bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thyrointegrin αVβ3 Antagonist: Implications in Acute Myeloid Leukemia
Authors: Darwish Noureldien , Glinsky Gennadi V. , Mousa Shaker A 
Published: November 2021

Abstract
Leukemic cells are able to receive and send several signals within bone marrow niche that play an important role in their survival. One of the important crosstalk is the interaction between the bone marrow microenvironment proteins (vitronectin, fibronectin, fibrinogen, and ostepontin) and thyrointegrin αVβ3 on leukemic cells, generating ligand-specific outside-in signals that are relevant to a variety of cell functions, including gene transcription, cell division, cell attachment, and motility
Our previous experiment using in vivo AML animal models with primary AML cells and cell lines have shown significant reduction of leukemic cell burden 74% and &gt;95% (P&lt;0.0001), respectively, after daily subcutaneous treatment with thyrointegrin αvβ3 antagonist fb-PMT (Ki 0.23 nM) at 3 and 10 mg/kg, for 3-4 weeks. In this study we focused on evaluations of the molecular effects of fb-PMT in leukemic cells. Acute myeloid leukemia cell lines (K562-Luc and KG1a cells) were cultured in 50 cm² cell culture flasks with 10 mL phenol red free RPMI media containing 10% fetal bovine albumin. The leukemic cells were treated (at 50% confluence) with 30 µM fb-PMT for 48 hours. Total RNA was immediately isolated from harvested cells using Triazole and used for microarray analysis. Overall, there were 370 significantly down-regulated gene expression records and 273 significantly up-regulated gene expression records, expression of which were changed at least 1.5-fold in fb-PMT-treated human leukemic cells. Significant examples of the fb-PMT-induced gene expression signatures (GES) of pathway's interference include SNAI, MYC, HIF1A, TWIST1, and TFAP2C (P&lt;0.05). Notably, inference of potential contribution to the fb-PMT anticancer activity of the interference with these pathways seems highly congruent with their known biological functions such as cell cycle control (MYC), survival and maintenance of stem cells (HIF1A, TFAP2C), and essential features of the malignant phenotype (TWIST1, SNAI) (Figure 1).
Consistently, examples of the fb-PMT-induced GES of transcriptional pathway's activation include RB1, IRF9, MAML1, RAP1A, and GATA4 pathways (P&lt;0.05), known biological functions of which appear highly consistent with the hypothesis that activation of these pathways might contribute to fb-PMT anticancer activity. Finally, we found that fb-PMT interfered with estrogen signaling in human AML cells. The fb-PMT was associated with decreased phosphorylation and nuclear enrichment of Erα (Figure 1).
Collectively, our in vivo study and genomic data have shown the key role thyrointegrin αvβ3 in leukemogenesis. The thyrointegrin αvβ3 antagonist fb-PMT demonstrated potent anticancer actions on human AML through the molecular interference mechanism with multiple signaling pathways supporting growth and survival of leukemic cells
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Early Transfusion with Mesenchymal Stem Cell Derived Extracellular Vesicles: A New Transfusion Strategy for Life-Threatening Hemorrhage and Traumatic Brain Injury
Authors: Dekker Simone E , Bambakidis Ted , Williams Aaron M , Biesterveld Ben , Bhatti Umar , Li Yongqing , Pickell Zachary , Buller Benjamin , Alam Hasan B 
Published: November 2021

Abstract
Background: Life-threatening hemorrhage and traumatic brain injury (TBI) have a significantly increasing global burden and remain leading causes of preventable deaths. Effective interventions may protect the brain against ongoing damage and improve the long-term outcomes. A growing area of interest is transfusion of cell-based therapies, particularly with bone marrow-derived mesenchymal stem cells (MSC). Transfusion using MSC derived extracellular vesicles (EVs) have shown to improve neurologic outcomes in animal models of life-threatening hemorrhage, stroke, and TBI. However, the precise mechanisms remain poorly characterized. In the present study, we aimed to elucidate some of the key cerebral genes, pathways, and networks that were modulated after transfusion of EVs in a porcine model of hemorrhagic shock (HS) and TBI.
Methods: Swine were subjected to HS (40% blood volume) and severe TBI (8-mm cortical impact). After 1 hour of shock, animals were randomized (n=4/group) to treatment with either lactated Ringer's (LR) or LR+EV. Both groups received fluid resuscitation after 2 hours of shock, and autologous packed red blood cells 5 hours later. After 7-days, brains were harvested and RNA-sequencing was performed. The transcriptomic data was imported into the iPathway pipeline for bioinformatics analyses.
Results: 5,273 genes were differentially expressed in the LR+EV group vs. LR alone (total 9,588 measured genes, Figure 1). Table 1 lists the top 10 genes exhibiting the greatest up- and down-expression based on fold change. Genes with the greatest up-regulation were involved in synaptic transmission and neuronal development and differentiation, while down-regulated genes were involved in inflammation. Gene Ontology terms experiencing the greatest modulation were involved in inflammation, brain development, and cell adhesion. Pathway analysis revealed significant modulation in the glutamatergic and GABAergic systems. Network analysis revealed down-regulation of inflammation (Figure 2), and up-regulation of neurogenesis, and neuron survival and differentiation.
Conclusions: In a porcine model of HS+TBI, EV transfusion was associated with an attenuation of cerebral inflammatory networks and a promotion of neurogenesis and neuroplasticity. These transcriptomic changes could explain the observed neuroprotective and neurorestorative properties associated with EV transfusion. EV transfusion reduces the hyper-inflammatory response and may have great promise in improving outcomes in concurrent life-threatening hemorrhage and severe TBI. Further testing of this novel strategy and its implications in transfusion medicine are warranted.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Analysis of Polymorphism in the Genes TLR3, TLR4 and TLR9 in Patients with Acute Myeloid Leukemia
Authors: Rybka Justyna , Bogunia-Kubik Katarzyna , Kuszczak Bartłomiej , Kalicińska Elżbieta , Łacina Piotr , Dratwa Marta , Wróbel Tomasz 
Published: November 2021

Abstract
Acute myeloid leukemia (AML) is a bone marrow malignancy characterized by uncontrolled proliferation of transformed myeloid precursors. It is the most common acute leukemia and is a highly heterogeneous disease. AML leads to impaired hematopoiesis, which may result in increased risk of infection. Toll-like receptors (TLRs) are major elements of the innate immune system which recognize pathogen-associated molecular patterns. They also have a role in autoimmune diseases and cancer through their recognition of endogenous danger-associated molecular patterns. Various TLRs were found to be expressed in AML cells, and their expression was found to affect growth, differentiation, and immunostimulatory capacity in AML.
We retrospectively analysed 90 patients (48 females and 42 males) with diagnosis of AML, hospitalized in Department and Clinic of Hematology, Blood Neoplasms and Bone Marrow Transplantation of Wroclaw Medical University. The median age of patients was 62 (range:18-81). Infectious complications after induction chemotherapy occurred in 62 patients. In the present study, we detected the effect of seven single nucleotide polymorphisms (SNPs) located in the genes coding for TLR3 (rs5743305, rs3775296, 3775291), TLR4 (rs4986791, rs4986790) and TLR9 (rs187084, rs5743836). DNA was isolated from samples of peripheral blood taken on EDTA. Melting curve genotyping with the use of LightSNiP asays(TIB MOLBIOL, Germany) was employed to determining TLR3, TLR4 and TLR9 genotypes, and the reactions were performed LightCycler 480 device (Roche Applied Science, Germany).Statistical analyses were performer using Real Statistics Resource Pack for Microsoft Excel.
We found that TLR3 rs5743305 AA homozygosity was more common in AML patients with infectious complications than in patients unaffected by infections (p=0.015). TLR4 rs4986791 C and TLR9 rs5743836 C were associated with lower levels of the inflammation marker CRP (p=0.015 and p=0.048, for rs4986791 and rs5743836, respectively), TLR4 rs4986791 C and was less frequently detected among patients with CRP&gt;5 (p=0.028). We also found TLR9 rs187084 C to be more common in patients with adverse cytogenetic risk according to ELN criteria (0.012). Additionally, a trend for higher prevalence of extramedullary disease was observed for carriers of TLR3 rs3775296 T (p=0.054), TLR3 3775291 GG (p=0.068), TLR4 rs4986790 G (p=0.078), as well as TLR9 rs187084 TT (p=0.053).
In conclusion, our results suggest that SNPs in the genes coding for TLR3, TLR4 and TLR9 may be implicated in clinical outcome of AML and is related with increased risk of infectious complications.

Disclosures
Wróbel: BMS: Honoraria; Roche: Honoraria, Research Funding, Speakers Bureau; Janssen: Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau; Takeda: Honoraria, Speakers Bureau; BeiGene: Honoraria.
Obesity and Oral Contraceptives Synergistically Increase Thrombotic Risk By Downregulating Protein S
Authors: Thangada Lekha , Kumar Manoj , Pilli Satish Sekhar , Majumder Rinku 
Published: November 2021

Abstract
Background:
The increased thrombotic effects of estrogen-based oral contraceptives and obesity have been documented independently. However, obesity and oral contraceptives combined are associated with a far greater thrombotic risk, but we have a poor understanding of the mechanism of this greater effect. Increasingly, women are using oral contraceptives, and the national obesity rate has been skyrocketing. Thus, it is imperative to explain how obesity and oral contraceptives work together to significantly elevate thrombotic risk. We know that hypoxia-inducible factor 1-ɑ (HIF-1ɑ) and the estrogen receptor (Erɑ) bind to the promoter of the Protein S gene; binding occurs at sites within ~450 nucleotides of each other, and the two transcription factors downregulate Protein S expression independently. We hypothesize that the two factors, by binding the promoter simultaneously, synergistically downregulate Protein S to a degree much greater than the downregulation mediated by each factor separately.
Aims:
The goal of this project is determining whether estrogen and obesity-induced hypoxia work synergistically to downregulate Protein S transcription and increase thrombotic risk.
Methods:
We measured the effects of obesity and oral contraceptives on hepatocarcinoma (HEP G2) cells because the liver is the major producer of Protein S. Estrogen of varying concentrations (25-150 μM) was used to mimic the effects of oral contraceptives, and cobalt chloride of varying concentrations (25-150 μM) was used to stimulate hypoxia and HIF-1ɑ expression. Cells were exposed to estrogen only, cobalt chloride only, and estrogen and cobalt chloride together for 24 hours, after which the cells were harvested and subjected to q-PCR and immunoblot blot analyses to measure Protein S transcription and protein expression. Although cobalt chloride is a reliable inducer of hypoxia and HIF-1ɑ expression, we also performed hypoxia experiments by incubating cells in a chamber with varying O 2 concentrations (20%, 15%, 10%, 5%, 1%).
Results and Conclusions:
Immunoblot analysis of cells treated with either CoCl 2 and estrogen supplementation revealed a ~20% reduction in Protein S levels compared to control conditions and a more significant reduction in (~60%) Protein S expression in cells treated with estrogen and CoCl 2 together. These results supported our hypothesis that obesity and estrogen-based contraceptives increase thrombotic risk by downregulating anticoagulant Protein S transcription and subsequently decreasing Protein S level.

Disclosures
No relevant conflicts of interest to declare.
Telemedicine in Hematology: Tool to Improve Access and Patient Satisfaction
Authors: Master Samip R , Ramadas Poornima , Mansour Richard Preston 
Published: November 2021

Abstract
Introduction: Telemedicine can transform the future of medicine, especially in rural settings by improving access to medical care. Here were present the patient satisfaction data from both telemedicine and regular visits in our Hematology & Oncology clinics. In Hematology, we have used tele medicine for sickle cell disease, benign hematology cases like iron deficiency, thrombocytopenia, leukocytosis etc., and in malignant hematology for new patient consults, toxicity checks, follow up of plasma cell disorders, chronic leukemias and low intensity chemotherapies.
Methods: We retrospectively collected patient satisfaction survey data from March 2020 to April 2021 for both telemedicine and regular visits. The patients were allowed to give a score from 1 to 99 regarding their experience at visit. We collected data on the following: likelihood of recommending, care provider, access and overall assessment.
Results: A total of 53 patient satisfaction survey were available for telemedicine and 462 for regular visits. Over 60 % of tele visits were non cancer related and 40% were cancer related. The average score for likelihood of recommending, care provider, access, and overall assessment for tele visit were 73.5, 41.5, 73.5 and 62 respectively. The average score for likelihood of recommending, care provider, access, and overall assessment for regular visit were 13.75, 7.5, 26.5, and 11 respectively.
Conclusion: The patient satisfaction scores for likelihood of recommending, care provider, access and overall assessment were significantly better for tele visits compared to regular visits.

Disclosures
Master: Blue Bird Bio: Current holder of individual stocks in a privately-held company.
PI3K/AKT Activation Mediates B-Cell Transformation By the "T" Splice-Variant of Fyn Kinase
Authors: Kumar Preet , Li Lingxiao , Schatz Jonathan H. 
Published: November 2021

Abstract
The SRC-family kinase FYN has key roles downstream of growth-factor receptor activation and is implicated in various cancers. Activating FYNutations were reported in angioimmunoblastic T-cell lymphoma (AITL), but its role in transforming lymphocytes is poorly characterized. FYN also may play a role in signaling downstream of the B cell Receptor (BCR) receptor in Diffuse Large B-Cell Lymphoma (DLBCL), and laboratory studies show inhibition of FYN and other SRC family kinases (SFKs) can block BCR-mediated lymphomagenesis. To investigate FYN's role in oncogenic singling, we introduced FYN alleles to IL3-dependent FL5.12 murine pro-B cells and tested for selective advantage upon cytokine withdrawal and ability to transform the cells to cytokine independence. FYN B and T variants result from alternate splicing of exon 7, with B originally detected in brain and T in hematopoietic cells, but both are expressed and may be deregulated associated with disease states in diffuse tissues. We employed wildtype (WT) alleles for both spliceoforms along with mutations detected recurrently in AITL (L174R and R176C), kinase active (Y528H), and kinase dead (K299A). No FYN B alleles showed any selective advantage in the prolymphocyte cells subjected to cytokine withdrawal. In contrast, all FYN T alleles other than kinase dead were strongly enriched through multiple rounds of IL3 withdrawal and rescue. Indeed, complete withdrawal of cytokine resulted in transformation to IL3-independent growth, generating stable cell lines addicted to FYN kinase activity for survival. The SFK inhibitor PP2 was toxic to all FYN-T transformed cells but not to baseline FL5.12 cells growing in cytokine. WT- and L147R-transformed cells were similarly sensitive to PP2, while the other AITL-derived mutant R176C demonstrated reduced sensitivity. Immunoblotting demonstrated PP2-sensitive phosphorylation of AKT and the MTORC1 target 4EBP1, suggesting PI3K/AKT activation as a key survival pathway downstream from transformation by FYN T alleles. Consistently, the AKT inhibitor ipatasertib and PDK1 inhibitor GSK2334470 were highly active against all FYN-transformed lines, and AKT phosphorylation was sensitive to PDK1 inhibition, showing convergence on AKT activation in all transformants. Pan-PI3K inhibition was active against FYN transformants, while the delta isoform-specific inhibitor idelalisib showed only partial activity, suggesting activation of multiple PI3K isoforms downstream of FYN activity. These results suggest that AKT, PI3K, and PDK1 are important targets downstream of FYN, and that the PI3K/AKT pathway is the key survival mechanism in lymphomagenesis mediated by FYN. We show that WT and activating mutants of the FYN T spliceoform specifically, but not FYN B, induce drug-sensitive B lymphocyte malignant transformation by activating PI3K/AKT/MTORC1 signaling.

Disclosures
No relevant conflicts of interest to declare.
Caplacizumab Is Effective in Treating Immune-Mediated Thrombotic Thrombocytopenic Purpura in Real-World Scenarios and May Lower Mortality: An Aggregative Analysis
Authors: Völker Linus A. , Balduin Gesa , Kühne Lucas , Kaufeld Jessica , Schönermarck Ulf , Menne Jan , Knöbl Paul , Brinkkoetter Paul 
Published: November 2021

Abstract
Introduction: The treatment of acquired thrombotic thrombocytopenic purpura has been a subject to change lately due to the introduction of caplacizumab as a novel, and potentially life-saving agent. Here, we compare patient-centered outcomes of a historic cohort of 119 aTTP-patients and an extended real-world caplacizumab-treated cohort of 113 aTTP-patients from Germany and Austria in the context of multiple recently published international cohorts of aTTP patients.
Methods: From October 2018 until May 2021, data from German and Austrian patients presenting with an acute episode of aTTP as defined by an ADAMTS13 activity below 10 % treated with caplacizumab were gathered retrospectively. We aggregated data from two published randomized controlled trials (TITAN, HERCULES), one single-arm prospective trial and two retrospective cohorts with a historical control cohort. In total, we report 846 aTTP cases, 396 of those treated with caplacizumab and 450 without. For data analysis, placebo groups from randomized trials and historical controls from retrospective analyses were combined.
Results: All cohorts combined reported 30 aTTP-related deaths, predominantly in the non-caplacizumab cohort. Mortality in the caplacizumab-cohort (8 cases total) was mainly driven by 5 reported deaths in the UK-cohort, which have been ascribed to a delayed use of caplacizumab by the authors. The number of daily PEX was significantly less in the caplacizumab-treated cohort (7.2 vs 10.2 days). In line, the number of hospital (14.0 vs 18.1) and ICU (4.7 vs 8.9) days differed significantly. Exacerbations (14% vs 39%) were significantly reduced in the caplacizumab-group. In view of all treatment-related outcomes, the data presented herein argues strongly in favor of an early and consistent use of caplacizumab in all instances of aTTP.

Disclosures
Völker: Sanofi-Genzyme: Honoraria, Other: counselling fees.
Effect of Hydroxyurea Therapy on the Incidence of Infections in Ugandan Children with Sickle Cell Anaemia
Authors: Namazzi Ruth , Conroy Andrea L. , Bond Caitlin , Goings Micheal J. , Datta Dibyadyuti , Ware Russell E. , Jang Jeong Hoon , John Chandy C. , Opoka Robert O. 
Published: November 2021

Abstract
Hydroxyurea is efficacious against sickle cell anaemia (SCA)-related complications in African children. Prior studies demonstrated conflicting results on the effect of hydroxyurea on risk of infection, the most common cause of morbidity and mortality in African children with SCA.
We evaluated the incidence of infections before and after starting hydroxyurea in 117 children aged 1-5 years with SCA enrolled in the Zinc for Infection Prevention in Sickle cell anaemia (ZIPS) clinical trial that received zinc or placebo treatment for one year (Clinicaltrials.gov, NCT03528434). Children were enrolled between March 2018 and November 2019 and initiated on hydroxyurea (20 mg/kg/day) if they met Uganda SCA guideline criteria for hydroxyurea treatment at any time during the study. We compared the incidence of infections before and after hydroxyurea therapy, adjusting for zinc treatment.
Overall, the mean duration on hydroxyurea was 223.8 (85.2) person days. The mean(SD) incidence of any severe/invasive infections (infections meeting strict clinical and laboratory or radiological diagnostic criteria) was 6.2(9.0) vs. 1.9(2.3) infections per child per year before and after hydroxyurea (incidence rate ratio [IRR]: 0.40, 95%CI: 0.29-0.54, p&lt;0.001), including a decrease in the incidence of bacteremia [IRR: 0.05] malaria, [IRR 0.28], cellulitis [IRR: 0.24], gastroenteritis [IRR: 0.41], pharyngitis [IRR: 0.59] and sinusitis [IRR: 0.38] (all p&lt;0.05). Similarly, the incidence of clinically defined infections (infections meeting clinical but not laboratory/radiological criteria) decreased after hydroxyurea, (IRR: 0.49, 0.41-0.59), influenced primarily by large reductions in the incidence of lower and upper respiratory tract infections (p&lt;0.001 for both). As expected, hydroxyurea treatment was also associated with a decrease in complications of SCA, including vaso-occlusive crises, hospitalizations and transfusions (all p&lt;0.001). There was no interaction between zinc and hydroxyurea therapy on risk of infection and SCA-related complications.
In Ugandan children with SCA, hydroxyurea therapy not only decreases the incidence of SCA-related complications, but also substantially reduces the incidence of infections. Research to understand the underlying mechanisms of protection from hydroxyurea against infection and exploration of its potential use for infection prevention is warranted.

Disclosures
Ware: Bristol Myers Squibb: Research Funding; Addmedica: Research Funding; Hemex Health: Research Funding; Nova Laboratories: Research Funding; Novartis: Other: DSMB Chair; Editas: Other: DSMB Chair.