bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of HACE1 Expression Enhances HDAC Inhibitor Lmk-235-Mediated Apoptosis in Diffuse Large B-Cell Lymphoma Cells
Authors: Li Xinyao , Ma Dan , Lu Tingting , Kuang Xingyi , Yu Kunling , Zhang Zhaoyuan , Wang Weili , Wang Jishi 
Published: October 2019

Abstract
Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma (NHL). It is a group of adult lymphomas with a large clinical presentation and prognosis heterogeneity. In order to improve the cure rate of diffuse large B-cell lymphoma, it is necessary to find a new DLBCL treatment plan that can control the progression of the disease and explore its unknown potential therapeutic targets. Epigenetic modification has become a new treatment for leukemia in recent years, and histone deacetylase inhibitor (HDACi) is a novel drug for the treatment of DLBCL in recent years. However, in addition to effective clinical efficacy, Pan-HDACis has side effects. In contrast, LMK-235 is well tolerated as a novel inhibitor of HDAC4 and HDAC5. HACE1 is a potential tumor suppressor gene located on chromosome 6q21, which is involved in the regulation of tumor cell growth and apoptosis by mediating autophagy and ubiquitination of Rac1. The potential target of HACE1 as an HDAC inhibitor plays an important role in the occurrence, invasion and metastasis of human malignant tumors, but its underlying mechanism of HDACi-mediated apoptosis is still unclear. In this study, we investigated the potential mechanisms by which HACE1 regulates LMK-235-mediated apoptosis in DLBCL cells. In vitro, LMK-235 mediates apoptosis in DLBCL cells in a dose-dependent manner, and up-regulation of HACE1 by lentiviral transfection promotes LMK-235-induced apoptosis. At the same time, high levels of HACE1 can inhibit the expression of HO-1 at mRNA and protein levels. As a previous study in our laboratory, low expression of HO-1 enhanced the apoptotic rate of DLBCL cells. However, the correlation between HACE1 and HO-1 has not been reported. Therefore, we regulated HACE1 levels and found that HACE1 regulates HO-1, which affects the sensitivity of DLBCL cells to LMK-235. Our data show that HACE1 plays a key role in LMK-235-induced apoptosis in DLBCL cells. This provides new hope for the treatment of diffuse large B-cell lymphoma and is a potential new target for the treatment of diffuse large B-cell lymphoma.

Disclosures
No relevant conflicts of interest to declare.
microRNA-22 and TET-2 Mutation in Myelodysplastic Syndrome Patients Treated with Hypomethylating Agents
Authors: Park Seong Kyu , Kim Se Hyung , Lim Sung Hee , Kim Chan Kyu , Won Jong-Ho , Hong Dae-Sik 
Published: October 2019

Abstract
Background: MicroRNAs are small RNA species that regulate gene expression post-transcriptionally and are aberrantly expressed in many cancers including hematological malignancies. Some reports suggested that aberrations in the miR-22-TET2 regulatory network are common in myelodysplastic syndrome (MDS) and leukemia, and its aberrant expression correlates with poor survival. We attempted to identify the clinical role of miR-22 and TET-2 in patients with myelodysplastic syndrome.
Materials and Methods: A total of 41 MDS patients who treated with hypomethylating agents were recruited. Real time RT-PCR was performed to assess the expression levels of miR-22 and TET-2 mutation in bone marrow samples at the time of diagnosis. And we investigated the relationship between its results and clinical outcomes.
Results: TET2 mutation frequency in the higher risk group based the IPSS and IPSS-R was lower than that of lower risk group (11.1% vs 38.1%). miR22 expression was also down-regulated in higher risk group (higher risk: 1.70 ± 0.96 vs lower risk: 3.14 ± 1.38, p=0.006). TET2 mutation seemed to be different according to the responsiveness to hypomethylating agents. TET2 mutation, IPSS, and IPSS-R were significantly associated with the risk of leukemic transformation. Patients with an decreased value in the consecutive assessment of miR-22 at the time of diagnosis and 3 months after initial treatment tended to be associated with poor survival outcome (survival rate at 3 years: 18.2% vs 35.1% for patients with an increase, p=0.168). Prognostic factors for survival included TET-2 mutation, cytogenetics, IPSS or IPSS-R, and leukemic transformation.
Conclusion: miR-22 expression and TET2 mutation had a clinical impact on outcomes in MDS patients treated with hypomethylating agents. And these biomarkers might have a potential as a prognostic factor for MDS patients.

Disclosures
No relevant conflicts of interest to declare.
Quantification of Non-Hodgkin Lymphoma Disease Burden Using FDG PET-CT Helps Predict the Severity of Cytokine Release Syndrome
Authors: Wang Jiasheng , Hu Yongxian , Zhang Yanlei , Wei Guoqing , Xu Huijun , Wu Wenjun , Liu Shaohui , Wang Kangyan , Guo Zhenxing , Chang Alex Hongsheng , Zhao Kui , Huang He 
Published: October 2019

Abstract
Background: CD19-targeting chimeric antigen receptor (CAR) T-cell therapy has shown great efficacy in patients with refractory/relapsed non-Hodgkin lymphoma (NHL), but was associated with serious adverse effects such as cytokine release syndrome (CRS). It has been speculated that NHL baseline disease burden might affect clinical outcome and CRS, but such assumption has not been explored in detail in previous studies. Metabolic tumor volume (MTV) and total lesion glycolysis (TLG), calculated using FDG PET-CT, are quantitative indicators of baseline tumor burden.
Methods: Utilizing FDG PET-CT, we calculated MTV and TLG at baseline and post CAR T-cell therapy in 15 patients with NHL.
Results: Among all the patients, the median MTV was 72 (range 0.02-3024.9) cm3 and the median TLG was 610.1 (range 0.011-13156.3). After a median follow-up of 6 months, the overall response rate (ORR) was 66.7% (95% CI 38.4-88.2%). The baseline MTV and TLG did not have significant difference in patients with or without response (p=0.271 and 0.95, respectively). Cox-regression analysis did not find lower baseline MTV and TLG significantly associated with better overall survival (p=0.67 and 0.45, respectively). Patients with mild and moderate CRS (defined as Grade 0-2) had significantly lower MTV and TLG than those with severe CRS (Grade 3, 4) (median MTV: 49.3 v.s. 1137.7 cm3, p=0.012; median TLG: 379.1 v.s. 9384, p=0.012, Figure A, B). The median MTV in patients received tocilizumab, an IL-6 antagonist for the treatment of severe CRS, was 963.4 cm3, which was significantly higher than the patients not receiving tocilizumab (58.1 cm3, p=0.037). The median TLG in patients received tocilizumab was 9187.1, compared with 610.1 in patients not receiving tocilizumab (p=0.053). Using FDG PET-CT, we also demonstrated that CAR T-cell therapy in NHL patients could associate with severe local complications such as local compression and local inflammation.
Conclusions: Low NHL baseline disease burden is not associated with better response rate or long-term outcome. Patients with higher baseline disease burden have more severe CRS
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
Triple-Negative Myeloproliferative Neoplasms Vs. Calr, JAK2 or MPL-Mutated Myeloproliferative Neoplasms: Distinct Molecular Characteristics
Authors: Yun Jiwon , Kim Jung-Ah , Hwang Byungjin , Park Hee Sue , Im Kyongok , Kim Sung-Min , Jeong Dajeong , Lim Kyu Min , Bang Duhee , Lee Dong Soon 
Published: October 2019

Abstract
Background: We compared the clinical, cytogenetic, molecular features, and telomere lengths of patients with triple negative MPN and MPN with any of CALR, JAK2 or MPL mutations.
Methods: Target capture sequencing of 87 genes and molecular cytogenetic studies were performed in 61 Korean patients with MPN. Also, we searched the newly reported mutations and novel mutations in triple negative MPN [JAK2-G335D (germline), JAK2-F556V, JAK2-G571S (germline), JAK2-V625F (germline), MPL-T119I, MPL-S204F/P, MPL-E230G, MPL-V285E (germline), MPL-R321W (germline), MPL-Y591D, MPL-S505N and MPL-W515R]. We compared clinical and molecular characteristics between two groups. Additionally, we performed telomere quantitative FISH for 48 patients' samples and measured telomere/centromere ratios of them.
Results: Among 61 patients, 13 patients showed mutations in CALR, 34 in JAK2, and 3 in MPL. All of JAK2 mutation and CALR mutation site were reported sites, but 2 among 5 mutation site of MPL were novel mutation [D128N, D261Y]. Twelve patients showed triple negative (7 of PMF 7, 2 of ET, and 3 of MPN-U) - they showed 8 different mutation sites among 6 different genes (ASXL1, DNMT3A, NPM1, POLG, SRSF2, and ZMYM3). NPM1, POLG, and ZMYM3 mutations were seen only in triple negative patients. NPM1 mutation was significantly higher in triple negative MPN (P=0.0301). In telomere length study, there was no statistical difference between triple negative group (T/C ratio mean 12.5) and CALR, JAK2 or MPL mutated group (T/C ratio mean 10.0). Although, MPN patients with telomere length shorter than normal control's lower 10% (7.04) showed poor prognosis (P=0.0045).
Conclusions: Patients with triple negative MPN are characterized by high frequency of NPM1 mutation and less number of somatic mutations. Since the mutational analysis for diagnostic purposes is limited to exons 14 of JAK2, exon 10 of MPL and exon 9 of CALR at present, search for JAK2 and MPL mutations in other sites are essential in triple negative MPNs.
Keywords: Myeloproliferative neoplasms, next-generation sequencing, triple negative MPN, chromosome, FISH, telomere

Disclosures
No relevant conflicts of interest to declare.
Hematologic Responses in Patients with Heavily Pretreated Light Chain Deposition Disease (LCDD) Receiving Daratumumab
Authors: Milani Paolo , Basset Marco , Foli Andrea , Merlini Giampaolo , Palladini Giovanni 
Published: October 2019

Abstract
Introduction: The monoclonal antibody daratumumab showed high response rates and a good safety profile in multiple myeloma and is being evaluated in clinical trials in AL amyloidosis. Light chain deposition disease (LCDD) is a rare monoclonal gammopathy of renal significance. Treatment directed against the underlying plasma cell clone can prevent renal progression. Bortezomib is commonly used upfront in these patients and daratumumab may represent a powerful novel option.
Methods: We report the outcome of six patients with refractory light chain deposition disease (LCDD) treated with daratumumab at the Amyloidosis Research and Treatment Center of Pavia. All patients gave written informed consent. Hematologic response was assessed according to the International Society of Amyloidosis criteria.
Results: Six patients (5 males and 1 female) received daratumumab intravenously at 16 mg/kg weekly for 8 weeks, followed by every other week infusions for 8 doses and then monthly infusions. Patients' clinical characteristics are reported in Table 1. All patients received daratumumab single agent except one who was treated with daratumumb and bortezomib combination (this patient received only 1 prior line of therapy). All patients were refractory to the last line of therapy. All patients received at least two months of therapy. All patients were previously treated with bortezomib, pomalidomide was used in 4 cases, lenalidomide, thalidomide and bendamustine in 2 cases each, and autologous stem cell transplant was performed in 4 subjects. The median time from LCDD diagnosis to daratumumab initiation was 8.3 years (range 8 - 147 months). Five of the 6 patients obtained hematologic response with at least a reduction of 50% of the dFLC value (partial response). Three patients obtained a very good partial response (dFLC <40 mg/L). The estimated glomerular filtration rate improved in one subject (from 30 to 45 mL/min per 1.73 m2) and in all the others remained stable. In 2 subjects treatment was temporarily discontinued due to pneumonia.
Conclusions: This is the first report of the use of daratumumab in LCDD. This antibody yielded rapid and significant hematologic responses in five of six heavily pretreated patients with this disease, preventing renal progression. Daratumumab represents a promising option for these patients and larger, international studies are warranted.

Disclosures
Merlini: Akcea: Consultancy; Ionis: Consultancy; Prothena: Consultancy; Millenium: Consultancy; Janssen: Consultancy; Pfizer: Consultancy. Palladini:Celgene: Other: Travel support; Janssen: Membership on an entity's Board of Directors or advisory committees; Prothena: Honoraria.
The NUP98-HOXD13 Fusion Oncogene Induces Thymocyte Self-Renewal Via Lmo2/Lyl1
Authors: Shields Benjamin J. , Slape Chris , Jackson Jacob T. , Vo Anh , Shi Wei , Curtis David J. , McCormack Matthew P. 
Published: October 2019

Abstract
T-cell Acute Lymphoblastic Leukaemias (T-ALL) cases include subfamilies that overexpress the TAL1/LMO, TLX1/3 and HOXA transcription factor oncogenes. Whilst it has been shown that TAL1/LMO transcription factors induce self-renewal of thymocytes, whether this is true for other transcription factor oncogenes is unknown. To address this, we have studied NUP98-HOXD13-transgenic (NHD13-Tg) mice, which overexpress HOXA transcription factors throughout haematopoiesis and develop both myelodysplastic syndrome (MDS) progressing to acute myeloid leukemia (AML) as well as T-ALL. We find that thymocytes from preleukemic NHD13-Tg mice can serially transplant, demonstrating that they have self-renewal capacity. Transcriptome analysis shows that NHD13-Tg thymocytes exhibit a stem cell-like transcriptional program closely resembling that induced by Lmo2, including Lmo2 itself and its critical cofactor Lyl1. To determine whether Lmo2/Lyl1 are required for NHD13-induced thymocyte self-renewal, NHD13-Tg mice were crossed with Lyl1 knockout mice. This showed that Lyl1 is essential for expression of the stem cell-like gene expression program in thymocytes and self-renewal. Surprisingly however, NHD13 transgenic mice lacking Lyl1 showed accelerated T-ALL and absence of transformation to AML, associated with a loss of multipotent progenitors in the bone marrow. Thus, multiple T-cell oncogenes induce thymocyte self-renewal via Lmo2/Lyl1, however NHD13 can also promote T-ALL via an alternative pathway.

Disclosures
Curtis: MERCK: Membership on an entity's Board of Directors or advisory committees; CRC Cancer Therapeutics: Patents & Royalties, Research Funding.
Pyrimido-Indole Derivative, UM171 Expands Distinct Types of Myeloid and Lymphoid Progenitors from Human Pluripotent Stem Cells
Authors: Mesquitta Walatta-Tseyon , Kang Hyunjun , Wandsnider Matthew , Brok-Volchanskaya Vera , Suknuntha Kran , Slukvin Igor 
Published: October 2019

Abstract
Human pluripotent stem cell (hPSCs) have created alternative platforms for producing blood cells for transfusion, immunotherapies, and transplantation. Advancing blood cell manufacturing from hPSCs and translating hPSC-based technologies to the clinic requires improving the scalabilty of blood cell production through enhancing hematopoietic differentiation from hPSCs, and increasing expansion of lineage committed hematopoietic progenitors. The pyrimido-indole derivative UM171 has been described as one the most potent small molecules agonists for HSC expansion in vitro. However, the effect and mechanism of UM171 action on hPSC-derived hematopoietic progenitors (HPs) has not been explored. It also remains unclear whether UM171 selectively expands the most primitive multipotential HPs with lin-CD34+CD43+HSC phenotype, or affects progenitors already committed to a particular hematopoietic cell lineage. In our studies, we evaluated the effect of UM171 on the expansion and differentiation of CD34+CD43+HPs that were generated in chemically defined, serum- and feeder-free conditions from hPSCs. We revealed that culture of hPSC-derived HPs in HSC expansion conditions (SFEM with added TPO, SCF, FLT3, IL3 and IL6) in the presence of UM171 selectively expanded HPs with a unique CD34+CD41loCD45+phenotype. Isolation of this population by FACS revealed that it mostly possesses myeloid potential and is highly enriched in granulocytic progenitors (G-CFCs). In contrast, in lymphoid cultures on OP9-DLL4 in presence of SCF, Flt-3 and IL7, UM171 predominately expands CD34+CD7+CD41a-lymphoid progenitors with NK cell potential and increases up to 10-fold NK cell output in NK differentiation cultures. NK cells generated with UM171 possess strong cytotoxicity, express perforin and upregulate IFNg production, following stimulation with K562 or PMA. As determined by annexin V immunostaining, UM171 treatment decreased the number of apoptotic HPs in expansion cultures. In addition, UM171 expansion of HPs was associated with increased proliferation, as determined by BrdU assay and Ki67 staining. Extending these observations, cell cycle analysis revealed that UM171 predominantly increases the proportion of HPs in the early S phase of the cell cycle. These studies should improve our understanding of the effect of UM171 on de novo generated HPs and facilitate development of protocols for robust granulocyte and lymphoid cell production from hPSCs for adoptive immunotherapies.

Disclosures
Slukvin: Cynata Therapeutics Limited: Consultancy, Equity Ownership.
Nuclear Factor I-X May Regulate a Myeloid-Biased Hematopoietic Stem Cell Population during Stress Hematopoiesis
Authors: Walker Megan , Hall Trent , Brown Scott A , McKinney-Freeman Shannon 
Published: October 2019

Abstract
Hematopoietic stem and progenitor cells (HSPC) are routinely exploited in the clinic to treat patients with cancers or other hematologic diseases. The transcription factor nuclear factor I-X (NFIX) has a proven role regulating migration, adhesion, differentiation and quiescence in neural stem cells. Importantly, Nfix was recently shown to be upregulated in FLT3-ITD+ acute myeloid leukemia (AML) patient samples and the NFI binding motif was protected during DNase hypersensitivity screening. We recently identified Nfix as a novel regulator of HSPC repopulating potential. Using our mouse models, Nfixflox/floxRosa26-CreERT2 and Nfix+/+Rosa26-CreERT2, we induced the deletion of Nfix and performed competitive transplants; here we observe a 50% increase in chimerism 20 weeks post-transplant accompanied by a 10% increase in reconstitution of peripheral blood (PB) B cells. Primary recipient whole bone marrow (WBM) was then transplanted into secondary recipients. Beginning four weeks post-transplant, a 2.5-fold loss of PB myeloid reconstitution was observed in secondary recipients of Nfix-deficient WBM. This effect persisted beyond 20 weeks post-secondary transplant, suggesting that a self-renewing myeloid-biased HSPC is functionally perturbed by the loss of Nfix. To identify genes transcriptionally regulated by NFIX, we recently validated an anti-NFIX monoclonal antibody that is currently being used for chromatin immunoprecipitation (ChIP) followed by paired-end sequencing. Future efforts will focus on investigating Nfix-deficient myeloid-biased HSC subsets. This study will illuminate the transcriptional control of specific HSC subsets.

Disclosures
No relevant conflicts of interest to declare.
Clinical Efficacy of Therapies for Transfusion-Dependent Non-Severe Aplastic Anemia: A Retrospective Cohort Study in Multiple Hospitals
Authors: Lin Shengyun , Yang Yan , Yin Hua , Huang Chang , Chen Jian , Wu Liqiang , Shen Jianping , Ye Baodong , Hu Zhiping , Shen Yiping , Zheng Zhiyin , Shen Wenyi , Lu Xingyu , Wu Xuemei , He Guangsheng , Li Jianyong 
Published: October 2019

Abstract
BACKGROUND: This study aims to evaluate the clinical efficacy and prognosis of currently popular treatments for patients with transfusion-dependent non-severe aplastic anemia (TD-NSAA).
METHODS: Medical records of patients with TD-NSAA were collected from three hospitals and divided into four groups based on their treatment. Treatment efficacy was evaluated at 6 months after treatment, with subgroup analysis based on patients' disease courses. Adverse events and 3-year survival status were also reviewed.
RESULTS: A total of 175 patients' records were collected in this study. The analysis of treatment efficacy indicated that the most effective treatment was allo-hematopoietic stem cell transplant (HSCT) with the response rate of 84.62%, which was followed by Anti thymocyte globulin and ciclosporin A (ATG + CsA) (68.52%), CsA + androgen (26.92%), and androgen (0%), respectively. The subgroup analysis showed that for patients with less than 6-month disease courses, ATG + CsA treatment could achieve similar response rate as allo-HSCT (84.00%). Besides, ATG + CsA showed better efficacy than CsA + androgen in all stratified arms (P<0.05). Three-year event-free survival rates were higher in patients treated with ATG + CsA (83.3%) than with CsA + androgen (64.4%) (P=0.013).
CONCLUSIONS: Therefore, we concluded that ATG+ CsA could achieve better efficacy to treat TS-NSAA than did CsA+ androgen.

Disclosures
No relevant conflicts of interest to declare.
Dynamins 2 and 3 Are Required for Human Megakaryocytes Directional Migration
Authors: Suraneni Praveen K , Corey Seth J. , Hession Michael , Ishaq Rameez , Awomolo Arinola , Hasan Shirin , Debili Najet , Crispino John D. , Eklund Elizabeth A. , Chen Yolande 
Published: October 2019

Abstract
Abstract:
Megakaryocytes (MKs) undergo directional migration from the proliferative osteoblastic niche within the bone marrow (BM) environment to the capillary-rich vascular niche for platelet production and release into the pulmonary circulation. This process is regulated, in part by dynamins, large GTPase proteins that regulate cellular functions such as endocytosis, vesicle transport and cell migration. Additional functions of dynamins include the formation of actin-rich structures, such as lamellipodia and dorsal membrane ruffles, invadopodia and podosomes. Previous studies have shown that mutations in Dynamin 2 (DNM2) cause thrombocytopenia in humans. To explore the function of dynamins in megakaryocyte migration and platelet production in more depth, we monitored the response of cells to chemotaxis SDF1α gradient signal by a microfluidic device-based approach. We observed an impaired directional migration by both human megakaryocytic cell lines and primary cells treated either with dynasore, a small molecule inhibitor of dynamins, or shRNA knockdown of Dynamin 2 and 3 (DNM2, DNM3). Since directional cell migration is tightly regulated by actin cytoskeletal rearrangements, we next measured actin polymerization and RhoA activity. We observed a profound decrease in the F-actin and Rho GTPase activity upon loss of DNM2 and DNM3 function. Next, since the response to chemoattractant signal is navigated by SDF1 through its receptor CXCR4, we explored the CXCR4 receptor response to ligand in dynamin defective megakaryocytes. Interestingly we observed an increase in CXCR4 expression in the dynasore treated primary human cells. Additionally, combined inhibition of DNM2 and DNM3 or over expression of dominant negative Dnm2-K44A or GTPase-defective DNM3 decreased the active β1- integrin (ITGB1) activity, which indicates a decrease in the integrin mediated endo/exocytic cycling during cell migration. Finally, to understand the role of dynamin in endosome recycling, we assayed the distribution of Rab11, a marker of recycling endosomes. We noticed an abnormal clustered staining pattern of Rab11 in dynasore-treated MKs which is indicative of a disruption in recycling pathways. This observation suggests decreased recruitment of the recycling pathway in dynasore-treated cells. Altogether, in this study we demonstrate that dynamins regulate MKs directional migration towards the SDF1α chemotaxis signal in the bone marrow and governs endocytosis and cell receptor trafficking.

Disclosures
Crispino: Scholar Rock: Research Funding; Forma Therapeutics: Research Funding.