bluuebunny/crossref_metadata_2025_split huggingface dataset:
12-HETrE, An Endogenous Inhibitor of Platelet Activation,
Authors: Yeung Jennifer , Ikei Kenneth , Vesci Joanne , Arnouk Alex , Holman Theodore R , Holinstat Michael 
Published: October 2019

Abstract
Abstract 3254
Platelet activation plays a pivotal role in thrombosis and hemostasis. Understanding the signaling events mediating this process is essential in preventing unwanted clot formation, which can lead to the development of heart attack, stroke, and venous thromboembolism. Fatty acids may play an important role in determining the level of platelet reactivity, however the mechanism(s) by which this occurs are not entirely clear. 12-lipoxygenase (12-LOX) has been shown to oxidize the fatty acid, arachidonic acid (AA), in order to produce the bioactive eicosanoid 12-HETE, which has recently been shown to play a role in tissue factor activation and subsequent thrombin generation in the platelet (Thomas et al., 2010, J Biol Chem; 285:6891–903). 12-HETE has also been shown to signal to cells, in part, through the G protein-coupled receptor GPR31 (Guo et al, 2011, J Biol Chem; epub). While research on 12-LOX-mediated eicosanoid regulation of platelets has primarily focused on 12-HETE, the potential for regulation of platelets by eicosanoids derived from other fatty acids has been overlooked. As the fatty acid content on the platelet membrane is extremely dynamic and fatty acid supplementation is correlated with a reduced risk for cardiovascular disease, we hypothesized that 12-LOX oxidation of another fatty acid may play a direct role in regulating platelet function. To test this hypothesis, the Ω-6 fatty acid dihomo-γ-linolenic acid (DGLA) as well as its eicosanoid derived from 12-LOX oxidation, 12-hydroxyeicosatrienoic acid (12-HETrE), were applied to washed platelets followed by stimulation with thrombin, PAR1-AP, or PAR4-AP. Platelets treated with either DGLA or 12-HETrE showed significant attenuation in platelet aggregation following stimulation with thrombin or PAR-AP. A number of biochemical intermediates were also tested in the presence of 12-HETrE or DGLA including Rap1 activation, αIIbβ3 integrin activation, α-granule secretion, and dense granule secretion. All endpoints tested were attenuated in the presence of DGLA or 12-HETrE relative to control. To confirm the regulation was unique to DGLA and its metabolite, the same endpoints were measured in the presence of AA or 12-HETE. Pre-treatment with either AA or 12-HETE did not attenuate any of the agonist-mediated platelet activation endpoints. Thus, our data supports a unique role for 12-HETE and 12-HETrE and that fatty acid regulation of platelet function may be highly dependent on the lipid content of the platelet. Shifting the ratios of fatty acids in the platelet through dietary supplementation or pharmacological intervention may be sufficient to induce a cardio-protective state, in part, through increasing 12-HETrE formation and subsequently inhibiting platelet activation.

Disclosures:
No relevant conflicts of interest to declare.
A Genome Wide Association Study Identifies Novel Susceptibility Genes for Pediatric Venous Thrombosis,
Authors: Arning Astrid , Hiersche Milan , Bidlingmaier Christoph , Stoll Monika , Nowak-Gottl Ulrike 
Published: October 2019

Abstract
Abstract 3336

Background:
Genome wide association studies (GWAS) are the current method of choice to dissect the genetic basis of common complex diseases. Up-to-date, studies in families with a known first onset of symptomatic arterial or venous thrombosis (VT) in early childhood are lacking. Methods: Here, we performed a GWAS in a large family-based study sample comprising 241 nuclear families with pediatric VT using the Illumina 660W Infinium SNP array. The average genotype call rate was >99.5%, and the genomic inflation factor was a= 1.012. Single point and haplotype association was assessed using the Transmission Disequilibrium Test (TDT) as implemented in PLINK and FBAT, respectively, and corrected for multiple testing using permutation testing. In addition, associations were corrected for age, gender, and in a subsequent analysis for Factor VLeiden. Results and Conclusion: Four SNPs exceeded the threshold for genome wide significance in this dataset as determined by permutation testing using 100.000 bootstrap permutations (p<10−5), and are likely true associations. Among these, 2 SNPs reside in a region on chromosome 6q13 comprising the gene for beta-1,3-glucoronyltransferase 2 (B3GAT2), a member of the human natural killer 1 (HNK1) carbohydrate pathway are associated with pediatric VT with p-values for rs1304029 (p=3.42×10−6) and rs2748331 (p=6.92×10−6). The corresponding haplotype association test resulted in a p=5.37×10−6 for the GA-haplotype, further underlining the robustness of the association. The SNPs rs636434 on chromosome 6q12 and rs1565242 on chromosome 15 both reside within hypothetical genes and are associated with VT with a p=2.70×10−6 and p=8.24×10−6, respectively. Additional SNPs exceeding a p<10−5 are included in subsequent analyses looking at gene networks and replication in independent study samples including our second GWAS on pediatric thromboembolic stroke (TS). Future studies using larger study samples are warranted to validate these findings and to characterize the genetic architecture underlying VT and TS in children.

Disclosures:
No relevant conflicts of interest to declare.
Non-Steroidal Anti-Inflammatory Drugs and Imatinib; Drug Interactions That May Impact Efficacy,
Authors: Wang Jueqiong , Hughes Timothy P , Kok Chung Hoow , Saunders Verity A , Frede Amity , GrootObbink Kelvin , Osborn Michael Philip , Somogyi Andrew A , D'Andrea Richard J , White Deborah L. 
Published: October 2019

Abstract
Abstract 3501

Aim:
The human organic cation transporter-1 (OCT-1) is the primary active influx protein for imatinib (IM) into target BCR-ABL-positive cells. The functional activity of the OCT-1 protein (OCT-1 activity; OA) is predictive of molecular response and event-free survival in de-novo chronic phase chronic myeloid leukemia (CP-CML) patients. Due to the common use of non-steroidal anti-inflammatory drugs (NSAIDs) among CML patients on IM to treat musculoskeletal pain, it is of particular interest to investigate the impact of NSAIDs on OCT-1 mediated IM uptake.

Methods:
12 NSAIDs were firstly examined for their impact on OA using the intracellular uptake and retention (IUR) assay and [14-C]-IM. The degree of in vitro kinase inhibition achieved was assessed using p-Crkl in the IC50imatinib assay. Viable cell numbers were determined by trypan blue exclusion method after 72 hours incubation with the co-administration of NSAIDs and IM. NSAIDs concentration was adjusted to median blood levels on standard doses.

Results:
The majority of NSAIDs (10 of 12) selected did not induce any significant change in OA in CML cell lines. Compared to control cells, increased OA were observed when K562 or KU812 cells were treated with diclofenac (10μM), and this resulted in a concomitant decrease in the IC50imatinib. In keeping with this, after 72 hours co-incubation with IM, diclofenac resulted in a significantly lower viable cell number compared with IM alone. In contrast, ibuprofen (130μM) led to a significant decrease in OA, resulting in an increased IC50imatinib and thus reduced the inhibition of IM on cell growth number (Table 1).
The effects of diclofenac and ibuprofen on IM efficacy were then further investigated using primary samples. In keeping with the results observed in cell lines, diclofenac resulted in significant increases in OA, which translated to a significant decrease in IC50imatinib. Ibuprofen induced significant decreases in OA in all patients and healthy donors (Table 1).

Conclusion:
These finding provide strong evidence for interactions between some NSAIDs and IM which may substantially impact the response of CML patients to IM. While drugs such as diclofenac are likely to have a positive influence on IM efficacy, the findings observed with ibuprofen suggest caution when administrating NSAIDs to CML patients on IM treatment.

Disclosures:
Hughes: Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; ARIAD: Honoraria, Membership on an entity's Board of Directors or advisory committees. Osborn:Novartis: Research Funding. White:Novartis Pharmaceuticals: Research Funding; BMS: Research Funding.
Chronic Lymphocytic Leukemia Associated with Autoimmune Hemolytic Anemia Is Characterized by a Distinctive miRNA Signature,
Authors: Ferrer Gerardo , Navarro Alfons , Hodgson Kate E , Colomer Dolors , Aymerich Marta , Baumann Tycho , Pereira Arturo , Monzó Mariano , Moreno Carol , Montserrat Emili 
Published: October 2019

Abstract
Abstract 3897
Chronic lymphocytic leukemia (CLL) is frequently associated with autoimmune hemolytic anemia (AIHA). However, the mechanisms governing the association between CLL and AIHA are poorly understood. MicroRNAs (miRNAs) are small 18–22 nucleotide long RNA molecules that regulate gene expression and play a key role in several biological processes. Importantly, deregulated miRNA expression has been implicated both in CLL and autoimmunity. This led us to speculate that patients with CLL who develop AIHA might have a different miRNA expression pattern as compared to those in whom this complication is not observed. We report here the first results of this study. We have evaluated the miRNA expression in purified CLL cells (CD19+, CD5+) from 14 patients who developed AIHA over the course of their disease and 19 sex-, age-, and clinical stage-matched controls who, after a comparable follow up time, did not develop this complication. The expression of 377 mature miRNAs was analyzed using TaqMan Human MicroRNA Arrays A v2.0 (Applied Biosystems) in an ABI 7900 HT sequence detection system. miRNA expression data was analyzed by the 2–ΔΔCt method, using RNU48 as endogenous control. Statistical analyses were performed with TiGR MultiExperiment Viewer, BRB-ArrayTools and R software. The unsupervised hierarchical cluster analysis identified two groups (CLL AIHA+ and CLL AIHA-). The supervised analysis revealed 7 miRNAs that were down-regulated in CLL AIHA+ patients compared to CLL AIHA- patients: miR-19a, miR-20a, miR-29c, miR-146b-5p, miR-324-3p, miR-340, miR-660. Interestingly, miR-146, which has previously been related to autoimmunity, regulates IL-1 receptor-associated kinase 1 and TNF receptor-associated factor 6, two key adapter molecules downstream of Toll-like and cytokine receptors. The prediction analysis of microarrays (PAM) was used to determine a set of miRNAs able to classify the samples into CLL AIHA+ and CLL AIHA-. We obtained a 122-miRNA model that classified CLL AIHA+ patients with a sensitivity of 69.4% and specificity of 68.4%. This model correctly classified 67% of the analyzed samples. In summary, CLL AIHA+ samples were characterized by a distinctive signature of 7 down-regulated miRNAs, one of which (miR-146) has previously been related to autoimmunity. Moreover, we identified 122 miRNAs that are able to predict AIHA in CLL patients. This is the first study establishing a relationship with CLL associated with AIHA and a distinctive miRNA signature, which should be useful for further studies aimed at unfolding the biologic complexity of AIHA in CLL.

Disclosures:
No relevant conflicts of interest to declare.
Association of Disparities in Known Minor Histocompatibility Antigens with Relapse-Free Survival and Graft-Versus-Host-Disease Upon Allogeneic Stem Cell Transplantation,
Authors: Hobo Willemijn , Broen Kelly , van der Velden Walter J.F.M. , Greupink-Draaisma Annelies , Adisty Niken , Wouters Yannick , Kester Michel G.D. , Fredrix Hanny , Jansen Joop H. , van der Reijden Bert A. , Falkenburg J.H. Frederik , de Witte Theo , Preijers Frank W.M.B. , Schattenberg Anton , Feuth Ton , Blijlevens Nicole N.A. , Schaap Nicolaas , Dolstra Harry 
Published: October 2019

Abstract
Abstract 4136

Purpose:
Allogeneic stem cell transplantation (allo-SCT) can induce remission in patients with hematological malignancies due to graft-versus-tumor (GVT) responses. This, however, is often accompanied by graft-versus-host disease (GVHD). Both the GVT effect and GVHD are mediated by minor histocompatibility antigen (MiHA)-specific T cells recognizing peptide products from polymorphic genes that differ between recipient and donor. Here, we evaluated whether MiHA mismatches are associated with clinical outcome after partial T cell depleted allo-SCT.

Patients and Methods:
We retrospectively analyzed the impact of MiHA mismatches in a cohort of 327 patients who received a partially T cell-depleted allo-SCT because of a hematological malignancy. MiHA allele genotyping was performed by fluorescence-based competitive allele-specific PCR. Subsequently, a multivariable statistical analysis of immunogenic MiHA disparity rates and association with clinical outcome was performed. In addition, development of MiHA-specific T cell responses was assessed by dual-color tetramer staining.

Results:
Statistical analysis revealed that an autosomal MiHA disparity on DNA level associates with increased relapse-free survival in sibling transplants, especially in patients transplanted for multiple myeloma. In addition, mismatches for the ubiquitous Y chromosome-encoded MiHA resulted in more acute GVHD (grade 3–4), while other MiHA mismatches, either ubiquitous or restricted to hematopoietic cells, were not associated with GVHD. Finally, we demonstrated considerable differences between MiHA in the capability to induce in vivo T cell responses post-transplantation.

Conclusion:
These data support that autosomal MiHA contribute to the induction of GVT immunity providing a rationale for MiHA-based post-transplantation immunotherapy to prevent and treat persistent and recurrent cancer following allo-SCT.

Disclosures:
No relevant conflicts of interest to declare.
Favorable Responses to Novel Agents for Multiple Myeloma in African American Patients,
Authors: Saraf Santosh , Patel Pritesh R. , Ozer Howard , Peace David , Nimmagadda Sangeetha , Quigley John G. , Rondelli Damiano 
Published: October 2019

Abstract
Abstract 4213
Multiple myeloma (MM) accounts for 10% of hematologic malignancies in the U.S. African Americans (AA) have twice the risk of developing multiple myeloma and, in previous studies, a higher mortality rate when compared to non-African Americans (non-AA). In recent years, clinical outcomes for patients with MM have improved as a result of new agents, such as immunomodulatory drugs (IMiDs) and bortezomib. However, the therapeutic impact of these new therapies in AA vs. non-AA patients has not been evaluated. In this study, 53 consecutive patients (23 AA, 30 non-AA) with newly diagnosed MM were retrospectively analyzed after induction treatment with thalidomide/dexamethasone (n=22), lenalidomide/dexamethasone (n=8), bortezomib/dexamethasone (n=5), bortezomib/thalidomide/dexamethasone (n=3), lenalidomide/bortezomib/dexamethasone (n=12), or an IMiD plus melphalan (n=3). AA and non-AA patients were comparable for age, immunoglobulin isotype, MM stage of disease, serum β2microglobulin and albumin levels, and cytogenetic abnormalities including del13 (27% vs. 38%, respectively, p=NS). When measured according to international uniform criteria, the response rate to induction therapy was not statistically different between AA and non-AA patients: complete remission (CR, 22% vs. 21.4%), very good partial remission (VGPR, 26% vs. 21.4%), or partial remission (PR, 43.4% vs. 32.1%). However, the rate of stable/progressive disease following induction therapy was significantly higher in non-AA patients (p = 0.03). Of 53 patients, 34 received high-dose chemotherapy followed by autologous stem cell transplant (ASCT). In this group, the CR + VGPR rate following ASCT was significantly higher in AA compared with non-AA patients (59% vs. 35%, p=0.0007). At a median follow up of 47 months, the relapse rate was 59% in AA and 46% in non-AA patients (p=NS) and the median time to progression (TTP) was 9.1 months in both groups. Five patients (9%) died. All deaths occurred in the non-AA cohort and in 4/5 cases death was due to disease progression. In conclusion, our findings demonstrate that despite their increased disease risk, AA patients with MM have a favorable outcome that is equivalent to that of non-AA patients when treated with IMiDs and/or bortezomib.

Disclosures:
No relevant conflicts of interest to declare.
A Retrospective Evaluation of the Effects of Severe Aplastic Anemia Treatment in Children with Horse and Rabbit ATG.Polish Pediatric Hematology Group
Authors: Pawelec Katarzyna , Matysiak Michal , Salamonowicz Malgorzata , Kowalczyk Jerzy , Balwierz Walentyna , Zaleska-Czepko Ewa , Chybicka Alicja , Szmyd Krzysztof , Szczepanski Tomasz , Bubala Halina , Wysocki Mariusz , Kurylak Andrzej , Wachowiak Jacek , Szprecht Dawid , Mlynarski Wojciech , Bulas Monika , Krawczuk-Rybak Maryna , Leszczynska Elzbieta , Urasinski Tomasz , Peregud-Pogorzelski Jaroslaw , Balcerska Anna , Kaczorowska Barbara 
Published: October 2019

Abstract
Abstract 4375
Antithymocyte horse globulin (hATG) or rabbit (rATG) is used to treat children with severe aplastic anemia (SAA) in the absence of a compatible bone marrow family donor’s. We presenting (viewing) retrospective evaluation of the effects of SAA therapy using hATG and rATG.
49 children (20 girls and 29 boys, aged 1.2–17) were treated between 1993 and 2000 - hATG (15mg/kg/day for 5 days with cyclosporine 5mg/kg/day for 12–24 mths). On day 180 day of treatment there was a complete response (CR) in 22 of 49 children (44.5%) and a partial response (PR) in 12 children (24.4%). 34 of 49 children (69,4%) responded to treatment. There was no remission (NR) in 15 of 49 children (30.6%). In the whole group there were 7 deaths during the first 6 months. There were no relapses within 5 years of observation. The 5-year probability of survival in this group was 84%.
55 children (22 girls and 33 boys, aged 3,5–17,7 years) were treated in the years 1996–2009 using rATG (3.75 mg/kg/day for 5 days and cyclosporine 5mg/kg/day for 12–24 mths). On day 180 of treatment 5 patients (9%) achieved a CR, 23 children (41.8%) had a PR. 28 of 55 children (50.9%) responded to treatment. NR was found in 27 children (49%).In the rATG group 15 deaths (27%) were recorded, 4 patients (7%) had relapses. In one child paroxysmal nocturnal hemoglobinuria was noted.The 5-year survival in the rATG.group was 63.54%.
Our results show the greater effectiveness of treatment with horse globulin than rabbit globulin in terms of hematological response and overall survival.

Disclosures:
No relevant conflicts of interest to declare.
Unexpected 5 Year Survival Benefit in Patients Given Oral Cryotherapy During Conditioning for Stem Cell Transplantation. A Prospective Randomized Study
Authors: Svanberg Anncarin , Öhrn Kerstin , Birgegard Gunnar 
Published: October 2019

Abstract
Abstract 4559
Patients receiving high dose chemotherapy (HDC) in connection with bone marrow transplantation (BMT) often are afflicted with severe oral mucositis (OM). OM may affect 75–99% of patients. Oral cryotherapy has been shown to alleviate symptoms of mucositis alleviating oral pain and inability to maintain nutritional status. In a randomised controlled trial we have shown that patients receiving oral cryotherapy had less mucositis, less use of i.v. opioides and fewer doses of total parenteral nutrition (TPN) than a control group receiving routine oral care. Adult patients scheduled for bone marrow transplantation were randomly assigned to experimental (EXP) or control (CTR) group. A stratified randomisation was used with regard to type of transplantation (autologous vs allogeneic/unrelated donor (URD)). Randomisation was performed between January 2002 and August 2004. The final sample consisted of 78 patients, (31 autologous BMT and 8 allogeneic/URD BMT), and 39 constituted the CTR group and received standard treatment (31 autologous BMT and allogeneic/URD BMT). Concern has been raised for a possible protection of tumor cells by cryotherapy which could increase the risk of relapse and reduce survival. Thee aim of the present study was to investigate any difference in survival and relapse rates 5 years post-BMT for the two treatment groups from the randomised study.
After 5 years, 25/39 (64%) of the cryotherapy patients were alive compare to 15/39 (38%) of the control group (odds ratio 0,35, 95 % CI 0,14–0,88, p = 0,025)(Figure 1). No significant difference could be found with regard to the relapse rate between the groups. Most of the deaths were due to relapse.
The study offers no support for the speculation about tumor protection from cryotherapy during high dose chemotherapy conditioning for stem cell transplantation. These data indicate that oral cryotherapy is a safe prophylactic treatment for mucositis during chemotherapy.
Unexpectedly, the cryotherapy group showed a significantly better 5-year survival. Further analyses are needed to explore the difference in survival rate.

Disclosures:
No relevant conflicts of interest to declare.
DNMT3A Mutations Are Connected with Lower DNA Methylation Levels and Numbers of Concurrently Hypermethylated Genes in AML Patients
Authors: Hajkova Hana , Markova Jana , Haskovec Cedrik , Cermak Jaroslav , Petrbokova Radka , Michalova Kyra , Schwarz Jiri 
Published: October 2019

Abstract
Abstract 4629
Recently, mutations in DNA methyltransferase 3A (DNMT3A) have been found in patients with acute myeloid leukemia (AML) and have been confirmed to be connected with adverse clinical outcome. As DNMT3A plays direct role in the process of DNA methylation by adding methyl group to the cytosine residue of CpG dinucleotides, the question is obvious: What is the impact of DNMT3A mutations on DNA methylation levels?
We examined 79 AML patients at diagnosis for the presence of aberrant DNA methylation of 12 tumor suppressor genes (TSG) (CDKN2B, CALCA, CDH1, ESR1, SOCS1, MYOD1, DAPK1, TIMP3, ICAM1, TERT, CTNNA1, EGR1) by methylation specific real-time PCR (MethyLight) and for mutations in the gene DNMT3A by direct sequencing. Next we studied methylation status of 24 HOX genes from all four clusters A-D using methylation-restriction endonucleases followed by RQ-PCR arrays in 10 AML samples compared to 4 healthy donor samples.
Sequencing of cDNA between amino acids 300 and 930 revealed that 32 of 79 AML patients had DNMT3A mutation. The reason for higher DNMT3A mutation incidence in our patients’ cohort consists in preferential selection of AML patients with a higher percentage of probability of DNMT3A mutation (it means normal karyotype and mutations in NPM1, FLT3 and/or IDH1/2 genes). MethyLight assessment of 12 TSG showed subsequent frequencies of hypermethylation: CDKN2B (47%), CALCA (43%), CDH1 (22%), SOCS1 (24%), MYOD1 (18%), ESR1 (14%). The remaining 6 genes were weakly methylated in less than 10 % AML patients at diagnosis and were therefore excluded from further analysis. DNA methylation arrays revealed a set of differentially methylated HOX genes (n=12): 11 HOX genes were hypermethylated (HOXA4, HOXA6, HOXB13, HOXB3, HOXB4, HOXB7, HOXB8, HOXC8, HOXD10, HOXD11, HOXD3) and HOXA5 was hypomethylated compared to healthy donor samples. Comparing overall cumulative DNA methylation levels and numbers of simultaneously hypermethylated genes to mutational status of DNMT3A gene, we did observe lower levels of DNA methylation (P<0.0001) as well as displaying lower numbers of concurrently hypermethylated genes (P<0.0001) in patients with DNMT3a mutations. We observed the same trend also in DNA methylation levels of HOX genes when compared mutant (n=4) versus wild-type (n=6) DNMT3A patients.
These results clearly show that numbers of simultaneously hypermethylated genes and DNA methylation levels of chosen tumor suppressor genes (TSG) as well as HOX genes differs between AML patients with wild type and mutant DNMT3A.
This study is part of the COST Action BM0801 (EuGESMA) and is supported by NS10632-3/2009, OC10042 and IHBT00023736.

Disclosures:
No relevant conflicts of interest to declare.