most differentially expressed genes could separate the responding (n=4) and non-responding (n=10) DLBCL patients into two distinct subgroups (Fig. 1A). Similarly, the responding (n=3) and non-responding (n=4) PTCL patients could be separated into distinct subgroups by supervised clustering with the ten most differentially expressed genes (Fig. 1B).
Conclusion: Molecular profiles of aggressive NHL are heterogeneous and may be utilized for predicting the treatment response. More detailed molecular analyses are currently ongoing.

Disclosures
Jørgensen: Gilead: Membership on an entity's Board of Directors or advisory committees; Roche: Membership on an entity's Board of Directors or advisory committees. d'Amore:Servier: Research Funding. Leppa:Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen-Cilag: Research Funding; Bayer: Research Funding; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Roche: Honoraria, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding.

OffLabel Disclosure:
combination of bendamustine and pixantrone for relapsed NHL
GRK6 Regulates the Hemostatic Response to Injury through Its Rate-Limiting Effects on GPCR Signaling in Platelets
Authors: Chen Xi , Gupta Shuchi , Cooper Matthew , Dehelian Daniel , Zhao Xuefei , Brass Lawrence F. , Naik Ulhas P. , Naik Meghna Ulhas , Benovic Jeffrey , Ma Peisong 
Published: November 2019

Inappropriate platelet activation remains a major cause of cardiovascular and cerebrovascular diseases. Most agonists activate platelets through G protein-coupled receptors (GPCRs). However, questions remain about mechanisms that provide negative feedback towards activated GPCRs to limit platelet activation and thrombus formation.
Here we provide the first evidence that GPCR kinase 6 (GRK6) serves this role in platelets, using GRK6-/- mice generated by CRISPR-Cas9 genome editing to examine the consequences of GRK6 knockout on GPCR-dependent signaling. Hemostatic thrombi formed in GRK6-/- mice are larger than in WT controls during the early stages of thrombus formation, with a rapid increase of platelet accumulation at site of injury. Platelet activation in the absence of GRK6 is enhanced, but in an agonist-selective manner. Responses to PAR4 agonist peptide or ADP stimulation in GRK6-/- platelets are increased compared to WT control littermates, while the response to TxA2 is normal. Underlying these changes in GRK6-/- platelets is an increase in Ca2+ mobilization, Akt activation, and granule secretion. Furthermore, deletion of GRK6 in human MEG-01 cells causes an increase in Ca2+ response and PAR1 surface expression in response to thrombin. Finally, we show that in human platelets, platelet activation in response to thrombin causes an increase in binding of GRK6 to PAR1, as well as an increase of the phosphorylation of PAR1. Deletion of GRK6 in MEG-01 cells causes a decrease in PAR1 phosphorylation.
Collectively, these observations, for the first time, show that 1) GRK6 regulates the hemostatic response to injury by thrombin and ADP, 2) it mediates platelet activation by reducing PAR1/4- and P2Y12-dependent signaling, and 3) GRK6 limits the rate of platelet activation during early stage of thrombus growth and helps prevent inappropriate platelet activation.

Disclosures
No relevant conflicts of interest to declare.
Dual Anticoagulant Facilitied Peripheral Blood Stem Cell Harvest from Childhood Donors with Low Weight and Low MCV
Authors: Yue Li , Bu Chaoke , Ma Zhirui , Li Chunfu , Wang Wenjun , Zhou Jun 
Published: November 2019

Background:Low weight children can sometimes be selected as donor in HSCT. Donor with thalassemia minor has low MCV, which affects peripheral blood stem cell (PBSC) harvest. It is worthwhile to explore a harvest program for donors with low body weight and low MCV.
Objective:The aim of this study is to find out an appropriate program of PBSC collection in children with low body weight and low MCV.
Methods:10 children, with median age of 2 (1-6) years, average weight of 13.4 (9.5- 20) kg and average MVC value of 63fl (54-78)) were hospitalized in our center from October 2018 to July 2019 for stem cell mobilization. Donor weight were less than recipients in 7 cases. 5 donors have 5 kg less than recipient, 3 have 10 kg less and 1 has 18 kg less, respectively. They received G-CSF of 5 mg/kg/d twice a day for 5 days before harvest. 500 ml ACD-A solution and 3000 IU heparin were mixed and given by the ratio of 1 ml mixed anticoagulant versus 24 ml whole blood. Blood-flow was 25-40 ml/min.
Results:More 20/ul CD34+ cells were harvested in 9 cases (90%), and 19.2/ul in 1 case (10%). Median nucleated cells were13.2(6.7-22.4)x108/kg in donor weightand 9.8 (8.3-13.9) x108/kg in recipient weight. The median CD34+ cells were 5 (1.9-10.2) x106/kg in donor weight and 3.4 (1.4-9.7) x106/kg in recipient weight. The coagulation function was normal in reexamination. PBSC were harvested only once in all donors. Average circulating volume was 2.48 (1.14-5.79) total body volume, average harvesting time was 2.75 (2-6) hours, and average PBSC volume was 89 (63-160) ml. Donor peripheral blood calcium and potassium ion concentration showed no obvious abnormality.
Conclusion:Combination of citrate and heparin can be well used to collect PBSC in donors with low body weight and low MCV. All children were well tolerance to the program, which was safe and effective for large volume collection of PBSC.

Disclosures
No relevant conflicts of interest to declare.
Outcome Among Adolescents and Young Adults with Acute Myeloid Leukemia at Pediatric Versus Adult Centers: A Population-Based Study Using the IMPACT Cohort
Authors: Gupta Sumit , Baxter Nancy , Pole Jason , Lau Cindy , Sutradhar Rinku , Nagamuthu Chenthila , Nathan Paul 
Published: November 2019

Background: Survival outcomes among adolescents and young adults (AYA) with acute myeloid leukemia (AML) remain poor. In AYA with acute lymphoblastic leukemia, outcomes differ between patients treated in pediatric vs. adult centers. This has not been well evaluated in AML. We therefore compared outcomes between AYA with AML treated at pediatric vs. adult centers using a population-based clinical database. In addition, we determined other predictors of outcome within this population.
Methods: The IMPACT Cohort comprises all Ontario, Canada AYA aged 15-21 years diagnosed with one of six common cancers (including AML) between 1992-2012. Detailed demographic, disease, treatment, and outcome data were collected through chart abstraction and validated by content experts. Locus of cancer care (LOC - pediatric vs. adult center) was determined based on where the majority of therapy was delivered in the first three months after diagnosis. Linkage to population-based health administrative data identified additional cancer events (second cancers, relapse, death). Event-free (EFS) and overall survival (OS) were determined using Kaplan-Meier methods. The impact of LOC on EFS and OS was determined using multivariable Cox proportional hazard models, adjusting for demographic, disease, and treatment variables. Events included disease progression, relapse, death, and second malignancies.
Results: Among 140 AYA with AML, 89 (63.6%) received therapy at an adult center. AYA treated in pediatric centers were younger than those treated at adult centers (median 16 years vs. 19 years; p<0.001) and were more likely to live in higher-income neighborhoods [37/51 (72.5%) vs. 47/89 (52.8%); p=0.02]. Disease markers such as presenting white blood cell count and AML subtype did not differ by LOC. The 5-year EFS and OS for the whole cohort were 35.0%±4.0% and 53.6%±4.2%. Neither EFS nor OS differed by LOC (Table 1). In multivariable analyses adjusting for disease characteristics, LOC was not predictive of either EFS [adult vs. pediatric center hazard ratio (HR) 1.3, 95thconfidence interval (CI) 0.8-2.2, p=0.27] or OS (HR 1.0, CI 0.6-1.6, p=0.97). AYA with AML living in rural areas however experienced significantly inferior outcomes as compared to their urban counterparts (EFS: HR 2.5, CI 1.3-4.7, p=0.005; OS: HR 2.0, CI 1.1-3.8, p=0.04).
Conclusions: In this population-based cohort, outcomes did not differ between AYA with AML treated at pediatric vs. adult centers, unlike what has been previously shown in AYA with acute lymphoblastic leukemia. However, rural AYA experienced substantially inferior outcomes than urban AYA, suggesting that even within a universal single payer system of healthcare, socioeconomic disparities persist in this population.

Disclosures
No relevant conflicts of interest to declare.