bluuebunny/crossref_metadata_2025_split huggingface dataset:
Resveratrol Induces Cell Cycle Arrest and Apoptosis in Malignant NK Cells Via JAK2/STAT3 Pathway Inhibition
Authors: Trung Ly Quoc , Espinoza Luis Jorge , Takami Akiyoshi , Yoshida Akiyo , Nakao Shinji 
Published: October 2019

Abstract
Abstract 1343
Natural killer (NK)-cell malignancies, particularly aggressive NK-cell leukemias/lymphomas, have poor prognoses. Although recent regimens that include L-asparaginase substantially improve outcomes, novel therapeutic approaches are needed to improve clinical responses. Recent reports have shown that the signal transducer and activator of transcription 3 (STAT3) pathway is critical for proliferation and survival of malignant NK cells. Resveratrol is a naturally-occurring polyphenol that has been extensively studied for its anti-inflammatory, cardioprotective, and anti-cancer activities. In this study, we investigated the potential anti-tumor activities of resveratrol against the NK cell lines KHYG-1, NKL, NK-92, and NK-YS. Resveratrol significantly suppressed cell proliferation in a dose- and time-dependent manner in these four cell lines. Flow cytometry analysis with annexin V/propidium iodide staining showed a variable but consistent induction of apoptosis in the four cell lines treated with resveratrol for 48 hours, ranging from 57.1±6.9% of apoptosis in the L-asparaginase resistant cell line KHYG-1 to 53.4±9.7%, 28.8±3.3%, and 51.7±6.7% in NKL, NK-92, and NK-YS cells respectively (Fig. 1a). Notably, the anti-tumor activity of resveratrol against NK cell lines was p53 independent as demonstrated by equal efficacy of resveratrol against NK cell lines pretreated with the p53 inhibitor Pifithrin-α. Immunoblot analysis to study intracellular signaling in resveratrol-treated cells showed suppression of constitutively active STAT3 in all four cell lines 24 hours after treatment. Remarkably, resveratrol inhibited JAK2 phosphorylation, but had no effect on other known upstream mediators of STAT3 activation such as PTEN and Tyk2 (Fig. 1b). Resveratrol induced robust G1 cell cycle arrest and down-regulation of two anti-apoptotic proteins, Mcl-1 and survivin, both of which are downstream effectors of the STAT-3 pathway. Furthermore, resveratrol enhanced the pro-apoptotic and anti-proliferative activities of L-asparaginase against NKL and NK-92 cells by 32% and 126% respectively.
These data indicate that resveratrol possesses a potent anti-tumor effect via inactivation of the STAT3 pathway in malignant NK cells. These mechanistic findings suggest that resveratrol may have therapeutic potential against NK cell malignancies. Our finding that resveratrol is a bonafide JAK2 inhibitor extends its therapeutic potential to other diseases with deregulated JAK2 signaling.

Disclosures:
No relevant conflicts of interest to declare.
AML Exosomes Regulate Stromal Bystander Cells and Provide a Novel RNA Biomarker Platform
Authors: Huan Jianya , Hornick Noah , Lenz Kyle , Goloviznina Natalya A , Skinner Amy M. , Tyner Jeffrey W , Roberts Charles T , Kurre Peter 
Published: October 2019

Abstract
Abstract 1378
Cell membrane-derived vesicles carrying protein and RNA cargo have recently emerged as potent mediators of cell-cell communication. These vesicles traffic from their tissue of origin into the blood stream, providing a serum-based and minimally invasive biomarker platform for cancer-specific RNA and protein signatures. We were interested in evaluating vesicle production by acute myeloid leukemia (AML) cells, with an emphasis on RNA content and cell-cell trafficking. Here we demonstrate for the first time that cell lines and primary cells from AML patients release exosome-sized vesicles (30–100nm) capable of transfer to bystander cells. Analysis of their non-protein content revealed the enrichment of many coding- and non-coding RNAs with relevance to AML biology, including prognostically important transcripts encoding FLT3, NPM-1, IGF-IR, CXCR4 and MMP9. In addition, we demonstrated the presence of a diverse group of transcription factor mRNAs (MYC, MEF2C, GATA1, SHIP1, ID1, E2F1, CEPBA CEPBB) in exosomes. PCR array studies also confirmed the broad representation of cellular miRNAs. Among the 234 miRNAs represented, the levels of several (miR −9, −155 and −223 and LET-7A) were enriched over the levels seen in donor cells (U6 RNA control). When examining cell-cell trafficking of AML exosomes, we noted rapid entry into co-cultured stromal cells, with maximum uptake occurring between 2 and 3 hours. AML-derived mRNA transcripts were transferred to bystander cells without direct cell-cell contact after Transwell co-culture or direct exposure to purified exosome preparations, or via vesicle-rich media. Results show that exosome trafficking induced changes in gene expression (OPN, ANGPT, SCF, SDF-1A) and altered the secretion of cytokines and angiopoietic growth factors by bone marrow stromal cells. Additional proof-of-concept studies provided evidence for the canonical function of transferred RNA. Specifically, we found that transfer of IGF-IR mRNA led to proliferative responses in co-cultured OP9 stromal cells that were abrogated by the IGF-IR specific inhibitor picropodophyllin. Further, exosomes derived from AML cell lines (HL60 and Molm14) altered c-FOS and VEGF expression in OP9 cells as well as improving viability and promoting migration toward SDF-1A by a hematopoietic progenitor cell line (Baf3), respectively. Our observations support the concept that AML exosomes exhibit regulatory potential in the leukemic bone marrow niche via direct cytoplasmic cell-cell transfer of transcripts encoding tyrosine kinase receptors and transcription factor as well as micro- RNAs. We propose that AML exosomes contribute to the extrinsic signaling cues in the microenvironment that promote disease resistance in AML and that they are rich in biomarkers to guide diagnosis and treatment.

Disclosures:
No relevant conflicts of interest to declare.
Oral Realgar-Indigo Naturalis Formula Achieved Comparably High Rate of Remission and Survival to Intravenous Arsenic Trioxide As a Front-Line Treatment in Newly Diagnosed Acute Promyelocytic Leukemia: A Multi-Center Randomized Trial APL07
Authors: Zhu Zhu-Hong , Wu De-Pei , Jin Jie , Li Jian-Yong , Ma Jun , Wang Jiangxiang , Jiang Hao , Chen Sai-Juan , Huang Xiao-Jun 
Published: October 2019

Abstract
Abstract 140

Background
Arsenic trioxide increases the rate of complete remission (CR) and prolongs long-term overall survival in patients with acute promyelocytic leukemia (APL). We demonstrated that an oral arsenic-containing Realgar-Indigo naturalis formula (RIF) could yield clear synergistic effects in an in vivo murine APL model as well as an in vitro APL cell lines. The efficacy and safety of this formula were confirmed by a CR rate of 96.7% and reasonable safety profile in a multicenter Phase II clinical trial in China. To find out whether oral RIF could achieve similar remission and survival with ATO, the Chinese APL Cooperative Group conducted a phase 3 randomized study (APL07) that compared oral RIF with ATO in both the induction and maintenance therapies.

Methods
In this phase 3 randomized trial, 242 newly diagnosed APL patients were randomly assigned to receive oral RIF (n=121, daily 60 mg per kg body weight) or ATO (n=121, daily 0.15 mg per kg body weight) as a part of induction therapy. All patients also received all-trans-retinoic acid (ATRA; daily 25 mg per square meter) as induction therapy, and were between the ages of 15 and 60 years. After achieving CR, all patients were given three courses of consolidation chemotherapy with anthracycline and cytosine arabinoside. The maintenance treatment included eight cycles of sequential use of ATRA and RIF or ATO for two years. The primary endpoint was disease-free survival (DFS). The secondary endpoints were the rate of CR and overall survival (OS) and safety.

Results
At a median follow-up of 28 months, no significant differences were noted between RIF group and ATO group with respect to CR rate (99.1% vs.97.2%, P=0.62), DFS (98.9% vs.97.64%, P=0.6) (Figure 1A) and OS at 3 years (99.1% vs.96.6%, P=0.18) (Figure 1B). The rates of adverse events were similar in the two groups. The rate of treatment-related grade 3/4 liver toxic effects was 9.6% in the RIF group and 12.0% in the ATO group(p=0.67). When on the arsenic-containing treatment, the plasma arsenic content was higher in the ATO group than in the RIF group (56.3 vs. 23.6 μg/L, p=0.0002). The median levels of arsenics concentration of plasm, urine, hair and nail samples after completing therapy 12 months were below the lower limit of the normal control.

Conclusions
Oral Realgar-Indigo naturalis formula yielded a comparable high remission and long-term survival with arsenic trioxide as front-line treatment of newly diagnosed APL.

Disclosures:
No relevant conflicts of interest to declare.
Reduced Intensity Conditioning Allogeneic Stem cell Transplantation (RIC-alloSCT) for Elderly Acute Myeloid Leukaemia (AML) Patients Over 60 Years: A “donor” Versus “no donor” Comparison
Authors: Peterlin Pierre , Raffoux Emmanuel , Chevallier Patrice , Guillaume Thierry , Gastinne Thomas , Le Gouill Steven , Moreau Philippe , Dombret Hervé , Mohty Mohamad , Delaunay Jacques 
Published: October 2019

Abstract
Abstract 2037
Acute Myeloid Leukemia(AML) in elderly patients is associated with a poor outcome even for patients able to receive intensive chemotherapy. Relapse rate remain elevated around 60 to 80% at 3 years especially in case of unfavourable cytogenetics. From more than 10 years, the approach of RIC-alloSCT has permitted to extend the age limit of transplant in this population. However, feasibility remains controversial for patient aged of 60 years or more because of more frequent co-morbidities translating into an higher risk of treatment related mortality (TRM). To address the question of feasibility and results of RIC-alloSCT in AML patients of 60 years or more in a intend to treat manner approach, we have conducted a retrospective bi-centric analysis based on a comparison “donor” versus “no donor” for patient in first complete remission (CR1).

Methods:
96 AML were included, all have received a 3+7 or equivalent induction regimen, between 2001–2009 in two different French centers and have benefited of an HLA typing in CR1. All patients of the “donor” group are considered for a RIC-alloSCT after one or two courses of consolidation.

Results:
The “donor” group is constitute of 30 patients with a median age of 65.5 years and with an HLA identical donor identified. Eighteen 60%) have been finally transplanted with a median period of 188 days from diagnosis to graft. The “no donor” group is constituted by 66 patients with a median age of 68.5. The two groups are similar considering cytogenetics risk and haematological features of the disease.
Median follow up is 48 months for alive patients. No difference of overall survival (OS) or Leukemia free survival (LFS) has been observed between the ”donor” and “no donor group” (respectively 51% vs 44%, p=0.7 and 35%vs 32%, p=0.27). When restricting analysis to the patient who could actually received the RIC allo-SCT, the transplant group (n=18) has a significant better outcome for both OS and LFS as compared to the “non transplant” (n=78) group (69% vs 32% and 52% vs 29% respectively). In the transplant group, no patient died from transplant-related mortality. Acute and extensive chronic graft versus host disease (aGVHD and cGVHD) occurred respectively in 5/18 and in 8/18 patients of the “transplant” group.

Conclusion:
In a intend to treat manner, RIC-alloSCT provides similar outcome as compared to chemotherapy alone in this series of 96 patients. However, both OS and LFS are significantly improved for patients who have actually received RIC-alloSCT. Toxicity appears to be limited in the “transplant” group. Recruiting patients in this study is still ongoing and final results will be presented at the congress.

Disclosures:
No relevant conflicts of interest to declare.
A Novel EGLN1/PHD2 High-Frequency Variant in Tibetans Protects Against Hypoxia-Induced Polycythemia.
Authors: V Felipe R Lorenzo , Huff Chad Daniel , Myllymäki Mikko , Swierczek Sabina , Salama Mohamed E , Semenza Gregg , Gordeuk Victor R. , Jorde Lynn , Koivunen Peppi , Prchal Josef T. 
Published: October 2019

Abstract
Abstract 2079
The hypoxic response, mediated by hypoxia inducible transcription factors (HIFs), is central to the control and development of many essential biological functions, including erythropoiesis. As a high altitude population, Tibetans are genetically adapted to the environmental stress of high altitude, having normal hemoglobin concentration both at sea level and at high altitude. Several haplotypes have undergone positive selection in Tibetans, including variations at or near the EGLN locus, which encodes prolyl 4-hydroxylase (PHD2). PHD2 triggers the degradation of hypoxia-inducible factors (HIFs) that mediate many physiological responses to hypoxia, including erythropoiesis.
We now describe two high-frequency Tibetan variants, PHD2D4E and PHD2C127S, that originated on the same haplotype about ∼6,000 years ago and contribute functionally to the Tibetan high-altitude phenotype. The PHD2D4E, C127S variant exhibits a lower Km value for O2, suggesting that it promotes HIF degradation under hypoxia. Because HIF directly stimulates erythropoiesis, the measured kinetic PHD2D4E, C127S properties were experimentally reproduced in native PHD2D4E, C127S erythroid progenitors that had decreased proliferation under hypoxic conditions, whereas wild-type progenitors have increased proliferation. Our results demonstrate that the Tibetan selected PHD2D4E, C127S variant contributes to protection from polycythemia at high altitude.
We describe the first high-altitude adaptive mutations in Tibetans and demonstrate that they abrogate hypoxia-induced HIF-mediated augmentation of erythropoiesis, explaining Tibetan protection from polycythemia at high altitude.

Disclosures:
No relevant conflicts of interest to declare.
Impact of Factor IXa Content On Function, Safety and Efficacy of Recombinant Factor IX Products.
Authors: Auer Wilfried , Rottensteiner Hanspeter , Schrenk Gerald , Hoellriegl Werner , Schiviz Alexandra , Scheiflinger Friedrich , Schwarz Hans Peter , Turecek Peter L , Muchitsch Eva-Maria 
Published: October 2019

Abstract
Abstract 2233
Baxter has developed a new recombinant factor IX (rFIX) product (BAX 326) for treating patients with hemophilia B. The aim of the presented studies was to evaluate the function, safety and efficacy of BAX 326 compared to a commercially available rFIX regarding differences in activated FIX (FIXa) content.
The FIXa concentration of BAX 326 was about 10 fold less than that of a commercial rFIX product. To address the influence of FIXa on functional FIX in vitro assays, the FIXa concentration of BAX 326 was increased. Samples were analyzed by one-stage clotting assay, non-activated partial thromboplastin time assay and thrombin generation assay. In all three assays, FIXa affected the potency determination for FIX, indicating that rFIX products should preferentially contain a low FIXa content.
The thrombogenic potential of BAX 326 was assessed in rabbits using a modified Wessler Test at 750 IU/kg (10-fold human clinical dose). No thrombogenic potential was observed with BAX 326 (individual scores of 0), whereas the mean score for the commercially available rFIX was 0.5. After increasing the FIXa concentration of BAX 326 to equalize it with the commercially available rFIX, a mean score of 0.42 (individual scores 0 – 0.5) was determined. These data strongly suggest that the differences in preclinical thrombogenicity were caused by the higher FIXa content of the commercially available rFIX, thereby confirming earlier findings [1].
Efficacy of the rFIX products was studied in hemophilia B (FIX ko) mice that received prophylactic treatment with 75 IU/kg of BAX 326 or the commercially available rFIX. Treated animals were analyzed in two primary pharmacodynamic models: a carotid occlusion model and one using thrombelastography. In both models, results for the two groups were similar, demonstrating that despite its lower FIXa content, BAX 326 was as efficacious as a commercially available rFIX at 75 IU/kg (p<0.0076).

Disclosures:
Auer: Baxter Innovations GmbH: Employment. Rottensteiner:Baxter Innovations GmbH: Employment. Schrenk:Baxter Innovations GmbH: Employment. Hoellriegl:Baxter Innovations GmbH: Employment. Schiviz:Baxter Innovations GmbH: Employment. Scheiflinger:Baxter Innovations GmbH: Employment. Schwarz:Baxter Innovations GmbH: Employment. Turecek:Baxter Innovations GmbH: Employment. Muchitsch:Baxter Innovations GmbH: Employment.
Factor V Leiden Mutation Increases the Risk of Venous Thromboembolism in Cancer Patients – Results From the Vienna Cancer and Thrombosis Study (CATS).
Authors: Pabinger Ingrid , Ay Cihan , Dunkler Daniela , Thaler Johannes , Reitter Eva-Maria , Marosi Christine , Zielinski Christoph , Mannhalter Christine 
Published: October 2019

Abstract
Abstract 2253

Background:
Patients with cancer are at an increased risk of venous thromboembolism (VTE). The risk varies markedly in different patient populations and improvement of the prediction of VTE would be of advantage for tailoring thrombosis prophylaxis. Factor V Leiden is the most common genetic risk factor for VTE and the impact of factor V Leiden on cancer-associated thrombosis is not yet fully elucidated.

Objective:
To study the impact of factor V Leiden on the risk of VTE in cancer patients.

Patients and Methods:
Nine-hundred-eighty-two patients with newly diagnosed cancer (n=745) or progression of disease after complete or partial remission (n=237) were included in the cancer and thrombosis study (CATS), a prospective observational single centre cohort study at the Medical University Vienna. Patients were followed for a maximum period of 2 years. Blood samples were collected at inclusion and factor V Leiden was determined by genotyping. The main outcome measure was symptomatic or lethal objectively confirmed VTE. All VTE events were adjudicated independently.

Results:
Of the 982 patients (median age 62 years, interquartile range (IQR) 52–68, 537 men, 445 women) factor V-Leiden was found in 72 (7.3%), 70 had a heterozygous and two a homozygous genotype. Ten of 72 (14%) patients with factor V-Leiden developed VTE, whereas this was the case in 69 of 910 (7.6%) patients without factor V-Leiden. Interestingly, both patients with homozygous factor V Leiden developed VTE. In multivariable analysis that included age, sex, different tumour types, newly diagnosed versus recurrence of disease and the treatment modalities (chemotherapy, radiotherapy and surgery) the hazard ratio (HR) for factor V Leiden was 2.04 (95% confidence interval (CI) 1.04–3.97)). In patients with newly diagnosed tumours the HR for factor V Leiden was 3.7 (95% CI 1.2–12.2) after 30 days. In Kaplan Meier analysis the probability for development of VTE after 6 months was 5.7% in those without and 13% in those with factor V Leiden, after one year the corresponding rates were 7.3% and 15%.

Conclusions:
Factor V Leiden is a genetically determined and thus disease-independent parameter, which is associated with VTE in cancer patients, especially shortly after cancer diagnosis, and could therefore be used for individual risk assignment.

Disclosures:
No relevant conflicts of interest to declare.
Immunogenicity of Blood Group Antigens in Female and Male Transfusion Recipients: Is It Possible to Avoid Immunization to RBC Antigens?
Authors: Szymanski Irma O. 
Published: October 2019

Abstract
Abstract 4377
In men blood group incompatibilities between a donor and blood transfusion recipient may stimulate RBC antibody production, whereas in women both transfusion and pregnancies play a role. Immunogenicities of various RBC antigens correlate with their ability to induce a specific antibody response. These parameters were calculated on data obtained previously in 711 patients found to be immunized in pre-transfusion tests using the method of Giblett (Transfusion 1961), and corrected for the fraction of antibody decline in time by the method of Torney and Stack (Blood 2009).
Immunogenicities of RBC antigens expressed as a fraction of that of the K antigen (Potency = 1) are shown in columns IV & VII (method of Giblett) and in V & VIII (method of Torney & Stack) of the following table.
I Antigen X II Anti-X, N III Anti-X, Females, N IV Females Potency vs. K V Females, Corrected Potency VI Males, Anti-X, N VII Males, Potency Vs. K VIII Males, Corrected Potency C 71 51 0.227 0.156 20 0.108 0.075 c 37 27 0.164 0.141 10 0.074 0.063 E 220 117 0.554 0.565 103 0.594 0.606 e 9 5 0.244 4 0.238 Fya 44 28 0.122 0.086 16 0.085 0.060 Fyb 3 1 0.007 0.004 2 0.017 0.012 Jka 60 41 0.225 1.083 19 0.127 0.611 Jkb 5 3 0.015 2 0.012 K 153 84 1 1 69 1 1 k 1 1 0.488 0 S 18 13 0.051 0.027 5 0.024 0.013 s 1 0 1 M 21 10 0.056 0.069 11 0.075 0.093
In males the corrected potencies of E and Jka were about 60% of that of K, but in females the corrected potencies of Jka and K were equal. The antigens C, c, Fya,Jka and S had higher corrected
potencies in females than in males, probably reflecting the additional role of pregnancies in immunization. Note the big difference in antibody response between allelic non-Rh antigens (e.g. Jka vs. Jkb).
Immunogenicity of the Rh D antigen was not calculated since Rh negative patients receive Rh negative blood and Rh negative pregnant women are given Rh immune prophylaxis. However, in this study 29.6 % of the patients were Rh negative. Over 70% of the susceptible patients had been immunized to the Rh D.
In conclusion, RBC antibodies interfere with the benefits of RBC transfusions and may also cause severe adverse effects. Introduction of molecular methods into Blood Bank setting could eventually minimize exposure and immunization of transfusion recipients to incompatible RBC antigens. Information about immunogenicity of various blood group antigens may point out those antigens that should not be given to patients who are lacking them. It is also critical to understand why immunization to the D antigen is still prevalent in order to pave the way to future multiple antigen matching.

Disclosures:
No relevant conflicts of interest to declare.