bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sickle Cell Education for Medical Professionals: A Pre and Post Test Analysis
Authors: Simmons Diana D , Lucia Robert , Saving Kay Linn , Alwan Nicole 
Published: October 2019

Abstract
Background and Objectives: Sickle cell disease is a severe inherited form of anemia caused by a genetic mutation. Polymerization of hemoglobin leads to a cascade of effects decreasing blood flow. This causes tissue hypoxia leading to acute and chronic damage to the organs and endothelial lining. This disease requires complex management that relies on comprehensive training and knowledge regarding the disease process. Often accurate knowledge of sickle cell disease and how to provide appropriate care in the general medical population is limited. The purpose of this project was to develop a sickle cell educational training module for medical professionals. Such a module could be used to guide the provision of accurate education regarding sickle cell disease and best practice when caring for this patient population.
Methods: Goals and learning objectives were created and current medical literature about caring for sickle cell disease was reviewed. A comprehensive PowerPoint presentation was produced along with a provider tip sheet and a pre and posttest. The presentation, tip sheet, and tests were reviewed by a board certified pediatric hematologist/oncologist along with the hospital's educational review committee in the Department of Professional Regulation. Once approved, the PowerPoint, tip sheet, and tests were combined into a learning module and uploaded onto an online learning system utilized by the hospital system. The module was sent to over 2,400 outpatient providers and staff and to all inpatient staff on units where sickle cell patients stay when admitted. The module consisted of the participant completing a 10 question pretest, then reviewing the PowerPoint presentation and tip sheet. Following the review of the PowerPoint and tip sheet, the participant completed a 10 question posttest and completed an evaluation of the module.
Analysis: There were 223 people who completed the Sickle Cell Disease Learning Module. A paired t-test was conducted to compare pre-test scores to post-test scores. There was a significant difference in the pre-test scores (M = 5.98, SD = 1.66) and post-test scores (M = 9.17, SD = 1.36); p = <0.0001.
Conclusion: The goal of this module was to increase baseline medical knowledge of sickle cell disease. The results indicate there was statistically significant improvement in baseline knowledge, based on pre and post data (p = <0.0001). While the results indicate statistically significant increases in performance, it would be important to see if improvements are sustained over time. Reassessment of participants one year after completion of module can be beneficial to see if learned knowledge has been retained.

Disclosures
No relevant conflicts of interest to declare.
Invariant Natural Killer T Cells from Donor Lymphocyte Infusions (DLI-iNKTs) Contribute to Anti-Tumor Immunity after Allogeneic Hematopoietic Cell Transplantation
Authors: Jahnke Simona , Schmid Hannes , Secker Kathy-Ann , Duerr-Stoerzer Silke , Keppeler Hildegard , Baur Rebecca , Schumm Michael , Handgretinger Rupert , Bethge Wolfgang , Kanz Lothar , Schneidawind Corina , Schneidawind Dominik 
Published: October 2019

Abstract

Allogeneic hematopoietic cell transplantation (HCT) is a curative treatment option for hematologic malignancies but relapse remains the most common cause of death. Infusion of donor lymphocytes (DLIs) can induce remission and prolong survival by exerting potent graft-versus-leukemia (GVL) effects. However, sufficient tumor control cannot be established in all patients. We showed previously that invariant natural killer T (iNKT) cells promote anti-tumor immunity in murine models of allogeneic HCT without exacerbating graft-versus-host disease (GVHD).
We therefore studied iNKT cells in DLIs and investigated how culture-expanded iNKT cells from such DLIs (DLI-iNKTs) could lyse leukemia cells. We analyzed 63 cryopreserved DLI samples by flow cytometry. iNKT cells were identified using the PBS57-loaded CD1d tetramer. Under steady state conditions, iNKT cells represent 0.04% (range, 0.01-0.6) of live donor lymphocytes and need to undergo ex vivo expansion for further experiments and clinical application. We established a two-week protocol resulting in a 300-fold expansion of functional DLI-iNKTs with a purity of 94%. Interestingly, we observed a preferential expansion of CD4+ DLI-iNKTs. This subset turned out to be critical for tolerance induction and prevention of GVHD after allogeneic HCT. For degranulation and leukemia cell lysis assays, culture-expanded DLI-iNKTs were co-cultured with leukemia cells. CD107a as a marker of degranulation and iNKT-cell activity was upregulated on DLI-iNKTs upon engagement with leukemia cells that were subsequently lysed in a dose-dependent manner. We also observed an increased release of cytokines like IFN-γ (85 vs. 7 pg/ml, p=0.04). Moreover, the cytotoxic effects of culture-expanded DLI-iNKTs were CD1d-dependent: blocking the interaction between the MHC-I-like molecule CD1d and the T-cell receptor of DLI-iNKTs abrogated iNKT-cell degranulation and efficient leukemia cell lysis.
Our results suggest that iNKT cells from DLIs are involved in anti-tumor immunity after allogeneic HCT and therefore may reduce the risk of relapse and improve progression-free and overall survival. Prior expansion of iNKT cells could promote beneficial effects on tumor control, immune tolerance and overall survival.

Disclosures
Handgretinger: Miltenyi Biotec: Patents & Royalties: Co-patent holder of TcR alpha/beta depletion technologies, Research Funding. Bethge:Miltenyi Biotec GmbH: Consultancy, Honoraria, Research Funding; Neovii GmbH: Honoraria, Research Funding.
Effect of Renal Impairment on the Pharmacokinetics and Safety of Rivipansel in Subjects with Renal Impairment and in Healthy Subjects
Authors: Tammara Brinda , Ryan Kelly , Plotka Anna , Shafer Frank E. , Wei Hua , Readett David , Fang Annie , Korth-Bradley Joan 
Published: October 2019

Abstract
Background: Rivipansel is a pan-selectin inhibitor in phase 3 development for treatment of sickle cell disease vaso-occlusive crises. Previous studies have shown almost complete elimination of unchanged drug in urine following an intravenous (IV) infusion. The objective of this study was to evaluate the effect of varying degrees of renal impairment on the pharmacokinetics (PK), safety, and tolerability of rivipansel.
Methods: A single 840-mg dose of open-label rivipansel was administered IV over 20 minutes to 7 subjects with mild, 7 with moderate, and 7 with severe renal impairment, and to 7 healthy subjects with normal renal function. Classification of renal impairment groups was based on the Cockroft-Gault estimated glomerular filtration rate (CGeGFR): 60-89 mL/min (mild), 30-59 mL/min (moderate), and <30 mL/min (severe). Normal renal function was CGeGFR ≥90 mL/min. Plasma and urine samples were collected for 96 hours postdose and analyzed by validated LC-MS/MS methods. Pharmacokinetic parameters were estimated using noncompartmental modeling. ANOVA was used to assess the effect of renal impairment on PK parameters.
Results: All 28 subjects completed the study. A summary of PK parameters is presented in Table 1. Overall rivipansel exposure was greater in subjects with mild, moderate, and severe renal impairment, with values 1.4×, 2.3×, and 5.5× that of subjects with normal renal function, respectively. Renal clearance decreased with decreasing renal function. Total clearance was lower by 31%, 56%, and 82% in the mild, moderate, and severe renal impairment groups, respectively, compared with the normal renal function group. Five treatment-emergent adverse events (TEAEs) were reported in 3 subjects in the mild renal impairment group, and 3 TEAEs were reported in 2 subjects in the severe renal impairment group. None of the TEAEs reported was considered to be treatment-related.
Conclusions: Greater rivipansel exposure and decreased clearance were observed in subjects with renal impairment compared with subjects with normal renal function. A single 840-mg IV dose of rivipansel was well tolerated in all groups.

Disclosures
Tammara: Pfizer Inc.: Employment. Ryan:Pfizer Inc.: Employment. Plotka:Pfizer Inc.: Employment. Shafer:Pfizer Inc.: Employment. Wei:Pfizer Inc.: Employment. Readett:Pfizer Inc.: Employment. Fang:Pfizer Inc.: Employment. Korth-Bradley:Pfizer Inc.: Employment.
Altering Glycosphingolipid Composition to Improve Multiple Myeloma Bone Complication
Authors: Leng Houfu , Ersek Adel , Morris Emma , Molina Beatriz Gamez , Edwards Claire M. , Horwood Nicole 
Published: October 2019

Abstract
Multiple myeloma (MM) is an incurable cancer of plasma cells (PC), with a median survival of 5-7 years. Osteolytic bone disease and skeletal complications occur in more than 80% of MM patients and significantly contribute to the morbidity and mortality of these patients. Glycosphingolipid (GSL), an essential constituent of the outer leaflet of the cellular membrane, is altered in MM and other hematological cancers. We previously reported that GM3, a subtype of GSL promotes osteoclastogenesis. On the other hand, the GSL synthase inhibitor N-butyl-deoxynojirimycin (NB-DNJ) reduces myeloma bone disease in the 5TGM1 mouse model of MM. Mechanistically, NB-DNJ prevents osteoclast (OC) development and activation by disrupting RANKL-induced localization of TRAF6 and c-SRC into lipid rafts and preventing nuclear accumulation of the transcriptional activator NFATc1.
Although NB-DNJ is an FDA-approved drug treating Gaucher's disease, it has many undesired off-target effects, such as inhibiting lysosomal and plasma membrane Beta-glucocerebrosidase and interfering with intestinal glucosidases which leads to gastrointestinal toxicities and severe weight loss. Therefore, more specific GSL inhibitors are required to minimize the side effects.
Here we report a novel GSL inhibitor called Genz112638 with comparable effects as NB-DNJ but reduced side effects. Genz112638 inhibits both OC formation (p < 0.01) and MM cell growth (p < 0.0001) in vitro in a dose-dependent manner. Moreover, compared to NB-DNJ, Genz112638 more significantly improved bone condition and potentially reduced MM burden, as evidenced by the amelioration of bone loss in the 5TGM1 model of myeloma, and a reduction in the proportion of MM within bone marrow and spleen without obvious adverse effects (n=6) (p < 0.01).
As excessive malignant PC in MM normally arise from germinal centre, we also checked the effects of Genz112638 on germinal centre reactions in wildtype mice. We found that Genz112638 suppresses the formation of germinal centre B cells in mouse spleen induced by sheep red blood cells (n=7). Thus, Genz112638 may affect the pathogenesis of MM disease at the initial stage.
Taken together, our data elucidate a novel specific GSL inhibitor as a promising candidate drug relieving two main features of MM: bone destruction and tumour burden with negligible side effects. In vitro, it decreases OC differentiation and proliferation, and meanwhile decreases MM viability and proliferation. In vivo, it may suppress B cell formation in germinal centre, ameliorate bone destruction, and potentially interfere with the vicious cycle between increased OC and susceptibility to MM. In short, we provide a preclinical platform for GSL inhibition as a new tool against MM and its related complications.
Figure. Figure.

Disclosures
Horwood: Genzyme: Research Funding.
Safety and Efficacy of Dose-Escalation Hydroxyurea Therapy in Infants and Young Children: A Retrospective Cohort Study
Authors: George Alex , Tran Jennifer Nguyen 
Published: October 2019

Abstract
The BABY HUG clinical trial of hydroxyurea in infants and young children with sickle cell disease (SCD) established the safety and clinical benefit of hydroxyurea therapy in this age group. The study did not establish, however, if escalation to maximum tolerated dose (MTD) could be safely accomplished and resulted in increased benefit in this age group.
We sought to determine the safety and clinical effectiveness of dose-escalation hydroxyurea therapy in children with SCD under the age of two years. We conducted a retrospective chart review of children with HbSS or HbS β0Thalassemia who initiated hydroxyurea therapy before the age of two years between 2011 and 2017 at the Texas Children's Hematology Center. We assessed the clinical and laboratory response to dose-escalation therapy as well as any adverse events associated with hydroxyurea therapy.
We identified 95 patients ranging from 5 to 24 months in age who were initiated on hydroxyurea therapy during the study period. MTD was achieved in 75 patients (76.5%), of whom 51 were still at MTD at end of monitoring. There were significant increases in hemoglobin concentration (Hb) and hemoglobin F (HbF) levels and decline in absolute reticulocyte count (ARC) on hydroxyurea therapy (Table 1). Additionally, the magnitude of increases in Hb and HbF were greater than those seen in the BABY HUG hydroxyurea treatment group both at the time of MTD and at end of assessment. The proportion of patients experiencing neutropenia or thrombocytopenia as well as the number of events per patient-year (PPY) over 193.9 patient-years were similar to those in the BABY HUG hydroxyurea treatment group (Table 2). The incidence of acute chest syndrome (ACS) and transfusions was similar between the two groups, while episodes of splenic sequestration were more common in patients advanced to MTD.
In summary, hydroxyurea therapy with dose escalation to MTD appears to be safe in infants and young children and results in more significant and sustained increases in hemoglobin concentration and %HbF than were seen with fixed dosing in the BABY HUG study. In the context of this retrospective study, though, hydroxyurea dose escalation did not clearly result in increased clinical benefit relative to fixed-dose treatment.

Disclosures
No relevant conflicts of interest to declare.
Attempt to Prove the Existence of Abnormal B Lymphocyte As Myeloma-Initiating Cells from B Cell-Derived Induced Pluripotent Stem Cells
Authors: Yanagi Aki , Tsuyama Naohiro , Yanai Yukari , Abe Yu , Sugai Misaki , Ota Akinobu , Sivasundaram Karnan , Shigemura Tomonari , Sasatani Megumi , Kamiya Kenji , Hanamura Ichiro , Ikezoe Takayuki , Onodera Masafumi , Sakai Akira 
Published: October 2019

Abstract
We previously established normal B cell-derived induced pluripotent stem cells (BiPSCs; BiPSC13 and MIB2-6). BiPSCs are known to maintain VDJ rearrangement of the IgH gene, and they can be induced by the tet-off system to express activation-induced cytidine deaminase (AID; BiPSC13-AID and MIB2-6-AID) and differentiate into hematopoietic progenitor cells (HPCs) (Scientific Rep, 2017). Using these BiPSCs, we attempted to prove the existence of abnormal B cells, which are thought to be myeloma-initiating cells, originating from mature B cells transformed by reprogramming. We speculated that BiPSCs could develop into myeloma-initiating cells that undergo chromosomal translocation or gain genetic abnormalities during redifferentiation into mature B cells. First, using the comet assay, we confirmed the DNA-damaging effect of AID in BiPSCs-AID. Secondly, we differentiated BiPSC13-AID into CD34+/CD38-/CD43-/CD45- cells by co-culture with stromal cells (mouse embryo cell line: 10T1/2), and we subsequently transplanted the cells into the bone marrow (BM) of immunodeficient NRG mice. The presence of CD34+ cells was still observed in mouse BM 4 months after transplantation; however, no differentiation into B cells was detected. Next, using the CRISPR/Cas9 system, we attempted to make BiPSCs with chromosomal translocation t(11;14); we succeeded in establishing a 293T cell line with t(11;14), then confirmed t(11;14) in MIB2-6-AID, and the clone is now being established. Furthermore, we established BiPSC13-Pax5, which can be induced by the tet-off system to express Pax5, and we then differentiated BiPSC13-Pax5 into CD34+/Pax5+/CD38-/CD43-/CD45- cells by co-culture with stromal cells (10T1/2). We expect that the HPCs or hematopoietic stem cells (HSCs) derived from BiPSCs will further differentiate into B cells due to the expression of Pax5 in the BM of NRG mouse. We also established BiPSC13-AID-p53-/-, in which p53 was deleted using the CRISPR/Cas9 system, and the cells differentiated into HPCs. Interestingly, we detected some CD43+/CD45+ cells among CD34+/CD38- cells after the co-culture of BiPSC13-AID with aorta-gonad-mesonephros-derived stromal cell (AGM-S3) instead of 10T1/2. Therefore, AGM-S3 may promote the differentiation of BiPSCs into cells that are more similar to HSCs. These CD34+ cells differentiated from BiPSCs will be transplanted into the BM of NRG mouse.

Disclosures
Hanamura: CHUGAI PHARMACEUTICAL CO., LTD.: Research Funding; Kyowa Hakko Kirin Company, Limited: Research Funding; Fujimoto Pharmaceutical Corporation: Research Funding; Takeda Pharmaceutical Company Limited.: Other: Lecture fee; Bristol-Myers Squibb: Other: Lecture fee, Research Funding; Celgene: Other: Lecture fee.
Resistance to Nutrient Deprivation Via Metabolic Re-Programming As Mechanism of 'Metastasis' Promotion in Multiple Myeloma
Authors: Savvidou Ioanna , Chandrasekara Sahan , Chenn Maoshan , Khong Tiffany , Spencer Andrew 
Published: October 2019

Abstract
Aim: Multiple Myeloma (MM) is a disease of the intra-medullary (IM) compartment but with progression myeloma cells can survive in nutrient deprived extramedullary (EM) sites. Autophagy mediates the degradation and re-cycling of intra-cellular proteins and may promote tumor resistance to metabolic stressors, including nutrient deprivation. Using the MM cell lines TK1 and TK2, contemporaneously propagated from bone marrow (IM) and peripheral blood (EM), respectively, of a MM patient we investigated the role of autophagy in resistance to glutamine (Gln) deprivation.
Methods & Results:TK2 but not TK1 exhibited up-regulated autophagy under both basal and Gln deprived conditions with higher LC3BII/I turnover on immunoblotting and autophagic vacuole formation on electron microscopy following chloroquine (CQ) exposure. Under conditions of Gln deprivation, TK2 was able to proliferate until day 14, whereas TK1 stopped proliferating at day 3. This proliferative advantage under Gln deprivation was abrogated by autophagy inhibitors (CQ or 3-MA) as determined by both viable cell enumeration (p=0.008 at day 7, inhibitor vs no inhibitor) and Ki67 expression (p=0.0017) only in TK2. Modulation of Gln concentration demonstrated an inverse correlation with the viability of TK1 but not TK2 - TK1 cell death increased from 36% at Gln 8mM to 56% at Gln 0mM, irrespective of autophagy inhibition. In contrast, while Gln deprivation had no significant effect on the viability of TK2, under Gln deprivation CQ exposure induced a 2-fold increase cell death measured by annexin V/PI positivity and 2.5-fold increase of cPARP expression (p=0.0045).
RNAseq and GSEA analysis showed enrichment of the Krige Amino Acid Deprivation Response (AADR) pathway (ES=0.083) for TK1 with up-regulated expression of Gln transporters (SLC7A5, SLC1A5, SLC38A5 and SLC7A11). In contrast, Glutamine Synthetase (GS) expression, which is responsible for Gln synthesis, was up-regulated in TK2 cells. Interestingly, immunoblotting analysis revealed that Gln deprivation was able to induce the expression of GS while further inducing autophagic flux (2.66 fold increase of LC3II expression at Gln=0mM) only in TK2, whereas it inhibited mTOR activity in both cell lines. Additionally, GS chemical inhibition under Gln deprivation induced cell death in TK2 but not TK1 (32.56% vs 2.5% increase, respectively) demonstrating that Gln synthesis enables TK2 to overcome Gln deprivation.
Conclusion. Metabolic reprogramming mitigating against nutrient deprivation may promote a more 'metastatic' phenotype in advanced MM.

Disclosures
Spencer: Celgene: Honoraria, Research Funding, Speakers Bureau; Janssen-Cilag: Honoraria, Research Funding, Speakers Bureau; Amgen: Honoraria, Research Funding; BMS: Research Funding; Takeda: Honoraria, Research Funding, Speakers Bureau; STA: Honoraria.
The Role of Nestin+ Mesenchymal Stem Cells (MSCs) in Bone Marrow Chronic Graft-Versus-Host Disease
Authors: Zhang Haiyan , Yang Kaibo , Chen Yanqiu , Jin Hua , Liu Qifa 
Published: October 2019

Abstract
Chronic graft-versus-host disease (cGVHD) is a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). cGVHD is a syndrome of variable clinical features resembling autoimmune, involving not only the epithelial target tissues (skin, lung, gastrointestinal tract and liver) but also any other organ system (musculoskeletal, joint, ocular and genital tissues). Bone marrow (BM) suppression without infection is often observed in patients undergoing allo-HSCT as GVHD symptoms appear, suggesting that BM could be a target of GVHD (Yusuke S. et. al. Blood 2010). However, the BM cGVHD is rarely reported and the mechanism for BM cGVHD is still unclear. Nestin+ mesenchymal stem cells (MSCs) are capable of differentiating into cells of multiple cell lineages and involved in the formation of fibrotic lesions (Miaohua M. et. al. Cell Mol Life Sci 2016). To determine whether Nestin+ MSCs play an important role in the development of BM cGVHD. We used two murine models of cGVHD, including MHC-matched BALB/c (H-2d) recipients given B10D2 (H-2d) grafts, and MHC-mismatched BALB/c (H-2d) recipients given C57BL/6 (H-2b) grafts. We observed that BM displayed a significant increase in collagen and reticulin fibers deposition around blood vessels in cGVHD group compared to non-cGVHD group. The decreased absolute numbers of monocytes and neutrophils in the BM indicated the broad BM suppression in the cGVHD group. We further demonstrated that Nestin+MSCs located in perivascular BM niche underwent tremendous expansion in cGVHD group (p<0.001). Nestin+ MSCs in cGVHD group acquired expression of alpha smooth muscle actin (a-SMA), indicating myofibroblast differentiation. Moreover, transforming growth factor- beta1 (TGF-β1) concentrations also increased in cGVHD BM microenvironment (p<0.0001). High concentrations of TGF-β1 induced formation of Nestin+ MSC clusters and more transition from Nestin+ MSCs to a-SMA+ cells (p<0.05). Besides, expanded Nestin+ MSCs were surrounded by many macrophages and expressed high level of TGF-β1 receptor (p<0.001 and p<0.005). Taken together, these results indicate that Nestin+ MSCs participate BM cGVHD development through transition to a-SMA+ cells induced by high TGF-β1 in cGVHD microenvironment.

Disclosures
Zhang: National Natural Science Foundation of China (No. 81600141, No. 81770190) and Natural Science Foundation of Guangdong Province (No. 2016A030310390): Research Funding. Yang:National Natural Science Foundation of China (No. 81600141, No. 81770190) and Natural Science Foundation of Guangdong Province (No. 2016A030310390): Research Funding. Chen:National Natural Science Foundation of China (No. 81600141, No. 81770190) and Natural Science Foundation of Guangdong Province (No. 2016A030310390): Research Funding.