bluuebunny/crossref_metadata_2025_split huggingface dataset:
Long-Term Outcomes of De Novo Acute Myeloid Leukemia (AML) Patients (pts) with Life Threating Complications
Authors: Bazhenov Aleksei , Parovichnikova Elena N. , Galstyan Gennadiy M. , Troitskaya Vera V. , Gribanova Elena O. , Savchenko Valery G. 
Published: October 2019

Abstract
Background. Induction of remission in pts with AML is often associated with the life-threating complication and ICU admission. The long-term outcomes of these AML pts discharged from ICU are unknown. The aim was to estimate the long-term outcomes of de novo AML pts required ICU admission due to life-threating complications during induction of remission.
Methods. All de novo AML pts, younger than 60 y.o, median age 34 (17-60 yo), from 2013 to 2016 were enrolled in the study. All pts were divided into 2 groups: pts who required ICU admission during induction chemotherapy due to life-threating complications and pts who didn't require ICU admission during induction chemotherapy. Overall survival (OS) and disease free survival (DSF) were assessed by the Kaplan - Meier method, log rank value p<0.05 consider as significant. Univariate analysis was performed with χ 2 tests or Fisher's exact tests for categorical variables to find an independent ICU mortality predictor.
Results In total, 73 AML pts included in study, 26 (36%) of them were admitted in ICU during induction of remission (tab. 1).
ICU pts had more advanced AML. Reasons for ICU admissions were acute respiratory failure (ARF) (62%); neurological events (16%); septic shock (SS) (16%); cardiac arrhythmia (7%). ICU survival rate was 69%. 8 pts died in ICU (5 SS and 3 ARF). Needs for mechanical ventilation, vasopressors and ≥2 organ dysfunctions were independent predictors (p<0.05) of ICU mortality. 18 of 26 (69%) pts received chemotherapy during their stay in ICU. Modifications of chemotherapy were required in 46% of ICU pts and in 36% in non-ICU pts (p>0.05). There were differences in 30-day OS between ICU and non-ICU pts (69% vs. 0%, p=0.001). However, a landmark analysis for long term OS for patients who survived the first 30 days of treatment were the similar in ICU and non-ICU groups (p=0.946)( picture1, A). DSF also were the similar in ICU and non-ICU groups (p=0.946) (picture 1,B)
Conclusion. During remission induction 36% of AML pts needed ICU admission due to life-threating complications. Mortality rate in ICU is high (31%). However, after discharging from ICU AML patents had the similar long-term outcomes as well as non-ICU AML pts.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
Platelet (hem)ITAM Signaling Mediated Cancer-Associated Venous Thrombosis
Authors: Wang Xia , Wu Jingchang , Gu Haidi , Qian Jun , Ruan Changgeng , Zhao Yiming 
Published: October 2019

Abstract
Background-Patients with cancer are at a high risk of venous thromboembolism (VTE) and underlying mechanisms are unclear. Recent studies have shown that podoplanin (PDPN) expressing highly in primary brain tumors is associated with increasingly risk of VTE by activate platelets. The C-type lectin-like receptor 2(CLEC-2) is a type II transmembrane protein expressed on the surface of platelets and PDPN is the only known endogenous ligand for it. CLEC-2 mediates platelet activation through a hem-immunoreceptor tyrosine-based activation motif ((hem)ITAM).
Objective: We tested the hypothesis that the platelet (hem)ITAM signaling is involved in the development of VTE in patients with malignant tumors, and targeted inhibitors may reduce thrombosis and improve prognosis in patients with cancer.
Materials and Methods-We selected two human malignant cell lines (NCI-H226 and C8161) expressing high levels of PDPN as models to study the role of the platelet (hem)ITAM signaling in cancer-associated venous thrombosis in vitro and in vivo.
Results and Conclusions-Our results showed that both types of cell lines expressing PDPN triggered platelet activation via CLEC-2 in vitro, which could be abrogated by an anti-PDPN antibody SZ-168 or syk inhibitor R406. Further, in vivo study showed that injection of NCI-H226 or C8161 in two mouse models of venous thrombosis activated platelets, increased platelet counts and enhanced thrombosis. These results suggested that platelet activation induced by PDPN was correlated with hypercoagulability, contributed to thrombosis in cancer patients. Importantly, PDPN enhanced thrombosis was reduced in mice treated with SZ168 or R406. Immunohistochemical staining using anti-PDPN antibody was also performed in pulmonary squamous cell carcinoma specimens of 166 patients. During up to 4-year-follow-up, 20 (12.05%) patients developed VTE. 105 tumor specimens stained positive for PDPN (27 high expression, 43 medium expression, 35 low expression). High PDPN expression was associated with an increased risk of VTE and poor prognosis, independently of age, sex and tumor type. In this study, we found that PDPN expression in tumors induced platelet aggregation and was associated with a high risk of VTE via platelet (hem)ITAM signaling. Our data also suggested that SZ168 inhibited PDPN-induced platelet aggregation in vitro and decreased the incidence of VTE in mice. In conclusion, our research demonstrated novel insights into the pathogenesis of cancer-associated venous thrombosis.

Disclosures
No relevant conflicts of interest to declare.
Novel MEIS1-FOXO1 Fusion Gene in a Case of Pediatric B-Cell Precursor Acute Lymphoblastic Leukemia
Authors: Jiang Chuang , Qian Jiabi , Hao Wenge , LIU Wei , Shen Shuhong , Jiang Hua , Zhang Hui 
Published: October 2019

Abstract
Background: Thanks to the total therapy and systemic basic-translation research, the overall survival rate in children with acute lymphoblastic leukemia (ALL) has dramatically improved to almost 90% over these past few decades. FOXO1 gene belongs to the forkhead family of transcription factors, which play roles in myogenic growth and differentiation. Translocation of FOXO1 with PAX3 has been reported in pediatric alveolar rhabdomyosarcoma. In B-cell precursor ALL, two cases with FOXO1 fusions have been identified already, while its function on ALL remains unknown. Here, we report a novel MEIS1-FOXO1 fusion gene in a case with B-ALL. Methods: Flowcytometery, karyotype, RT-PCR and fluorescence in were employed, MEIS1-FOXO1 was identified as novel fusion gene in a case of pediatric BCP-ALL. Using IL-3 dependent BaF3 cells as study model to test the leukemia transformation potential of MEIS1-FOXO1. Results: A novel MEIS1-FOXO1 fusion was identified in one cease of pediatric B-ALL. Panel next generation sequencing (NGS) showed that the leukemia clone had concurrent NRASG12D, TP53R273H, WHSC1E1099K, ABCC1R1166X, PHGR1H37P, HOXA3P219L and DSTP4606L somatic mutation. This patient was enrolled in CCCG-ALL2015 clinical trial (ChiCTR-IPR-14005706) and achieved completed remission and low minimal residual disease (MRD) level (MRD<0.01%) at day 19 from induction therapy. Functional study showed that MEIS1-FOXO1 fusion gene can potentiate BaF3 cells growth independent of IL3 supplement, as compared to those without MEIS1-FOXO1 fusion transduction. In the meanwhile, we have found that MEIS1-FOXO1 fusion gene can drive cells into S-phase with concurrent decreased G0/G1 phase, which might be its oncogenic role in leukemogenesis. Using qPCR methods, we have found that MEIS1-FOXO1 fusion gene altered the cell cycle related genes expression. Conclusions: Integrating the FOXO1-fusion reports, our data have added more evidence to underline the role of FOXO1 deregulation in the pathogenesis of acute lymphoblastic leukemia. Novel fusion of MEIS1-FOXO1 can potentiate B-ALL via cell cycle entry. Detailed mechanisms involved into the MEIS1-FOXO1 should be further investigated.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
Platelets Promote Activation of the Complement System in Ovarian Cancer
Authors: Gonzalez Pagan Omayra , Cho Min Soon , Afshar-Kharghan Vahid 
Published: October 2019

Abstract
Platelets promote metastasis and growth of ovarian cancer. We have shown that platelets extravasate into the tumor microenvironment (TME) and increase proliferation and epithelial-mesenchymal transition (EMT) in ovarian cancer cells. We have also shown that activation of the complement system in TME of ovarian cancer enhances tumor growth. Ovarian cancer cells secrete complement proteins that upon activation in the TME increase proliferation of cancer cells and promote EMT via an autocrine pathway. The activators of the complement system in the TME have not been identified. We have demonstrated that upon activation platelets activate the complement system on their surface. In the current study, we examined whether extravasated platelets inside tumors contribute to the complement activation in the TME.
1) We examined the effect of antiplatelet reagents on platelet extravasation into TME, using murine models of ovarian cancer. Tumors induced by injection of ovarian cancer cells into the peritoneum of Nu/Nu mice were resected after 6-8 weeks and the number of extravasated platelets was determined by immunostaining tumor sections and counting the number CD42 (GPIb) positive cells that were outside the blood vessels (CD31 positive). We found that platelet extravasation is an active process and platelet inhibition by aspirin or ticagrelor reduces the number of extravasated platelets. Furthermore, P2Y12 deficient platelets extravasate less than normal platelets. In all of these experiments, the number of extravasated red blood cells were significantly less than extravasated platelets and was not affected by the inhibition of platelets.
2) We examined the effect of platelet inhibition on the activation of the complement system in the TME. We immunostained resected tumors from aspirin- or ticagrelor-treated tumor-bearingmice and from P2Y12-deficient tumor-bearingmice for the endproductof complement activation (C5b-9 or membrane attack complex). Inhibition of platelet function by aspirin or ticagrelor,or the presence of hypoactive platelets in P2Y12 deficient mice reduced the amount of C5b-9 deposited in the tumors induced in murine models of ovarian cancer.
Our result showed that complement activation in the TME is at least partially dependent onextravasated platelets. We propose that platelets in addition to directly increasing proliferation of ovarian cancer cells, also enhance tumor growth by activating the complement system in the vicinity of cancer cells. Our study links platelets, complement activation, and ovarian cancer growth; and raises the possibility of using antiplatelet reagents and complement inhibitors as novel synergisticanti-tumor reagents in ovarian cancer.

Disclosures
No relevant conflicts of interest to declare.
Functional Characterization of Epigenetic Modifiers of Demethylating Therapy in AML
Authors: Redhaber Desiree Melanie , Phely Laurent , Ehrenfeld Sophia , Veratti Pia , Schäfer Silvia , Mitschke Jan , Beckert Jessica , Shoumariyeh Khalid , Duyster Justus , Miething Cornelius 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) is the most common type of acute leukemia in adults, and remains a difficult to treat disease, especially in elderly patients. AML arises in hematopoietic stem and progenitor cells and frequently carries alterations in genes involved in epigenetic regulation. Furthermore, azacitidine (AZA) and decitabine (DAC), two nucleoside analogs inhibiting DNA methylation (DNMTi), have shown clinical efficacy in the therapy of myelodysplastic syndrome (MDS) and AML, highlighting the importance of epigenetic regulation in AML. Up to now, the mechanism of action of AZA and DAC in AML has not been fully elucidated, and reliable biomarkers of therapy response to these drugs do not yet exist.
To identify epigenetic response modifiers towards AZA and DAC treatment, we conducted a pooled shRNA-based screen in three human leukemia cell lines using an inducible custom shRNA library targeting more than 660 different genes involved in epigenetic pathways. After selection of retrovirally infected cells, shRNA expression was induced and a fraction of cells were treated with either AZA, DAC or left untreated. 10 days after shRNA induction, genomic DNA was extracted and deep-sequenced to identify genes conferring a selective advantage/disadvantage under these conditions. By comparing shRNA representation changes between the differentially treated groups, we identified multiple candidate genes sensitizing or protecting leukemic cells to/from AZA and DAC treatment. Notably, the DNMT1 gene itself expectedly emerged as an important sensitizer to DNMTi, as did other CXXC domain-containing genes binding to (un-)methylated CpG domains. Another prominent set of genes in the pooled analysis belonged to the DNA damage pathway, including genes such as PARP1 and BRCA1. The identified candidate genes were validated and confirmed in more than 80% when tested in single shRNAs competition assays. Their impact on DNA methylation and gene expression is currently being assessed, with ongoing experiments focusing on the mechanistic role of some of the validated target, as well as the clinical potential for combinatorial therapies. Thus, using an unbiased functional genetic approach, we identified multiple genes including CpG binding proteins and members of the DNA damage pathway as important modulators of DNMTi response. Our results will increase our understanding of the molecular mechanisms driving DNMTi efficacy, and may help to identify new biomarkers and novel targets for combinational therapy in AML.

Disclosures
No relevant conflicts of interest to declare.
Endothelial Cell-Derived Extracellular Vesicles Mitigate Radiation-Induced Hematopoietic Injury
Authors: Piryani Sadhna O. , Jiao Yiqun , Kam Angel Y.F. , Liu Yang , Vo-Dinh Tuan , Chen Benny J. , Chao Nelson J. , Doan Phuong L. 
Published: October 2019

Abstract
Hematopoietic stem cells (HSCs) reside in specialized microenvironments and adjacent to endothelial cells (ECs), from which they receive regulatory cues. Following injury from radiation or chemotherapy, transplanted ECs can facilitate hematopoietic regeneration. Since these ECs do not engraft in the marrow, we hypothesize that ECs facilitate hematopoietic regeneration through secretion of extracellular vesicles (EVs). EVs are becoming recognized as a mode of cell-to-cell communication through transfer of their cargo of nucleic acids and proteins. Using ultracentrifugation, we visualized these EVs by transmission electron microscopy and measured their size to be a mean size of 160 nm (range 121-217 nm, n=18). Using flow cytometric analysis with Syto13, we demonstrate that these EVs contain nucleic acids in 30% of the particles. Quantitative real-time PCR show that these EVs express CD31 and VECadherin, and not Fibroblast Specific Marker 1 (FSP1), indicating EVs bear EC markers. At day 7 following 300 cGy, C57BL/6 cKit+Sca-1+Lineage- (KSL) cells cultured with EVs demonstrated total cell expansion and colony-forming unit cells (CFCs) compared to cultures with cytokines alone (*p< 0.0001 and 0.01, respectively, n= 3-6/group). Following myelosuppressive, sub-lethal radiation dose of 500 cGy, C57BL/6 mice treated with EVs on days 1-4 display increased marrow cellularity (*p= 0.003, n= 2-3/group), preserved vascular endothelial cells by mouse endothelial cell antigen immunohistochemistry (*p=0.03, n=2-4/group), CD150+CD48-KSL cells (*p= 0.03, n= 5/group), and increased CFCs (*p= 0.03, n= 6/group) compared to saline-treated mice. No differences were detected in these parameters when EV-treated mice were compared to mice treated with primary, cultured C57BL/6 marrow ECs. Following lethal-dose irradiation of 800 cGy, mice that received EVs on day 1-4 display prolonged survival compared to saline-treated mice (*p= 0.008 by Log-rank Analysis, n= 10/group). At day 30 following irradiation, 5 of 10 mice were alive in the EV-treated group compared to 0 of 10 in the saline-treated group. No differences in survival were observed between EV-treated and EC-treated mice (p= 0.4). Treatment of irradiated C57BL/6 hematopoietic stem/progenitor cells (HSPCs) with EVs generated from a genetically distinct strain (BALB/c mice) showed similar levels of cell expansion and CFCs compared to treatment of HSPCs from syngeneic EVs. Our findings show that syngeneic or allogenic EVs could be cell-derived therapy to deliver physiologic doses of nucleic acids and growth factors to hematopoietic cells to accelerate hematopoietic regeneration.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
Exon Skipping of AIMP2 and Lymphomagenesis
Authors: Kim Doyeun , Song Seulki , Kim Dae-Yoon , Kim Dongchan , Kwon Nam Hoon , Kim Byung-Su , Lee Hyun Jung , Kim Hyo Jung , Lee Jung Lim , Yoon Sung-Soo , Jeon Yoon Kyung , Kim Sunghoon , Koh Youngil 
Published: October 2019

Abstract
ARS-interacting multifunctional protein 2 (AIMP2), the component of aminoacyl-tRNA synthase complex, acts as a potent tumor suppressor in conjunction with p53. The second exon deficient AIMP2 generated by alternative splicing mechanism (AIMP2-DX2) induces loses its important role as a signaling modulator. Previously, it predisposes mouse to increase lung cancer, but it did not induce tumorigenesis by itself.
To further validate the oncogenic role of AIMP2-DX2 in mouse model, we generated inducible transgenic mouse which express DX2 in doxycycline dependent manner. Unexpectedly, we report the late onset spontaneous lymphoma development instead of lung cancer in this model. Polyclonal expansion of pre-B cells and infiltration to multi-organ, especially kidney, was major events. More importantly, switching off the transgene by doxycycline removal reduced pre-B cell hyperplasia by inducing apoptotic cell death. Destabilization of p53 and dysregulation of cell cycle gene by DX2 overexpression underlined the increased lymphoid cell proliferation. Coincident with the results, in International Cancer Genome Consortium (ICGC) data analysis, AIMP2-DX2 expression was the highest in Burkitt lymphoma (BL) and diffuse large B cell lymphoma (DLBCL) among various cancer subtypes. When analysis was performed using RNA sequencing data of various B-cell lymphoid malignancies, germinal B-cell origin (GCB) DLBCL, B-acute lymphoid leukemia and BL showed completely different gene expression pattern according to the AIMP2-DX2 expression status suggesting a distinct disease entity. AIMP2-DX2 expression in DLBCL cell line and tissue samples was validated and regulation of DX2 in lymphoma cell line affected cell viability as well as p53 stability. Lastly, we confirmed expression of AIMP2-DX2 in FFPE tissue of 35 lymphoma patients using RNA in situ hybridization (ISH) with specifically designed probes that capture the exon 1-3 junction of AIMP2 transcript.
In summary, our data demonstrate the causality of AIMP2-DX2 in lymphoma oncogenesis and provide evidence that AIMP2-DX2 is a potential therapeutic target especially in DLBCL and BL.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
LSD1 Inhibitors Induce Fetal Hemoglobin in Primary Human Erythroid Cells
Authors: LE Cuong , Myers Greggory , Habara Alawi , Chui David H.K. , Steinberg Martin H. , Engel James Douglas , Cui Shuaiying 
Published: October 2019

Abstract
Significant amount of clinical literatures have supported that high level of fetal hemoglobin (HbF) improves the disease pathophysiology of β-globinopathies [sickle cell disease (SCD) and β-thalassemia]. Thus, we certainly can treat β-globinopathies by increasing the HbF level in adult erythroid cells.
We originally reported that the lysine-specific histone demethylase 1 (LSD1) plays an important role in the regulation of the fetal γ-globin genes. Inhibition of LSD1 by using RNAi and monoamine oxidase inhibitor tranylcypromine (TCP) in primary human erythroid progenitor cells induces HbF to therapeutic levels. Furthermore, LSD1 inhibitor RN-1 treatment of SCD mice results in increased HbF synthesis and leads to effective improvement of many aspects of the disease pathology normally associated with SCD.
Most recently, we examined thein vivo effects of some additional, publically available small molecule chemical inhibitors of LSD1 (including GSK-LSD1, LSD1-C12, LSD1-C76, OG-L002, and S2101) on HbF synthesis and erythroid physiology in SCD mice. There was a statistically significant increase in the percentage of HbF positive cells after 4 weeks of treatment with GSK-LSD1 or OG-L002 in SCD mice. Here, we report the effects of these two inhibitors in primary human erythroid cell derived from peripheral blood CD34+ cells. We isolated CD34+ cells using MACS column and cultured them using the two-phase-culture system. After seven days in expansion phase (phase 1) and three days in differentiation phase (phase 2), cells were treated with different doses of GSK-LSD1 and OG-L002 LSD1 inhibitors along with controls (DMSO, hydroxyurea and TCP) for 3 or 5 days in phase 2 culture. Flow cytometric assays showed that the percentage of HbF positive cells were significantly high when CD34+ cells treated with OG-L002 LSD1 (~50% at 0.1 µM) or GSK-LSD1 (~30% at 0.1 µM) as compared to control DMSO (~20%) after 5 days. These results suggest that GSK-LSD1 and OG-L002 could be two new promising HbF inducers based on LSD1 inhibition. These findings provide additional evidence to support that LSD1 comprises a useful molecular target for possible therapeutic intervention in treating SCD. Further study will be necessary to address the potential therapeutic effects of the compounds in SCD patients.

Disclosures
No relevant conflicts of interest to declare.