bluuebunny/crossref_metadata_2025_split huggingface dataset:
The ATP Competitor INNO-406 (NS-187) Inhibits Proliferation and Survival of Mouse Cells Expressing Several Immatinib Mesylate Resistant Bcr/Abl Mutants and Cells from CML Patients.
Authors: Tao Wenjing , Sun Tong , Craig Adam R. , Cortes Jorge E. , Ottmann Oliver G. , Hochhaus Andreas , Reddy Premkumar E. , Kantarjian Hagop M. , Arlinghaus Ralph B. 
Published: October 2019

Abstract
Imatinib mesylate (IM) resistance during the therapy of chronic myeloid leukemia (CML) patients is associated with point mutations in the Bcr/Abl kinase, which strongly interferes with IM binding to Abl kinase domain. INNO-406 (NS-187) is reported to be a new specific inhibitor of the Bcr/Abl and Lyn kinase, and is a candidate for treatment of IM resistant CML patients (Kimura et al, Blood2005, 106:3948). We tested INNO-406 on both IM resistant mouse cell lines and CML patient samples. The MTT assay was used to detect the anti-proliferation effects of INNO-406 on Bcr/Abl wild type cells (32Dp210 wt) and Bcr/Abl mutants (L387M, H396R, Q252R, F311L, T315I and Y253H) after 72 hours treatment. IM or dasatinib treatment was included as controls.
The MTT results showed 32Dp210 wt and four of the six Bcr/Abl mutants were sensitive to INNO-406, with the IC50 values of 0.6 nM (wild type), 15 nM (L387M ), 35 nM (H396R), 0.8 nM (Q252R) and 8.2 nM (F311L). The IC50 values of those four cell lines treated with IM were much higher than with INNO-406 treatment. The IC50 values for IM were: 100 nM (wild type), 500 nM (L387M), 1220 nM (H396R), 100 nM (Q252R) and 100 nM (F311L). But neither INNO-406 nor IM exhibited inhibitory effects on the proliferation of T315I and Y253H mutants (IC50 > 10000 nM). These results indicated that INNO-406 suppressed the growth of Bcr/Abl mouse leukemia cell lines expressing wt Bcr/Abl and Bcr/Abl mutants except T315I and Y253H. We note that the Y253F mutant was sensitive to INNO-406 (IC50 64 nM, Nippon Shinyaku Co. Ltd.). Annexin V/PI apoptosis assays were conducted on cells from CML patient samples after 72 hours treatment with various concentrations of INNO-406. Flow cytometry results showed that INNO-406 (10–20 nM) induced apoptosis in blood/marrow cells from two blast crisis patients, one of which had the G250E mutation. INNO-406 also induced apoptosis in marrow cells from two patients in IM induced remission. One of these samples with a Bcr/Abl mutation at M244V showed a weak response to IM but a strong apoptosis with INNO-406 (20 nM), indicating that NS-187 worked more efficiently in the M244V mutant. Based on these results, INNO-406 (NS-187) appears to be a candidate for the treatment of some forms of imatinib resistant CML. A phase I study of INNO-406 has been initiated in patients with CML who have failed prior imatinib therapy and the first cohort has been safely treated.
Hsp-90 Inhibition Decreases Survival of BCR-ABL T315I Leukemic Stem Cells in Mice.
Authors: Peng Cong , Brain Julia , Hu Yiguo , Kong Linghong , Goodrich Ami , Grayzel David , Pak Roger , Read Margaret , Li Shaoguang 
Published: October 2019

Abstract
Although advances have been made in the development of novel molecularly targeted drugs, a major therapeutic challenge in the treatment of patients with Philadelphia chromosome positive (Ph+) leukemia includes understanding how to target the leukemic stem cell. We used the bone marrow transplant (BMT) model of chronic myelogenous leukemia (CML) to study effects of imatinib mesylate and the novel, orally active heat shock protein 90 (Hsp90) inhibitor, IPI-504, on leukemic stem cells, based on our observation that unlike imatinib, IPI-504, prolongs survival in a murine model of drug-resistant T315I BCR-ABL-induced CML. We first identified BCR-ABL-expressing hematopoietic stem cells (HSCs) (Lin-c-Kit+Sca-1+) in mouse bone marrow as CML stem cells, as these cells sorted out by FACS from primary CML mice are sufficient to confer leukemia in recipient mice. We then investigated the effects of imatinib and IPI-504 on survival of leukemic stem cells from BCR-ABL T315I induced CML. Bone marrow cells from mice with T315I-induced CML were cultured under conditions that support survival and growth of stem cells, with or without IPI-504 or imatinib. FACS analysis of GFP+Lin-c-Kit+Sca-1+ cells showed that imatinib treatment did not lower the percentage and the number of leukemia stem cells, whereas IPI-504 treatment had a dramatic inhibitory effect on this population (p<0.001) at therapeutically achievable doses. To determine whether IPI-504 attenuates development of leukemia by specifically inhibiting stem cell survival, GFP+Lin-c-Kit+Sca-1+ cells were sorted from bone marrow of mice with BCR-ABL T315I-induced CML, and cultured with a placebo or IPI-504. When these cells were transferred into lethally-irradiated recipient mice, FACS analysis showed that myeloid leukemia cells were present in mice receiving the placebo-treated leukemic stem cells but not in mice receiving the IPI-504 treated leukemic stem cells. To examine whether IPI-504 inhibits leukemia stem cells in vivo, mice with BCR-ABL-T315I-induced CML were treated with a placebo, imatinib, or orally administered IPI-504 for six days. Bone marrow cells were analyzed by FACS for GFP+Lin-c-Kit+Sca-1+ cells. Consistent with the in vitro results, imatinib treatment did not lower the percentage and number of leukemia stem cells, as compared with the untreated group, whereas IPI-504 treatment had a dramatic inhibitory effect on the stem cells. Analysis of bone marrow from non-leukemic mice treated with IPI-504 for two weeks showed no change in levels of Lin-c-Kit+Sca-1+ cells, indicating that IPI-504 treatment did not inhibit survival of normal HSCs. These results provide a rationale for use of an Hsp90 inhibitor as a first-line treatment to inhibit leukemia stem cells and prevent emergence of imatininb-resistant clones in patients.
Immunomodulatory Effects of STI571 in Chronic Myeloid Leukaemia.
Authors: Bund Dagmar , Yang Ting , Buhmann Raymund , Kolb Hans-Jochem 
Published: October 2019

Abstract
Background: The tyrosine kinase inhibitor imatinib (imatinib, STI571, Glivec, and Gleevec) is highly effective in the treatment of chronic myeloid leukaemia (CML) and has already been shown to be effective in the setting of allogeneic stem cell transplantation. But until now, less is known with respect to its immunomodulating effects.
Objective: In the present survey we investigated, whether imatinib could modify the antigen-presenting capacity of myeloid cells and in turn affects the cellular immune response.
Method: For this purpose, patient derived chronic myeloid cells were incubated with different concentrations of imatinib (0, 1, 2, or 5microM), characterized for their antigen-presenting profile and their stimulatory capacity in the context of HLA-matched and mismatched T-cells. After 5 days, the proliferative immune response was evaluated in presence or absence of different concentrations of imatinib and altered effector-to-target ratios. Thereby, proliferation was detected via a CFDA, SE (5,6-carboxyfluorescein diacetate succinimidyl ester) based assay.
Result: The proliferative capacity of the T- cells (allogeneic, HLA-mismatched) was inhibited by imatinib in a dose-dependent manner. Also, the expression of the activation markers was reduced in the presence of the different STI571 concentrations. Moreover, myeloid blasts were sensitized for T cell mediated effector functions by pre-treatment with increasing concentrations of imatinib.
Conclusion: Taken together, imatinib can interfere with the T cellular immune response in vitro, and its impact on graft-versus-leukemia (GvL) and graft-versus-host (GvH) reactions will be further investigated.
SPAN-XB Core Promoter Sequence Is Regulated in Myeloma Cells by Specific CpG Dinucleotides Associated with MeCP2 Protein.
Authors: Wang Zhiqing , Zhang Jian , Zhang Yana , Srivenugopal Kalkunte , Lim Seah H. 
Published: October 2019

Abstract
SPAN-Xb is a spermatid-specific protein encoded by the SPAN-XB gene on chromosome Xq27.1. It is a novel Cancer-Testis antigen in multiple myeloma. We recently demonstrated that SPAN-Xb expression in myeloma cells is regulated through promoter methylation and could be upregulated by IL-7 and GM-CSF. In this present study, we set out to investigate the mechanism of SPAN-XB expression and the promoter association with the methyl-CpG binding protein, MeCP2. Elucidation of these interactions is likely to shed light on potential therapeutic strategies to upregulate antigen levels that could be used to improve the outcome of SPAN-Xb-based tumor vaccines.
We previously showed that the putative SPAN-XB promoter resides within exon 1 and contains 433 bp, starting from position −546 to position −104 upstream of the translational start site. Using a panel of truncated promoter constructs generated from the 3′ and 5′ ends of the promoter gene, we localized the core sequence of SPAN-XB promoter to the 73 base pairs at the 3-end of the promoter, a region that lacks CpG dinucleotides within the full length promoter. There are 11 CpG dinucleotides within the putative SPAN-XB promoter. We previously found that DNA methylation provides the primary regulatory mechanism for SPAN-XB gene expression.
We also identified that hypomethylation at positions −310, −307, −299 and −221 strongly predicted for SPAN-XB expression, suggesting the involvement of these four CpG dinucleotides in the regulation of SPAN-XB gene expression through DNA methylation. In the present study, using reporter gene expression assays, we found that the core promoter function is significantly modulated by these adjacent CpG sequences so that mutation of the CpG dinucleotides outside the core sequence resulted in changes in the promoter function. We also previously demonstrated by bisulfite conversion and sequence analysis the association between methylation at specific CpG dinucleotides with repression of SPAN-XB gene. Here, we extended our study to determine whether or not the methylated cytosine binding protein, MeCP2, interacts with the SPAN-XB promoter gene in myeloma cells. Chromatin immunoprecipitation assays revealed a specific association of the MeCP2 with SPAN-XB promoter, and MeCP2 binding strongly correlated with repression of the SPAN-XB gene in myeloma cell lines and CD138-enriched fresh myeloma cells. Reactivation of the SPAN-XB gene by 5-azacytidine treatment resulted in the loss of MeCP2 from this site. We, therefore, conclude that SPAN-XB promoter consists of a core sequence and a regulatory element; the core element resides within the 73 base pairs at the 3′ end of the full length promoter. SPAN-XB gene expression by the core sequence is regulated in myeloma cells by specific CpG nucleotides and associated with MeCP2 binding to the promoter sequence.
PU.1 Is a Downstream Target of SOX4 in Myeloid Cells.
Authors: Aue Georg , Du Yang , Dunbar Cynthia E. , Jenkins Nancy A. , Copeland Neal G. 
Published: October 2019

Abstract
Mice that express 20% the normal levels of the Ets transcription factor PU.1 develop AML, unlike mice that express 50% or 80% the normal levels, indicating that PU.1 is a dosage-sensitive tumor suppressor gene. In addition, 3 of 13 AMLs induced by transplanting mice with cells transduced with a Sox4 oncogene-containing retrovirus were found to carry a Sox4 retroviral integration in one PU.1 allele, suggesting that downregulation of PU.1 may cooperate with Sox4 in AML induction. Since the other PU.1 allele remains intact in these AMLs and a 50% decrease in PU.1 expression is not sufficient to induce AML, we hypothesized that Sox4 might further downregulate PU.1 expression in these AMLs. To test this hypothesis, we transfected HL60 promyelocytes with an expression vector carrying both GFP and Sox4 cDNAs or a GFP vector control. Transfected GFP+ cells were purified by flow cytometry and PU.1 mRNA levels were analyzed by real-time RT-PCR. PU.1 mRNA levels were consistently downregulated 4 to 10 fold in cells transfected with Sox4 cDNA compared to cells transfected with the vector control, while β-actin mRNA levels were maintained constant, confirming that overexpression of Sox4 downregulates PU.1 expression in myeloid cells. The decrease of PU.1 mRNA was observed as early as 8 hours after Sox4 transfection, further suggesting that Sox4 may directly repress the PU.1 promoter in myeloid cells. Consistent with this, analysis of 2 published microarray databases comprising 401 de novo AML patient samples showed that SOX4 expression is significantly negatively correlated with PU.1 expression (coefficient: −0.337, P-value: 1E-07). Interestingly, AML FAB M1 and M2 subtypes were associated with statistically significant higher SOX4 expression levels compared to AML FAB M3, M4, M5. In order to confirm that downregulation of PU.1 cooperates with Sox4 in AML induction, we infected wild type or PU.1 heterozygous knockout bone marrow cells with the Sox4 retrovirus and then monitored the time of AML development in transplanted mice. Early results show accelerated leukemogenesis in mice transplanted with Sox4-infected PU +/− bone marrow (115 days) compared to mice receiving Sox4-infected wild type marrow (160 days). We are currently trying to identify Sox4 binding sites in the PU.1 promoter, or in an upper regulatory element that may be responsible for mediating the repression of PU.1.
Target Proteins of C/EBPa-p30 in AML: A Critical Role of Ubiquitin-Conjugating Enzyme (Ubc9) in Modulating C/EBPa-p42 Functions.
Authors: Geletu M. , Balkhi M. , Zada A. Peer , Trivedi A. , Pulikkan J. , Christopeit M. , Tenen Daniel G. , Behre G. 
Published: October 2019

Abstract
The mutations in the transcription factor CCAAT/enhancer binding protein-a (C/EBPa) occur in 10% of patients with acute myeloid leukemia which often leads to the expression of an N-terminal truncated 30KDa isoform of C/EBPa. The mutated 30KDa isoform has been shown to be dominant negative over wild type isoform that affects most of its biological functions. In the present study, we applied a global proteomics approach to identify the target proteins of C/EBPap30. This analysis revealed that C/EBPap30 modulates the expression of 60 different proteins notably, ubiquitin-conjugating enzyme (Ubc9), Peptidylprolyl isomerase (pin1), hnRNP A2/B1, Cacyclin binding protein, calgranulin B, and Tubulin beta5. Further, we show that in AML patients with C/EBPap30 mutations Ubc9 was predominantly upregulated. We discovered that this ligase was responsible for the enhanced sumolyation of C/EBPap42 on a lysine residue (K161) which results in its reduced transactivation in a promoter assay. When lysine (K) residue at 161 of C/EBPa-p42 was mutated to argenine residue(R), transcriptional activity of C/EBPap42 was restored. This finding was further validated by silencing the expression of Ubc9 expression which completely abrogates the sumoylation at K161 residue and further restored the transactivation and differentiation capacity of C/EBPap42. Our results demonstrate that increasing expression of Ubc9 by C/EBPap30 enhances sumolyation of C/EBPap42 and thereby inhibits its transactivation and differentiation capacity.
Identification and Characterization of Novel SET Domain-Containing Histone Lysine Methyltransferase and Implications of Histone Methylation in Acute Leukemia.
Authors: Kook Hoon , Choe Nakwon , Kee Hae-Jin , Kim Sung-Mi , Kim Ji-Young , Han Dong-Kyun , Baek Hee-Jo , Lee Kun-Soo , Hwang Tai-Ju , Seo Sang-Beom , Kook Hyun 
Published: October 2019

Abstract
Evolutionally conserved SET domains, which methylate histone lysine residues, and thereby methylation of histones have been implicated in diverse malignancies including leukemias or multiple myelomas. Here we describe novel SET domain-containing proteins with histone methyltransferase (HMTase) activity and their characteristics. Using bioinformatics for homology screening, SET-domain containing proteins named WHISTLE, WHSC1-like 1 isoform 9 with methyltransferase activity to lysine, and RE-IIBP, interleukin-5 response element II binding protein, were identified. By mass spectrometric and immunoblot analysis, we demonstrated that WHISTLE dimethylates H9K4 and di-, and tri-methylates H3K27, while RE-IIBP methylates only H9K27. WHISTLE and RE-IIBP repressed transcription of the SV40 and IL-5 promoter activity and they recruited histone deacetylase. Chromatin immunoprecipitation analysis revealed that shRNA-mediated knockdown of RE-IIBP reduces histone methylation on the IL-5 promoter. Both proteins induce apoptosis in leukemic cells via caspase-3 activation. In acute lymphoblastic leukemia patients, the expression of RE-IIBP and WHISTLE was increased, which was accompanied with increase in the methylation histone 3. These data illustrate the important regulatory role of novel SET domain proteins with HMTase activity in transcriptional regulation and apoptosis, thereby pointing to the critical role in leukemogenesis.
Association of cis-Acting rs530 of the ETS2 Transcriptional Factor Gene with High-Risk Acute Myelogenous Leukemia (AML) and Allelic Expression Imbalance Assessment.
Authors: Lee Il-Kwon , Choi Jeong-Hwa , Kim Hee Nam , Kim Yeo-Kyeoung , Park Kyeong-Soo , Lee Je-Jung , Shin Myung-Geun , Choi Chan , Kook Hoon , Ju Hwang Tai , Park Moo-Rim , Jo Deog-Yeon , Won Jong-Ho , Kim Hyeoung Joon 
Published: October 2019

Abstract
We have previously implicated ETS2 in the etiopathogenesis of acute myeloid leukemia through case-control study by revealing that polymorphisms of ETS2, a hematopoietic transcription factor gene is associated with increased risk to acute myelogenous leukemia (AML). Two SNPs out of 7 genotyped sites, rs711 and rs530 were shown to be associated with increased risk to AML with the odds ratio (OR) of 1.76 (95% C.I. 1.2–2.5, p=0.0019) for rs711 and 1.67 (95% C.I. 1.3–2.2, p=0.0003) for rs530 relative to wild type genotypes respectively. Haplotype and linkage disequilibrium (LD) map was estimated, but haplotype association was not found with statistical significance. Korean LD structure was similar to Japanese LD, but least similarity was shown with LDs from African (Yoruba in Ibadan, Nigeria). Since these two SNPs are located in the 3′ UTR region we simulated the change in secondary structure in silico of the 3′ UTR region with two variants introduced in the sequence. Most dramatic change in the secondary structure was observed in the rs530 containing domain suggesting this variant of being cis-acting genetic variant. Real time Q-PCR and western blot analysis showed that expression of ETS2 decreased in individuals with heterozygous or mutant homozygous genotypes, showing most abundant expression with two wild type copies of rs530, less expression with the rare homozygous or heterozygous genotype. Sequencing cDNA of 55 heterozygous AML patients revealed mRNA expression imbalance in 13 cases (24%) effectively reverting heterozygous genotype to homozygous wild type mRNA species. The detection of a discrepancy between the mRNA alleles of rs530 clearly proves cis-acting effect of rs530. However there was not a case in 51 healthy control samples suggesting differing transcript levels derived from the two alleles of an autosomal gene is disease-specific phenomena. Taken together, our results suggest that two polymorphic variants in the 3′ UTR region predispose individual to high-risk AML by inducing change in mRNA expression as a cis-acting variant.
Integrated CGH- and Epression Array Profiling of Mantle Cell Lymphoma.
Authors: Schraders Margit , Jares Pedro , Bea Silvia , Schoenmakers Eric F.P.M. , Van Krieken Joannes H.J.M. , Campo Elias , Groenen Patricia J.T.A. 
Published: October 2019

Abstract
Mantle cell lymphoma (MCL) is characterized by the translocation t(11;14)(q13;q32), which leads to the overexpression of cyclin D1. The cyclin D1-overexpression by itself is however not sufficient for lymphoma development. To identify other genes that are deregulated in MCL, we have performed integrated profiling of high-resolution array-based comparative genomic hybridization (array-CGH) with RNA-expression array analysis on the same set of MCL cases. DNA-alterations of MCL cases were assessed using a 3.6k BAC array with a resolution of 800 kb. On the same cases, RNA-expression array analysis was performed using the GeneChip® Human Genome U133 Plus 2.0 Array from Affymetrix. Integration of genomic-and transcription profiling data was performed using statistical tools. With array-CGH, we identified regions of chromosomal gain or loss and determined minimal common regions (mcr’s) by identifying the smallest region of overlap present in at least 36% of the cases. Integration of array-CGH and expression profiling was performed for genes located within the mcr’s and unsupervised clustering of gene-expression values within each mcr was performed. For several mcr’s, e.g. 1p22.1–31.1, 6q23.2–27 and 11q22.3–23.3, the clustering tree showed two groups, one with the chromosomal aberration and one without. Also, we observed that only a subset of genes located within a cytogenetic anomaly has a concomitant change in mRNA expression level. These genes are regulated by DNA-copy number (gene dosage). Amongst these, we identified several “hypothetical genes” and genes, which encode proteins involved in mitochondrial protein synthesis, the DNA damage repair pathway and the cAMP regulated pathway. This study shows the potential of the integrated profiling approach for identifying genes that are regulated by gene dosage (DNA-copy number). We anticipate that the genes we identified are important for MCL and it’s characteristic features like the low apoptosis rate and the chemotherapy-resistance.
Mouse B Cell Lineage Neoplasms: Comparative Analyses of Gene Expression Using High Throughput Real Time RT-PCR and Oligonucleotide Microarrays.
Authors: Shin Dong-Mi , Shaffer Daniel J. , Roopenian Derry C. , Morse Herbert 
Published: October 2019

Abstract
Gene expression profiling of human and mouse B cell lineage neoplasms using microarrays has provided a powerful means for:identification of previously unrecognized subsets of a single histologically-defined disease category;relating a specific disease to a previously unrecognized cell of origin; andestablishing disease subsets defined by characteristic host-tumor interactions.
Other anticipated returns including the rapid identification of new therapeutic targets have fallen short of expectations. In addition, discrepancies among results obtained using different platforms pose a major obstacle to bringing this approach into common clinical practice. Quantitation of gene expression by qPCR provides an attractive alternative approach but has been limited by the lack of high-throughput methodologies. We have approached this issue by developing qPCR methods that assay 384 “cancer genes” in a single plate, covering a wide range of targets from pathways implicated in the development of hematopoietic as well as solid tumors. We used this system to compare RNA prepared from normal spleens, mouse PCT and two lymphoma types with strikingly similar cytologies but differing origins - high-grade splenic marginal zone lymphomas (MZL) and centroblastic DLBCL (CBL). Terminally differentiated PCT were readily distinguished from MZL and CBL and MZL from CBL. Statistical analyses identified subsets of genes that distinguished each normal or tumor set from all the others and these are being validated against new groups of each tumor type. The same initial cases were also evaluated using oligonucleotide microarrays, providing instructive similarities and differences in measures of shared genes. From these comparisons, we have identified potentially important signaling pathways, such as Notch, Wnt and IL6, as well as drugable targets. These are being evaluated in studies of cultured cell lines and can be extended to in vivo tests of efficacy in mice bearing transplanted or primary tumors.