bluuebunny/crossref_metadata_2025_split huggingface dataset:
MLL Is Essential for NUP98-HOXA9-Induced Leukemia
Authors: Shima Yutaka , Yumoto Minori , Kitabayashi Issay 
Published: October 2019

Abstract
NUP98, a component of the nuclear pore complex (NPC), plays an important role in molecular trafficking between the cytoplasm and the nucleus. NUP98 has two Phe-Gly (FG) repeat domains, which are characteristic of NPC proteins. The NUP98 gene is rearranged and fused to several partner genes, such as HOXA9 and DDX10, in acute myeloid leukemia and myelodysplastic syndromes. Leukemia with NUP98 rearrangement is associated with poor prognosis. However, the molecular mechanism of leukemogenesis induced by NUP98-fusions remains unclear.
To investigate the role of NUP98 of NUP98 fusion proteins in leukemogenesis, we generated a set of deletion mutants of NUP98-HOXA9 lacking each FG repeat domain. These mutants were transduced into normal murine progenitor cells and their activity in the immortalization of murine cells was examined by colony-formation assays. Deletion of the second FG domain resulted in a dramatic delay in leukemia induction when cells were transplanted into mice, suggesting that the second FG repeat domain is crucial for immortalization in vitro and leukemogenesis in vivo. qPCR and ChIP analyses showed that the FG repeat domain is necessary for Hoxa gene activation and recruitment to the Hoxa gene locus. To identify interacting proteins of the FG repeat domain of NUP98-HOXA9, protein complexes containing full-length and mutant NUP98-HOXA9 were purified. Mass spectrometry analysis showed that full-length NUP98-HOXA9, but not the mutant lacking the second FG repeat, interacted with MLL. ChIP analysis also showed co-localization of NUP98-HOXA9 and MLL at the Hoxa gene locus in NUP98-HOXA9-expressing cells. Furthermore, analysis of Mll null mice showed that MLL is essential for the recruitment of NUP98-HOXA9 to the Hoxa gene locus and for NUP98-HOXA9-induced immortalization and leukemogenesis. These results suggest that NUP98 fusion proteins interact with MLL to activate HOXA genes during leukemogenesis.

Disclosures
Kitabayashi: Daiichi Sankyo Co., Ltd.: Research Funding.
Clinical Significance of Co-Existence of PHF6 and NOTCH1 mutations in Adult T-Cell Acute Lymphoblastic Leukemia
Authors: Ge Zheng , Li Min , Xiao Lichan , Zhang Run , Li Jianyong 
Published: October 2019

Abstract
Objective: T-cell acute lymphoblastic leukemia (T-ALL) is caused by collaboration of multiple genetic abnormalities in the transformation of T-cell progenitors. PHF6 is founded as a new key tumor suppressor and mutated in T-ALL. The clinical significance of PHF6 mutations has not been fully determined in adult T-ALL. This study aimed to screen the PHF6 mutations in adult T-ALL and explore the associations of PHF6 mutations with other genetic lesions, as well as their clinical relevance in adult T-ALL patients.
Methods: We amplified the exons of PHF6, NOTCH1, FBXW7, PTEN and JAK1 following by DNA sequencing to identify the genomic mutations and examined the PHF6 mRNA level by qPCR in adult T-ALL patients. We also analyzed the correlations of PHF6 and NOTCH1 mutations with clinical features using a χ2 test and survival curve using the Kaplan-Meier method.
Results: The 27.1% (16/59) PHF6 mutations including 10 novel mutations were detected in Chinese adult T-ALL. Six of 16 (37.5%) were frame-shift mutations, which could result in the deletion of the protein. We also observed PHF6 expression was significantly lower in T-ALL patients with PHF6 mutations compared with wide type cases (0.00423 vs. 0.06464, P=0.035) , indicating PHF6 mutations could be loss of function. Moreover, PHF6 mutation was significantly associated with NOTCH1 mutation(P=0.035). We further analyzed the domains involving co-existence mutations of NOTCH1 with PHF6. The most commonly mutated domains in NOTCH1 co-existed with PHF6 were HD-N only 6/12 (50.0%), followed by HD-C only 2/12(16.7%), PEST only 2/12(16.7%), HD-C+PEST 1/12(8.3%) and HD-N+HD-C 1/12(8.3%), indicating that HD domain (especially HD-N) of NOTCH1 may contribute to the synergistic effect on oncogenesis of the two genes. Furthermore, the patients with co-existence of PHF6 and NOTCH1 mutations had lower hemoglobin and higher incidence of splenomegaly or lymphadenopathy compared to that without co-existence of the mutations (95.0 vs 122.0, P=0.007; 81.8% vs 38.3%, P=0.009; 90.9% vs 44.7%, P=0.006). Importantly, the patients with co-existence of mutations in PHF6 and NOTCH1 (PHF6mutNOTCH1mut) had significant shorter event-free survival (EFS) compared with that without co-existence (non-PHF6mutNOTCH1mut)(2.0 months vs. 12.0 months, P=0.027).
Conclusion: PHF6 is inactivated in T-ALL due to its low expression and mutations. PHF6 mutation is co-existed with NOTCH1 mutations, and the patients with PHF6mutNOTCH1mut had a poor prognosis. Our results indicated synergistic effect of PHF6 and NOTCH1 mutations on leukemogenesis and PHF6mutNOTCH1mut may be potential prognostic marker in adult T-ALL.

Disclosures
No relevant conflicts of interest to declare.
Predictors of Early Death and Survival Among Children, Adolescents and Young Adults with Acute Myeloid Leukemia in California, 1988-2011: A Population-Based Study
Authors: Abrahão Renata , Keogh Ruth H , Lichtensztajn Daphne Y , Marcos-Gragera Rafael , Medeiros Bruno C. , Coleman MIchel P , Ribeiro Raul C. , Keegan Theresa HM 
Published: October 2019

Abstract
Background A better understanding of factors associated with early death and survival may guide future health policy aimed at improving outcomes among children, adolescents and young adults with acute myeloid leukemia (AML).
Methods We examined trends in early death and survival among 3,935 patients aged 0-39 years diagnosed with de novo AML in California during 1988-2011. We estimated overall survival and applied logistic and Cox regression to evaluate the association between sociodemographic and selected clinical factors with early death and survival.
Results There was a trend towards decline in early death from 9.7% in 1988-1995 to 7.1% in 2004-2011 (P=0.062). Survival improved substantially over time, but 5-year survival was still only 50% (95% CI 47%-53%) even in the most recent calendar period. Overall, the main factors associated with poor outcomes were older age at diagnosis, treatment at hospitals not affiliated with National Cancer Institute (NCI)-designated cancer centers, and black race/ethnicity. For patients diagnosed during 1996-2011, survival was lower for patients without health insurance and those who did not receive a hematopoietic stem cell transplant (HSCT). The association between survival and sociodemographic and clinical factors were stronger among patients aged 30-39 years.
Conclusions Mortality after AML remained strikingly high and increased with age. To improve outcomes, strategies may include wider insurance coverage, treatment at hospitals affiliated with NCI-designated cancer centers and access to HSCT for high-risk patients. Collaborative clinical trials aimed to evaluate the efficacy and toxicity of novel agents for each subtype of disease are warranted.

Disclosures
No relevant conflicts of interest to declare.
Alvocidib Potentiates the Activity of Azacytidine in an MCL-1-Dependent Fashion
Authors: Kim Wontak , Soh Katherine K. , Bearss Jeremiah J. , Lee Ye Sol , Peterson Peter , Whatcott Clifford J. , Siddiqui-Jain Adam , Weitman Steven , Bogenberger James M , Tibes Raoul , Bearss David J. , Warner Steven L. 
Published: October 2019

Abstract
Despite significant efforts, the clinical mechanism of action of hypomethylating agents such as 5-azacytidine (5-aza) is still poorly understood. 5-aza is currently indicated for the treatment of patients with myelodysplastic syndrome (MDS). While 5-aza has achieved good single-agent activity in acute myeloid leukemia (AML), complete response rates remain low when used as a single agent. In a recent report aimed at identifying rational therapeutic combinations with 5-aza, Bogenberger and colleagues identified multiple BCL-2 family member/BH3-containing therapeutic targets, which synergize with 5-aza when inhibited genetically or pharmacologically. The CDK9 inhibitor, alvocidib, has achieved significant improvement in complete response rates of newly diagnosed AML patients when administered before cytarabine and mitoxantrone (FLAM regimen) in a randomized multi-center Phase 2 trial when compared to 7+3 standard of care treatment. Recent reports suggest that the transcriptional repression of key anti-apoptotic proteins (eg., MCL-1) mediated by alvocidib's CDK9 inhibition, drive the pro-apoptotic activity of alvocidib in the FLAM regimen. We, therefore, hypothesized that alvocidib and 5-aza would synergize therapeutically in the treatment of AML by means of transcriptional repression of MCL-1 and sensitization to 5-aza. In this report, we demonstrate that treatment of AML cell lines with alvocidib inhibits both mRNA and protein expression of MCL-1 in a time and concentration-dependent fashion. Pre-treatment of cells with alvocidib, to repress MCL-1 expression prior to 5-aza treatment, reduced the 5-aza cell viability EC50 more than 2.5-fold, from 1.8 µM to 0.6 µM in MV4-11 cells. The alvocidib/5-aza combination also resulted in synergistic increases in caspase activity relative to either single agent within the combination, at multiple dose levels. Therefore, following reports suggesting inhibition of BCL-2 family members including MCL-1, sensitizes cells to 5-aza, our data suggest that the alvocidib/5-aza combination may constitute a viable therapeutic regimen. We also conclude that a CDK9 inhibitor/5-aza combination may be an effective clinical approach for the treatment of AML.

Disclosures
Kim: Tolero Pharmaceuticals: Employment. Soh:Tolero Pharmaceuticals: Employment. Bearss:Tolero Pharmaceuticals: Employment. Lee:Tolero Pharmaceuticals: Employment. Peterson:Tolero Pharmaceuticals: Employment. Whatcott:Tolero Pharmaceuticals: Employment. Siddiqui-Jain:Tolero Pharmaceuticals: Employment. Weitman:Tolero Pharmaceuticals: Employment. Bearss:Tolero Pharmaceuticals: Employment. Warner:Tolero Pharmaceuticals: Employment.
The Highly Potent Bromodomain (BRD) Inhibitor FV-281 Displays Preclinical Efficacy in Acute Myeloid Leukemia (AML)
Authors: Wang Zezhou , Dove Peter , Shamas-Din Aisha , Hurren Rose , Wang Xiaoming , MacLean Neil , Gronda Marcela , Bo Feng , Li Janessa , Skorupski Clarissa , Hadri Wissam , O'Neill David , Aman Ahmed , Al-awar Rima , Minden Mark D. , Slassi Malik , Schimmer Aaron D. 
Published: October 2019

Abstract
Recent studies have demonstrated the therapeutic potential of targeting BRD2, BRD3 and BRD4 in hematological malignancies including AML. Here, we report a novel orally bioavailable BRD inhibitor, FV-281, that displays preclinical efficacy in AML in vitro and in vivo.
In a binding screen assay of 32 bromodomains (BromoScanTM), FV-281 selectively bound BRD2, BRD3 and BRD4 with Kd values of 2.7 to 7.3 nM and was 2-5 fold more potent than the leading bromodomain inhibitor, OTX015. In AML cell lines (OCI-AML2, Tex, HL-60 and MV4-11), FV-281 reduced cell growth and viability after 72 hour incubation with IC50 values < 50 nM and was 6-fold more potent than OTX015. FV-281 was not cytotoxic to normal hematopoietic cells (n=4) with all IC50 values > 10 µM. In contrast, FV-281 induced cell death as measured by Annexin V/PI staining in 3 of 4 primary AML samples with IC50values < 850 nM.
Within 4 hours of incubation with FV-281, c-Myc mRNA and protein level in OCI-AML2 cells were reduced by >95% and >80%, respectively. Following FV-281 washout, c-Myc protein expression returned to baseline within 20 hours with no loss of cell viability observed. Thus, the FV-281 is a reversible bromodomain inhibitor and sustained target inhibition is required to induce cell death. Likewise, after 6 and 48 hours of incubation with OCI-AML2 cells, FV-281 reduced Bcl-2 mRNA and protein level by >60% and >90%, respectively.
To assess in vivo efficacy and toxicity, MV4-11 AML cells were engrafted subcutaneously in NOD-SCID mice (n=10). Once tumors were palpable, mice were treated with FV-281 orally (30mg/kg/d x 2 weeks) or vehicle control. FV-281 suppressed tumor growth without evidence of overt toxicity. In addition, primary AML cells were engrafted into the femurs of NOD-SCID mice (n=10). Three weeks after implantation, mice were treated with FV-281 orally (30mg/kg, 5 of 7 days, x 4 weeks) or vehicle control. FV-281 decreased AML engraftment in the injected and non-injected femur without evidence of overt toxicity (% human CD45+/CD19-/CD33+ in non-injected femur treated vs control: 20% vs 60%, p<0.0001).
Thus, FV-281 is a novel oral reversible bromodomain inhibitor with significant in vivo activity in murine models of AML. These data support evaluation of this agent in upcoming phase I clinical trials.

Disclosures
Wang: Fluorinov Pharma Inc.: Employment. Dove:Fluorinov Pharma Inc.: Employment. Hadri:Fluorinov Pharma Inc.: Employment. O'Neill:Fluorinov Pharma Inc.: Employment. Slassi:Fluorinov Pharma Inc.: Employment.
Extensive Chronic Graft-Versus-Host-Disease Significantly Increases the Risk of Severe and Multifocal Genital Tract HPV Disease in Long-Term Survivors of Allogeneic Stem Cell Transplantation
Authors: Shanis Dana L , Anandi Prathima , Grant Caitlin , Pophali Priyanka A , Koklanaris Eleftheria , Savani Bipin N. , Barrett Austin John , Battiwalla Minoo , Stratton Pamela 
Published: October 2019

Abstract

Background: High rates of cervical HPV disease in women after allogeneic stem cell transplantation (SCT) have been reported, but risk factors related to severe, multifocal, including vaginal and vulvar, HPV disease are not defined.
Objective: To determine rates and risk factors for multifocal and severe HPV disease in post-transplant women.
Methods: In a prospective long-term study after SCT, gynecologic history and assessment, cervical cytology and HPV testing were obtained with follow-up colposcopy and surgery as indicated for abnormal results. Prior HPV disease, genital graft-versus-host-disease (gGVHD), chronic GVHD (cGVHD) and immunosuppression treatment (IST) >3years were assessed for their association with extent and severity of genital HPV disease. Logistic regressions were used for multivariate analysis.
Results: Sixty five long term (>3 year) SCT survivors were studied prospectively on protocol. Patients received allogeneic transplantation from HLA-identical sibling donors with most undergoing myeloablative total body irradiation (94%) and T lymphocyte-depleted peripheral blood stem cells in 91% Of 65 women, 62 had gynecologic assessment with 8 (13%) having prior history of HPV disease; 16 (26%) had gGVHD. 20 women (32%) had acute GVHD, 46 (74%) had cGVHD; extent was limited in 23(37%) and extensive in 23(37%). 26(42%) had cGVHD requiring IST >3years. Of 21(34%) women with HPV disease after transplant, 12 required surgery and 7 had multifocal disease. Extensive chronic GVHD (but not acute GVHD) was found to significantly impact occurrence (OR=3.5, p=0.038), high-grade severity (OR=7.1, p=0.024) or multifocal HPV disease (OR=14.6, p=0.017).
Conclusion: Women who have undergone SCT have an increased risk of genital HPV disease, with highest rates in women with extensive cGVHD. Likely as a result of chronic immune dysregulation and the temporal nature of HPV, these women are at high risk of severe, multifocal disease, which if untreated may progress to genital cancer. Thus, gynecologic assessment as well as possible post-transplant HPV vaccination are critical aspects of care for women with significant GVHD post-transplant.
Support: Intramural programs of NHLBI, Clinical Center and NICHD, NCT00106925

Disclosures
Stratton: Allergan: Research Funding.
Impairment of IL-17 Immunity to Candida and IFN-g Immunity to Mycobacterium in Humans with Bi-Allelic Rorc mutations
Authors: Okada Satoshi , Markle Janet , Kobayashi Masao , Bustamante Jacinta , Casanova Jean-Laurent 
Published: October 2019

Abstract
Inborn errors of human IL-17 immunity underlie chronic mucocutaneous candidiasis (CMC), which is characterized by infections of the skin, nails, oral and genital mucosae with Candida albicans. Inborn errors of human IFN-γ immunity underlie Mendelian susceptibility to mycobacterial disease (MSMD), a rare congenital disorder characterized by susceptibility to infections by poorly virulent intracellular pathogens such as Mycobacterium bovis Bacille Calmette-Guérin vaccines (BCG) and non-tuberculosis Mycobacterium. Rarely, patients may be affected by both candidiasis and mycobacteriosis, including some patients with IL-12p40 and IL-12Rβ1 deficiencies that impair IFN-γ immunity and IL-17 immunity. We studied seven patients from three unrelated consanguineous kindreds with this unusual combination of infectious diseases without known genetic disorder. We combined whole exome sequencing and genome wide linkage analysis, and discovered bi-allelic loss-of-function mutations in RORC, which encodes the transcription factors RORγ and RORγT. All of the seven patients suffered from severe mycobacterial infections, and six also exhibited mild CMC. RORγT is the key transcription factor of Th17 cells, which produce IL-17 and IL-22. Therefore, as predicted by the mouse model, RORγT deficiency prevented the development of IL-17-producing T cells, which accounts for the patients' CMC. Consistent with the phenotype of Rorc-/- mice, these patients presented with mild T cell lymphopenia, small thymus, lack of palpable axillary and cervical lymph nodes, and absence of MAIT and iNKT cells.
The patients' severe infections with mycobacteria were not predicted by previous studies of Rorc-/- mice. To explain this unexpected phenotype, we focused on IFN-γ immunity. Leukocytes from RORC-/- patients showed impaired IFN-γ production in response to a mycobacterial challenge, and this defect is probably attributable to the functional impairment of CD4+ CCR6+ CXCR3+ αβ T cells, γδT cells, or both. These findings also suggested that IFN-γ treatment may be beneficial for RORC-/- patients. Moreover, this phenotype does not seem to be human-specific, as we also found that Rorc-/- mice deficient were susceptible to mycobacterial infection. We thus discovered that human RORC is essential not only for the development of IL-17-producing lymphocytes protecting the mucocutaneous barriers against Candida, but also for the activation of IFN-g-producing T cells, and for systemic protection against Mycobacterium.

Disclosures
No relevant conflicts of interest to declare.
Unstaged Diffuse Large B Cell Lymphoma in the United States: Predictors and Patient Outcomes
Authors: Shah Binay K. , Bista Amir , Sharma Sandhya 
Published: October 2019

Abstract
Background: Treatment and prognosis of diffuse large B cell lymphoma (DLBCL) depends on the stage of lymphoma. We conducted this study to examine unstaged DLBCL in the United States.
Methods and methodology: We used Surveillance Epidemiology and End Result (SEER) 18 registries to select patients with DLBCL diagnosed during January 2000 to December 2012. We used LRD Summary stage 2000 was used to determine stage of the disease - localized, regional, distant or unstaged. We used Logistic regression to investigate factors associated with unstaged DLBCL. We used Cox Proportional Hazard model to compare survival outcomes.
Results: Among 67765 patients, 3194 (4.71%) were unstaged. Age (60+years), "Others" and Caucasian races, single or single/divorced/widow marital status, metropolitan residence, median household income> $50,000, lymph node as the primary site and cased with other primaries before diagnosis of DLBCL were the factors associated with unstaged cases (Table 1). The 5- year relative survival rate for unstaged patients was inferior to those with localized and regional disease, and superior to those with distant disease (HRs of 0.58, 0.66 and 1.24 for localized, regional and distant respectively when compared to unstaged cases).
Conclusion: In this large population-based study, 4.71% patients with DLBCL had unstaged disease. Patients with unstaged DLBC had significantly inferior survival rates compared to patients with localized and regional stage.
Table 1. Factors associated with unstaged DLBCL cases Parameters Unadjusted OR (95% CI) P value Adjusted OR (95% CI) P value Age (60+ Vs. <60 years) 1.478 (1.363 - 1.602) <0.001 1.458 (1.335 - 1.592) <0.001 Sex (Female Vs. Male) 1.063 (0.990 - 1.141) 0.093 0.983 (0.911 - 1.059) 0.646 Race Caucasians Reference Reference African American 0.804 (0.691 - 0.935) 0.005 0.835 (0.715 - 0.974) 0.022 Others 1.109 (0.976 - 1.261) 0.112 1.257 (1.104 - 1.431) 0.001 Marital Status Married Reference Reference Single 1.026 (0.927 - 1.135) 0.662 1.208 (1.086 - 1.345) 0.001 Single/divorced/widow 1.249 (1.152 - 1.355) <0.001 1.185 (1.087 - 1.291) <0.001 Rural/Urban Rural Reference Reference Urban 0.878 (0.651 - 1.183) 0.393 0.896 (0.661 - 1.214) 0.479 Metropolitan 0.882 (0.667 - 1.165) 0.882 1.028 (0.767 - 1.379) 0.852 Median annual household income Upto 25,000 Reference Reference >25,000-50,000 01.009 (0.753 - 1.354) 0.951 0.927 (0.675 - 1.271) 0.636 >50,000 0.758 (0.563 - 1.021) 0.068 0.673 (0.486 - 0.933) 0.017 Sequence (Not first or only primary vs. first or only primary) 1.261 (1.156 - 1.377) <0.001 1.219 (1.115 - 1.334) <0.001 Site of primary Lymph nodes Reference Reference Extra-lymphatic 0.760 (0.704 - 0.821) <0.001 0.748 (0.693 - 0.808) <0.001 Unknown primary 6.295 (4.569 - 8.672) <0.001 6.727 (4.865 - 9.300) <0.001

Disclosures
No relevant conflicts of interest to declare.
ATP11C Encodes a Major Flippase in Human Erythrocyte and Its Genetic Defect Causes Congenital Non-Spherocytic Hemolytic Anemia
Authors: Arashiki Nobuto , Takakuwa Yuichi , Ogura Hiromi , Utsugisawa Taiju , Miyano Satoru , Ogawa Seishi , Kojima Seiji , Ohga Shouichi , Mohandas Narla , Kanno Hitoshi 
Published: October 2019

Abstract
Flippases are members of the P-IV ATPase family of proteins, and contribute to localization of phosphatidylserine (PS) in inner leaflet by its ATP-dependent active transport from outer to inner leaflet of lipid bilayer of erythrocyte membranes. It is critical that erythrocytes maintain PS in the inner monolayer to ensure their 120-day survival in circulation since externalization of PS will be recognized as an eat-me signal, resulting in phagocytosis by splenic macrophages. In the present study, we identified that ATP11C gene encodes a major flippase in human erythrocytes. A 13 years-old boy was referred to our hospital for consultation for work up of an undiagnosed congenital hemolytic anemia. Since extensive biochemical and molecular analysis failed to identify hemoglobin, erythrocyte membranes and enzyme abnormalities for the pathogenesis of hemolysis, we performed the whole exome analysis by massively parallel sequencing. We identified that the proband is hemizyogous, and the mother is heterozygous for a missense mutation of ATP11C, c.1253C>A, corresponding to a single amino acid substitution, p.Thr418Asn. Flipping activity as measured by PS internalization was decreased to 10% in the red cells of the proband compared to a normal control, clearly demonstrating that ATP11C encodes a major flippase in the human erythrocyte membranes. The PS-positive erythrocytes were not significantly increased in the whole blood but only in the most dense senescent cells, suggesting that PS exposure did not occur until very late stages of lifespan. We showed that PS exposure mediated by Ca2+-stimulated phospholipid scrambling was not different between red cells of the proband and control. Taken together, our findings imply that suppressed scrambling activity rather than flippase activity is the major contributor to maintainance of PS in inner leaflet of normal red cells during their 120-day lifespan, and that PS exposure to cell surface as an 'eat-me' signal depends primarily on scramblase activity at the end of lifespan. Importantly, our study has enables us to identify the major flippase of human erythrocyte membrane.

Disclosures
No relevant conflicts of interest to declare.