bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow Cytometric Analysis of Platelet Function in Patients on Antiplatelet Therapy and Suspected Thrombocytopathy
Authors: Huskens Dana , Roest Mark , Florin Lisa , De Laat Bas , Devreese Katrien 
Published: October 2019

Abstract
Background: Light transmission aggregometry (LTA) is still the most used test for the identification and diagnosis of platelet function defects (PFD). Disadvantages of LTA include its laborious nature, the large volumes of blood required and the relative insensitivity to small changes in platelet function. We investigated if the flow cytometry-based whole blood platelet activation test (WB-PACT) correlates with LTA or could be used as a complimentary test.
Methods: WB-PACT quantifies αIIbβ3 activation and P-selectin expression in response to 3 agonists and VWF binding to platelets in response to ristocetin. In total, 212 patients (51 on dual antiplatelet therapy (acetylsalicylic acid and ADP receptor antagonist), 161 suspected for bleeding diathesis) were tested. For WB-PACT parameters, the 2.5th, 10th and 25th percentiles were determined in 56 healthy donors to score the patient samples. For LTA, maximal aggregation, disaggregation and prolongation of the lagtime in response to 4 agonists and ristocetin was scored. Patients with at least one parameter lower than the 10th percentile measured with WB-PACT in healthy donors or at least one value deviating from the normal range measured with LTA were diagnosed with PFD. A bleeding score (BS) was calculated with the ISTH/SSC bleeding assessment tool (Rodeghiero et al., JTH, 2010).
Results: A moderate correlation between LTA versus WB-PACT was found with a R of 0.60 (Figure 1). In total, 95 patients were diagnosed with PFD by both tests (κ=0.40, Table 1) and BS were recorded for 28/95 patients. Of these 28 patients, 25% had an elevated BS (adapted according to gender and age: >5 in women, >3 in men and >2 in children) and 36% had a BS>3. BS were recorded for 30/38 and 22/24 patients who were diagnosed with PFD according to WB-PACT only and LTA only, respectively. Interestingly, 27% of patients with PFD according to WB-PACT had an elevated BS and 40% had a BS>3. For LTA, only 13% of patients with PFD had an elevated BS and 13% had a BS>3.
Conclusions: Flow cytometric analysis of platelet function by WB-PACT gives additional value to LTA to determine PFD.

Disclosures
No relevant conflicts of interest to declare.
How KEAP1 Deletion in Single Cell Mitigates Sickle Cell Pathology
Authors: Lukwete Nadine Keleku , Suzuki Mikiko , Panda Harit , Otsuki Akihito , Katsuoka Fumiki , Saitou Ritsumi , Saigusa Daisuke , Uruno Akira , Yamamoto Masayuki 
Published: October 2019

Abstract
Sickle cell disease (SCD) is one of the most frequent blood disorders caused by a point mutation of β-globin gene. In the pathology of SCD, we often encounter contributions of vascular and cellular components. The activation of vascular endothelial cells promotes adhesion of inflammatory cells and vaso-occlusion responsible for low blood flow, hypoxemia and organ damages. Hemolysis triggers activation and recruitment of myeloid cells to the inflammation site, further sustaining the inflammatory process. Recent progress has created the need to examine targeted induction of the KEAP1-NRF2 system as a potential therapeutic tool for the SCD. We have demonstrated that genetic as well as pharmacological induction of NRF2 in SCD prevent organ damages and suppress inflammation, although it still remains unclear how the NRF2 induction modifies the pathology or which type of cells contribute to the NRF2-mediated improvement of SCD. To address these questions, we conducted targeted deletion of Keap1 gene and induced expression of Nrf2 gene in endothelial cells and macrophages/monocytes using Tie1-Cre and LysM-Cre system, respectively. We found that SCD::Keap1F/F::Tie1-Cre mice (NRF2-increase in endothelial cells) showed low hematocrit and low platelet counts. While severe infiltration of monocytes, lymphocytes and macrophages was observed in the lungs of SCD mice, such phenotype was diminished in the SCD::Keap1F/F::Tie1-Cre mice. In the compound mutant mice, livers showed fewer necrotic zones and less to none accumulation of free iron. On the contrary, imaging MS (iMScope) analyses revealed decrease of free heme in livers and spleens in SCD::Keap1F/F::LysM-Cre mice (NRF2-increase in macrophages/monocytes). Furthermore, upregulations of most known Nrf2-dependent pathways and other yet-to-be-defined pathways are found in an RNA-sequencing analysis of the mice. Importantly, NRF2 induction enhanced heme catabolism and decreases the plasma level of free heme and the quantity of accumulated free heme in the SCD::Keap1F/F::LysM-Cre mouse tissues, whereas the change is not obvious in SCD::Keap1F/F::Tie1-Cre mice. On the contrary, vascular leakage of fluids was decreased in the SCD::Keap1F/F::Tie1-Cre mice. Thus, our results demonstrate that cell-lineage-specific activation of Nrf2 does inhibit the inflammation and organ injury with some particularities to each genotype.

Disclosures
No relevant conflicts of interest to declare.
Grb10 Is a Tumor Suppressor in Human Acute Myeloid Leukemia
Authors: Chai-Ho Wanxing , Roos Martina M , Li Michelle , Amara Pang , Zhang Yurun , Fang Tiancheng , Termini Christina , Chute John P 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) is a common and potentially fatal hematologic malignancy. Allogeneic stem cell transplantation is the only curative therapy for most subtypes of AML, but carries a significant risk of transplant-related mortality. The development of novel therapies to eradicate AML remains a substantial area of unmet medical need. Growth factor receptor bound protein 10 (Grb10) is a member of the family of imprinted genes. Our laboratory demonstrated that in normal hematopoietic system, deletion of the maternal allele of Grb10 significantly increased hematopoietic stem cell long-term repopulating capacity (Yan et al. Cell Rep 2016). Grb10 has been shown to bind the intracellular domain of various tyrosine kinase receptors, e.g. KIT, FLT3 and RET, as well as low-density lipoprotein receptor-related protein 6, a negative regulator of the Wnt/β-catenin pathway. Analyzing RNAseq data from the Leucegene Project, we found that Grb10 is expressed on the majority of patient AML samples regardless of leukemia mutation profile. Silencing of Grb10 expression via Grb10 shRNA increased the proliferation and colony forming capacity of human AML cell lines, Kasumi-1, THP-1 and OCI-AML3 in vitro (p<0.0001 and p<0.01). Conversely, overexpression of Grb10 suppressed human AML cell growth (p<0.05). In order to determine the role of Grb10 in regulating AML growth in vivo, we transduced bone marrow lineage negative cells from mice with Grb10 maternal allele deletion (Grb10 m/+) and wild type (Grb10 +/+) mice with HoxA9-Meis1-neo-MSCV (gift from G. Savageau) and transplanted the progeny into congenic mice. Primary and secondary mice transplanted with Grb10m/+ HoxA9-Meis1 leukemia cells displayed significantly decreased survival compared to mice transplanted with Grb10+/+ HoxA9-Meis1 cells (p<0.001 and p<0.05). Furthermore, leukemia cells with Grb10 maternal allele deletion displayed an increase cell cycle progression and increased leukemia colony forming capacity. RNAseq analysis of Grb10 m/+ leukemia cells from diseased mice revealed significant dysregulation of the canonical Wnt/β catenin signaling pathway compared to Grb10 +/+ mice. RT-qPCR analysis confirmed that Wnt/β-catenin target genes, including MYC, CCND1, and SOX2 were significantly up-regulated in Grb10 knockdown human AML cells lines. Taken together, these data suggest that Grb10 is a powerful tumor suppressor in human AML, and represents a novel mechanistic target for the development of new therapies for human AML.

Disclosures
No relevant conflicts of interest to declare.
Intravesicular Cidofovir in the Treatment of BK Virus-Associated Hemorrhagic Cystitis (BKV-HC) Following Allogeneic Hematopoietic Stem Cell Transplant (HSCT)
Authors: Tooker Graham , Badros Ashraf Z. , Nishioka Jennifer , Riedel David 
Published: October 2019

Abstract
Background: BKV HC is a well-known complication following allo-SCT. Supportive care with bladder irrigation and blood transfusions were the only available treatment. Since our initial report (Bridges B et al. Am J Hematol 2006;81:535), several studies confirmed that intravesicular cidofovir is a potential effective treatment for BKV HC. In this study, we report a large series of consecutive patients who developed BKV HC following allo SCT and received intravesicular cidofovir.
Methods: We conducted a retrospective review of allo SCT patients who developed BKV HC and were prescribed intravesicular cidofovir from 2012 to 2017.
Results: 33 patients were diagnosed with BKV HC. The median age was 50 years (range=23-73), and 18 (55%) were male. Acute myeloid leukemia (n=12, 35%) was the most common diagnosis followed by non-Hodgkin lymphoma (n=7, 21%) and B cell acute lymphoblastic leukemia (n=4, 12%). Conditioning regimens were myeloablative (n=19, 58%) or reduced-intensity (n=14, 42%); 15 (45%) patients received cyclophosphamide, and 22 (67%) received total body irradiation. The median time to onset of HC symptoms following SCT was 37 days (range: 8-178); 17 (52%) patients had acute graft vs. host disease. HC symptom severity ranged from grade 0-4 (median=2). The median BK urine viral load pre-treatment was 100,000,000 IU/ml. Patients received a median of 2 intravesicular treatments (range=1-7) at a dose of 5 mg/kg. Four patients (12%) were also treated concurrently with intravenous cidofovir. 19 (59%) patients demonstrated complete clinical resolution of symptoms, 9 (28%) demonstrated partial response to treatment, and 4 (13%) had no change in symptoms following treatment. These improvements in clinical status were independent of viral load, though most had reductions in the viral load. The median time to symptom resolution was 17 days (range=7-53; n=28). 82% of patients had no recurrent symptoms of HC. The main side effect of intra-vesicular instillation was increased discomfort and bladder spasms; severe in 3 patients (9%). No patient had impaired renal function directly attributable to intra-vesicular cidofovir. At 12 months after BKV HC diagnosis, 26 (79%) patients were alive.
Conclusions: To our knowledge this is the largest study of intravesicular cidofovir for BKV HC reported to date; 77% of patients with BVK HC achieved clinical improvement of symptoms with minimal side effects. Clinical trials of intravesicular cidofovir could provide further evidence for adding intravesicular cidofovir as a standard tool for the treatment of BKV HC.

Disclosures
Badros: Celgene: Consultancy, Research Funding; Karyopharm: Research Funding; GSK: Research Funding.
Socioeconomic Disparities Affect Survival of Classical Hodgkin Lymphoma in a Statewide Cancer Registry Analysis
Authors: Myint Zin W. , Shrestha Runa , Chen Quan , Huang Bin , Goldberg Amit , Ramlal Reshma , Monohan Gregory , Fleischman Roger A. , Herzig Roger H. , Hildebrandt Gerhard Carl , Saeed Hayder 
Published: October 2019

Abstract
Introduction:
Low socioeconomic status (SES) has been shown to shorten survival in classical Hodgkin lymphoma (cHL). We sought to determine if differences in outcome of cHL varied according to SES and geographic setting in a population-based analysis.
Methods:
All adult patients diagnosed in 2000-2014 with cHL were collected through Kentucky Cancer Registry (KCR). Baseline SES and clinical variables were collected. Pearson Chi-square, log-rank, and cox regression tests were utilized. To minimize selection bias, cases with less than 6 months survival after cancer diagnosis were excluded.
Results:
A total of 1075 cHL patients were included, of which 605 (56.3%) were early stage vs. 405 (37.7%) were advanced stage. Most patients were under age 50 (63.2%) with a male predominance (55.4%). Nodular sclerosis was the most common histology (57.6%). Therapy consisted of chemotherapy alone in 729 (67.8%) patients, while 222 (20.7%) had received both chemotherapy and radiation. 633 (58.8%) lived in metro areas. Metro-area residents were more likely to come from non-Appalachian counties (93.7%), have more advanced education (76.8%) and have private insurance (60.1%) than non-metro-area residents (41.2%) (p<0.0001). There were no differences in age, gender, stage and histology at presentation according to geographic setting. Patients living in metro-area had a better OS by bivariate analysis (p=0.01) (Figure 1). A multivariate analysis identified age <50, early stage, privately insured, non-lymphocyte-depleted histology, and treatment with combined chemo-radiation as independent factors favoring an improved OS.
Conclusion:
Our study shows that despite the absence of differences in presenting clinical variables based on SES or geographic setting, significant discrepancies in outcome are observed. This disparity in outcomes may have been affected by differences in access to or quality of care delivery and may widen with the introduction of more costly novel interventions.

Disclosures
No relevant conflicts of interest to declare.
The Middle-Aged Bone Marrow Microenvironment Drives Hematopoietic Stem Cell Aging through Altered Production of Insulin-like Growth Factor-1 (IGF1)
Authors: Young Kira , Loberg Matthew , Eudy Elizabeth , Bell Rebecca , Trowbridge Jennifer J. 
Published: October 2019

Abstract
Globally, lifespan continues to increase. A growing challenge is how to maintain a healthy aging population. In hematopoiesis, healthspan is limited by a decline in regenerative capacity and overproduction of myeloid cell types at the expense of lymphoid cell types. These alterations contribute to decreased immune function, poor response to vaccination and increased susceptibility to myeloid neoplasms. Methods to extend hematopoietic healthspan are unclear, in part due to our relatively poor understanding of the factors controlling aging.
To address this gap in knowledge, we utilized mouse models to examine at what age did impaired regenerative capacity and myeloid-biased hematopoiesis begin to occur. We find that these phenotypes become significant by 9-12 months of age, representing middle age in mice. Through reciprocal transplantation studies of purified long-term hematopoietic stem cells (LT-HSCs) between young and middle-aged mice, we find that the middle-aged bone marrow (BM) microenvironment is sufficient to cause impaired regenerative capacity and myeloid-biased hematopoiesis from young LT-HSCs. These studies clearly implicate components of the BM microenvironment at middle age as causing hematopoietic aging.
To uncover the factors altered in the middle-aged BM microenvironment that cause hematopoietic stem and progenitor cell aging, we utilized single cell RNA-Seq and ELISA analysis. Together, these identified a reduction in Insulin-Like Growth Factor-1 (IGF1) in the BM microenvironment as the top candidate regulator of hematopoietic stem and progenitor cell aging. Expression of the receptor for IGF1, IGF1R, was dynamically altered on hematopoietic stem and progenitor cell populations with aging, suggesting that these respond to altered local levels of IGF1. In vitro colony-forming unit studies demonstrate that stimulating hematopoietic stem and progenitor cells isolated from middle-aged mice with exogenous IGF-1 ligand resulted in a dose-dependent suppression of myeloid cell overproduction, rescuing the aging phenotype. Finally, direct restoration of IGF1 levels in the BM microenvironment by short-term intrafemoral injection, at middle age, suppressed myeloid cell overproduction and restored B cell production in the BM and peripheral blood.
In summary, this study reveals a novel role for IGF1 in hematopoietic aging. We show that decline in local levels of IGF1 in the BM microenvironment at middle age cause hematopoietic aging. In addition, we demonstrate that short-term restoration of IGF1 at middle age can rescue hematopoietic aging. IGF1 thus represents an attractive candidate molecule to restore hematopoietic regenerative capacity and balanced lineage cell production to extend hematopoietic healthspan into older age.

Disclosures
No relevant conflicts of interest to declare.
Extravesicular Mir-7977 Released from Leukemic Cells Inhibites the PCBP1-Mediated mRNA Stabilization and Hippo-YAP Signaling Pathway in Bone Marrow Mesenchymal Stromal Cells
Authors: Yoshida Masahiro , Iyama Satoshi , Horiguchi Hiroto , Goto Akari , Kikuchi Shohei , Murase Kazuyuki , Takada Kohichi , Miyanishi Koji , Ikeda Hiroshi , Maeda Masahiro , Kato Junji , Kobune Masayoshi 
Published: October 2019

Abstract
[Background] We and others have revealed that various abnormalities of the bone marrow (BM) environment such as aberrant cytokine expression and impaired microRNA (miRNA) biogenesis are observed in patients with myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). However, underlying mechanisms to induce BM stromal dysfunction have not yet been clarified. Recently, we have found that abundant extracellular vesicles including exosomes, were detected in the BM interstitial fluid. Furthermore, we have shown that exosomal miR-7977 was highly released from AML cells (GSE64029) and was taken up by BM mesenchymal stem cells (MSCs). Based on these findings, we hypothesized that exosomal miR-7977 could be involved in the alteration of BM stromal function of MDS/AML.
[Materials and Methods] To gain an insight into the function of exosomal miR-7977 in BM, we firstly performed transduction of a miR-7977 mimic into BM MSCs and compared transcriptomes between control-transduced and miR-7977-transduced MSCs (GSE108186). Subsequently, we selected the differentially expressed genes (DEGs) and conduced gene set enrichment analysis (GSEA).
[Results] Regarding DEGs, we focused on poly(rC) binding protein 1 (PCBP1), since PCBP1 was predicted to be a target gene of miR-7977 by an online database for miRNA target prediction using miRDB. The reductions in PCBP1 mRNA and protein were confirmed in MSCs after miR-7977 transfer by real time PCR. Because PCBP1 is known to be involved in splicing, and the expression levels of several genes, we analyzed the expression of several hematopoietic factors and found that expression level of stem cell factor (SCF), angiopoietin-1 (ANGPT1) and Jagged-1 (JAG1) were remarkably decreased. Regarding the pathway analysis, GSEA showed that the gene sets of Yes-associated protein 1 (YAP1)_up were significantly enriched (p<0.001, q<0.25), suggesting that miR-7977 modulates the Hippo-YAP signaling pathway. Visualization of pathway and network showed that miR-7977 significantly reduced the expression of Hippo core kinase, STK4. The transfer of miR-7977 mimic inactivated the Hippo-YAP signaling pathway as proven by GFP-tagged YAP nuclear trans localization and TEAD reporter assay. The miR-7977-transduced MSCs showed elevated saturation density and enhanced entry into the cell cycle.
[Conclusion] Collectively, these results indicated that miR-7977 is an important factor that inhibits normal hematopoiesis via PCBP1 reduction and up-regulate the growth of functionally-disturbed MSCs via inactivation of Hippo-YAP signaling pathway in MDS/AML.

Disclosures
No relevant conflicts of interest to declare.
Direct Oral Anticoagulants and Mortality in Atrial Fibrillation
Authors: Ellis Martin , Ruslan Sergienko , Ariel Hammerman , Greenberg-Dotan Sari , Battat Erez , Hamburger-Avnery Orly , Greenberg Dan , Ronen Arbel 
Published: October 2019

Abstract
Background
Direct oral anticoagulants (DOACs) are at least as effective as vitamin K antagonists (VKAs) for stroke prevention in patients with non-valvular atrial fibrillation (NVAF). DOACs are associated with less intracranial hemorrhage and are easier to administer and therfore have become standard of care for stroke prevention in NVAF patients. Nevertheless, many eligible patients still receive no anticoagulation. The effect on overall mortality in routine practice of DOAC therapy compared to no anticoagulation in NVAF patients is unknown.
Methods
We identified all newly diagnosed, anticoagulant naive NVAF patients eligible for DOAC therapy from 2011 to 2016 in Clalit Health Services, the largest HMO in Israel. We created a matched cohort, using 1:1 propensity score matching of DOAC -treated versus non-anticoagulant treated NVAF patients. The primary outcome was overall mortality rate in the two cohorts. Secondary outcomes were rates of stroke, major cardiac and bleeding events.
Results
28,195 newly diagnosed CHADS2 ≥2 NVAF patients were identified. Of these 8 298 received a DOAC and 10 603 received no anticaogulation. The remainder received a VKA and were not included in this study. Patients recieving DOAC therapy were younger (77.4 vs 78.0 years), had a higher socioeconomic status (5.6 vs. 5.3), had a female predominance (53.7% vs 52.0%), had a higher BMI (29.6 vs 28.3) and had a greater prevalance of accompanying cardiovascular morbidities namely congestive heart failure (CHF) (29% vs 27%) and stroke (33% vs 30%) (P<0.001 for all variables) . 5,657 patients who received DOAC therapy were matched with 5,657 patients not receiving an anticoagulant using propensity score matching. DOAC therapy was associated with significantly lower risk for death (hazard ratio 0.66, 95% CI 0.60-0.71, P<0.001). Rates of stroke and major cardiac were significantly lower in the DOAC-treated than in the non-anticoagulated patients. DOAC therapy was associated with a significant survival benefit across all patient subgroups.
Conclusions
In this cohort of newly diagnosed NVAF patients treated in routine clinical practice, DOAC therapy was associated with a lower risk for death compared to no oral anticoagulation. Our findings provide further evidence for the importance of DOAC therapy in NVAF patients.

Disclosures
No relevant conflicts of interest to declare.
D-Dimer Levels and Risk of Recurrence Following Provoked Venous Thromboembolism: Findings from the Riete Registry
Authors: Ellis Martin , Mar Martin , Manuel Monreal , Hamburger-Avnery Orly , Bura-Riviere Alessandra , Barillari Giovanni , Mazzolai Lucia , Mahe Isabelle , Marchena Pablo Javier , Verhamme Peter 
Published: October 2019

Abstract
Background. Patients with venous thromboembolism (VTE) secondary to transient risk factors or cancer may develop VTE recurrences after discontinuing anticoagulant therapy. Identifying at-risk patients could help to guide the ideal duration of anticoagulant therapy in these patients.
Methods. We used the RIETE database to assess the prognostic value of d-dimer testing after discontinuing anticoagulation to identify patients at increased risk for recurrences. The proportion of patients with raised d-dimer levels was determined and the hazard ratio (HR) for VTE recurrences compared to those with normal levels was calculated. Univariate and multivariate analyses of factors associated with VTE recurrence were performed.
Results. 3 606 patients were identified in the database in April 2018: 2 590 had VTE after a transient risk factor and 1016 had a cancer. D-dimer levels were measured after discontinuing anticoagulation in 1 732 (67%) patients with transient risk factors and 732 (72%) patients with cancer-associated VTE and these patients formed the cohort in which recurrent VTE rate was calculated. D-dimers and were elevated in 551 (31.8%) of patients with a transient risk factor and were normal in 1181 (68.2%). In the cancer-associated group, d-dimers were elevated in 398 (54.3%) and normal in 334 (45.7%) patients. The adjusted hazard ratio for recurrent VTE was: 2.32 (95%CI: 1.55-3.49) in patients with transient risk factors and 2.23 (95%CI: 1.50-3.39) in those with cancer.
Conclusions. Patients with raised d-dimer levels after discontinuing anticoagulant therapy for provoked or cancer-associated VTE are at increased risk for recurrent VTE and death. Future studies could target these patients for extended anticoagulation.

Disclosures
No relevant conflicts of interest to declare.