bluuebunny/crossref_metadata_2025_split huggingface dataset:
Greater Need for FVIII Replacement in Patients of Type Non-O Blood Groups with Moderate Hemophilia A
Authors: Lima Magaly BPLV , Siqueira Lúcia Helena , Saraiva Ana Suely , Montalvão Silmara Aparecida , Machado Tânia de Fátima Gomes Siegl , Trento Thais , Oliveira-Filho Aldemir Branco , Ozelo Margareth C , Lemos José Alexandre Rodrigues 
Published: October 2019

Abstract
Abstract 4390
Hemophilia A is a bleeding disorder of variable expressivity caused by mutations in clotting factor 8 gene located on Xq28 chromosome. In Brazil, treatment of such patients consists of replacement therapy with FVIII concentrate derived from human plasma, which is administered either to control an episode of hemorrhage or prophylactically in order to prevent bleeding and its consequences, such as arthropathy. Recent studies have demonstrated that the half-life of FVIII administered to patients of the type O blood group with severe hemophilia A is shorter than that in patients of Type non-O groups. A likely explanation for this fact would be the greater plasma concentration of von Willebrand factor (vWF) in individuals with type non-O blood. In this study, was investigated the association between the blood group and the annual consumption of FVIII concentrate (UI/Kg/year) in the last three years among 99 patients (68 with moderate and 31 with mild hemophilia A) treated at the two blood centers in Brazil (Hemocentro do Pará and Hemocentro/UNICAMP). Most cases were treated for episodes of hemorrhage. Determinations of antigen levels of von Willebrand factor (vWF:Ag), activity of ristocetin cofactor (vWF:RiCof) and FVIII clotting activity (FVIII:C) were performed to investigate the correlation between these variables and immunophenotypes of the ABO blood system. The Mann-Whitney test was used for the comparison of FVIII consumption (UI/Kg/year) between O type and non-O type hemophiliacs, with the level of significance set to 5% (p<0.05). Unexpectedly, significantly greater FVIII consumption was found among patients of the non-O type group (p=0.017). In the evaluation of patients with moderate hemophilia A, plasma levels of vWF:RiCof were significantly higher in non-O type patients in comparison to those with type O blood (p=0.045). No significant differences were found between blood groups in the evaluation of patients with mild hemophilia A. The present findings lead one to conclude that the higher level of plasma vWF:RiCof (which could exert an influence on the increased half-life of FVIII) in patients of the non-O type group with moderate hemophilia A was not enough to impede the need for greater FVIII replacement and lessen the frequency of bleeding in this group. The authors hypothesize that other variables, such as mutations in moderate hemophilia A, affect the stability of the FVIII molecule, which has antigen A and B determinants in individuals of non-O type groups. Further studies involving a larger sample are needed to corroborate the findings of the present investigation.

Disclosures:
No relevant conflicts of interest to declare.
The Average Baseline BCR-ABL Levels Are Significantly Higher in Patients with Resistance to Dasatinib As First-Line Treatment for Early Chronic Phase Chronic Myeloid Leukemia
Authors: Tomasetti Cristian , Kantarjian Hagop M. , Ramchandani Ritesh , Jabbour Elias , Quintás-Cardama Alfonso , Parmigiani Giovanni , Cortes Jorge E. 
Published: October 2019

Abstract
Abstract 4436

Background:
The development of drug resistance remains a problem for some chronic myeloid leukemia (CML) patients treated with tyrosine kinase. Point mutations are the main mechanism behind drug resistance, and they can occur prior to therapy. Thus, there may be a positive correlation between the baseline BCR-ABL levels and the occurrence of drug resistance.

Methods:
We define a “relapse/no-response” as a BCR-ABL/ABL transcript ratio of >10% (international scale) at 6 months or later during continuous dasatinib treatment. Statistical analysis is performed an all early chronic phase CML treated with dasatinib as first-line therapy (n=102) in a prospective clinical trial at the M.D. Anderson Cancer Center. The probability of having drug resistant leukemic cells already present at diagnosis is also estimated.

Results:
Seven of the 90 (7.8%) evaluable patients had a “relapse/no-response”. The mean (median) BCR-ABL/ABL transcript ratio at baseline for the “relapse/no-response” group was 23.3% (20.7%), while for the rest of the patients was 14.1% (9.9%), a statistically significant difference. The incidence rates of “relapse/no-response” for patients with a BCR-ABL/ABL transcript ratio at baseline equal to or larger than 20% was 13.8%, versus 7.8% overall for the entire dataset. Our estimates suggest that if the therapy were to be started on average one month earlier, the probability of a “relapse/no-response” would decrease from 7.8% to 6.9%.

Conclusions:
The BCR-ABL transcript levels at baseline are significantly higher for those patients later incurring in a “relapse/no-response”. Moreover, our estimates suggest that starting the treatment as soon as possible after diagnosis may be critical for reducing the number of patients developing drug resistance.

Disclosures:
Kantarjian: Novartis: Research Funding; Pfizer: Research Funding; Ariad: Research Funding; Bristol-Myers Squibb: Research Funding. Jabbour:Pfizer: Honoraria; Novartis: Honoraria; BMS: Honoraria. Cortes:Novartis: Consultancy, Research Funding; Bristol-Myers Squibb: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Ariad: Consultancy, Research Funding.
Prognostic Significance of Immunocytometric Count of Medullar Blasts in Myelodysplastic Syndromes
Authors: Molteni Alfredo , Speziale Valentina , Riva Marta , Nichelatti Michele , Greco Rosa , Cesana Clara , Scarpati Barbara , Morra Enrica 
Published: October 2019

Abstract
Abstract 4945

Background:
bone marrow blast count is a crucial parameter to determinate the prognostic score according to IPSS in myelodysplastic syndromes. However even among the major experts the agreement about detection of a blast cells by optical microscopy is less than 90% (Mufti et al: Haematologica 2008). The aim of this study is to evaluate prognostic significance of immunocytometric count of medullar blasts compared to cytological count.

Method:
in a retrospective analysis of 104 patients with a minimum follow-up of three years, IPSS was calculated replacing cytological count of blasts with the flow cytometry one. Blasts were expressed as percentages of total cellularity, and identified by the combination of CD45/SSC with/without CD33, CD34 and CD117 antigens. The monoclonal antibodies used were CD34 FITC, CD117 PE, CD45 PerCp, CD33 APC. Acquisition of information on 1×105 stained cells corresponding to the whole bone marrow cellularity was assessed on a dual-laser FACSCalibur flow cytometer using the CellQUEST software (Becton Dickinson, San José CA USA). Survival of “low + intermediate-1 risk” (“IPSS1+2”) patients was compared to “intermediate-2 + high risk” ones (“IPSS3+4”) using Kaplan Meier method followed by logrank test. The comparison of ability among all blast count models to distinguish the probability of survival of the two groups (“IPSS 1+2” vs “IPSS3+4”) was based on χ2 value obtained by the logrank test.

Results:
considering IPSS obtained from any of the immunocytometric markers considered (CD33, CD34, CD117 and CD45dim/SSC), “IPSS1+2” group had always an extremely better survival than “IPSS 3+4” group with a level of statistical significance similar to survival study of “standard IPSS”. Of all markers, CD45dim/SSC was the most capable to distinguish survival of “IPSS1+2” group versus “IPSS3+4” group (χ2 = 42. 5). Among others, CD117 and CD33 had a better discriminating power than cytological count (respectively χ2 = 38, 1 and χ2 = 22, 3 versus χ2 18, 2); only CD34 was worse than cytological count (χ2 =14, 2).

Conclusion:
the immunocytometric markers considered are demonstrated to be even better than cytological count as parameter in IPSS score. Their use has to be encouraged, especially where there is little experience in optical microscopy.

Disclosures:
No relevant conflicts of interest to declare.
Targeting Mir-548m-HDAC6 Axis Circumvents Stroma- Mediated Drug Resistance (CAM-DR) and Clonogenicity in Non-Hodgkin B-Cell Lymphomas
Authors: Lwin Tint , Zhao Xiaohong , Cheng Fengdong , Zhang Xinwei , Zhang Yizhuo , Huang Andy , Moscinski Lynn , Dalton William S. , Sotomayor Eduardo M. , Tao Jianguo 
Published: October 2019

Abstract
Abstract 5094
A dynamic interaction occurs between the lymphoma cell and its microenvironment, with each profoundly influencing the behavior of the other. Here, we demonstrate that adhesion of mantle cell lymphoma (MCL) and other non-Hodgkin lymphoma cells to lymph node stromal cells, follicular dendritic cells (FDCs), confers drug resistance, enhances lymphoma cell clonogenicity, and is associated with induction of histone deacetylase 6 (HDAC6). Furthermore, stroma down-regulated miR-548m contributes to HDAC6 up-regulation, and HDAC6 is a key determinant for FDC-mediated lymphoma cell survival and colony formation. We further showed that stroma-mediated drug resistance and clonogenic growth are reversed by enforced expression of miR-548m and inhibition of HDAC6. The HDAC6-selective inhibitor tubastatin significantly enhances cell death, abolishes cell adhesion-mediated drug resistance, and suppresses clonogenicity and lymphoma growth suppression ex vivo and in vivo. Together, these data suggest that the lymphoma–stroma interaction in lymph node microenvironment directly impacts the biology of lymphoma through epigenetic regulation of miRNA and HDAC, with HDAC6 as a potential therapeutic target to overcome drug resistance in MCL and other B-cell lymphomas.

Disclosures:
No relevant conflicts of interest to declare.
Molecular Diagnosis of One Pedigree with Protein S Deficiency and Antithrombin Deficiency
Authors: Zhou Rong-Fu , Tao Hong , Ouyang Jian , Zhang Xian , Yang Yonggong , Zhang QiGuo , Xu Jingyan , Chen Bing 
Published: October 2019

Abstract
Abstract 5140

Objective
To identify gene mutations for one patient and his family members with protein S and antithrombin deficiency.

Methods
ELISA were used to detect protein S (PS), protein C (PC) and antithrombin (AT) activities for the proband and family members, respectively. The genomic DNA was extracted from the peripheral blood of proband and family members. All exons and their flanks of protein S gene and antithrombin gene were amplified by polymerase chain reaction (PCR). The PCR products were sequenced directly. The mutation-related exons of his famliy members were amplified by PCR and sequenced directly.

Results
The proband was a 49-year-old male. He presented with sudden left lower extremity swelling and pain without casues. Regular examination revealed that his APTT, PT, and TT were all in normal levels, but D- dimmer was 5. 62mg/L, Color doppler ultrasonography showed thrombosis in his left femoral vein. The activity of PS for his family members was ‡1 0%, ‡2 0%, ‡3 0%, ‡4 130. 8%, ‡5 8. 4%, ‡1 0%, ‡2 0%, and that of AT was ‡1 129. 1%, ‡2 51. 9%, ‡3 73.2%, ‡4 119. 1%, ‡5 136. 2%, ‡1 65. 5% and ‡2 60. 1%, respectively. The sequencing analysis showed that a heterozygous missense mutation G68395T (NG_009813. 1) was detected in Exon 4 of PS gene leading to the substitution of Arg90 by Leu (NP_000304. 2) for the propositus. The heterozygous mutation (Arg90Leu) was also found in other family members. A heterozygous (nonsense) mutation G12444A (NG_012462. 1) was detected in Exon 4 of AT gene leading to Trp257Ter (NP_000479. 1) for the propositus. The mutation (Trp257Ter) was found in other family members with reduced activity of AT. These two mutations (G68395T in PS gene and G12444A in AT gene) were not reported before and were thus novel ones.

Conclusion
The novel mutation G68395T in PS gene and G12444A in AT gene might be the causes of deficiency of PS and AT for the family.

Disclosures:
No relevant conflicts of interest to declare.
Noncanonical Wnt Signaling Maintains Hematopoietic Stem Cells in Different Zones
Authors: Sugimura Ryohichi , Li Linheng 
Published: October 2019

Abstract
Abstract 639
Hematopoietic stem cells (HSCs) are maintained in a balance between quiescent state and proliferating state. While proliferating HSCs are critical for supporting the routine blood production, quiescent HSCs are essential for long term maintenance and can also be activated to replenish lost proliferating or active HSCs. How the different states of HSCs are regulated is a fundamental question. Accumulated evidence supports a model that quiescent HSCs are located in the endosteal zone and active HSCs are in the perivascular zone. The underlying signaling to regulate the quiescence and activation in different niches remains largely unknown. To address this question, we have analyzed the expression profile of Wnt receptors, Frizzleds, in HSCs. We found that noncanonical Wnt signaling via receptor Frizzled8 (Fz8) and co-receptor Flamingo presents in and functionally maintains quiescent HSCs in the endosteal zone (Sugimura, et al., Cell 2012). However, it has not been clear whether and how active HSCs in the perivascular zone are regulated by Wnt signaling.
Recently, we detected that another noncanonical Wnt receptor, Frizzled5 (Fz5), is expressed in metabolically active (indicated by Mitotracker) HSCs and also in Nestin-GFP+ mesenchymal stem cells (MSCs) in the perivascular zone of central marrow. Fz5 expresses neither in H2B-GFP label-retaining quiescent HSCs nor in endosteal cells and nor in sinusoidal cells as well. Using an Mx1-Cre:Fz5 knockout mouse model, we found a 60% decrease of HSCs isolated from central marrow, but no change in the number of HSCs isolated from endosteum. Functionally, hematopoietic reconstitution was not affected in the primary transplantation, but was substantially decreased (by 80%) in the secondary transplantation compared to the control.
This indicates that Fz5 maintains HSCs in the perivascular zone. To examine the role of Fz5 in Nestin+ MSCs for HSC maintenance, we examined the Nestin-Cre:Fz5 model. We observed a large loss of CD49hiHSCs that were reported to represent intermediate (IT) HSCs. We further found a correlation of the quiescent vs. active HSCs respectively to long term (LT) HSCs vs. IT-HSCs with the latter population sensitive to 5FU treatment. Mechanistically we observed that Fz5 inactivation also led to a loss of Cdc42 polarity in the HSCs residing in the perivascular niche. The results suggest that Fz5-mediated noncanonical Wnt signaling regulates polarity of active HSCs in the perivascular zones. Future study is required to see whether the Fz5-Cdc42 mediated polarity in HSCs is associated with symmetric vs. asymmetric division.
We propose that noncanonical Wnt signaling maintains quiescent and active HSCs reside respectively in the endosteal zone and the perivascular zone. In these zones, Fz8 and Fz5 are differentially expressed and mediate noncanonical Wnt signaling for HSC maintenance in the endosteal niche and to regulate active HSC action in the perivascular niche.

Disclosures:
No relevant conflicts of interest to declare.
Low RPS14 Expression in MDS without 5q- Aberration Is Associated with Increased Apoptosis of Erythrocytes and Predicts Prolonged Survival and Possible Response to Lenalidomide in Lower-Risk Patients
Authors: Wu Lingyun , Li Xiao , Xu Feng , Zhang Yan , He Qi , Chang Chunkang 
Published: October 2019

Abstract
Abstract 698

Purpose:
Haploinsufficiency of the ribosomal protein S14 (RPS14) has been identified as a causal factor in 5q- syndrome. This study was carried out to investigate RPS14 expression in myelodysplastic syndrome (MDS) without 5q deletion and the role under-expressed RPS14 plays in pathogenesis and the prognosis and lenalidomide-response predicting of non-5q-MDS.

Patients and Methods:
The expression level of RPS14 transcription was detected in 156 MDS patients without 5q-. The apoptosis index of bone marrow (BM) erythrocytes was also analyzed. Furthermore, patient prognosis with low or normal RPS14 expression was analyzed and the role of RPS14 expression in lenalidomide-response prediction was elevated.

Results:
The reduced expression of RPS14 occurred in 83 of 156 (53.2%) patients. Patients with RPS14 under-expression presented higher platelet counts in the peripheral blood compared to RPS14 normal patients (p=0.012). RPS14 expression status was inversely corrected with the apoptosis index of erythrocytes from BM (r=-0.54, p=0.013). Patients with low RPS14 expression have a higher two-year survival probability than normal RPS14 cases in the international prognosis scoring system (IPSS) lower-risk group (90.8% vs. 71.7%; p = 0.018). A multivariate analysis showed RPS14 expression status was an independent predictor for survival in lower-risk MDS patients without 5q deletion. Twelve patients were treated with lenalidomide. Five of seven patients achieved an erythroid response in the low RPS14 expression group (5/7, 71.4%), compared to zero responses in the five normal RPS14 patients (p=0.018).

Conclusion:
Low RPS14 expression is common in MDS patients without 5q deletion and is associated with increased apoptosis of erythrocytes in lower-risk MDS. Additionally, low RPS14 predicts prolonged survival and possible response to lenalidomide in lower-risk MDS patients.

Disclosures:
No relevant conflicts of interest to declare.
TAS-117, a Novel Selective Akt Inhibitor Demonstrates Significant Growth Inhibition in Multiple Myeloma Cells in Vitro and in Vivo
Authors: Mimura Naoya , Ohguchi Hiroto , Cirstea Diana , Cottini Francesca , Gorgun Gullu Topal , Minami Jiro , Suzuki Rikio , Shimomura Toshiyasu , Utsugi Teruhiro , Hideshima Teru , Anderson Kenneth C. 
Published: October 2019

Abstract
Abstract 942
The PI3K/Akt pathway mediates multiple myeloma (MM) cell growth and drug resistance, and targeting this molecule is a promising therapeutic option. In this study, we examined anti-MM activities of TAS-117 (TAIHO PHARMACEUTICAL CO., LTD., JAPAN), a selective potent Akt inhibitor in MM cell lines including MM.1S, MM.1R, OPM1 and H929 cells with high level of baseline Akt phosphorylation. TAS-117 induced significant growth inhibition in these cell lines, associated with downregulation of phosphorylation (Ser473 and Thr308) of Akt and downstream molecule FKHR/FKHRL1, without cytotoxicity in normal peripheral blood mononuclear cells. TAS-117 triggered G0/G1 arrest followed by apoptosis, evidenced by increased annexin V-positive cells, in both MM.1S and H929 cell lines. Apoptosis was further confirmed by cleavage of caspase-8, -3 and PARP. Interestingly, TAS-117 also induced: autophagy, evidenced by increased LC3-II; as well as endoplasmic reticulum (ER) stress, confirmed by induction of phospho-eIF2α, phospho-IRE1α and a molecular chaperone BiP/GRP78. Since the bone marrow (BM) microenvironment plays a crucial role in MM cell pathogenesis including drug resistance, we further examined the effect of TAS-117 in the presence of BM stromal cells (BMSCs). TAS-117 induced significant cytotoxicity in MM cells even in the presence of BMSCs, associated with downregulation of phospho-Akt. Importantly, TAS-117 inhibited secretion of IL-6 from BMSCs, and exogenous IL-6 and IGF-1 did not block cytotoxicity induced by this agent. We have previously shown the bortezomib activates Akt, and that Akt inhibition with bortezomib triggers synergistic MM cell cytotoxicity. TAS-117 enhanced bortezomib-induced cytotoxicity in MM.1S cells, associated with increased CHOP followed by PARP cleavage, suggesting that TAS-117 augments bortezomib-induced ER stress and apoptotic signaling. TAS-117 also enhanced cytotoxicity induced by other therapeutic agents (ie, rapamycin, dexamethasone, 17-AAG) in MM.1S cells. Finally, we examined anti-MM activities of TAS-117 in a xenograft murine model. Oral administration of TAS-117 for 14 days significantly inhibited growth of H929 plasmacytoma and was well tolerated. Taken together, the novel and selective Akt inhibitor TAS-117 blocks MM cell growth in vitro and in vivo, providing the preclinical framework for clinical evaluation of this agent to improve patient outcome in MM.

Disclosures:
Shimomura: TAIHO PHARMACEUTICAL CO., LTD.: Employment. Utsugi:TAIHO PHARMACEUTICAL CO., LTD.: Membership on an entity's Board of Directors or advisory committees. Anderson:Celgene, Millennium, BMS, Onyx: Membership on an entity's Board of Directors or advisory committees; Acetylon, Oncopep: Scientific Founder, Scientific Founder Other.
A Novel ENU-Induced Intronic Mutation of Ank1 Causing Quantitative Defect of Ankyrin-1 Models Hemolytic Hereditary Spherocytosis in Mouse
Authors: Du Xin , Huang Hua , Krieg Lara 
Published: October 2019

Abstract
Abstract 989
The ability of red blood cells (RBCs) to maintain the surface area and deformability is vital for their survival. The maintenance of membrane surface is dependent upon the strong cohesion between the lipid bilayer and the skeletal network, achieved by vertical linkages between transmembrane proteins and spectrin tetramers. Mutations causing functional deficiencies in these proteins have been identified in various hemolytic anemias. Here we reported a mild hereditary spherocytosis (HS) and hemolytic anemia phenotype in mouse, named hema6, induced by N-ethyl-N-nitrosourea (ENU) mutagenesis. Hema6 phenotype is transmitted as a semidominant trait as heterozygous mice are less severely affected than homozygotes. The causal mutation was traced to a single nucleotide transition in the deep intronic region of intron 13 of gene Ank1, encoding the anchorage protein ankyrin-1 in RBC. In vitro minigene assay revealed two abnormally spliced transcripts in addition to wild-type mRNA. The wild-type Ank1 transcript was detected in the homozygous mutant mouse with 30% reduction in expression level compared to that in wild type mouse. The aberrant transcripts presumably encoded a 509 amino acids protein, which lacks beta-spectrin binding domain and C-terminal regulatory region. The truncated protein was not detected by western blotting using currently available antibodies against full-length ankyrin-1 in homozygous hema6 erythrocyte ghosts, whereas the wild-type Ankyrin-1 are present with reduced quantity. Employing biochemical and cell biology assays, we characterized the mechanism by which Ank1hema6 mutation causes hemolytic hereditary spherocytosis in mouse. Hema6 strain provides a novel tool to study ankyrin-1 and its pathogenesis role in HS.

Disclosures:
No relevant conflicts of interest to declare.