bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of NOTCH1 in Aggressive NK-Cell Leukemia.
Authors: Makishima Hideki , Ota Masao , Suzuki Takefumi , Nakayama Kozo , Ishida Fumihiro 
Published: October 2019

Abstract
Back ground: Aggressive NK-cell leukemia (ANKL) is a malignant NK-cell proliferative disorder resistant to most chemotherapies and frequently shows rapidly progressive clinical course. ANKL is characterized with hepatosplenomegaly, hemophagocytic syndrome, and strong association with EBV, and is mostly observed among Asian populations. Oncogenic mechanism of ANKL is not clear. NOTCH1 is a gene encoding a transmembrane receptor that regulates T-cell development and is a major oncogene in T-cell lymphoblastic leukemia (T-ALL). Recently, the potential involvement of NOTCH signaling in the development of NK-cells has been reported. We discuss here that the NOTCH1 protein might play a significant role in the pathogenesis of ANKL.
Patients, materials, and methods: 6 ANKL cases, 7 Chronic NK-cell lymphocytosis (CNKL) cases, and 3 NK tumor cell lines (NKL, NK-92, and KHYG) derived from ANKL patients were enrolled. Chromosomal translocation (7;9) was not found in any subjects. The expression levels of NOTCH1 intracellular subunit (NOTCH-IC) were examined with flowcytometry and western blotting. NK-cell lines were cultured with g-secretase inhibitor (GSI) (L-685,458) for evaluation of the effect. Cell proliferation and apoptosis were measured with XTT assay and annexinV positivity, respectively. The mutation analysis of NOTCH1 genome was performed with nested PCR and direct sequencing. mRNA expression of NOTCH1 gene was measured with real time PCR using the corresponding TaqMan probes.
Results: NOTCH-IC was detected in CD2+CD3− fractions from healthy donors, CNKL patients, ANKL patients, NK-cell lines, and Cos7 cells transfected with NOTCH1 gene coding NOTCH-IC, but not in CD2-CD3− fractions. The expression levels were significantly high in ANKL patients, compared to healthy donors (P < 0.05). With western blotting, cleaved form (110kDa) of NOTCH1 protein was recognized in all NK-cell lines. NOTCH-IC expression was reduced with the treatment of GSI in NKL and NK-92, but not in KHYG. Cell proliferation was inhibited with GSI in NKL and NK-92, whereas the inhibitory effect was not seen in KHYG. There was no significant difference in the increase ratio of apoptotic cells after the treatment of GSI among 3 NK-cell lines. Mutations in HD, TAD, or PEST domain, where frequent mutations are detected in T-ALL, were not found in NK-cell lines, and no enhanced expression of NOTCH1 mRNA in ANKL cells, compared with healthy donor NK-cells.
Conclusion: These results indicate that NOTCH-IC was increased in ANKL cells and might be an important role in the proliferation. Genomic or transcriptional dysregulation would not be the cause for enhanced expression of NOTCH1 in ANKL contrast with T-ALL. Further investigation is required for the relationship with NOTCH1 ligands. In the future, NOTCH1 might be a novel therapeutic target in ANKL.
FLT3 Mutation in Patients with Acute Promyelocytic Leukemia Has No Effect on Therapy Response.
Authors: Xue Meng Xing , Qiu Hui Ying , Feng Yu Feng , Pan Jin Lan , Chen Su Ning , Wu De Pei 
Published: October 2019

Abstract
Fms-Like tyrosine kinase 3 (FLT3) mutations are one of the most frequent genetic changes in acute myeloid leukemia (AML) and are related to poor prognosis. However, in acute promyelocytic leukemia (APL) the prognostic significance of this mutation is not firmly established. We investigated FLT3 internal tandem duplications (FLT3/ITD) or point mutation of the activation loop domain (FLT3/ALM) in initial marrow samples in 160 APL patients, also studied the impact of FLT3 mutations on disease characteristics and clinical outcome. Some of FLT3/ITD+ samples were further examined to determine the ITD allelic ratio(ITD-AR) using Genescan analysis. FLT3/ITD and FLT3/ALM were detected in 31 (19.4%) and 17(10.6%) of the patients, 2(1.25%) showed both ITD and ALM mutations. 19 of 31 FLT3-ITD+ patients were examined to determine ITD-AR, which varied from 0.11 to 1.79, with a median of 0.92. ITD-AR in 8 patients were greater than 1.0. Both mutations were associated with higher white blood cell(WBC) count at presentation(P<0.05); Among patients with WBC counts greater than 10×109/L, FLT3/ITD and FLT3/ALM rates were 49.1%(26/53) and 22.6% (12/53) respectively, were both higher than those of patients with WBC counts lower than 10×109/L (P<0.01). For clinical outcome, the complete remission(CR) of FLT3-ITD+ patients was lower than that of FLT3-ITD− patients(64.5% vs 91.5%, P<0.01), however the CR rates were not significantly different between patients with and without FLT3-LAM(84.6% vs 100%, P>0.05). Six originally FLT3-ITD+ patients were not detectable mutation after remission; so were six patients with originally FLT3-LAM+; however FLT3-LAM in one originally FLT3-LAM+ patient reappeared after relapse. In conclusion, FLT3 mutations (FLT3/ITD or FLT3/ALM) were frequently identified in patients with newly diagnosed APL, and both mutations were associated with higher WBC count at presentation. FLT3/ITD is more frequent than ALM mutation, and predicts a poorer prognosis because of the lower CR rates, while, FLT3/ALM mutation did not show the same unfavorable prognostic effect. In our study, ITD-AR were not found significantly difference between CR and no remission patients (P>0.05)
Wilm’s Tumor 1 Gene Expression Is a Useful Marker for Minimal Residual Disease in Acute Myeloid Leukemia.
Authors: Hsu Jack W. , Liu Hong , Jamieson Katarzyna , Hou Wei , Chang Myron N. , Ahrens Kim P. , Braylan Raul C. , Wingard John R. 
Published: October 2019

Abstract
One of the difficulties in managing patients with acute myeloid leukemia (AML) is the detection of minimal residual disease. Only a fraction of AML patients have a distinct phenotype on flow cytometry or a genetic abnormality which can be followed as a sensitive marker for minimal residual disease (MRD). The Wilm’s tumor - 1 gene (WT1) is overexpressed in leukemic blasts, but not in normal blood or marrow cells, and thus can be used as a marker for MRD in leukemia patients who otherwise do not have a characteristic immunophenotypic or molecular marker that can be monitored. We prospectively measured serial peripheral blood WT1 levels by RQ-PCR in 54 patients with AML. Of the 54 patients, 23 patients were newly diagnosed. The remaining 31 patients were in complete remission prior to a planned allogeneic transplant. Subsequent WT1 levels were obtained during routine evaluation both during and up to three years after completion of either chemotherapy and/or transplant. Using regression analysis, we found that a normalized WT1 level greater than 10−4 (as compared to a level of 1 in K562 cells) indicates a higher likelihood of the patient having active leukemia with a sensitivity of 87.5% and a specificity of 89.1%. The positive and negative predictive values are 71.19% and 95.89% respectively. We also compared WT1 levels with peripheral blood and bone marrow blasts to determine whether a correlation existed. We found a strong correlation between WT1 expression and bone marrow blasts with a correlation coefficient of 0.77 (p< 0.0001). A weak correlation was found with peripheral blood blast count (correlation coefficient = 0.35, p<0.0001). Our data implies WT1 can be a useful marker for minimal residual disease in patients with acute myeloid leukemia, especially in patients who do not have an unusual phenotype or molecular marker to monitor MRD.
Prognostic Implications and Biological Roles of RhoH in Acute Myeloid Leukemia.
Authors: Iwasaki Toshihiro , Katsumi Akira , Kiyoi Hitoshi , Tanizaki Ryohei , Kobayashi Miki , Abe Akihiro , Matsushita Tadashi , Naoe Tomoki 
Published: October 2019

Abstract
The Rho family of small GTPases including Rho, Rac, and Cdc42 has been well characterized as molecular switches to transduce signals from plasma membrane to the downstream effectors. Rac1 and Rac2 are known to regulate engraftment and mobilization of hematopoietic stem cells. RhoH, a member of the Rho family, is specifically expressed in hematopoietic cells, and has been reported to inhibit the cell adhesion through regulating Rac and αLβ2 integrin. As RhoH is GTPase deficient and constitutively active, GTP-bound form, the activity of RhoH is directly related to the level of expression. Previous reports demonstrated the aberrant somatic hypermutation of RhoH gene as a novel mechanism of genetic lesion in diffuse large B-cell lymphoma, possibly through the deregulated expression although the role of the RhoH on leukemia is largely unknown. Here we have screened for the expression level of RhoH gene in the bone marrow samples from 90 previously untreated acute myeloid leukemia cases by using a real-time fluorescence detection method. The expression level of RhoH was neither related to the FAB classification, CR rate, nor WBC counts. In addition, the RhoH expression was not associated with the known gene mutations such as N-Ras, FLT3, and p53. However, the multivariate analysis demonstrated that low expression of RhoH was the independent unfavorable prognostic factor for overall and disease free survival (p=0.0028 and 0.003, respectively). RhoH did not affect the affinity modulation of α4β1 integrin, however, RhoH negatively regulate Rac activation in our system, suggesting that RhoH might work as a proapoptic molecule through Rac deactivation. Further investigations would be required to clarify the biological roles of RhoH on leukemic cells.
Retrospective Analysis of Post-Remission Modalities in AML Patients with Normal Karyotype.
Authors: Chae Yee Soo , Sohn Sang Kyun , Kim Jong Gwang , Cho Yoon Young , Moon Joon Ho , Ahn Byung Min , Kim Shi Nae 
Published: October 2019

Abstract
Background: Cytogenetics is considered as the most important factor to determine type of post-remission treatment in patients with AML However, there are controversy regarding to select consolidation modality in the intermediate risk group because of heterogeneity. We compared the outcome of patients with normal karyotype according to the different consolidation modalities.
Methods: AML patients with normal karyotype who achieved complete remission (CR) and did not have any contraindication for SCT, were included. The outcomes were retrospectively analyzed according to consolidation treatments.
Result: Among 51 patients who achieved CR, 35 patients underwent stem cell transplantation (SCT; 27 HLA-matched allogeneic and 8 autologous SCT), while 16 patients received high-dose cytarabine chemotherapy (HDAC) for consolidation therapy. No differences in the patient characteristics were observed. The estimated relapse rate at 3 years was significantly lower in the SCT group (38.6 vs. 75.6%, p=0.010). The DFS was significantly higher in the SCT group (HR=0.360; 95%CI=0.161–0.870; p=0.013), while OS was insignificantly higher in the SCT group (HR=0.489; 95%CI=0.199–1.205; p=0.120). There were no differences in DFS and OS between allogeneic and autologous SCT (p=0.714 and 0.515, respectively)
Conclusion: SCT showed a better survival outcome than chemotherapy alone as post-remission therapy in AML patients with normal chromosome. Considering late complications like GVHD, auto-SCT might be a feasible option for post-remission therapy in patients with normal karyotype.
Expression of p14ARF in De Novo AML with Normal Karyotype. Implication on Drug Resistance and Survival.
Authors: Paul Esbjorn O. , Deneberg Stefan , Lehmann Soren , Bengtzén Sofia , Nahi Hareth 
Published: October 2019

Abstract
The p53 protein exerts a significant role in growth control of cells and impaired function of p53 by mutation or otherwise is regarded to be important for development of many human cancers. The p53 protein is strongly regulated by the E3-ligase HDM-2 that specifically binds to p53 and causes proteosomal degradation. P14ARF encoded from the INK 4a/ARF gene locus of chromosome 9p, activates the p53 pathway by binding to and inhibiting HDM-2. The objective was to study if the level of mRNA for p14ARF in leukemic cells from patients with AML was related to in vitro drug resistance and clinical outcome. Leukemic cells (>90% pure) isolated from 46 adult patients with normal karyotype de novo AML were incubated with antileukemic drugs (daunorubicin, mitoxantrone, etoposide and Ara-C) cultured for 4 days. The effect was determined by bioluminiscence measuring of intracellular ATP compared to an untreated control. We also tested the activity against PRIMA (p53-dependent reactivation and induction of massive apoptosis), a novel, small molecule shown to activate down regulated p53. mRNA for p14ARF was determined by real time PCR.
Results: Patients whose leukemic cells expressed high level of p14ARF mRNA in the leukemic cells (≥0.2 compared to the housekeeping gene) had a significantly better survival compared to those with low level (<0.02). Median survival not obtained compared to 9 months. The mean activity of all tested conventional antileukemic drugs was higher on leukemic cell samples expressing p14ARF mRNA ≥0.2 compared to those with low levels. In contrast, PRIMA exerted significantly higher in vitro effect on leukemic cell samples with low levels of p14ARF mRNA. We conclude that low levels of p14ARF in leukemic cells from patients with normal karyotype AML is a marker for poor prognosis, which may depend on impaired p53 activity causing resistance against conventional antileukemic drugs. Treatment with p53 targeting drugs as PRIMA may be a future possibility to improve the outcome for these patients.
Predictive Value of Flow Cytometric MRD Analysis in Childhood Acute Lymphoblastic Leukaemia at the End of Remission Induction Therapy.
Authors: Motwani J. , Jesson J. , Sturch E. , Eyre L. , Short P. , Williams M.D. , Darbyshire P.J. , Hill F.G.H. , Lawson S. 
Published: October 2019

Abstract
Patients with acute lymphoblastic leukaemia (ALL) in morphological remission may still have up to 1010 residual malignant cells. Detection of minimal residual disease (MRD) at the end of induction therapy allows better estimation of the leukaemic burden and can help selection of appropriate therapeutic strategies. Flow cytometric (FC) detection of MRD is based on the identification of immunophenotypic combinations expressed on leukaemic cells but not on normal hematopoietic cells - leukaemia associated immunophenotypes (LAIPs). We prospectively analysed bone marrow samples from 77 patients who presented with ALL to our unit between 1999–2003 and attained morphological remission. These patients were treated on a standard protocol. Multiparameter FC identification of LAIPs was performed at various time points, as dictated by the treatment protocol. Our results show that flow cytometric MRD at the end of induction therapy is an independent and the most significant predictor of relapse, both on univariate and multivariate analysis. The relapse risk was 4% if day 28 MRD was <0.01% and 50% if day 28 MRD was >0.01% (p<0.05). We conclude that flow cytometric based MRD assays can be used to assess early response to treatment and predict relapse in a similar way to molecular MRD analysis at the end of induction therapy. Flow cytometric analysis of MRD offers the advantages of being cheaper, more widely available and has quicker turnaround times.
Influence of Maternal Folic Acid Supplementation on Hematopoietic Factors in Offspring.
Authors: Johnson Kimberly J. , Largaespada David A. , LeBien Tucker W. , Allaei Raha , Ross Julie A. 
Published: October 2019

Abstract
Observational studies of childhood acute lymphoblastic leukemia (ALL) have generally supported a protective role for maternal intake of foods or vitamins rich in folic acid during pregnancy. Using C57BL/6J mice, we evaluated the effect of dietary folic acid intake in the perigestational period on surface marker and gene expression in B-lineage cells as a potential model to further elucidate the role of maternal diet in the etiology of childhood ALL. Thirty-six female mice were randomized to one of three folic-acid varied diets (low=0.3, control=2.0, high=8.0 mg/kg) one month prior to mating and maintained on the diet until pup weaning at three weeks. Reproductive characteristics and offspring red blood cell (RBC) folate were recorded, and bone marrow lymphocytes were isolated for surface marker characterization by flow cytometry. For gene expression analyses, CD19 positive B-lineage bone marrow cells were purified from three pups per dietary group using anti-CD19 microbeads. Gene expression levels were interrogated using the Agilent mouse whole genome microarray. No statistically significant differences were found among dietary groups with respect to mating success, litter size, estimated 21 day offspring weights, or the percentage of male offspring. However, there were significant differences among the 3 diets in offspring survival to the end of the experiment (39%, 84%, and 30% for the 0.3, 2.0, and 8.0 mg/kg groups, respectively). Pup RBC folate values were positively correlated with dietary folic acid dose with mean values of 428, 615, and 724 ng/ml, respectively. Mice were euthanized at 3 weeks. There was no significant difference in the expression of CD19, IgM, or kappa light chains on bone marrow lymphocytes in the offspring among dietary groups. ANOVA analysis, using Genedata Expressionist Pro software, showed differential expression of 267 genes using a Benjamini and Hochburg false discovery rate cutoff of 10%. Three main patterns of gene expression between pups of different dietary groups were identified through unsupervised hierarchical clustering analysis; 152 and 86 genes were up- and down-regulated, respectively, in both the high and the low folic acid groups relative to the control group. Twenty-seven genes showed increasing expression with increasing dietary folic acid dose. In conclusion, dietary folic acid given to dams prior to conception through weaning had a significant effect on offspring survival and gene expression. Additional analyses are underway to determine the relevance of genes found to be differentially expressed with respect to risk of B-cell lineage leukemias followed by validation of relevant gene candidates through quantitative RT-PCR. These data will be used to help inform future etiologic studies of childhood ALL.
A Mouse Strain Which Responds Poorly to Angiogenic Stimuli Has a Higher Incidence of Irradiation-Induced Hematological Malignancies.
Authors: Osorio Catarina , Fragoso Rita , Igreja Catia , Dias Sergio 
Published: October 2019

Abstract
Genetic heterogeneity of murine strains to vasculogenic/ angiogenic stimuli has been suggested to correlate with the susceptibility to carcinogenesis and subsequent tumor growth. A more “angiogenic” strain would, likely, be more prone to carcinogenesis/ tumorigenesis stimuli. To study the importance of the endothelial compartment (vasculogenesis/ angiogenesis) during irradiation-induced hematopoietic malignancies (leukemia and lymphoma) we administered 3 cycles of irradiation (sub-lethally; 3.5 Gy), with one month of interval between them, to 2 angiogenically different mouse strains (12 high angiogenic Balb/c mice vs 12 low angiogenic C57BL-6 mice). Every month after the last irradiation, the levels of circulating endothelial progenitor cells (EPC: Flk-1+Sca+ cells) in peripheral blood were quantified by flow cytometry. At the end of the assay (∼6 months after last irradiation) the incidence of hematopoietic disease among the 2 mouse strains was scored and the levels of EPC (Flk-1+Sca+ cells), endothelial cells (Flk-1+ cells) and HSC/HPC (Sca+c-Kit+ cells) were measured in BM (bone marrow) samples of sick and healthy mice. Interestingly, the less angiogenic C57BL-6 mouse strain displayed a significantly higher leukemia and thymic lymphoma incidence (50% - 2x-fold higher than Balb-c). As determined by FACS analysis of BM cells, in both mouse strains, sick mice showed a reduction in the levels of HSC/HPC (hematopoietic progenitor cells) compared to their healthy counterparts. Nonetheless, this reduction was more pronounced in the less angiogenic C57BL-6 mouse strain (2.1 reduction in C57BL-6 vs. 1.4 reduction in Balb-c). Beside the difference in the hematopoietic compartment, C57BL-6 leukemic mice also evidenced a dramatic change in the BM endothelial compartment, where a significant decrease in EPC and endothelial cells was observed compared to leukemic Balb-c and healthy mice. In contrast, C57BL-6 leukemic mice showed a higher number of circulating EPC compared to healthy mice. Taken together, our data suggests that a higher incidence of hematopoietic malignancies observed in the less angiogenic mouse strain may be associated with a deficiency in the BM vascular compartment. Since it is known that whole body irradiation induces the turnover and the mobilization of HSC/HPC inside the BM, we hypothesize that the vascular compartment (EPC/ endothelial cells) regulates HSC/HPC turnover during irradiation-induced carcinogenesis. As such, BM vascular deficiency/ depletion after BM irradiation (particular evident in low angiogenic genetic backgrounds) may promote the turnover of HSC/HPC and increased susceptibility to the irradiation effects. These results may be particularly relevant in the context of irradiation/ chemotherapy-induced secondary tumors in myelodysplastic/ leukemia patients.
Detection and Production of Haptoglobin within Acute Myeloid Leukemic Blasts.
Authors: Lingamfelter Daniel , Yu Na , Quinn Tim , Williams Casey , Guan Xiuqin , McGuirk Joseph , Chen Yayan , Abhyyankar Stella , Arora Navanshu , Gaur Rakesh , Wiemen Jeffery , Yan Ying 
Published: October 2019

Abstract
While a directly proportional relationship between serum haptoglobin (Hp) levels and acute myeloid leukemia (AML) has been documented, the exact source of this elevated haptoglobin remains in dispute. Furthermore, no reported studies thus far have analyzed haptoglobin-targeted immunohistochemistry as a potential diagnostic tool for AML. In this study, we examined Hp expression in patients with AML. Previously collected blood samples were obtained, with informed consent, during routine diagnostic blood studies from twenty-one patients with AML (13 males, 8 females; age range 4–73 years). Cytospin samples were prepared, and immunohistochemistry was performed using a polyclonal anti-haptoglobin antibody. Total RNA was extracted from 19 AML patient samples as well as from a K562 leukemic cell line for the RT-PCR assay. Cytospin specimens derived from all twenty-one patients with confirmed AML showed consistent anti-haptoglobin staining of blast cells in a granular, cytoplasmic pattern. The K562 cell line, used as a positive control, stained in a similar fashion to the patient samples. Immunohistochemical co-labeling of blasts was achieved with the AML anti-podocalyxin marker in conjunction with the anti-haptoglobin antibody. All of the AML blasts as well as the K562 cells expressed Hp by the RT-PCR assay. The ratio index (RI) of Hp band density against β-actin serving as an internal control was used to compare the expression level among the AML cases. The RIs ranged from 0.89 to 0.02, indicating different Hp transcription levels among the AMLs. We conclude that haptoglobin is produced by AML blasts and stored within the cytoplasm. This may partly explain the increased serum level of haptoglobin observed in AML patients. Furthermore, immunohistochemistry consistently detects these intra-blastic haptoglobin stores, suggesting a potential for a novel diagnostic tool for AML.