bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytochromes P450 and Glutathione S-Transferases Polymorphisms: Can They Predict Efficacy of Tyrosine Kinase Inhibitors in Chronic Myeloid Leukemia?
Authors: Mukry Samina N. , Shahni Aneeta , Zaidi Uzma , Shamsi Tahir S. 
Published: November 2019

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm characterized by over production of cells due to expression of an abnormal BCR/ABL1 kinase gene. Tyrosine kinase inhibitors (TKI) are the treatment option to achieve deep and prolonged molecular response in CML patients. Resistance or sub-optimal response to TKI such as imatinib has been reported in 5-20% of the patients carrying wild type of Cytochromes P450 (CYP) and glutathione S-transferases (GST). This study was planned to evaluate possible association of presence or absence of polymorphic variation in CYP1A1, GSTT1 and GSTM1 with response to nilotinib in CML patients
A total of 33 CML patients receiving nilotinib treatment (ELN 2013 guidelines) at National Institute of blood Diseases and Bone Marrow Transplantation were enrolled. Informed consent was obtained as per declaration of Helsinki. The presence (+) or absence (-) of CYP 1A1 (A2455G), GSTT1 and GSTM1 was determined by RFLP and multiplex PCR respectively. Treatment response was evaluated by quantification of BCR ABL1 transcripts by real time qPCR. The chi square test was used to compare the genotype distribution between patients and control. A p value less than 0.05 was considered as statistically significant. Odds ratio (OR) with 95% confidence interval (CI) calculated using statistical package SPSS version 22.
The male:female ratio for the study group was 1.75:1 with a mean age of 37±12 years. About 25, 5 and 3 patients were diagnosed with CML in chronic, accelerated and blast crisis phase respectively. There were 25 responders, 5 partial responders and 3 non-responders to nilotinib treatment altogether. The GSTT1- genotype was most frequent in responders (72%) followed by GSTM1+ and CYP 1A1- (p= <0.05). Interestingly, GSTT1-/GSTM1- and heterozygous CYP 1A1+ were less frequent (8% and 20%) but observed only in responders.
This is the first study on nilotinib response and CYP1A1, GSTT1, GSTM1 polymorphisms in CML from Pakistan. Null or GSTT1-, GSTM1+ and CYP 1A1- seemed to be frequent in nilotinib responders only and may have influence on drug response.

Disclosures
No relevant conflicts of interest to declare.
Myeloid Tumor-Related Somatic Mutations in 293 Children and Adults with Acquired Aplastic Anemia in Chinese
Authors: He Hailong , Yin Hua , Liu Xiaoqing , Cheng Lang , He Guangsheng , Li Jianyong 
Published: November 2019

ABSTRACT
Objectives and Methods:
It was postulated that the subset of aplastic anemia ( AA ) with somatic mutations typically seen in myeloid malignancies were at risk of clonal evolution[1,2]. But the response rate to immunosuppressive therapy (IST) was different or not between patients with or without mutations, remain unclear[1,2]. Age of patients with AA is a strong predictor of outcome for IST, and the presence of somatic mutations also rise appreciably in frequency with age[3]. Here we checked myeloid tumor-related somatic mutations from 293 children and adult patients who have acquired AA. We excluded Fanconi anemia, mutations of telomere maintenance, and a family history of BM failure (BMF) or cancer. We looked for somatic mutations by identifying previously characterized single-nucleotide variants in 25 genes that are recurrently mutated in myeloid malignancies in the peripheral blood cells by second-generation gene sequencing. And clinical significance of mutations in different ages was also annalyzed.
Results :
Somatic mutations in myeloid tumor-related genes are present in 6.48% of the patients. The the variants occurred in ten genes: ASXL1 (n = 2 ), KRAS (n = 1), PIGA (n = 2 ), TP53 (n = 2 ), BCOR (n = 2 ), TET2 (n = 5 ), SF3B1 (n = 1), DNMT3A (n = 2 ), SH2B3 (n = 1), MPL (n = 1) (Figure 1). The majority of the variants occurred in epigenetic genes: TET2, DNMT3A, ASXL1. Among patients of children and teen agers' group, adult group, the clonal mutations were observed in 2.82%, 9.93% (P= 0.013) (Figure 2). The presence of a somatic mutation was not associated with gender ( male or female, 3.87% vs 9.42%, P=0.054), severity ( non severe AA or severe AA, 7.87% vs 4.35%, P=0.233). Patients with somatic mutations had a simillar response rate, compared with patients without mutations in children and teen agers' group, or adult group. Somatic mutations in AA patients were not associated with high risk of transformation to MDS (P > 0.5 ).
Conclusions:
Age-related clonal hematopoiesis rise along age in patients with AA in chinese, but somatic mutations in AA patients were associated with efectiveness of IST, and risk of clonal evolution.
References
1 Kulasekararaj AG, Jiang J, Smith AE, et al. Somatic mutations identify a subgroup of aplastic anemia patients who progress to myelodysplastic syndrome. Blood. 2014; 124(17):2698-2704).
2 Yoshizato T, Dumitriu B, Hosokawa K et al. Somatic Mutations and Clonal Hematopoiesis in Aplastic Anemia. N Engl J Med. 2015 Jul 2; 373(1):35-47.
3 Jaiswal S, Fontanillas P, Flannick J, et al. Age-Related Clonal Hematopoiesis Associated with Adverse Outcomes. N Engl J Med 2014; 371:2488-98

Disclosures
No relevant conflicts of interest to declare.
Potential Metabolites with Diagnostic Value in Plasma for Angioimmunoblastic T-Cell Lymphoma By LC-MS Based Untargeted Metabonomics Study
Authors: Yang Haiyan , Lei Tao , Li Cong , Yu Haifeng , Chen Zhongjian 
Published: November 2019

Background: Angioimmunoblastic T-cell lymphoma (AITL) is an uncommon subtype of mature peripheral T-cell lymphoma (PTCL). and its characteristics is not fully understood. Because it is a rare form of lymphoma, there is difficulty in diagnosis based on current traditional method. Metabolic reprogramming is a major cancer hallmark, which provides cancer cells with vital energy and metabolites. Mass spectrometry-based metabolomics has emerged as an informative technique for profiling metabolic features associated with cancers.
Objective: This study aim to discover metabolites with potential diagnostic value in plasma of AITL patient.
Methods: Plasma was collected from 27 patients newly diagnosed with AITL in Zhejiang Cancer Hospital and 30 age- and gender- matched healthy controls. An aliquot of 50 μL plama was added with 150 μL of chilled acetonitrile, vortexed and centrifuged. the supernatant was dried, reconstituted for metabolomics. Mass spectrometry analysis was performed on a UPLC coupled with a Q Exactive Orbitrap mass spectrometer. "XCMS" and "MetaX" were used to analyze LC-MS data. Significant features were selected as candidate for identification by matching their MS and MS/MS data with online databases HMDB and METLIN.
Results: Eighteen altered metabolites (choline, betaine, L-leucine, creatine, 2-butyl-1H-benzimidazole, hypoxanthine, methionine sulfoximine, L-carnitine, 2-hydroxycinnamic acid, L-tyrosine, D-tryptophan, indoleacrylic acid, lysoPC(P-18:0), 5-aminopentanoic acid, uric acid, sucrose, L-lactic) were indentified (VIP value>1, p<0.05) in AITL patient plasma. The altered metabolites contributed to different metabolic pathways including glycine, serine and threonine metabolism (P=0.003), glycerophospholipid metabolism (P=0.024), Phenylalanine metabolism (P=.030).
Conclusion: This study revealed metabolic reprogram and potential diagnostic metabolites in plasma for AITL, and gave more metabolic information for carcinogenesis of AITL.

Disclosures
No relevant conflicts of interest to declare.
The SQSTM1-NUP214 Fusion Protein Cooperates with Crm1 to Activate Hoxa Genes and Drives Leukemogenesis in Mice
Authors: Aumann Waitman K. , Sze Sei-Gyung K. , Gupta Veerain , Ripple Katelyn , Port Sarah , Kehlenbach Ralph , Wechsler Daniel S. , Lavau Catherine P. 
Published: November 2019

Background: The NUP98 and NUP214 nucleoporins (NUPs) are recurrently fused to heterologous proteins in leukemia. The resulting chimeric oncoproteins retain the NUP phenylalanine-glycine (FG) repeat motifs that mediate interaction with the nuclear export receptor Crm1. NUP fusion leukemias are characterized by HOXA gene upregulation; however, their molecular pathogenesis remains poorly understood. To investigate the role of Crm1 in mediating the leukemogenic properties of NUP chimeric proteins, we studied the SQSTM1-NUP214 fusion. Methods: We synthesized a SQSTM1-NUP214 fusion protein which retains only a short C-terminal portion of NUP214 containing FG motifs that mediate interaction with Crm1, and then introduced point mutations targeting these FG motifs (SQSTM1-NUP214FGmut). We compared the activity of these two fusion proteins using co-immunoprecipitation with CRM1, methylcellulose colony assays, murine transplantation, RT-qPCR, and chromatin immunoprecipitation. Results: We found that the ability of the SQSTM1-NUP214FGmut protein to interact with Crm1 was reduced by more than 50% compared with SQSTM1-NUP214. Further, mutation of FG motifs affected transforming potential: while SQSTM1-NUP214 conferred robust colony formation to transduced hematopoietic progenitors in a serial replating assay, the effect of SQSTM1-NUP214FGmut was greatly diminished. Moreover, SQSTM1-NUP214 caused myeloid leukemia in all transplanted mice (6/6), whereas none of the SQSTM1-NUP214FGmut reconstituted mice developed leukemia (0/7). These oncogenic effects coincided with the ability of SQSTM1-NUP214 and SQSTM1-NUP214FGmut to upregulate the expression of Hoxa and Meis1 genes in hematopoietic progenitors. Indeed, chromatin immunoprecipitation assays demonstrated that impairing the interaction of SQSTM1-NUP214 with Crm1 reduced binding of the fusion protein to Hoxa and Meis1 loci. Conclusions: These findings highlight the importance of Crm1 in mediating the leukemogenic properties of SQSTM1-NUP214, and suggest a conserved role of Crm1 in recruiting oncoproteins to their effector genes.

Disclosures
No relevant conflicts of interest to declare.
Creating and Testing a CD207+ Langerhans Cell Histiocytosis-like Cell Line
Authors: Velazquez Jessica , Sengal Amel , Allen Carl E. , Chakraborty Rikhia 
Published: November 2019

Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplasia characterized by pathological CD207+ dendritic cells (DCs) with persistent mitogen-activated protein kinase (MAPK) activation. Investigations into LCH have historically been challenged by the small percentage of pathologic CD207+ DCs. No cell lines with morphology or function representing LCH lesion CD207+ DCs currently exist. We utilized four strategies to generate cell line(s) mimicking differentiated LCH pathogenic cells. First, CD207+ cells were isolated from the lesion of an LCH patient and cultured in a cytokine cocktail. The cells maintained CD1a and CD207 expression for two weeks, after which there were significant changes in cell morphology and progression to cell death. When a similar approach was implemented to isolate and culture skin CD207+ cells, the cells were viable for only three days, supporting the potential role for somatic activating MAPK mutations in LCH lesion DCs to prolong viability. CD207+ cells were then isolated from HLA-DR+ (lineage negative) cells from the lymphoid stroma of healthy tonsils (tCD207+). The tCD207+ cells were transduced with a lentivirus encoding human telomerase reverse transcriptase (hTERT). These cells, also lacking MAPK activating mutations, died two weeks post-transduction. A fourth approach has been more successful in which tCD207+ cells were cultured in a cytokine cocktail which provided MAPK pathway stimulation. The cells retained CD207 expression and survived in culture for over two weeks. The cells were then immortalized using a lentivirus encoding HOXA9. Immortalized cells maintained CD207 expression. Allele specific MAPK pathway mutations (BRAF and MAP2K1) are being generated by class II CRISPR/Cpf1 genome editing as Cpf1 has been shown to have robust activity to induce specific disruption of only mutant, but not wild-type, BRAF allele. The phenotypic and genomic characteristics of the immortalized cells expressing the different MAPK pathway mutations will be assessed by RHG-banding cytogenetic analysis, fluorescence in situ hybridization, gene expression analysis, and immuno‐cytochemistry and results will be compared to cells isolated from LCH lesions to confirm whether the established cell line may be a viable in vitro mimic of the LCH lesion DC.

Disclosures
No relevant conflicts of interest to declare.
Venous Thromboembolism in Lower Extremity Amputees: A Systematic Review of the Literature
Authors: Alsheef Mohammed Abdullah , Schulman Sam , Donadini Marco , Zaidi Abdul Rehman Z. 
Published: November 2019

Patients undergoing lower extremity amputation (LEA) are at risk of developing deep venous thrombosis (DVT) and pulmonary embolism (PE), but no generally accepted prevention guidelines exist. This systematic review aimed at understanding the incidence of VTE with or without thromboprophylaxis in adult patients with major lower extremity amputation (LEA). Primary outcomes were onset of DVT, PE, or mortality. Secondary outcomes were any major adverse events due to treatment. We searched English language full-text papers in multiple databases using keywords, including amputation/adverse effects, amputation/complications, venous thromboembolism, deep vein thrombosis, and pulmonary embolism. Twenty-eight studies providing observations for 4,841 patients were selected. The fatal PE risk was 2.6% without prophylaxis and significantly decreased to a non-zero residual risk of 0.9% with VTE prophylaxis. Above-knee amputees were at greatest risk of VTE and subsequent complications. The risk was not confined to the amputated stump and can involve the contralateral limb. The role of compression ultrasonography screening in asymptomatic patients remains controversial in various populations at risk for VTE. All patients undergoing major LEA should be considered at high risk for the development of VTE, even after discharge from hospital. We recommend prophylactic anticoagulation (if not contraindicated) and clinical surveillance in all patients undergoing LEA and further studies to determine the optimal prophylactic strategy.

Disclosures
No relevant conflicts of interest to declare.
Quality Improvement Project to Evaluate Fever Management and Appropriateness of Lab Draws in Hospitalized Sickle Cell Disease Patients in a Tertiary Care University Teaching Hospital
Authors: Jafri Syed Imran Mustafa , Hook Karen 
Published: November 2019

Introduction: Sickle cell disease is the most common inherited blood disorder that affects almost 100,000 people in the U.S. Infection is the most common cause of death in these patients. Early identification and initiation of antibiotics given functional asplenia is the key to appropriate management of hospitalized sickle cell disease patients. Approximately 4-5 billion laboratory studies are ordered every year in hospitals. Unnecessary blood tests lead to frequent blood draws and worsening anemia in this already anemic population as well as contribute towards increasing cost of hospital stay. Total national health expenditures in the U.S. is $3.3 trillion (2016) with percent of national health expenditures for hospital care: 32.4%.
Objective: We reviewed inpatient admissions of sickle cell disease patients to assess for appropriate work up and timely antibiotic initiation in case of fever. Additionally, we looked up the number of blood draws as well as number of days labs were drawn during the inpatient stay.
Methods: This was a retrospective chart review of all patients with sickle cell disease who were admitted to John Dempsey hospital at University of Connecticut with length of stay of at least 3 days between April 2018 to Feb 2019.
Results: 57 inpatient admissions were reviewed. Out of 57 admissions, 14 had fever either at the time of presentation or during the hospital stay. Mean time to first blood culture was 56 minutes with 92% (13/14) drawn within 1 hour of fever. Mean time to first antibiotic was 4.58 hours. Antibiotics were initiated in 7% (1/14) in less than 1 hour and 50% (7/14) within 3 hours. Urine culture was checked 78% and chest X-ray was checked 100% of the time in case of fever. On review of 57 patient admissions, labs were checked every single day of hospital stay in 35% (20/57) and if the day of discharge was excluded in 50% (29/57) of the patients. Overall, the labs were checked 82.33 percent of the days patients were hospitalized. On average, labs were drawn 1.14 times per day on each patient.
Conclusion: Our project addresses safety, efficiency and timeliness with identification of areas of improvement in febrile sickle cell disease patients. It also addresses patient-centeredness raising awareness for decreased needle sticks for our sickle cell disease patients. There is continuing need to educate our care providers on how to manage fever in sickle cell disease for appropriate work up and early initiation of antibiotics. Also, we need to be more considerate when ordering labs on our patients especially sickle cell disease patients with baseline anemia. This will lead to decreased number of needle sticks and iatrogenic anemia as well as better healthcare resource utilization.

Disclosures
No relevant conflicts of interest to declare.
SOX17 Is Essential for Integration of Arterial and HOXA Programs in Hemogenic Endothelium
Authors: Jung Ho Sun , Uenishi Gene I. , Park Mi Ae , Liu Peng , Raymond Matthew , Thomson James , Ong Irene , Slukvin Igor 
Published: November 2019

Recent advances in understanding the major bottlenecks in derivation of engraftable HSCs and lymphoid cells from pluripotent stem cells (PSC), have identified deficiencies in NOTCH and HOXA signaling as contributing factors to the observed functional deficits of PSC-derived hematopoietic progenitors. However, little is known about the mechanisms that are essential for establishing these pathways during PSC differentiation. Here, we revealed the critical role of SOX17 in linking HOXA and NOTCH-mediated arterial programs in hemogenic endothelium (HE) and specification of definitive lympho-myeloid hematopoiesis. Using SOX17-knockout (SOX17-/-) and SOX17 DOX-inducible (iSOX17) hESCs, we found that SOX17-deficiency substantially reduces formation of CD144+CD43-CD73-DLL4+CXCR4+/- arterial HE and definitive lympho-myeloid hematopoiesis, while SOX17 upregulation at mesodermal stage of development causes the opposite effect. Molecular profiling of HE generated from iSOX17 hESCs in DOX+ and DOX- conditions using RNAseq, SOX17 ChIPseq and ATACseq, revealed that SOX17 overexpression upregulates 522 genes enriched in NOTCH, TGFb, HEDGEHOG and WNT signaling, including DLL1, DLL4, NOTCH4, LFNG, WNT5a, WNT5b, GLI3, and genes associated with HSC development, CXCR4,KITLG and ALDH1A2. In addition, we noted significant upregulation of HOXA7,HOXA9, HOXA10, HOXB8, HOXC4 and CDX2 homeobox genes in SOX17-induced cultures, with no expression of HOXA genes observed in HE from SOX17-/- cells. ChIPSeq analysis revealed DOX+ specific SOX17 binding at transcriptional start sites (TSS) of 316 significantly upregulated genes, including ALDH1A2, CDX2, DLL1, DLL4, HEY1, HOXA7, HOXB8, HOXC4 and KITLG, suggesting that upregulation of these genes could be explained by their direct activation by SOX17.
Since ALDH1A2 and CDX2 are known to play a role in the activation of HOXA genes, we investigated whether SOX17's effect on HOXA expression could also be mediated by ALDH1A2 and CDX2. We found that adding ALDH1 inhibitor to DOX+ cultures had no effect on arterial HE development and HOXA expression. In contrast, transfection of iSOX17 hPSCs cultures with CDX2 shRNA significantly decreased arterial HE formation and downregulated HOXA7, HOXA9, and HOXA10 expression. Overall, our studies indicate that SOX17 plays a critical role in the activation and integration of arterial and HOXA programs in HE, which is mediated by CDX2. These findings will be important for designing a strategy for direct HSC fate programming from hPSCs.

Disclosures
Uenishi: Casebia Therapeutics: Employment. Slukvin:Cynata Therapeutics: Consultancy, Other: Founder and Stockholder.
Tumor Suppressor Function of Dnmt3b in Hematologic Malignancies Critically Depends on Its Catalytic Activity
Authors: Lopusna Katarina , Nowialis Pawel , Haney Staci L. , Abraham Ajay , Opavska Jana , Opavsky Rene 
Published: November 2019

DNA methylation regulates gene transcription and it is involved in various physiological processes in mammals including development and hematopoiesis. It is catalyzed by several DNA methyltransferases, including DNMT1, DNMT3A and DNMT3B that not only mediate methylation but also interact with a number of repressive proteins, thereby contributing to transcriptional repression in methylation-independent manner. Dnmt3b is critical gene in mouse embryogenesis whose loss results in embryonic lethality around E13.5. It also functions as a tumor suppressor in lymphoid and myeloid malignancies in mice. The extent to which Dnmt3b's catalytic activity is involved in these processes is unclear.
To address the role of catalytic activity in embryogenesis and tumorigenesis, we generated mice carrying a catalytically inactive allele of Dnmt3b (Dnmt3bCI) by introducing a double amino acid substitution in enzyme active center (P656V; C657D) in endogenous Dnmt3b locus. By interbreeding Dnmt3b+/CI mice we found that homozygous mutant Dnmt3bCI/CImice survived embryonic development and had a long life span. Such results demonstrated that catalytic activity of Dnmt3b is, surprisingly, dispensable for both pre- and post-natal development. The presence of catalytically inactive Dnmt3b rescued most methylation changes observed in the absence of Dnmt3b in embryogenesis suggesting that Dnmt3b plays a role of accessory protein for methylation mediated by other Dnmts. In contrast, the absence of Dnmt3b's catalytic activity promoted MYC/Ras-induced transformation of mouse embryonic fibroblasts in vitro, and accelerated both MYC-induced T-cell lymphomagenesis and MLL-AF9-induced acute myeloid leukemia. Global gene expression and methylation profiling revealed a number of deregulated events specific for MYC;Dnmt3bCI/CI, resulting in aberrant activation of IL7-R and STAT3 and downregulation of putative tumor suppressor genes including Cdkn2c and Dusp6.
Altogether, our data suggest that Dnmt3b's methyltransferase activity is dispensable for mouse development but critical to prevent hematologic malignancies by controlling events involved in cellular transformation.

Disclosures
No relevant conflicts of interest to declare.