bluuebunny/crossref_metadata_2025_split huggingface dataset:
Resolvin D1 and Resolvin D2 Protect Against Hypoxia/Reoxygenation Induced Lung and Kidney Damage in a Sickle Cell Mouse Model of Acute Vaso-Occlusive Crisis
Authors: Mattè Alessandro , Recchiuti Antonio , Serhan Charles , Siciliano Angela , Federti Enrica , Weinberg Olga , Kalish Brian , Brugnara Carlo , De Franceschi Lucia 
Published: October 2019

Abstract
Sickle cell disease (SCD) is characterized by hemolytic anemia in association with acute and chronic life-threatening clinical complications. Acute vaso-occlusive crisis (VOCs) are the main cause of hospitalization for SCD patients. In VOCs, amplified inflammatory response plays a key role in acute organ damage. Pro-Resolving lipid mediators such as resolvins (Rv) accelerate resolution of acute inflammation in different models, indicating that stimulation of endogenous resolution of inflammatory processes may be an additional strategy in limiting tissue damage. Recent data suggest beneficial effects of Rvs in hypoxia/reoxygenation (H/R) related tissue injury. Here, we study the effects of Rvs on a model of acute VOCs using humanized SCD mice (Hbatm1(HBA)Tow Hbbtm2(HBG1,HBB*)Tow). We treated SCD and control healthy mice (AA, Hbatm1(HBA)Tow Hbbtm3(HBG1,HBB)Tow) (n =6-7 animals in each group) with RvD1, 7S, 8R, 17S- trihydroxy-docosa-4Z, 9E, 11E, 13Z, 15E, 19Z-hexaenoic acid, 17R- RvD1, 7S, 8R, 17R- trihydroxy-docosa-4Z, 9E, 11E, 13Z, 15E, 19Z-hexaenoic acid; RvD2, 7S, 16R, 17S-trihydroxy-docosa-4Z, 8E, 10Z, 12E, 14E, 19Z-hexaenoic acid. Mice were treated with RvD1 and RvD2 at the dose of 2.5 ug/Kg by gavage 1 hour (hr) before H/R stress (10 hrs 8% oxygen followed by 3 hrs reoxygenation), which we have used in the past to mimic acute VOCs. We found that RvD1 and RvD2 significantly reduced the H/R-induced (i) increase in neutrophil count; (ii) lung inflammatory cell infiltrate, mucus and thrombi formation; (iii) glomerular inflammatory cell infiltration, glomerular sclerosis and thrombi formation. In the lung of H/R SCD mice, RvD1 prevented the H/R induced up-regulation of (i) cytokines such as MCP2, IL-6 and ET-1; (ii) vascular endothelial activation markers (VCAM-1 and ICAM-1; (iii) cytoprotective systems such as Prx6 and HO-1. In the kidney of H/R SCD mice, RvD1 significantly reduced H/R induced expression of IL-6 and ET-1 as well as HO-1. Our data indicate that RvD1 and RvD2 modulating inflammatory responses related to H/R in SCD, protect sickle cell target organs, and foster resolution. Thus, RvD1 and RvD2 might represent a novel therapeutic approach for acute VOCs in SCD.

Disclosures
No relevant conflicts of interest to declare.
Placenta Growth Factor Links the IL-13 Response and the Leukotriene Pathway to Augment Airway Hyper-Responsiveness
Authors: Eiymo Mwa Mpollo Marthe-Sandrine , Brandt Eric , Shanmukhappa Shiva Kumar , Arumugam Paritha , Tiwari Swati , Loberg Anastacia , Rizvi Tilat , Lindsey Mark , Jonckx Bart , Carmeliet Peter , Kalra Vijay , Lecras Timothy , Ratner Nancy , wills-Karp Marsha , Khurana hershey Gurjit , Malik Punam 
Published: October 2019

Abstract
Airway hyper-responsiveness (AHR) affects 55-77% of children with sickle cell disease (SCD) and occurs even in the absence of asthma. While asthma increases SCD morbidity and mortality, the underlying mechanisms of this highly prevalent AHR in a hemoglobinopathy remain unknown. We hypothesized that Placenta Growth Factor (PlGF), an erythroblast-secreted factor that is elevated in SCD, mediates this AHR. We studied AHR in allergen-exposed PlGF-/- and SCD mice. Sickle mice genetically deficient in PlGF were not viable. PlGF augmented AHR, lung inflammation and blood/lung eosinophilia via IL-13 (a Th2-cytokine) and increased 5-lipoxygenase (a leukotriene synthetic enzyme) expression. AHR, or pulmonary inflammation and leukotriene levels were blunted in PlGF-/- mice or in PlGFWT mice treated with anti-PlGF-Ab, and was rescued by intratracheal administration of leukotrienes to PlGF-/- mice. Notably, Th2-mediated Stat-6 activation further increased PlGF expression from lung epithelium, eosinophils and macrophages, linking the PlGF-leukotriene pathway in a positive feedback loop. Th2 response is classically seen in asthma, and indeed, expression of PlGF and its downstream genes was increased in respiratory epithelial cells of asthma patients. Like patients with SCD, sickle hematopoietic chimeras developed increased AHR and had higher leukotriene levels; and both were abrogated by anti-PlGF-Ab or zileuton (5-lipoxygenase inhibitor) phenocopying the blunted AHR in PlGF-/- mice, and indicating that sickle erythroid-cells/RBC potentiate AHR. Overall, PlGF exacerbates AHR and uniquely links the leukotriene and Th2 pathways in asthma. Both zileuton, licensed for asthma and anti-PlGF-Ab, in clinical trials for cancer, could be promising therapies to reduce the pulmonary morbidity in SCD.

Disclosures
Jonckx: ThromboGenics: Employment.
Factor XIII and Red Blood Cells in Venous Thrombosis
Authors: Wolberg Alisa S. 
Published: October 2019

Blood coagulation culminates in the thrombin-mediated conversion of fibrinogen to fibrin and production of a fibrin network that stabilizes the clot. The transglutaminase factor XIII(a) [FXIII(a)] crosslinks fibrin and increases the clot's resistance to fibrinolysis and mechanical disruption. Consequently, fibrin formation and crosslinking are essential for hemostasis. However, the formation of clots with abnormal fibrin network structure and/or stability (resistance to mechanical disruption or pharmacological dissolution) is a risk factor for both arterial and venous thrombosis. Improved understanding of fibrin formation and crosslinking during coagulation is essential for understanding the pathophysiologic mechanisms that promote thrombotic disease. We have shown that elevated levels of fibrinogen accelerate vessel occlusion and enhance thrombus stability in mice, demonstrating etiologic contributions of fibrin(ogen) to thrombosis. Recently, we discovered that FXIIIa transglutaminase activity also mediates venous thrombosis via its ability to promote red blood cell retention during clot contraction in whole blood. Genetic deletion of FXIII or pharmacologic inhibition of FXIIIa activity reduces red blood cell retention in clots and consequently, reduces thrombus size in vitro and in vivo. Furthermore, FXIII zymogen binding to soluble, circulating fibrinogen is necessary for normal FXIII activation and activity. The fibrinogen motif that mediates FXIII binding involves C-terminal residues in the fibrinogen gamma chain (gamma 390-396); mice that express a variant fibrinogen that has mutations within these residues show reduced binding of FXIII zymogen to fibrinogen, and delayed activation of FXIII. In venous thrombosis models, these mice phenocopy FXIII-deficient mice, producing smaller venous thrombi with reduced red blood cell content compared to thrombi from wild-type mice. Intriguingly, these mice do not exhibit bleeding or poor wound healing normally associated with FXIII deficiency. Collectively, these findings provide new insight into the pathophysiology of venous thrombosis, and expose the fibrin(ogen)-FXIII axis as a central determinant of venous thrombus formation and composition.

Disclosures
No relevant conflicts of interest to declare.
Choice of Better T-cells for Adoptive Immunotherapy
Authors: Busch Dirk Hans 
Published: October 2019

Abstract
Adoptive transfer of primary (unmodified) or genetically engineered antigen-specific T cells has demonstrated astonishing clinical results in the treatment of infections and some malignancies. The definition of optimal targets and antigen receptors as well as the differentiation status of transferred T cells are emerging as crucial parameters for generating cell products with predictable efficacy and safety profiles. Our laboratory has demonstrated that defined subsets within the memory CD8+ T cell compartment fulfill all key characteristics of adult tissue stem cells and are essential for robust and long-term maintained responses upon adoptive transfer. We have developed clinical multi-parameter enrichment technologies to purify these memory stem cells for clinical applications. In my presentation I will report on the status of ongoing clinical trials using such purified cell products either as a primary T cell population for the treatment of infections upon allogeneic stem cell transplantation or after genetic modification with a CD19 CAR for the treatment of malignancies (collaboration with Stan Riddell, FHCC/Seattle). Infusing small numbers of T cells within a memory stem cell product can be highly effective therapeutically, but bears some risk of toxicity. Therefore, safeguards that allow selective depletion of transferred cells in the case of un-tolerable side effects may be needed to further improve adoptive immunotherapy. I will present results exploring the capacity of a truncated version of EGFR (EGFRt) co-expressed with T cells expressing a CD19-CAR. In pre-clinical mouse models we demonstrate that application of Cetuximab, which binds to EGFRt, confers selective depletion of adoptively transferred CAR-T cells in vivo. Long-term B cell aplasia, which is a main side effect of CD19-CAR T cell therapy, can be completely reverted with this strategy. Vaccination studies upon B cell recovery demonstrate full functionality of antigen-specific antibody formation. EGFRt co-expressing CD19-CAR T cells have been successfully transferred into first human patients, providing the option to test for the first time in a clinical setting whether treatment of B cell aplasia after long-term leukemia remission can be achieved by selective depletion.

Disclosures
Busch: STAGE cell therapeutics: Other: I was share holder of STAGE cell therapeutics, a company that was recently bought by Juno therapeutics.. Off Label Use: CD19 CAR T cells.
Clonal Diversity and Mutated Genes in Myelodysplastic Syndromes
Authors: Ebert Benjamin L 
Published: October 2019

Myelodysplastic syndromes (MDS) are driven by somatic genetic mutations that drive clonal explansion of neoplastic cells. The identification of these lesions has enabled large-scale studies to assess the association of genetic lesions with clinical and biological phenotype. Single mutations in driver genes in isolation, without concurrent mutations in additional driver genes, commonly do not cause overt disease, but rather cause an expanded population of blood cells without cytopenias, a condition that we have termed clonal hematopoiesis of indeterminate potential (CHIP). CHIP is common in the general population and is associated with age, risk of hematologic malignancy, and overall mortality. In contrast, patients with MDS generally have mutations in multiple genes as well as the disease-defining morphologic dysplastia and peripheral blood cytopenias. The particular mutations that drive MDS in an individual patient are powerfully associated with clinical phenotype, including cytopenias, specific morphologic abnormalities, blast percentage, overall survival, and response to therapy. Clonal architecture adds an additional layer of complexity as disease progression or response to therapy may be associated with shifts in the dominant genetic clone in the bone marrow. As the availability of genetic testing for panels of genes becomes widespread as a clinical assay, current endeavors are to move validated research findings into routine clinical use and to employ deep genetic analyses to evaluate response to investigational therapies in clinical trials.

Disclosures
Ebert: Genoptix: Consultancy, Patents & Royalties; H3 Biomedicine: Consultancy; Celgene: Consultancy.
The Role of the Tumor Microenvironment in Lymphoid Malignancies
Authors: Steidl Christian 
Published: October 2019

Lymphoid cancers represent a heterogeneous group of neoplasms composed of malignant lymphoid cells with variable infiltration by non-neoplastic, mostly immune cells (tumor microenvironment). For some subtypes of lymphoid cancers, the contribution of the microenvironment to the histological appearance is widely recognized and used for pathological classification. Although microenvironment-related biology in lymphoid cancers has been primarily explored in a core set of lymphoma subtypes, the number of entities studied has recently accelerated. The pathogenic evolution of tumor microenvironments and in particular their composition and spatial distribution can be perceived as a complex function of 1) genetic alterations within the malignant cell population, 2) the extent and dependence on the molecular crosstalk involving cyto- and chemokines, and 3) host-specific factors. As a result, three major patterns of microenvironmental architecture can be distinguished termed "Re-education", "Recruitment" and "Effacement". Hodgkin lymphoma can serve as a paradigm for an extensive crosstalk between tumor cells and a quantitatively dominant tumor microenvironment. Importantly, related prognostic implications of tumor microenvironment composition (e.g. macrophage content, representation of T cell subsets) have been extensively studied in this disease. With focus on B cell lymphomas, this talk will highlight the emerging literature about genetic alterations in malignant lymphoma cells that provide the foundation for somatically acquired immune privilege and evasion from immune surveillance. The genomic changes discussed in this talk can be broadly categorized according to the effect that they exert on the tumor microenvironment: 1) Loss or down-regulation of (surface) molecules leading to decreased immunogenicity of tumor cells (e.g. mutations of B2M, CIITA); 2) Increased expression of surface molecules suppressing immune cell function (e.g. structural genomic changes of PDL1, PDL2); 3) Recruitment or induction of a regulatory cellular milieu (e.g. mutations in JAK-STAT and NFκB signaling pathways). The discovery of gene mutations underlying immune privilege, properties of the altered molecules, downstream functional consequences and clinical rationales for therapeutic intervention will be presented in the context of specific lymphoma subtypes. It will be discussed how precise description of genomic and molecular alterations underlying immune privilege might accelerate effective targeting of microenvironment-related biology in the clinical setting. Moreover, the development and clinical implementation of predictive biomarkers will be outlined that harbor the potential to inform on companion diagnostic approaches to accompany therapies such as immunological checkpoint inhibition.

Disclosures
No relevant conflicts of interest to declare.
Secondary Polycythemia and the Risk of Venous Thromboembolism (VTE) Among Hospitalized Patients in the United States
Authors: Giri Smith , Mehta Kathan Dilipbhai , Bhatt Vijaya R. 
Published: October 2019

Abstract
Introduction:The prevalence of secondary polycythemia is on the rise due to increasing prevalence of obesity, sleep apnea and other chronic cardiopulmonary diseases. While polycythemia vera is well known to increase the risk of VTE, the impact of secondary polycythemia on the development of VTE is less clear.
Methods: We utilized the Nationwide Inpatient Sample (NIS) database to identify all adults ≥18 years with secondary polycythemia (ICD-9-CM code 289.0) diagnosed during 2011. NIS is the largest all-payer inpatient database in the US that captures about 20% of all US hospitalizations. The outcome of interest was acute VTE identified using corresponding ICD codes for DVT or PE. We also identified the presence of the following confounding factors: malignancy, venous catheterization, long bone fracture, infection, trauma, mechanical intubation, congestive heart failure (CHF), pregnancy, length of hospital stay >5 days, stroke and prior history of VTE. Multivariate logistic regression was used to assess the impact of secondary polycythemia on the occurrence of VTE controlling the above confounding factors. Multicollinearity was assessed using variance inflation factor (VIF). All p-values were 2 sided, and the level of significance was chosen at 0.05. Statistical analysis was done using STATA 13.0 (StataCorp, College Station, TX) taking into account the complex sampling design of the database.
Results: A total of 3,972 hospitalizations with a primary or secondary diagnosis of secondary polycythemia were identified out of a total of 6,840,854 hospitalizations. Concurrent diagnosis of VTE was seen in 4.8% of patients with secondary polycythemia (n=190) as opposed to 2.3% of patients without secondary polycythemia (n=161,122) (odds ratio, OR 2.07; 95% confidence interval, CI 1.75-2.45; p value <0.01). In a multivariate analysis, secondary polycythemia continued to have a significantly greater risk of concurrent VTE (adjusted OR 1.87; 95% CI 1.58-2.22; p value <0.01) after adjusting for confounders including age, stroke, malignancy, infection, history of VTE, long bone fracture, trauma, CHF, pregnancy, mechanical intubation, nephrotic syndrome and hospital stay >5 days. No evidence of multicollinearity was noted in the final regression model (maximum VIF of 1.13).
Conclusion:
Within the limitations of the study design, our retrospective study indicates an independent association between secondary polycythemia and VTE in patients admitted in the hospital, however, this does not imply causality. Further, our study does not answer whether the use of therapeutic phlebotomy reduces the risk of VTE in patients with secondary polycythemia.

Disclosures
No relevant conflicts of interest to declare.
A Clinic-Genetic Score for Risk Assessment of Recurrent Venous Thrombo Embolism
Authors: Gerotziafas Grigorios T. , Onambele Manuella , Benzerara L , Mokrani H , Ketatni Hela , Planche V , Delassasseigne C , Elalamy Ismail 
Published: October 2019

Abstract
Background
Current risk assessment models to predict the risk of recurrent venous thromboembolism (VTE) in an individual patient (i.e Vienna Pedicitive Model, DASH score) include clinical and biochemical parameters and have showed discrimination values ranging from 0.64 to 0.68. Genetic variants associated to thrombophilia have showed to play a relevant role in predicting primary VTE. Combination of those genetic variables together with clinical parameters may improve the predictive capacity of current models for predicting recurrent VTE.
Aim
To analyze the predictive capacity of a new clinic-genetic score for predicting recurrence of VTE, Thrombo inCode-Recurrent (TiC-Reccurent).
Methods
A population of 55 patients with a first VTE and none or one or more recurrent VTE (21 males, 34 females; 47.8±14.7 years old) and 39 controls with only the first VTE event (13 males, 26 females; 40.2±13.2 years old) and the same follow-up was used.
A regression model was constructed including clinical variants (age, gender, diabetes status, pregnancy status, contraceptive hormonal therapy, smoker status and family history of VTE) and genetic variants (F5 rs6025/rs118203906/rs118203905, F2 rs1799963, F12 rs1801020, F13 rs5985, SERPINC1 rs121909548, and SERPINA10rs2232698 plus the A1 blood group (rs8176719, rs7853989, rs8176743, rs8176750).
The predictive capacity was assessed by calculating the c-statistic (AUC-ROC); sensitivity, specificity Positive and Negative Likehoood ratios and odds ratio of a positive test result.
Informed consent was obtained and the study was approved by the Institution Ethics Committee. This study complies with the guiding principles for experimental procedures found in the Declaration of Helsinkiof the World Medical Association.
Results
In an univariate analysis, none of the clinic or genetic variables showed a significant difference between the patient with or without recurrences. TiC-Recurrent score showed a predictive capacity for recurrent VTE measures as AUC-ROC: 0.738, p<0,0001. For TiC-Recurrent the clinical sensitivity was 81.82 and the specificity was 33.33. The predictive capacity measures as LR+, LR- were 1.23 and 0.55, respectively. The odds ratio for TiC-recurrent was 2.7.
Conclusions
TIC-Recurrent score showed a good predictive capacity to identify subject at high risk of suffering a recurrence of VTE. Although a direct comparison to current scores for predicting the risk for a recurrence of VTE was not possible in the present study if the observed predictive capability for TiC-recurrent is confirmed it will improve the capacity of current scores. Our study suggests that the use of a clinical-genetic algorithm could be an aid in the treatment of patient with a first VTE and contribute to the prevention of VTE recurrences.

Disclosures
No relevant conflicts of interest to declare.
Functional and Molecular Alterations of Bone Marrow Mesenchymal Stem and Progenitor Cells in Patients with Myelodysplastic Syndrome with Ring Sideroblast
Authors: Dolinska Monika , Xiao Pingnan , Johansson Anne-Sofie , Sandhow Lakshmi , Kondo Makoto , Bouderlique Thibault , Ungerstedt Johanna , Hellstrom-Lindberg Eva , Qian Hong 
Published: October 2019

Abstract
Myelodysplastic Syndrome with Ring Sideroblast (MDS-RS), a clonal hematopoietic cell neoplasm, is low risk MDS, characterized by anemia, hyperplastic ineffective erythropoiesis and marrow ring sideroblasts. Mouse studies have shown that bone marrow niche, including endothelial cells, osteoblasts, adipocytes and mesenchymal stem cells (MSCs), contribute to progression of various hematological disorders. However, in vivo contribution of the different bone marrow stromal cells to the progression of MDS-RS in patients remains largely unknown.
To investigate this, we have phenotypically, molecularly and functionally characterized the BM native stromal cell subsets including MSCs freshly isolated by multi-color fluorescence activated cell sorting (FACS) from bone marrow of MDS-RS patients and age-matched healthy donors. We found: 1) the MDS-RS MSCs, estimated by colony forming unit-fibroblast (CFU-F), shared similar immunophenotype with normal MSCs (CD45-CD235a-CD31-CD44-, most of which were CD271+CD146+CD106+); 2) the frequency of CFU-Fs was significantly increased in the phenotypically defined MSCs of MDS-RS bone marrow compared to that of age-matched healthy controls (p=0.005); 3) multi-lineage differentiation assay revealed impaired osteogenic differentiation potential, but enhanced adipogenic differentiation potential of MDS-RS MSCs; 4) FACS analysis showed increased frequency of the adhesion receptor integrin α4 (ITGA4) in the CD44- MSCs from MDS-RS bone marrow (p=0.013); 5) Correspondingly, RNA-sequencing of the freshly isolated bone marrow MSCs and endothelial cells revealed altered gene expression profile of these cells in MDS-RS patients. Among those, ITGA4, ITGA11, ITGAE and ITGB1 are upregulated in the MDS-RS MSCs, indicating potential abnormal adhesive interaction of the MSCs with hematopoietic stem cells in the patients. In addition, the cell cycling gene KI67 is upregulated whereas cell cycle negative regulators, like CDKN1A and CDKN1C are downregulated in the MDS-RS MSCs, which is consistent with their increased CFU-F activity. Interestingly, we detected abnormal expression of hematopoietic growth factors such as downregulation of ANGPTL4 in the MDS-RS MSCs and upregulation of ANGPT1 in the MDS-RS endothelial cells. The functional relationship between the stromal cell alterations and the abnormal hematopoiesis as well as the underlying molecular mechanisms are currently under investigation.
Taken together, our data provide new evidence for phenotypic, functional and molecular alterations of bone marrow mesenchymal cells in MDS-RS patients. The molecular pathways mediating bone marrow niche alteration could be potential therapeutic targets for new treatment of MDS-RS.

Disclosures
No relevant conflicts of interest to declare.