bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hhex Is a Critical Gene In The Development Of Normal and Malignant Lymphoid Cells
Authors: Goodings Charnise , Smith Stephen B. , Mathias Elizabeth , Smith Elizabeth , Tripathi Rati , Elliott Natalina E , Cleveland Susan , Dave Utpal P. , Du Yang 
Published: October 2019

Abstract
Hematopoietically expressed homeobox (Hhex) is a T-cell oncogene. It is frequently deregulated in murine retroviral insertional mutagenesis screens and its enforced expression induces T-cell leukemia in bone marrow transduction and transplantation experiments. We discovered that HHEX is a direct transcriptional target of an LIM domain Only-2 (LMO2)-associated protein complex. HHEX clusters with LMO2-overexpressing T-ALLs and is especially overexpressed in Early T-cell Precursor (ETP) – ALL where it is a direct transcriptional target of LMO2. To further understand Hhex's function, we induced a conditional knockout in floxed Hhex mice with the Vav-iCre transgene. Mice were viable and showed normal blood cell counts with highly efficient deletion of Hhex in all hematopoietic tissues. Thymocytes from conditional knockouts showed a normal pattern of development. Most impressively, Hhex conditional knockout markedly prolonged the latency of T-ALL onset in CD2-Lmo2 transgenic mice (figure 1). Hhex conditional knockouts (Hhex cKOs) also had a significant decrease in mature B cells in the spleen and bone marrow. Interestingly, hematopoietic stem and progenitor cells plated on OP9-GFP or OP9-DL1 stromal cells showed proliferative defects and incomplete differentiation towards both B and T lineage. Also under stress conditions such as sublethal irradiation and competitive bone marrow transplants, Hhex conditional knockouts show a marked defect in both B and T lineages but an increase in early progenitor populations. Our experiments show that Hhex is a critical transcription factor in lymphoid development and in LMO2-induced T-ALL.Figure 1Hhex conditional knockout markedly prolonged the latency of T-ALL onset in CD2-Lmo2 transgenic miceFigure 1. Hhex conditional knockout markedly prolonged the latency of T-ALL onset in CD2-Lmo2 transgenic mice

Disclosures:
No relevant conflicts of interest to declare.
Cytidine Deaminase Status As a Predictive Marker Of Response and Toxicity In Patients With Haematological Malignancies Treated With Azacytidine Or Cytarabine
Authors: Mercier Cédric , Fanciullino Raphaelle , Serdjebi Cindy , Ciccolini Joseph , Pourroy Bertrand , Milano Gerard , Ouafik L’Houcine , Costello Régis T 
Published: October 2019

Abstract

Background
Cytarabine and azacytidine are mainstays for treating haematological malignancies. As most nucleosidic analogs, both azacytidine and cytarabine are metabolized in the liver by an exclusive enzymatic step driven by cytidine deaminase (CDA). CDA expression polymorphism has been associated with clinical outcome in patients treated with gemcitabine.

Methods
We determined CDA levels in 39 adult patients (16F, 23M, mean age 77 years), treated mainly for AML and myelodysplastic syndromes with either aza-cytidine or a cytarabine-containing regimen. Response and treatment-related toxicities were monitored following current standards. In addition, impact of CDA status on azacytidine tolerance was evaluated in mice with or without CDA deficiency, as a proof of concept for the actual implication of metabolic deregulations in the toxicities observed in patients.

Results
In patients, mean CDA activity was 3.7 +/-2.8 U/mg (min: 1, max: 14.8 U/mg). Ten out of 39 patients (i.e., 25%) had low CDA activities and were considered as poor metabolizers (PM). Conversely, 8 patients (i.e., 20%) displayed high CDA activities (i.e., > 6U/mg) and were considered as high metabolizers (HM). PM showed severe toxicities, including two toxic-deaths. Conversely, HM showed little efficacy when treated with either azacytidine or cytarabine. In mice with CDA-deficiency, standard dose azacytidine led to profound and long-lasting neutropenia, as compared with normal mice. Drug monitoring confirmed that individuals with low CDA activity and toxicities showed higher concentrations of azacytidine as compared with normal individuals.

Conclusions
this study suggests that CDA status could be a relevant marker for predicting clinical outcome in patients treated with either azacytidine or cytarabine. CDA status could be further used as a covariate to tailor drug dosage so as to ensure an optimal efficacy/toxicity balance in patients with haematological malignancies.

Disclosures:
No relevant conflicts of interest to declare.
Second Primary Malignancies In Adult Acute Lymphoblastic Leukemia
Authors: Shah Binay K. , Ghimire Krishna B 
Published: October 2019

Abstract

Background
Pediatric acute lymphoblastic leukemia (ALL) patients have higher rates of second primary malignancies. There is limited data on second primary malignancies (SPM) among adult patients with ALL. This study was conducted to evaluate SPM in adult ALL patients using US Surveillance, Epidemiology and End Results (SEER) cancer registry database.

Methods
We analyzed the SEER 13 Registries using multiple primary standardized incidence ratio (MP-SIR) session. We analyzed secondary cancer rates among adult ALL patients during the period 1992 - 2010. We used SEER*Stat software provided by national cancer institute for statistical analysis.

Results
There were 3,259 adult (age ≥20 years) ALL patients reported in SEER database during 1992-2010. Among them, 65 ALL patients developed 75 second primary malignancies. Fifty-nine ALL patients developed 1 SPM each, 3 ALL patients developed 2 SPM each, 2 ALL patients developed 3 SPM each and 1 ALL patient developed 4 SPM. All site cancers were significantly higher among adult ALL patients compared to general population with observed/expected ratio (O/E): 1.47, p value< 0.05, an absolute excess risk of 24.43 per 10,000 populations. Similarly, oral cavity cancer, respiratory system cancer and hematological SPM were significantly higher in ALL patients than expected in general population. (Table)

Conclusions
Adult patients with ALL have higher rates of second primary malignancies compared to general population.

Disclosures:
No relevant conflicts of interest to declare.
Differentiation Induction In Acute Myeloid Leukemia Using Site-Specific DNA-Targeting
Authors: Palchaudhuri Rahul , Ang Kwan-Keat , Saez Borja , Sykes David B. , Verdine Gregory L. , Scadden David T. 
Published: October 2019

Abstract
Hoxa9 and Meis1 are overexpressed in >70% of acute myeloid leukemia (AML) and associated with poor prognosis and survival. Hoxa9 and Meis1 interact with DNA and PBX to achieve transcription of differentiation-blocking genes. We tested transcriptional repression at Hoxa9-PBX-Meis1 genomic binding sites to induce differentiation in a model of human AML
We designed a DNA-recognition strategy based on the known structure of the Hoxa9-PBX-DNA complex by fusing the DNA binding helices of Hoxa9 and PBX to create concise homeodomain fusion proteins that target the Hoxa9-PBX DNA recognition sequence. To confer transcription-repressing properties to the proteins, we attached a transcriptional repressor (sin3 interacting) domain and ectopically expressed this protein in Hoxa9-Meis1 immortalized murine progenitors. Introduction of this transcription repressor protein significantly enabled cell differentiation versus control (51.2% Mac-1high Gr-1high cells versus 11.3% for control). Multiple gene transcripts indicative of differentiation, such as GCSFR, myeloperoxidase, neutrophil elastase, and the calcium binding protein, S100A8, were also elevated in repressor-expressing cells. Furthermore, direct transcriptional targets of Hoxa9 (e.g. SOX2, CD34, FOXP1, FLT3R, DNAJC10) were down regulated in repressor-expressing cells. Importantly, a mutant repressor lacking the DNA-interacting amino acids did not affect transcription of Hoxa9 targets, demonstrating on-target specificity. Repressor-expressing cells also exhibited lower surface expression of c-Kit and Flt3 receptors and when transplanted into mice resulted in a significant increase in disease latency with a 94 day median latency versus 62 day latency for the control group (p value = 0.002).
Our results demonstrate that site-specific DNA-targeting using homeodomain fusion proteins can enable AML cell differentiation and significantly increase disease latency.

Disclosures:
Scadden: Fate Therapeutics: Consultancy, Equity Ownership.
CD34+CXCR4(CD184)+ Cells Differentiate Into Myeloid Dendritic Cell Progenitors
Authors: Donahue Robert E. , Jin Ping , Uchida Naoya , Bonifacino Aylin C. , Tian Yu , Klinman Johanna I. , Mattapallil Joseph J , Metzger Mark E. , Williams Ann , Malide Daniela , Tisdale John F. , Stroncek David 
Published: October 2019

Abstract
Dendritic cells (DCs) play a central role in innate immunity and adaptive immunity. DCs are antigen presenting cells capable of inducing primary T cell responses or facilitating self-tolerance. Myeloid DCs (mDC) express CD11c. They are further divided into Type 1 myeloid DCs (mDC1) which are CD1c+CD141+ and Type 2 mDCs (mDC2) which are CD1c-CD141+, and plasmacytoid DC which are CD11c- and express CD303. Plasmacytoid DCs are the main source of type 1 interferon upon infection. CD34+ cells are a heterogeneous population of cells which contain both hematopoietic progenitors and stem cells. The microarray signature of CD34+CXCR4 (CD184)+ cells in both human and non-human primates suggest that this cell population plays a role in innate immunity. The signaling pathway for the Triggering Receptor Expressed on Myeloid Cells 1 (TREM1) is the most up-regulated pathway in both human and non-human primate sorted CD34+CD184+ cells. The TREM family is involved in the amplification and regulation of inflammatory responses. Upon sorting both human and rhesus CD34+ cells into CD34+CD184+ and CD34+CD184-, we observe that CD34+CD184+ cells are both adherent and non-adherent in nature (Figure 1) and while maintained in Flt3-L, IL-3, and SCF form progeny which appear dendritic in nature (Figure 2). In addition, we have found the CD34+CD184+ subpopulation to be resistant to lentiviral vector transduction. Upon culturing in defined, serum free media supplemented with cytokines, the progeny of the CD34+CD184+ population using both flow cytometry and confocal microscopy are CD11c+ and contain both CD1c+ and CD1c- subpopulations (Figure 3) with a predominately CD80(B7-1)+, CD123(IL-3R)+, CD197 (CCR7)+ , and HLA-Dr+ immunophenotype, having variability in CD86(B7-2) +/-, CD141(BDCA-3/Thrombomodulin)+/- , and CD144 (VE-cadherin)+/- expression, and being negative for CD303(BDCA-2)and CD309(VEGFR). Of special interest is that the CD34+CD184+ progeny contain both CD1c-CD16+ and CD1c+CD16- subpopulations of mDC. CD1c-CD16+ mDC have been shown to have strong pro-inflammatory activity, whereas CD1c+CD16- mDC are mainly inducers of chemotaxis. The progeny of the CD34+CD184+ cells can be stimulated by LPS and IFNγ to produce IL-12 based on an enzyme-linked immunosorbent assay. These results demonstrate the importance of the CD34+CXCR4+ progenitor in mDC development and allow one to speculate on how this mDC progenitor might prove of therapeutic benefit in vaccine development and cancer therapy.Figure 1Human CD34+CD184+ Sorted PopulationFigure 1. Human CD34+CD184+ Sorted PopulationFigure 2Cultured Progeny CD34+CD184+Figure 2. Cultured Progeny CD34+CD184+Figure 3Non-Human Primate CD34+CD184+ Confocal MicroscopyFigure 3. Non-Human Primate CD34+CD184+ Confocal Microscopy

Disclosures:
No relevant conflicts of interest to declare.
Characterization Of a Weakly Expressed KIR2DL1 Allele
Authors: Li Hongchuan , Huehn Andrew R , Fellow Postbaccalaureate , Wright Paul W , Biologist Research , Cooley Sarah , Miller Jeffrey S. , Anderson Stephen K 
Published: October 2019

Abstract
The variegated expression of the human KIR family of class I MHC receptors provides an interesting model system for the study of stochastic activation of gene transcription. Previous studies have linked distal KIR promoter transcription to the initiation of KIR expression from the proximal promoter. In order to identify novel genetic alterations associated with decreased KIR expression, a group of 182 donors was characterized for KIR gene content, KIR transcripts, and FACS analysis of KIR surface expression. An individual was discovered that possessed a single copy of the KIR2DL1 gene but had a low level of gene expression by either FACS or Q-PCR. Complete sequencing of the KIR2DL1 gene confirmed the presence of an intact coding region. Analysis of promoter elements revealed a cluster of three single nucleotide polymorphisms (SNPs) in the distal promoter approximately 1 kb upstream from the start of KIR2DL1 translation. These SNPs are also found in the distal promoter region of the non-transcribed KIR2DL5*002 allele as well as the KIR3DP1 pseudogene. One of these SNPs creates a functional binding site for the ZEB1 transcription factor. Individuals possessing the ZEB1 site in their KIR2DL1 promoter produce high levels of a non-translatable distal KIR2DL1transcript that inhibits transcription from the proximal promoter, resulting in weak expression of this allele.
This research was supported in part by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. Funded by NCI Contract HHSN261200800001E.

Disclosures:
Miller: Coronado Biosciences: Scientific Advisory Board Other.
Oncomirnas and Tumor Suppressors In Microvesicles From Four Types Of Cancer
Authors: Li Wenying , Xiong Wei , Chen Xiaomei , Huang Shiang , Huiyu Li 
Published: October 2019

Abstract

Objectives
Microvesicles (MVs) are small vesicles that are shed from almost all cell types including cancer cells into their surroundings. These secreted MVs bear surface receptors/ligands and miRNA of the original cells and play a role in cancer. Our aim in this study was to identify MVs miRNAs that might play important roles both in solid and non-solid tumors.

Methods
The MVs miRNA expression from 4 kinds of cancer cell lines (SMMC-7721, K562, Nalm6 and Jurkat) and healthy peripheral blood cells were analyzed by miRNA microarray. Quantitative real-time polymerase chain reaction (qRT-PCR) was further performed to verify the expression of 4 dysregulated miRNAs.

Results
The miRNA microarray revealed aberrantly expressed miRNAs of MVs between the four types of cancer cells and normal cells. In the MVs from cancer groups, 73 miRNAs were up-regulated and 148 were down-regulated in all the four types of cancer cell lines, indicating that solid and non-solid tumors shared some dysregulated MVs miRNAs. qRT-PCR verified statistically consistent expression of four selected miRNAs with microarray analysis. The top six shared upregulated MVs miRNAs in the four cancer cells lines were miR-1290, miR-1268, miR-1246, miR-125a-3p, miR-1305, miR-1226*. The top seven shared downregulated MVs miRNAs in the four cancer cells lines were miR-335, miR-1, miR-363, miR-122, miR-223, miR-338-3p and miR-340.
The general roles of the upregulated MVs miRNAs were oncomiRNAs in multicancer. MiR-1290 and its potential targets are associated with characteristics of estrogen receptor α-positive breast cancer. Up-regulations of miR-1290 impaired cytokinesis and affected the reprogramming of colon cancer cells. miR-1246 might serve as a likely link of the p53 family with Down syndrome and proposed a new p53-miR-1246-DYRK1A-NFAT pathway in cancer.
The functional roles of the downregulated MVs miRNAs were tumor suppressors in many kinds of cancer. MiR-335 acted as a candidate tumor suppressor and inhibited tumor reinitiation and represented an invasion suppressor gene by targeting Bcl-w in many kinds of cancer. HBx regulated miR-338-3p in HCC and miR-338-3p inhibited proliferation by regulating CyclinD1, and miR-338-3p suppressed the invasion of liver cancer cell by targeting smoothened gene in liver cancer. miR-340 inhibited of breast cancer cell migrations and invasions through targeting of oncoprotein c-Met and may play an important role in breast cancer progression.

Conclusion
Our study identifies oncomiRNAs and tumor suppressors in MVs from four types of cancer. These MVs miRNAs may contribute to the developments of solid and non-solid tumors and should be further evaluated as a novel biomarker for cancers and may provide potential targets for novel therapeutic strategies.

Disclosures:
No relevant conflicts of interest to declare.
Src Family Kinase Inhibitor PP2 Enhances Differentiation Of Acute Promyelocytic Leukemia Cell Line Induced By Combination Of ATRA and Arsenic Trioxide
Authors: Kim Kyoung Ha , Cheong Hee-Jeong , Kim Sook-Ja , Yoon Jina , Kim Han Jo , Kim Se Hyung , Kim Hyun Jung , Lee Sang-Cheol , Bae Sang Byung , Kim Chan-Kyu , Lee Namsu , Lee Kyu Taeg , Park Sung Kyu , Hong Dae Sik , Park Hee Sook , Won Jong-Ho 
Published: October 2019

Abstract
All-trans-retinoic acid (ATRA) and arsenic trioxide (ATO) combination yields a high quality remission and survival in newly diagnosed acute promyelocytic leukemia. For subsequent similar data, NCCN guidelines indicate that ATRA plus ATO is an alternative for patients who cannot tolerate anthracycline therapy. We demonstrated that SFK (Src Family Kinase) inhibitor PP2 enhanced acute promyelocytic leukemia (APL) cell differentiation when combined with either ATRA or ATO with difference in activation of RA-induced genes. In this study, we investigated SFK inhibitor PP2 could enhances the differentiation of NB4 APL cells when combined with ATRA and ATO and the changes in the expression of intercellular adhesion molecule-1 (ICAM-1) derived from the retinoic acid receptor (RAR) target gene.
Treatment of NB4 cells with 1 nM of ATRA, 0.5 uM of ATO, or 10 uM of PP2 for 72 hours induced expression of CD11b-positive cells by 13.01%, 11.53% or 13.28%, respectively. However, the combination of ATRA and ATO and the combination of three agents (ATRA, ATO, and PP2) led to a significant higher expression of CD11b-positive cells (30.96% and 63.17%, respectively). The synergistic effect of the combination of three agents was more significant than the combination of ATRA and ATO. These results were confirmed by NBT staining. These effects were not related with apoptosis. Annexin-V-fluorescene staining revealed that combination of ATRA and ATO and combination of three agents did not induced apoptosis in NB4 cells. The expression of ICAM-1 was markedly increased in cells treated with the combination of three agents.
These findings suggest that the SFK inhibitor can enhances differentiation of APL cells combined with ATRA and ATO. FDA approved SFK inhibitors, such as dasatinib and bosutinib, may be beneficial for the treatment of APL in combination with ATRA and ATO.

Disclosures:
No relevant conflicts of interest to declare.
Induced Apoptosis Of Multidrug Resistance Leukemia K562/A02 Cells By Magnetic Nanoparticle Of Fe3O4 Copolymerizating JNK1 shRNA Expression Vector and Daunorubicin
Authors: Zhang Wei , Chen Baoan , He Xiangfeng , Fu Rong 
Published: October 2019

Abstract
Currently, serious toxicity and poor antitumor efficacy resulting from the induction of multidrug resistance (MDR) in hematological malignancies and solid tumors are common hindrances to successful systemic chemotherapy. One of the important mechanisms of tumorigenesis and resistance to anticancer drugs was blockade of the apoptosis-inducing pathway. In order to overcome tumor MDR while maintaining good antitumor activity, biomaterial combine with gene therapy provides a novel strategy for the treatment of MDR.The aim of this study was to explore the induction of apoptosis by magnetic nanoparticle of Fe3O4 (MNPs-Fe3O4), a novel delivery system for chemotherapeutic regimen that is chemically stable, environmentally friendly, and noncytotoxic, copolymerizating JNK1shRNA expression vector with DNR, which could overcome siRNA technique instability and enhance DNR chemotherapeutic efficiency and provide a new field to reverse tumor MDR. JNK1shRNA expression vector was constructed and screened the most effective plasmid and transfected the cells. Typical apoptotic characteristics and apoptosis effect of MNPs-Fe3O4 and PGM-3 with DNR were investigated by DAPI staining and FCM assay, respectively. Additionally, drug accumulation and apoptosis relative-genes were evaluated in K562/AO2 leukemia cells.The results showed that PGCsilencer-U6-neo-GFP-GV102 shRNA/JNK1 was successfully constructed, which was confirmed by sequencing that expression of JNK1 gene in mRNA and protein level that had the less in PGM-3 than that of PGM-1 or PGM-2. Meanwhile, MNPs-Fe3O4 and PGM-3 with DNR (DNR/PGM-3/MNPs-Fe3O4) could synergistically induced typical apoptotic characteristics of chromatin condensation and apoptotic body with DAPI staining, and a higher rate of apoptosis were detected in K562/A02 cells treated with DNR/PGM-3/MNPs-Fe3O4. Intracellular DNR was higher in K562/A02 cells treated with DNR/PGM-3/MNPs-Fe3O4 than that of DNR/MNPs-Fe3O4 by FCM analysis. Further study demonstrated that both DNR/PGM-3/MNPs-Fe3O4 and DNR/PGM-3 reduced the gene transcriptions and protein expressions of bcl-2, enhanced that of bax and caspase-3, decreased survivin in protein level, and has no different change in mRNA level by qRT-PCR and Western blotting analyses, respectively. These findings showed that the combination of MNPs-Fe3O4 with JNK1 shRNA expression vector and DNR could induce apoptosis of K562/A02 cells through elevating the ratio of bax/bcl-2, activating caspase-3, and decreasing survivin in protein level.

Disclosures:
No relevant conflicts of interest to declare.
CUDC-907, a Dual HDAC and PI3K Inhibitor, Potentially Targets Cancer Cells and The Microenvironment In Hematological Malignancies
Authors: Wang Jing , Pursell Natalie , Ma Anna , Atoyan Ruzanna , Samson Maria , Borek Mylissa , DellaRocca Steven , Yin Ling , Wang Da-gong , Zifcak Brian , Xu Guang-xin , Voi Maurizio , Lai Cheng-Jung 
Published: October 2019

Abstract
Histone deacetylase (HDAC) and phosphatidylinositol 3-kinase (PI3K) inhibitors have each emerged as promising anti-cancer therapeutics that may target not only cancer cells but also their microenvironment. We have developed a strategy to simultaneously inhibit HDAC- and PI3K-dependent signaling with a single small molecule, CUDC-907. In this study, we report that CUDC-907 induces apoptosis and G2/M arrest in hematological cancer cells. CUDC-907 also impairs cytokine and chemokine production, in association with inhibition of AKT and STAT phosphorylation. Moreover, we demonstrate that CUDC-907 reduces hypoxia-induced HIF-1a expression in cancer cells and VEGF-induced tube formation in endothelial cells, suggesting potential anti-angiogenic activity. With its integrated HDAC and PI3K inhibitory activity, CUDC-907 may thus offer improved therapeutic benefit through simultaneous suppression of cancer cell proliferation and perturbation of their protective microenvironment. These studies provide support for the clinical development of CUDC-907 in hematological malignancies. A Phase I study in patients with lymphoma or multiple myeloma is ongoing.

Disclosures:
Wang: Curis Inc.: Employment. Pursell:Curis Inc.: Employment. Ma:Curis Inc.: Employment. Atoyan:Curis Inc.: Employment. Samson:Curis Inc.: Employment. Borek:Curis Inc.: Employment. DellaRocca:Curis Inc.: Employment. Yin:Curis Inc.: Employment. Wang:Curis Inc.: Employment. Zifcak:Curis Inc.: Employment. Xu:Curis Inc.: Employment. Voi:Curis Inc.: Employment. Lai:Curis Inc.: Employment.