bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hypermethylation and Expression of Rassf1a in Childhood Acute Lymphoblastic Leukemia (ALL).
Authors: Yaniv Isaac , Betzer Hadar , Naumov Inna , Stark Batia , Avrahami Gali , Luria Drorit , Kodman Yona , Avigad Smadar 
Published: October 2019

Abstract
Epigenetic modifications including promoter hypermethylation and histone acetylation are emerging as one of the factors controlling the expression of many tumor suppressor genes. One such gene is the Ras effector RASSF1A (Ras association domain family 1). Promoter hypermethylation of Rassf1a is already well established and clinically relevant in many adult cancers and also many pediatric solid tumors. However not much is known about its status in childhood acute lypmphoblastic leukemia (ALL). The aim of this research was to study RASSF1A methylation and expression and also evaluate mutations in ras oncogene in childhood ALL and correlate it with clinical outcome. Bone marrow samples at diagnosis were obtained from 147 patients diagnosed and treated at the Schneider Children’s Medical Center of Israel. The cohort consisted of 101 and 46 B-lineage and T-cell ALL, respectively. The median age at diagnosis was 6 years (range 0.3–18.8 years). Nineteen (13%) of the patients responded poorly to steroids at day 8 and relapse occurred in 41 (28%) patients. Median follow up was 80.5 months (range 5–253 months). DNA from BM was bisulfite modified and subjected to methylation specific PCR (MSP) analysis using primers specific for RASSF1A. Direct sequencing was done for N, H and K ras mutations. RASSF1A expression was detected by RT-PCR. Methylation of RASSF1A was detected in 30% of patients. There was no correlation of gene methylation to known clinical or biological features of ALL. A total loss of expression of RASSF1A was seen only in 7/27 (26%) methylated samples. Furthermore 11/53 (21%) unmethylated samples also showed a loss of expression for RASSF1A.There could be two possible explanations for these findings; firstly unlike the scenario in solid tumors, RASSF1A hypermethylation may not be a major operant mechanism in childhood ALL. Secondly, a more precise correlation of RASSF1A methylation to its expression could be established by quantitative real time analysis. We also detected mutations in ras oncogene in 16 (11%) patients (14 in N-ras and 2 in K-ras). However no correlation between mutations and clinical parameters was found. Thus a more comprehensive epigenetic analysis using a wider panel of genes/genome wide study should be designed to generate a clinically relevant “Methylotype” for childhood ALL.
MiRNA Profiles in Acute Lymphoblastic Leukemia: Rather a Characteristic for the Leukemia Subtype Than a Reflection of Its Differentiation Status.
Authors: Schotte Diana , Broekhuis Mathilde J.C. , Jansen Mieke W.J.C. , Pieters Rob , Chen Chang-Zheng , Den Boer Monique L. 
Published: October 2019

Abstract
MicroRNAs (miRNAs) are noncoding RNAs that regulate processes involved in proliferation and differentiation. Since abnormal proliferation and differentiation is the hallmark of cancer, miRNAs may have a role in the pathogenesis of cancer. Here we examined the miRNA expression in various subtypes of Childhood Acute Lymphoblastic Leukemia (ALL) including MLL-rearranged ALL (n = 16), T-ALL (n = 10) and precursor B-ALL patients positive for TEL-AML1 (n = 10), BCR-ABL (n = 10), E2A-PBX (n = 8) or hyperdiploidy (more than 50 chromosomes, n = 10) and other precursor B-ALL patients that are negative for all major genetic abnormalities in ALL (n = 19). We have found that miR-196b is highly expressed only in MLL-rearranged ALL and its expression level is ∼400 to 700-fold (P < 0.05) higher than the levels in all other precursor B-ALL subtypes and T-ALL cases. Moreover, all precursor B-ALL subtypes including TEL-AML1, BCR-ABL, E2A-PBX and hyperdiploid positive cases have a 250- to 6500- fold upregulation (P ≤0.01) of miR-708 compared to MLL-rearranged ALL and T-ALL patients. Interestingly, it is known that MLL-translocated ALL arises in an earlier stage of B-lineage development compared to other precursor B-ALL subtypes negative for MLL rearrangements, thus subtype-specific expression of miR-196b and miR-708 could reflect a difference in maturation status. However, miR-196b and miR-708 expression levels do not seem to correlate with the differentiation status of the leukemia cells based on the immunoglobulin (Ig) and T-cell receptor (TCR) rearrangements in case of precursor B-ALL and EGIL classification in case of T-ALL. Furthermore, miR-196b is located between the HOXA9 and HOXA10 genes, which are known to be upregulated in MLL-translocated patients. Here we find a positive correlation between miR-196b expression and HOXA9/HOXA10 expression within MLL- rearranged and T-ALL patients. Further research has to reveal whether high expression of miR-196b actively contributes to leukemogenesis like HOX genes or whether it is an epiphenomenon of HOXA upregulation in MLL-rearranged ALL patients. All together, our present study suggests that the expression level of miR-196b and miR-708 reflects differences between genetic subtypes rather than differences in differentiation status and emphasizes the need to further investigate the role of these miRNAs in leukemogenesis.
Identification of Genes Associated with Risk to Aplastic Anemia (AA) Using Array-Based Genotyping.
Authors: Lee Il-Kwon , Kim Hee Nam , Kim Yeo-Kyeoung , Park Kyeong-Soo , Yang Deok-Hwan , Lee Je-Jung , Shin Myung Geun , Choi Chan , Kook Hoon , Hwang Tae Ju , Kim Hyeoung-Joon 
Published: October 2019

Abstract
In this study we tested whether 3104 SNPs in 200 candidate genes were associated with risk to aplastic anemia (AA). Genes were considered potential candidates for their known or suspected roles in DNA repair system, pharmacogenomics and transcriptional regulation in hematopoiesis or putative pathways related to development of AA. Selection of SNPs was performed using dbSNP and HapMap project databases, with emphasis on non-synonymous SNPs or haplotype tagging SNPs. To discover potential SNPs responsible for AA susceptibility, a case-control study using Affymetrix targeted genotyping 3K array was designed. We applied 3K array to analyze the samples from 132 AA patients and 382 healthy controls. Genotyping were processed using GeneChip scanner 3000 TG(Affymetrix) and analyzed with GCOS software(Affymetrix). In total more than 160,000 SNPs were genotyped for this study. Samples with suboptimal call rates were excluded. Statistical testing were carried out using χ2, Cochran-Armitage trend, Fisher’s exact, odds ratio, haplotype estimation, LD block definition. Here we report that 19 SNPs in 14 genes are associated with elevated or reduced risk to AA. Three associated genes map to chromosome 11 and 16 respectively. Some of associated genes were transcription factors such as RARB and ZNF233. Among those associated SNPs, one was located in coding region while the rest of SNPs were located in introns of associated genes. In conclusion, we identified SNPs responsible for AA susceptibility by candidate gene-based SNP array approach. These promising data when supported by further molecular validation would greatly enhance the current understanding of AA predisposition and diseases progression. Implication of polymorphic variants in AA etiopathogenesis will be presented and discussed.
Autophagy Induction of Acute Myeloid Leukemia (AML) Cells after Treatment with Low Dose-Cytarabine (Ara-C) and Enhanced Cytocidal Effect on AML Cells by Combined Treatment with Ara-C Plus Lenograstim.
Authors: Miyazawa Keisuke , Toyotake Juri , Naito Munekazu , Itoh Masahiro , Ohyashiki Kazuma 
Published: October 2019

Abstract
Autophagy fundamentally refers to a process where cells enclose their own cell organelles with isolated membrane and deliver them to lysosomes for degradation, servicing as a nonspecific degradation mechanism of autologous proteins. Autophagy is induced above basal levels in response to nutrient deprivation or trophic factor withdrawal to sustain metabolism by producing catabolites. In this context, autophagy acts as a cytoprotective mechanism for cells to protect themselves from extracellular stresses. However, in a culture using cells in which autophagy-related genes were suppressed by siRNA and to which an autophagy inhibitor, 3- methyladenine (3-MA), was added, cytotoxic effects of some anticancer agents on cancer cells were diminished. Based on these results, the concept of caspase-independent cell death via autophagy, which is designated as type II programmed cell death (PCD) in contrast to apoptosis as type I PCD, has been receiving increased attention. We here report that treatment of AML cells with low does-Ara-C induces autophagy and enhances its cytocidal effect on AML cells by combined treatment with G-CSF. Treatment of AML cell line U937 cells with 1 to 50 ng/ml of Ara-C induced cell growth inhibition in a dose-dependent manner. Exposure to the higher concentrations of Ara-C (more than 10 ng/ml) induced apoptosis in U937 cells along with formation of apoptotic bodies and caspase-3 activation. However, treatment with the lower concentration of Ara-C (2.5 to 7.5 ng/ml) induced autophagy as assessed by electron microcopy and the punctuated distribution pattern of cytoplasmic microtubule-associated protein 1 light chain 3B (LC3B) by fluorescent immunocytochemistry. Low dose-Ara-C inducing -cell death and -autophagy both were suppressed when the cells were cultured in the presence of 3-MA. This suggests that the autophagic cell death but not the cytoprotective autophagy was induced in response to Ara-C. Combined treatment with G-CSF (lenograstim; 100 ng/ml) plus 2.5 to 7.5 ng/ml of Ara-C, but not plus higher concentrations of Ara-C, resulted in the enhanced cytocidal effect in U937 cells in vitro. These data indicate that combined treatment of Ara-C plus G-CSF has a therapeutic benefit for the AML patients by enhanced cytocidal effect which appears to be mediated via induction of autophagy. In addition, our data appear to explain in part the high response rate of CAG therapy in refractory AML, which consists of low-dose Ara-C in combination with G-CSF and aclarubicin.
A Novel Target of Histone Deacetylase Inhibitor, Involves in Mitochondria Membrane Protein Ubiquitination and Regulates Apoptosis.
Authors: Wu Peng , Cao Yang , Tian Yuan , Hong Zhenya , Sun Lishi , Gui Lingli , Zhang Yicheng , Ma Ding , Zhou Jianfeng 
Published: October 2019

Abstract
Inhibitors of histone deacetylases (HDACIs) are a new generation of anticancer agents that selectively kill tumor cells. However, the molecular basis for their tumor selectivity is not well understood. we employed a genetic technique, named suppression of mortality by antisense rescue technique(SMART), to understand the molecular mechanism of apoptosis induced by HDACI and identify the genes that participate in this process. We identified several novel genes and demonstrated that HDACIs promote apoptosis through activation ubiquitin/proteasome by inducing ubiquitin B not ubiquitin C gene expression. According to degrading the mitochondria membrane protein Mcl-1 by activated ubiquitin/proteasome system, the mitochondria potential collapse and cell apoptosis. Further observation demonstrated that depletion of MCL-1 by RNA interference (RNAi) sensitizes Hela cells to TSA mediated mitochondria membrane potential collapse and apoptosis, and overexpression of MCL-1 confers apoptosis resistance.
Prospective Evaluation of the NIH Staging Criteria in Chronic GVHD and Correlation to Quality of Life - Results of a German Multicenter Validation Trial.
Authors: Wolff Daniel , Herzberg Philipp Y. , Heussner Pia , Mumm Friederike , von Harsdorf Stephanie , Rieger Katrin , Greinix Hildegard , Hahn Joachim , Holler Ernst , Freund Mathias 
Published: October 2019

Abstract
The NIH staging and response criteria offer for the first time the chance for uniform documentation and evaluation of physical functioning and quality of life aspects of chronic graft-versus-host disease (cGVHD) as well as its response to immunosuppressive treatment. Here we present the interim analysis of a prospective Germany multicenter study on the NIH staging criteria in cGVHD.
Methods: Seventy-six patients (median age 46 years, range 19–64) after allogeneic hematopoetic stem cell transplantation (HSCT) for hematologic malignancies were evaluated according to the NIH criteria based cGVHD activity assessment, the Lee Chronic GVHD Symptom-Scale, FACT-BMT, human activity profile (HAP), SF 36, Berliner Social Support Scale (BSSS), 24 item Adjective Measure (24 - AM), Hospital Anxiety and Depression Scale (HADS) and the NCCN-Distress-Thermometer. Enrolment occurred between day 100 and 1 year after HSCT or in the presence of active cGVHD also at later time points. Follow-up surveys were conducted at 1, 2, 3, 5, 8, 12 and 18 months after baseline survey. At all time points disease status, comorbidities and medication were documented. Additionally the NIH-Symptom-Scale was applied for clinician rating of cGVHD symptoms.
Results: Fifty-one patients had cGVHD (mild n=15, moderate n=32, severe n=4) while 25 patients did not have cGVHD. The cGVHD NIH consensus grading as well as the 10 point scale correlated with impairment of physical functioning (p<0.01, r=.48), the FACT-G total score (p=0.02, r=.33) and mental health (p<0.01, r=.41). The HAP maximum activity score correlated inversely with severity of cGVHD (p=0.028, r=.44), with the physical functioning (p<0.01, r=.53) and with role limitation due to physical problems (p<0.01, r=.43) evaluated by the SF36. In addition an impaired physical functioning correlated with role limitations due to emotional problems (p<0.01, r=.52) and with impaired mental health (p<0,01, r=.63). Moreover the HAP maximum activity score correlated with emotional well being as evaluated by the FACT (p<0.01, r=.39). Depression as measured by the HADS depression score correlated with pain (p<0,01, r=.35) and impaired physical functioning (p=0.03, r=.26) while the HADS anxiety score did not. The results demonstrate, that severity of cGVHD as assessed by the NIH consensus grading and the 10 point scale correlate with impairment of physical functioning as well as daily activities and quality of life. Moreover reduction of daily activities lead to impairment of emotional well being.
Inhibition of the Redox Function of Ape-1/ref-1 in Myeloid Leukemia Cells Results in Enhanced Sensitivity to Retinoic Acid Induced Differentiation and Apoptosis.
Authors: Robertson Kent A. , Colvin Edwin S. , Kelley Mark R. 
Published: October 2019

Abstract
Ape-1/ref-1 is a multifunctional base excision DNA repair protein that is involved in the repair of abasic sites in DNA. However, it also has a distinct role in the redox regulation of a variety of cellular proteins, such as Fos, Jun, p53, NFκB, PAX, HIF-1α, HLF, and others. Ape-1/ref-1 maintains these proteins in a reduced state thereby facilitating their DNA binding and transcriptional activation capability. HL-60 cells are known to respond to retinoic acid (RA) with terminal granulocytic differentiation and apoptosis, which is mediated through the RA receptors. Previous experiments suggested that elevated Ape-1/ref-1 expression is related to differentiation and apoptosis. To further define the role of the redox function of Ape-1/ref-1 in this relationship, redox function was blocked using two techniques. First, we used retroviral gene transduction to over-express a redox-inactive C-65 mutant of Ape1/ref-1 in HL-60 myeloid leukemia cells and examined the response to retinoic acid. In a second set of experiments we used an Ape-1/ref-1 specific small molecule inhibitor to pharmacologically block the redox function and again examined the response to retinoic acid. Differentiation was evaluated by morphologic change in differential cell counts and expression of CD11b by flow cytometry. Apoptosis was assayed by annexin-PI staining on flow cytometry and cell cycle analysis was examined with propidium iodide flow cytometry.
Results:
HL-60 cells expressing high levels of C-65 Ape-1/ref-1 responded to retinoic acid with a significantly higher level of differentiation and a moderate increase in apoptosis. Pharmacologic blockade of Ape-1/ref-1 redox function resulted in a profound increase in differentiation and a moderate increase in apoptosis compared to controls. dose dependent studies with retinoic acid demonstrated a similar degree of differentiation (CD11b expression) in cells treated with 10 μmolar retinoic acid and those treated with the redox inhibitor + 0.1 μmolar retinoic acid; alllowing HL-60 cells in the presence of the redox inhibitor to give a similar response to a 100 fold lower dose of retinoic acid.
The redox inhibitor alone did not induce differentiation and induced only a minimal amount of apoptosis but did increase the number of cells in S phase significantly. In conclusion, our data supports the contention that redox function of Ape-1/ref-1 may be important for controlling RA-induced myeloid differentiation and programmed cell death. The implication of these findings is that myeloid leukemia cells may be sensitized to retinoids by manipulation of the redox status of Ape-1/ref-1.
Importance of the Polymorphisms of the Thiopurine S-Methyltransferase Gene (TMPT) Determination in Mexican Mestizo Patients with Acute Lymphoblastic Leukemia (ALL).
Authors: Candelaria Myrna , Taja-Chayeb Lucia , Vidal Silvia , Gutierrez Olga , Dueñas-Gonzalez Alfonso 
Published: October 2019

Abstract
Background: Polymorphisms at the thiopurine S-methyltransferase coding gene (TPMT) determine enzyme activity and consequently, the development of toxicity secondary to thiopurines.
Methods: A total of 108 DNA samples from volunteer donors and 39 from patients with acute lymphoblastic leukemia (ALL) were analyzed. Genomic DNA from peripheral blood leukocytes was isolated by standard methods. Known (wild-type and polymorphic) sequenced polymerase chain reaction (PCR) fragments of the TPMT gene were used as controls. TPMT gene fragments were amplified for exons 5, 7 and 10. Thereafter, PCR products were analyzed by denaturating high performance liquid chromatography (DHPLC) for the most frequent mutant TPMT alleles, according to the method developed by Schaeffeler et al. on an analysis system from Transgenomics.
Results: No elution profiles on DHPLC analysis, different from those previously reported, were documented. Frequency of allele polymorphisms was 27.3% in healthy and 19.7% in ALL population. However, only 17.6% of all polymorphisms found are considered functional, the most frequent being *3A (n = 13; 4.4%), followed by *3B (n = 5; 1.7%), *3C (n = 5; 1.7%), and *2 (n = 3; 1.0%). From 39 ALL patients, 22 were treated with thiopurines, and particularly one patient, who was referred to our Hospital to investigate relapse, after developing pancytopenia while on treatment with thiopurine, had a TPMT*3C allele. He required broad spectrum antibiotics, platelets transfusions, as well as G-CSF. This example illustrates that TPMT polymorphisms must be determined before thiopurines are indicated.
Conclusions: DHPLC is a highly sensitive, rapid and efficient method to identify relevant TPMT gene mutations which allows the screening for genetic variability in the TPMT gene. This is the first analysis of the polymorphisms at this gene in Mexican population. The frequency of known polymorphisms was similar to those reported in other regions worldwide. Since a direct relation has been documented within functional polymorphisms and enzyme activity, and this methology is feasible to realize in any phase during the treatment of ALL patients, the routine typing of TPMT polymorphisms in the patients with ALL has been set in our Institution.
Table 1. Frequency of TPMT polymorphisms in mexican population Population TPMT*1 TPMT*1S TPMT*2 TPMT*3A TPMT*3B TPMT*3C * Acute Lymphoblastic Leukemia Healthy n(%) 50 (72.7) 42 (19.4) 2 (0.92) 7 (3.24) 5 (2.3) 3 (1.4) ALL* n(%) 24 (80.3) 5 (6.9) 2 (2.7) 6 (7.6) -- 2 (2.5)
Genetic Polymorphisms of DNA-Repair- and Detoxification Enzymes in Therapy-Related AML and MDS.
Authors: Fischer Stephanie , Seedhouse Claire , Schulz Thomas , Germing Ulrich , Truemper Lorenz H. , Haase Detlef 
Published: October 2019

Abstract
The most serious long-term complications of anti-tumour therapy are secondary malignancies. After treatment of haematological malignancies such as NHL, Hodgkin’ disease and multiple myeloma as well as solid tumours like breast cancer, lung cancer, soft tissue cancer and ovarian carcinoma therapy-related leukaemia occur in 0.5 to 10% of pts. Parameters which might allow an estimation of the individual risk to develop a therapy-induced neoplasia are urgently needed. Referring to recent findings defects in the DNA-repair-system and in detoxification genes are candidate predictive factors. We have examined 125 cases of t-AML/MDS in a retrospective study. The primary neoplasias were: breast cancer (33%), NHL (22%), M. Hodgkin (7%), multiple myeloma (8%) or other miscellaneous (30%). Cytogenetic data of therapy-induced malignancies were available in 116 of 125 pts. in 66% (n=78) of whom clonal abnormalities were detected. The most frequent single abnormalities were -7/7q- (15%), inv(16)(12%), +8 (8%), t(15;17) and del(20q) (5% each). Aiming to identify possible risk factors we used this patient cohort to study genetic polymorphisms influencing the activity of DNA-repair- and detoxification enzymes. For this purpose analyses of XRCC3, RAD51, GSTT1/M1/A1/P1 and NQO1 were performed. Our patient cohort (n=125) attracted attention that patients with a deficiency for GSTT1 or with combined defects of GSTT1 and M1 had a 1.7-fold (p=0.03625) and a 2.4-fold (p=0.00921) risk resp. of developing t-AML/MDS as compared to healthy controls (n=381). Noticeable is as well that the inactivating polymorphism of GSTA1*B was significantly more frequent in the control group (p=0.01847) in comparison to our patient cohort. Subsequently we generated different categories for statistical analysis according to the primary malignancy: As yet, focusing on GSTT1/M1 revealed an overrepresentation (p<0.0005) (OR=4.0) of double null genotypes for GSTT1/M1 in pts. with t-AML/MDS after adjuvant treatment for breast cancer (n=41). Preliminary results suggest that patients with tAML/MDS following NHL (n=22) who have a defect for XRCC3 have a 4.2-fold risk to develop a tAML/MDS (p=0.02960). The polymorphism of GSTA1*B was significant only in our control group in comparison to patients who showed NHL as a primary malignancy (n=22). We suppose that in this cohort the GSTA1*B polymorphism might protect against t-AML/MDS. This could be an explanation for the overrepresentation of GSTA1*B-defect in our control group as compared to our patient cohort. Other statistical analyses did not show any significance. Taken together polymorphism of GSTA1*B and XRCC3 might be associated with the risk to develop t-AML/MDS after NHL. Defects of GSTT1 and M1 can be taken as a first hint to a risk factor for t-AML/MDS following breast cancer.