bluuebunny/crossref_metadata_2025_split huggingface dataset:
Long Noncoding RNA HOXA-AS2 As a Predictor of Acute Myeloid Leukemia: Clinical Association between HOXA-AS2 Expression and Its Role in Leukemic Cell Growth
Authors: Yan Xiaojing , Qu Yi 
Published: October 2019

Abstract
Long noncoding RNAs (lncRNAs) have important roles in diverse cellular processes and carcinogenesis. Homeobox (HOX)A cluster antisense RNA 2 (HOXA-AS2) is a 1048-bp lncRNA located between human HOXA3 and HOXA4 genes whose overactivation was previously found to promote the proliferation and invasion of solid tumors. However, its biological roles in acute myeloid leukemia (AML) remain unclear. This study showed that HOXA-AS2 was overexpressed in patients with AML. In addition, the increased HOXA-AS2 expression level correlated with higher white blood cell and bone marrow (BM) blast counts, unfavorable karyotype classification, earlier death, and inferior survival of patients with AML. Moreover, the results of an in vitro study showed that the silencing of HOXA-AS2 significantly inhibited the growth of leukemic cells through inducing G1/G0 phase arrest and apoptosis. In conclusion, these findings suggested that HOXA-AS2 functioned as an oncogene and might be a useful biomarker for diagnosis and prognostic prediction in patients with AML, providing a promising therapeutic target for AML.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
Role of N6-Methyladenosine (m6A) RNA Modification in Multiple Myeloma
Authors: Bach Christian , Leffler Magdalena , Flamann Cindy , Kronke Jan , Mougiakakos Dimitrios , Mackensen Andreas , Bruns Heiko 
Published: October 2019

Abstract
Multiple myeloma (MM) is considered a chronic and incurable disease due to its highly complex and heterogeneous molecular abnormalities. In recent years, integrating proteasome inhibitors and immunomodulatory drugs into MM frontline therapy has significantly improved treatment efficacy with a median overall survival (OS) being prolonged from 3-4 to 7 years. Despite this progress, patients refractory to the aforementioned agent classes display a median OS of only 9 months. Thus, the clinical necessity for developing novel therapeutic alternative approaches is self-evident.
Methylation of N6-adenosine (m6A) is known to be important for diverse biological processes including gene expression control, translation of protein, and messenger RNA (mRNA) splicing. m6A regulatory enzymes consist of "writers" METTL3 and METTL14, "readers" YTHDF1 and YTHDF2, and "erasers" FTO and ALKBH5. However, the functions of m6A mRNA modification and the specific role of these enzymes in MM remain unknown.
Here we report that METTL3, a key component of the m6A methyltransferase complex, is highly expressed in MM cell lines and in isolated patient's MM cells. In contrast, we found no significant differences in the expression of the m6A demethylases FTO and ALKBH5. Accordingly, compared to plasma cells from healthy donors, global PolyA+ RNA showed a significant increase in m6A content in patient's MM plasma cells. In MM cell lines, global m6A profiling by methylated RNA-immunoprecipitation sequencing revealed m6A peaks near the stop codon in mRNAs of multiple oncogenes including MAF and CCND1. Cross-linking immunoprecipitation showed that METTL3 bound to the m6A peak within MAF and CCND1 mRNA. Depletion of METTL3 by shRNA had little effect on global mRNA levels, but specifically reduced protein levels of c-Maf and Cyclin D1. Moreover, downregulation of METTL3 in several MM cell lines results in cell cycle arrest and apoptosis.
Together, these results describe a role for METTL3 in promoting translation of a subset of oncogenes in MM and identify this enzyme as a potential therapeutic target for multiple myeloma.

Disclosures
No relevant conflicts of interest to declare.
Inhibition of Unc-51-like Kinase 1 (ULK1) with Novel Small Molecular Inhibitor MRT68921 Preferentially Induces Apoptosis and Autophagy in FLT3-ITD- Mutated Acute Myeloid Leukemia
Authors: Jang Jieun , Jeung Hoi-kyung , Seol So-Young , Chung Haerim , Kim Yu Ri , Cheong June-Won , Min Yoo Hong 
Published: October 2019

Abstract
In normal karyotype acute myeloid leukemia (AML), FLT3-ITD mutation is associated with dismal prognosis with early relapse even after allogeneic stem cell transplantation. Unfortunately, to date small-molecule inhibitors of FLT3 have resulted in only partial and transient clinical responses with residual leukemic blasts acquiring resistance to FLT3 inhibitors. Therefore, elucidation of novel molecular targets should be necessary for effective eradication of FLT3-ITD AML cells. Evidences are accumulating on the functional roles of autophagy in the initiation and maintenance of AML as well as the development of drug resistance. Unc-51-like kinase 1 (ULK1) is a conserved serine-threonine kinase that plays a central role in the initiation of autophagy. Our group demonstrated that ULK1 is potentially involved in the development of resistance of AML leukemia stem cells to BET inhibitor, JQ1. The fact that ULK1 is the only conserved serine/threonine kinase in the autophagy cascade makes it a very attractive target for therapeutic development. However, the role of Ulk1 in FLT3-ITD AML remains unclear. In this study, we observed that MRT68921, a potent inhibitor of both ULK1 and ULK2, induced apoptotic cell death in FLT3-ITD-mutated AML cell lines (MV4-11, Molm13, U937/FLT3-ITD-muated) in a dose-dependent manner. However, apoptosis-inducing effect of MRT68921 was significantly lower in FLT3-WT AML (HL-60, U937). Cell death was accompanied with cleavage of caspases and PARP, which were partially blocked with caspase inhibitor z-VAD-fmk, indicating the caspase-dependent mechanism exists. MRT68921 treatment led to a notable decrease in the levels of phosphorylated (p) ATG13 (Ser 318) as well as total ULK1 and p-ULK1 (Ser 555). Interestingly, MRT68921 induced LC3-II lipidation, autophagosome, and GFP/LC3 punta formation, indicating autophagy was paradoxically activated in FLT3-ITD-mutated AML cells. AMPKa phosphorylation (T712) was increased in MTR68921-responsive cells. In contrast, autophagy induction was negligible to modest in FLT3-WT AML cells. Treatment of FLT3-ITD cells with autophagy inhibitors, 3-MA, bafilomycin A1, and hydroxychloroquine, markedly enhanced the MRT68921-induced apoptosis, strongly suggesting that prosurvival autophagy activation occurred with MRT68921 in FLT3-ITD cells. Reduction in the levels of total FLT3 and p-FLT3 protein were observed concurrently with downregulation of p-STAT5 in FLT3-ITD cells. Endoplasmic reticulum stress-associated proteins, p-PERK and p-eIF2a were also downregulated with MRT68921 in FLT3-ITD cells. Taken together, targeting the ULK1 pathway could be an effective therapeutic strategy for combating FLT3-ITD AML. Inhibition of prosurvival autophagy pathway could enhance the anti-leukemia effects of MRT68921.

Disclosures
No relevant conflicts of interest to declare.
Androgen Therapy for Lower-Risk Myelodysplastic Syndrome
Authors: Choi Eun-Ji , Lee Je-Hwan , Park Han-Seung , Lee Jung-Hee , Seol Miee , Lee Young-Shin , Kang Young-Ah , Jeon Mijin , Woo Ji Min , Lee Kyoo Hyung 
Published: October 2019

Abstract
Background
Improvement of cytopenia is one of the primary treatment purposes for patients with the lower-risk myelodysplastic syndrome (MDS). Androgens have been used for the treatment of aplastic anemia, immune thrombocytopenia, and telomere diseases. In this retrospective study, we aimed to evaluate the efficacy of androgen therapy in lower-risk MDS.
Methods
We analyzed the data of 139 patients who received androgens (danazol or oxymetholone) for treatment of cytopenia between February 1997 and May 2018. All patients had the international prognostic scoring system low or intermediate-1 risk at the time of androgen therapy. The assessment of hematologic improvement (HI) was based on the international working group response criteria for MDS.
Results
Androgens (oxymetholone for 83 patients and danazol for 56) were given as first (n=108, 77.7%) or over second (n=31, 22.3%)-line treatment for MDS (Table 1). The time interval between diagnosis and androgen treatment was median 1.3 months (range, 0-240.6), and 75 patients (54.0%) were red blood cell (RBC) transfusion-dependent before treatment. The dose intensity of oxymetholone and danazol was 79 and 385 mg/day respectively, and the median treatment duration was 5.8 months (range, 0.9-92.2). Seventy-nine patients (56.8%) achieved HI at any lineage: 29.0% for erythroid (HI-E), 51.9% for platelet (HI-P), and 60.5% for neutrophil (HI-N). The median time to HI following androgen therapy was 4.1 months (range, 0.6-124.5) for HI-E, 1.7 (range, 0.4-40.4) for HI-P, and 1.8 (range, 0.2-8.4) for HI-N. In univariate analysis, presence of RBC transfusion-dependence (46.7% vs. 68.8%, P=.009), pre-treatment low hemoglobin (45.2% vs. 74.5%, P=.001), high platelet count (46.5% vs. 73.6%, P=.002), and high neutrophil count (4.94% vs. 67.2%, P=.036) were associated with lower HI rate (Table 2). In multivariate analysis, pre-treatment low hemoglobin, high platelet count, and high neutrophil count remained as significant factors for lower HI rate (Table 2). During the median follow-up duration of survivors of 40.8 months (95% confidence interval [CI], 38.0-67.5), the estimated 5-year overall survival (OS) and acute myeloid leukemia-free survival was 68.8% and 67.7%, respectively. Achievement of HI was associated with longer OS (hazard ratio, 0.346; 95% CI, 0.174-0.688). There were no significant differences in HI and OS rates between danazol and oxymetholone.
Conclusion
Our data suggest that androgen can be a reasonable treatment option for lower-risk MDS patients with significant cytopenia. Prospective studies are warranted to investigate the efficacy of androgen therapy in lower-risk MDS.

Disclosures
No relevant conflicts of interest to declare.
Treatment of Relapsing/Refractory Hodgkin's Disease
Authors: Gorenkova Liliya , Kravchenko Sergey K. , Klebanova Elizaveta , Moiseeva Tatiyana , Mangasarova Yana K. , Magomedova Aminat U. , Savchenko Valery G. 
Published: October 2019

Abstract
Relapses of Hodgkin's disease (HD) occur in 10-15% of patients with a local tumor and in 20-40% of patients with advanced stages. A relapse of HD is early in half of the patients whereas a second remission can be achieved in only 50% of cases. Autologous hematopoietic stem cell transplantation (auto-SCT) is considered to be the standard of consolidation therapy for relapsing and primary-resistant HD. In patients with relapses of HD, progression-free survival and overall survival are 50% to 60% and 50% to 80%, respectively.
Objective. To assess the efficacy of different second-line therapies with subsequent autologous stem cell transplantation in the treatment of relapsing or refractory HD.
Materials and methods: from 2000 to 2017, 76 patients with relapsing/refractory HD treated with at least first-line therapy were involved in the study with most of the patients (70%) having received various modifications of BEACOPP regimen. The median age of patients was 27 years. A majority of participants had advanced stages of the disease: 23 (30%), 14 (18%), and 35 (46%) of patients have stage I-II, III, and IV, respectively.
Results: DHAP-regimen was given as a second-line treatment in 34 patients resulting in progression, an absence of effect, and overall response in 11 (32%), 13 (38%), and 9 of the patients, respectively. Of them, a complete remission was achieved in only 4 patients, who further underwent autologous stem cell transplantation as a consolidation treatment.
Dexa Beam regimen was given as a second-line treatment in 53 patients including those with progression after DHAP-courses. As a result, partial/complete remission, progression, and stable disease were achieved in 35 (66%), 7, and 11 patients, respectively.
Auto-SCT was carried out in 32 patients of the overall response group; 28 (88%), 3, and 4 patients have a complete long-term remission, progressive disease, and early relapses, respectively.
In the general study population, overall and event-free survival rates were respectively 86% and 71% with a follow-up median of 83 months.
Thus, Dexa Beam given as an induction treatment before auto-SCT showed an advantage in the overall response in relapsing/refractory HD. Implementation of auto-SCT as consolidation treatment after complete/partial remission has been achieved allows a long-term positive response to be attained.

Disclosures
No relevant conflicts of interest to declare.
Treatment for X-Linked Agammaglobulinemia with Allogeneic Umbilical Cord Blood Stem Cell Transplantation from HLA-Matched Sibling Confirmed Healthy through Prenatal Diagnosis
Authors: Wan Dingming , Zhang Ran , Wang Meng , Li Li , Cao Weijie , Zhang Suping , Sun Kai , Chen Xiaona , Zhang Na 
Published: October 2019

Abstract
Background X-linked agammaglobulinemia (XLA) is a rare congenital humoral immunodeficiency characterized by marked reduction in all classes of serum immunoglobulins and the near absence of mature CD19+ B-cells which was caused by mutations in Bruton's tyrosine kinase (BTK). We herein describe the first case of XLA transplanted from HLA-matched sibling allogeneic umbilical cord blood donor.
Method The patient was an 8-year-old boy with XLA who previously had life-threatening infectious episodes and diagnosed by direct sequencing of BTK gene mutations (c.992-923delGAN(NM-000061)) when he was 3 years old. He was treated with the regular replacement of immunoglobulin G (IgG) products. Taking into consideration significant infectious complications and the prompt availability of HLA-matched sibling umbilical cord blood, he underwent HLA-matched sibling UCBT. The FC conditioning regimen consisting of Flu (30mg/m2/d×4 days) and CTX (30mg/kg/d×4 days) was provided for the patient. The source of stem cells was umbilical cord blood of his younger brother who was confirmed completely healthy through prenatal diagnosis with villi at the fetus period of 4 months. The number of infused total nucleated and CD34+ cells was 6.51×108/L and 2.67×106/L, respectively. He received a combination of CsA, short-term MTX and MMF for GVHD prophylaxis.
Result Hematopoietic engraftment was rapidly achieved, with an absolute neutrophil count of >0.5×109/L on day 15 and platelet count of >20×109/L on day 20. The patient was discharged on day 45 and has been followed as an outpatient without any infectious episodes for more than 180 days. No expected toxicities were observed. Regarding immune constitution, CD19+ cells, IgA and IgM, which were undetectable before transplantation, started to increase in number 10 days after transplantation and continued to increase for more than 3 months. The patient did not exhibit any GVHD manifestations. He is currently alive and doing well at six months post-transplant, and he is free of any infection episodes.
Conclusion The above results provide strong evidence that HLA-matched sibling UCBT may be safe and technically feasible for the treatment of XLA when an appropriately matched related or unrelated donor is unavailable. The umbilical cord blood could be considered being frozen, especially in the era of "the second child" in China.
Keywords X-linked agammaglobulinemia; HLA-matched sibling donor; allogeneic umbilical cord blood stem cell transplantation; immune recovery.

Disclosures
No relevant conflicts of interest to declare.
Clinical and Hematological Risk Factors for Postpartum Hemorrhage: A 2 Years Population Based Cohort Study in France
Authors: Salomon Claire , De Moreuil Claire , Hannigsberg Jacob , Trémouilhac Christophe , Drugmanne Guillaume , Gatineau Florence , Falchier Frédérique , Briend Delphine , Abgrall Jean-François , Le Moigne Emmanuelle , Lacut Karine , Pan-Petesch Brigitte 
Published: October 2019

Abstract
Objective: To determine clinical and hematological risk factors for immediate postpartum hemorrhage (PPH).
Design: A cohort study.
Setting: Obstetric unit of Brest University Hospital (France).
Population: All women who underwent a vaginal delivery between April 1, 2013 and May 29, 2015.
Methods: Clinical data were collected by obstetricians or midwives during antenatal care visits, labor and delivery, and recorded by trained research assistants in a standardized electronic case report form. Hematological variables, including immature platelet fraction, were measured from a blood sample systematically performed at the entrance in the delivery room.
Main Outcome Measures: Postpartum hemorrhage (PPH), measured with a graduated collector bag, was defined as blood loss of at least 500ml.
Results: 2742 women were studied. PPH occurred in 141 (5%) women. Seven clinical factors were independently associated with PPH: pre-eclampsia (OR 5.85, 95%CI 2.02-16.90), multiple pregnancy (OR 3.28 (1.21-8.91), assisted reproduction (OR 2.75, 95%CI 1.45-5.20), ante partum bleeding (OR 2.15, 95%CI 1.24-3.73), post-term delivery (OR 1.93, 95%CI 1.17-3.17), obesity (OR 2.95, 95%CI 1.76-4.93) and episiotomy (OR 2.51, 95%CI 1.63-3.74). Three hematological factors were additionally identified as potential predictive factors for PPH: platelets < 150 000 Giga/L (OR 2.98, 95%CI 1.63-5.46), fibrinogen < 4.5 g/dL (OR 1.86, 95%CI 1.21-2.87) and APTT ratio ≥ 1.1 (OR 2.16, 95%CI 1.31-3.57).
Conclusion: Besides classical risk factors, this study highlighted the role of assisted pregnancy and obesity in the PPH occurrence, and identified simple hematological predictive risk factors for PPH. How these parameters could be integrated in a helpful prediction score of PPH remains to be determined.

Disclosures
Pan-Petesch: LFB: Other: Investigator.
Asparaginase Immune Complexes Detectable after Asparaginase-Induced Hypersensitivities Activate Basophils Via FcγRIII
Authors: Rathod Sanjay , Finkelman Fred D , Fernandez Christian 
Published: October 2019

Abstract
Background: Asparaginase (ASNase) is an important chemotherapeutic agent for the treatment of acute lymphoblastic leukemia (ALL). However, the development of anti-ASNase IgG antibodies are frequent, can neutralize the enzyme activity of ASNase, and can increase the risk of ALL relapse. Nevertheless, the role of anti-ASNase IgG antibodies on the onset of ASNase-induced hypersensitivity reactions is unknown, but likely require the formation of ASNase immune complexes (ICs) to activate cells involved in hypersensitivities due to the low binding affinity of the Fcγ receptor for the IgG immunoglobulin.
Objective: To detect plasma ASNase ICs after the onset of ASNase hypersensitivities and to demonstrate that binding of ASNase ICs to peripheral blood cells, and the subsequent immune cell activation, is FcγRIII-dependent.
Methods: Mice were sensitized to native E.coli ASNase using aluminum hydroxide adjuvant. ICs prepared using the plasma of sensitized mice were precipitated using polyethylene glycol to remove free ASNase and anti-ASNase IgG. ICs were detected by flow cytometry using APC labeled ASNase and protein G beads, characterized by dynamic light scattering and TEM, and the binding properties of ASNase ICs or free ASNase were determined by flow cytometry using non-sensitized and sensitized peripheral murine blood cells and the anti-IgE and/or anti-FcγRIIβ/III blocking mAb. Activation of basophils, which can occur in an IgE- or FcγRIII-dependent manner, was determined ex vivo by measuring changes in CD200R1 or CD200R3 expression after incubating peripheral blood cells with ASNase or ASNase ICs.
Results: ASNase ICs were detected after the onset of ASNase hypersensitivities in mice. ASNase ICs formed ex vivo at high anti-ASNase IgG1 concentrations, bound to naïve peripheral blood cells (e.g., neutrophils macrophage/monocytes, and basophils), and activated basophils in an FcγRIII-dependent manner. In contrast, basophil binding and activation by free ASNase was IgE- and not FcγRIII-dependent.
Conclusion: Our data indicate that ASNase sensitized mice form ICs after drug administration and can activate basophils via the FcγRIII receptor in mice, whereas the binding and activation by free ASNase occurs via cell-associated IgE. Our study suggests that ASNase ICs can contribute to the onset and severity of ASNase hypersensitivity.

Disclosures
No relevant conflicts of interest to declare.
Anti-CD30 Chimeric Antigen Receptor T Cell Therapy for CD30+ Relapsed/Refractory Hodgkin Lymphoma and Anaplastic Large Cell Lymphoma Patients
Authors: Wang Di , Xiao Yi , Li Chunrui , Wang Na , Huang Liang , Zhang Yicheng , Zhang Tongcun , Zhou Jianfeng 
Published: October 2019

Abstract
Chimeric antigen receptor T (CAR-T) cell therapy had shown remarkable efficacy in B-cell acute lymphoblastic leukemia and several B-cell lymphomas using CD19/22 antigen as target. Similarly, CD30 was found over-expressed on all stage of the cells in classical Hodgkin lymphoma (HL) and anaplastic large cell lymphoma (ALCL). Given the encouraging clinical success achieved by CD30 monoclonal antibody brentuximab vedotin, targeting CD30 with CAR-T cells would be a promising therapeutic approach for these populations. Here we conducted an open-label, single-center and single-arm pilot study, aimed to evaluate the feasibility and safety of anti-CD30 CAR-T cell therapy relapsed/refractory in HL and ALCL patients. This trial is registered with ChiCTR, number ChiCTR-OPN-16009069.
From August, 2016 to May, 2018, 4 HL and 2 ALCL patients with a median age of 26.5 years (range, 22-55 years) were enrolled in this study. Disease diagnosis and CD30expression was confirmed again by pathology and immunohistochemistry at enrollment. All the patients had relapsed or primary refractory disease, after at least two lines of systemic therapy. A pre-conditioning FC (fludarabin 25mg/m2, cyclophosphamide 20mg/kg for 3 days) regimen was give before CAR-T cell infusion. A total dose of 0.7-2.45×107 (median 1.175×107) per kg of CAR-T cells was infused over 3 to 7 consecutive days. The CAR transgene copy, representing CAR-T cells expansion, reached the peak at about 1 week after first administration, and then decreased. CAR transgene copy was still above 100 per μg genomic DNA in peripheral blood even 6 months after infusion. 5/6 (83.3%) patients had grade 0-1 cytokine release syndrome (CRS), with fever lower than 39℃ and duration less than 2 week. Levels of cytokines, including interleukin 6, ferritin and C-reactive protein were monitored. The maximum fold change of the cytokines from baseline was less than 10. No sign of neurotoxicity, bone marrow suppression or other adverse events were observed. The toxicity was generally tolerable, except for one HL patient, who experienced grade 4 CRS, eventually died from pneumorrhagia at the 20th day of infusion. The other 5 patients all achieved complete remission (CR) successfully. PET scan showed a CR status as early as the first month after infusion. Although relapse was observed in one HL patient 2 months after infusion, there were 3 of them remained CR state for over 1 year according to PET scan. The median progression free survival was 7 months.
In conclusion, our work demonstrated that CD30 CAR-T cell therapy is efficient and safe for HL and ALCL patients, providing a new option in treating CD30+ relapsed/refractory lymphomas, while a lager study scale was further demanded in our trial to obtain more robust support.

Disclosures
No relevant conflicts of interest to declare.