bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of the Transcription Factor PAX5/BSAP in Childhood Acute Leukemia Cells and Haematological Tumor Cell Lines.
Authors: Zhang Bei , Tie Li-Jun , Ye Qidong , Zhao Yanxia , Gu Long-Jun , Tang JingYan , Yuan Xiangliang , Shen Li-song 
Published: April 2020

Abstract
PAX5 gene is a paired-box PAX gene family member,and encodes the transcription factor BSAP(B-cell specific activator protein) which is a key regulator of B-cell development and differentiation.Dysregulation of PAX5 gene function may contribute to tumorigenesis in lymphoid malignancies.But up to now,a detailed examination of PAX5/BSAP expression in acute leukemia(mainly acute B-lineage lymphoblastic leukemia) has not been reported.In this study,a real-time RT-PCR assay for the relative quantitation of PAX5 and CD19 mRNA expression was developed.It was applied on 6 haematological tumor cell lines and bone marrow cells of 6 normal children,58 previously untreated and 4 relapse acute leukemic children,including 39 cases of B-ALL,10 cases of T-ALL,and 13 cases of AML.PAX5 and CD19 mRNA expression were detected in B-cell lines tested,but almost not in other T- and myeloid cell lines.Among clinical samples,expression of PAX5 mRNA in B-ALL was significantly higher than that in T-ALL and AML(P=0.029 and P=0.013,respectively).PAX5 expression was significantly lower in T-ALL and AML than normal controls.The mRNA levels of PAX5 between T-ALL and AML had not any difference.Individual difference of PAX5 mRNA expression levels in children with B-ALL was great.Because binding sites for BSAP have been identified in the promoters of CD19,the study found that in B-ALL,there was clear correlation between the level of PAX5 expression and that of CD19,which was also studied by real-time RT-PCR.BSAP expression by Western Blotting analysis was also performed in haematological tumor cells,including 6 haematological tumor cell lines and 4 clinical samples(2 cases of B-ALL,1 case of T-ALL,and 1 case of AML).The results of Western Blotting analysis showed a 52-KD BSAP band in B lineage cells,but not in T- and myeloid lineage cells.The intensity of BSAP bands was in accordance with PAX5 mRNA expression level detected by real-time RT-PCR.It was concluded that PAX5 transcripts are readily detectable and quantified in clinical materials with B-ALL by real-time RT-PCR.The strong PAX5 mRNA expression in some B-ALL can be considered to be particularly interesting for further analysis.
Proteomic Analysis of Targeted Therapeutic Agents in Multiple Myeloma (MM).
Authors: Ghobrial Irene M. , Eckel Jeanette E. , Hoering Antje , Timm Michael , Leontovich Alexey A. , Fonseca Rafael , Greipp Philip R. , Witzig Thomas E. 
Published: April 2020

Abstract
The objective of this study was to analyze protein changes that occur a the molecular level with novel therapeutic agents used in MM in order to identify potential combinations for clinical trials. We employed antibody protein microarrays (BD Clonetech, CA) to measure changes in the patterns of protein expression in Kas 6/1 MM cell line after treatment with bortezomib, dexamethasone, CC-5013, and 17-AAG. The antibody array is a new technique that assays protein differences directly by hybridizing fluorescently labeled protein mixtures from cell extracts onto glass slides spotted with 512 different monoclonal antibodies specific for human proteins. Induction of apoptosis was assessed by Annexin/Propidium iodide FACS analysis at 24 and 48 hours using different concentrations of the inhibitors. Protein arrays were performed at concentrations and treatment times that did not induce more than 25% apoptosis to ensure adequate analysis of early changes in signaling pathways. Bortezomib 5nM at 10 hours, dexamethasone 100mM, CC-5013 250nM, and 17-AAG 1mM at 24 hours were used. Control cells were treated with vehicles ETOH or DMSO at the same concentrations as the inhibitors. To assess differential protein expression, the mean of the Cy5/Cy3 ratios for each sample and its control were analyzed using the Clontech software and an internally normalized ratio (INR) was calculated. INR values &gt;1.3 or &lt;0.77 were considered as valid changes in protein abundance for this study. All inhibitors induced the upregulation of apoptosis proteins including BAK, BIK, SMAC/DIABLO, and caspase 14. Several proteins were inhibited by all 4 inhibitors indicating common pathways affected by these agents. These included members of the PI3K pathway (PI3K-p85a, p70S6kinase, and cyclin dependent kinases), MAPKinases (ERK1 and 2), antiapoptotic protein Bclx, heat shock proteins Hsp70 and HSP60, and ubiqutin/NFkB pathway (Ubch6, IKKalpha, and IKBe). These results indicate that inhibitors of the PI3K pathway such as mTOR inhibitors, IKK inhibitors or MAPkinase inhibitors may be used in the future in combination with bortezomib, dexamethasone, CC-5013 or 17-AAG. Fatty acid synthetase was downregulated by bortezomib and dexamethasone indicating potential use of fatty acid synthase inhibitors in combination with bortezomib or dexamethasone in future clinical trials. These results suggest that antibody protein arrays may be used in the rational design of combinations of targeted therapies in MM.
T Cell Activation by t(9;22) Acute Lymphoblastic Leukemia-Derived Dendritic-Like Cells Is Associated with Increased Expression of Antigen Processing Machinery Components.
Authors: Claus Jonathan A. , Brady Michael A. , Lee Jaewoo , Donohue Kathleen A. , Ferrone Soldano , Wetzler Meir 
Published: April 2020

Abstract
We have recently shown that dendritic-like cells derived from t(9;22) acute lymphoblastic leukemia (ALL) cells can activate T cells, while the original unmodified leukemic blasts cannot. To define the molecular basis of this functional difference, we have compared the level of antigen processing machinery components in unmodified blasts and ALL-derived dendritic-like cells, since they play a major role in antigen presentation. Six patient samples and one t(9;22) ALL cell line (Z119) were studied.
Dendritic-like cells were generated by in vitro culture with CD40 ligand, interleukin (IL)-1β, IL-3, IL-7, stem cell factor and tumor necrosis factor-α for six days. Maturation was verified by morphology and CD80/CD86 expression. Cells were studied for HLA class I and class II antigen expression. Furthermore, antigen processing machinery component expression was analyzed by intracellular staining with a unique panel of monoclonal antibodies which recognize the constitutive proteasome (Z, MB1, delta) and immunoproteasome (LMP2, LMP7, LMP10) subunits, the transporter subunits TAP 1 and 2 and the chaperones calnexin, ERp57, calreticulin and tapasin. HLA Class I antigen and HLA-DR/DQ/DP antigen expression was significantly up-regulated on the dendritic-like cells. LMP2, LMP7 and tapasin expression was significantly up-regulated in all t(9;22) ALL-derived dendritic-like cells, in comparison to the unmodified blasts. No significant difference was detected in the expression of the other antigen processing machinery components. These results suggest that T cell activation by t(9;22) ALL-derived dendritic-like cells is associated with increased expression of some, but not all, of the antigen processing machinery components. Whether t(9;22) ALL-derived dendritic-like cells can be used as a cellular vaccine for adoptive immunotherapy of ALL is being investigated.
Amphiregulin Is Produced by Myeloma Cells and Is Involved in Tumor-Environment Interactions in Multiple Myeloma.
Authors: Mahtouk Karène , Hose Dirk , Reme Thierry , De Vos John , Jourdan Michel , Moreaux Jérome , Fiol Geneviève , Raab Marc , Jourdan Eric , Moos Marion , Goldschmidt Hartmut , Rossi Jean-François , Cremer Friedrich W. , Klein Bernard 
Published: April 2020

Abstract
Multiple myeloma (MM) is characterized by the accumulation of clonal malignant plasma cells in the bone marrow. One of the hallmarks of plasma cells is the expression of the heparan-sulfate proteoglycan syndecan-1. In epithelial cells, syndecan-1 plays a major role as a coreceptor for heparin-binding growth factors and chemokines. This stresses that heparin-binding growth factors may play a major role in the biology of MM cells. Recently we have demonstrated that heparin-binding EGF-like growth factor (HB-EGF), one of the ten members of the Epidermal Growth Factor (EGF) family, is produced by the tumor microenvironment and is able to trigger myeloma cell growth. As amphiregulin (AREG) is another member of the EGF family that also binds heparan-sulphate chains, we investigated its role in MM.
We looked for AREG expression on a panel of 7 normal plasmablastic cells (PPCs), 7 normal bone marrow plasma cells (BMPCs), purified MM cells from 65 patients and 20 myeloma cell lines (HMCLs), with Affymetrix U133A+B microarrays. We showed that primary MM cells overexpress AREG compared to normal BMPCs and PPCs. We then investigated the expression of the ErbB receptors with real-time RT-PCR. Myeloma cells variably expressed the 4 ErbB receptors. Normal BMPCs also expressed ErbB1 and ErbB2 unlike PPCs that did not express any ErbB receptors. We demonstrated that the high AREG expression by primary myeloma cells may have a dual effect. On the one hand, AREG stimulated IL-6 production and growth of bone-marrow stromal cells that highly express the AREG ErbB1 receptor. On the other hand, AREG could promote HMCL proliferation, suggesting that a functional autocrine loop involving AREG and ErbB receptors is involved in MM cell growth. Finally, we looked for the effect of ErbB inhibitors on MM cells of 14 patients cultured for 6 days together with their bone marrow environment. A pan-ErbB inhibitor (PD-169540, Pfizer) and an ErbB1-inhibitor (IRESSA, Astrazeneca) induced strong MM cell apoptosis in respectively 71% of patients (10 of 14) and 29% of patients (4 of 14). Of major interest, when PD169540 or IRESSA were combined with dexamethasone, they induced a dramatic myeloma cell death (respectively 92% and 69% inhibition of MM cell survival), while non-myeloma cells were unaffected. Thus ErbB activation is critical to trigger MM-cell survival in short-term culture.
In conclusion, our findings provide evidence for a major role of AREG and HB-EGF in the biology of multiple myeloma and identify ErbB receptors as putative therapeutic targets. These data emphasize the interest of clinical evaluation of specific-ErbB-inhibitors in patients with MM, either used alone or in combination with dexamethasone.
BCR-ABL Activates STAT3 Via MEK Kinase 1 in Embryonic Stem Cells.
Authors: Nakamura Yukinori , Yujiri Toshiaki , Nawata Ryouhei , Tagami Kozo , Tanizawa Yukio 
Published: April 2020

Abstract
BCR-ABL oncogene, the molecular hallmark of chronic myelogenous leukemia, arises in a primitive hematopoietic stem cell that has the capacity for both differentiation and self-renewal. Its product, Bcr-Abl protein, has been shown to activate STAT3 and to promote self-renewal in ES cells, even in the absence of leukemia inhibitory factor (LIF). MEK kinase 1 (MEKK1) is a 196-kDa mitogen-activated protein kinase (MAPK) kinase kinase involved in Bcr-Abl signal transduction (Oncogene22:7774, 2003). To investigate the role of MEKK1 in Bcr-Abl-induced transformation of ES cells, p210 Bcr-Abl was stably transfected into wild type (WT+p210) and MEKK1−/− (MEKK1−/−+p210) ES cells. Bcr-Abl enhanced both MEKK1 expression and activation in ES cells, as it does in other Bcr-Abl-transformed cells. In the absence of LIF, WT+p210 cells showed constitutive STAT3 activation and formed compact colonies having strong alkaline phosphatase activity, a characteristic phenotype of undifferentiated ES cells. MEKK1−/−+p210 cells, by contrast, showed less STAT3 activity than WT+p210 cells and formed large, flattened colonies having weak alkaline phosphatase activity, a phenotype of differentiated ES cells. These results indicate that MEKK1 plays an essential role in Bcr-Abl-induced STAT3 activation and in the capacity for LIF-independent self-renewal, and may thus be involved in Bcr-Abl-mediated leukemogenesis in stem cells.
All Trans-Retinoic Acid (ATRA) in Combination with PPAR-g Ligand Synergistically Enhances Inhibition of Cell Growth and Induction of Tumor Suppressor PTEN in Leukemic Cells.
Authors: Park Jinny , Lee Young-Rae , Shim Hyun-Jaung , Kwak Jae-Young , Yim Chang-Yeoul , Song Eun-Kyung , Han Myung-Kwan , Park Jin-Woo , Rho Hye-Won , Lee Sang-Young , Kim Jong-Suk 
Published: April 2020

Abstract
Peroxisome proliferator-activated receptor-γ (PPAR-γ), a transcriptional factor that plays an essential role in mediating the pharmacologic actions of PPAR-γ ligands, is highly expressed in normal monocytes, various leukemias, and epithelial malignancies. PPAR-γ ligands have been developed to induce differentiation, growth arrest, and apoptosis. PPAR-γ must form a heterodimer with the retinoic acid receptor (RAR) to bind DNA, and its transcriptional activity is thought to be maximal in the presence of both PPAR-γ ligand and RAR ligand. Recently, it has been shown that activated PPAR-γ and PPAR-γ ligand may influence tumor growth, directly and indirectly, through regulation of the tumor suppressor gene PTEN, a lipid phosphatase that plays a significant role in several cellular functions, including survival and proliferation, by antagonizing phosphatidylinositol 3-kinase (PI 3-kinase)-mediated signaling pathways. The human leukemia cell line HL60 displays controlled differentiation along the granulocyte or monocyte lineages in response to various inductive signals. Retinoic acid receptor ligands are inducers of HL60 granulocytic differentiation. PPAR-γ ligands and RAR ligands induced growth arrest and oxidative burst capacity in HL60 cells. Previously, we reported the induction of PTEN gene expression by the PPAR-γ ligand ciglitazone in HL60 cells in the presence of retinoic acid using northern and western blotting. Here, using RT-PCR and an AdenoVector system to examine the contribution of transcriptional control in PTEN regulation and for PTEN overexpression, we show that PPAR-g ligand and RAR ligand can synergistically upregulate PTEN in human promyeloid leukemia cells, resulting in inhibition of cell growth and cell cycle progression of acute leukemic cells. Adenovirus-delivered PTEN overexpression resulted in arrests of both cell growth and the G1 phase of the cell cycle in HL-60 cells, suggesting that the growth-suppressive effect of PTEN is associated with its ability to induce cell cycle arrest. The PPAR-γ ligand, ciglitazone, and the RAR ligand, retinoic acid, increased PTEN expression of HL-60 cells in a time- and dose-dependent manner, respectively. Upregulation of PTEN was significantly enhanced by a combination of both ciglitazone and retinoic acid. Moreover, these compounds synergistically induced arrests of both cell growth and the G1 phase of the cell cycle. These findings suggest that the activation of the PPAR-γ: RAR heterodimer represents a novel regulatory pathway for HL60 cells and there may be a possible role for PPAR-γ and RAR ligand in prophylactic and therapeutic approaches for controlling leukemia through the upregulation of PTEN.
Mutations in the AML1 Gene Define an Unfavorable Subgroup in Acute Myeloid Leukemia with FAB M0.
Authors: Dicker Frank , Klaus Mirjam , Haferlach Torsten , Kern Wolfgang , Hiddemann Wolfgang , Schoch Claudia , Schnittger Susanne 
Published: April 2020

Abstract
The AML1/RUNX1 gene is the most frequent target for chromosomal translocations in leukemia. Recently point mutations in the AML1 gene have been demonstrated as another mode of genetic aberration. AML1 mutations have been reported in de novo MDS and AML, as well as in therapy related MDS and AML. The AML M0 subtype has been found to be most frequently affected by sporadic AML1 gene mutations. We analysed AML1 gene mutations in a cohort of 49 M0 patients. Mutation screening was performed either with SSCP (n=21) and/or denaturating High Performance Liquid Chromatography (dHPLC) (n=33), 5 cases were analyzed by both methods. SSCP screening of exons 3–5 of the AML1 gene was carried out at the genomic level. These exons cover the socalled Runt domain, which is most frequently mutated. Fragments with aberrant mobility were sequenced. With this method 5 cases were found to be mutated. Subsequently, to improve the screening efficiency an assay using dHPLC was established. Hereby, we screened the cDNA of patient samples for mutations in amino acid codons 1–277 of the AML1b transcript, where the Runt domain is located between codons 49 and 178. All 5 cases detected by SSCP were confirmed by dHPLC. Nine mutations were detected in the cohort of 28 cases (32%) which had not been analyzed by SSCP. In total, 14 of the 49 samples (29%) tested were identified to be mutated, which is a slightly higher frequency than previously reported. In the cohort of 35 AML1 non-mutated cases 20 (57%) had a normal karyotype and 15 (43%) an aberrant karyotypes, whereas only 6 of the 14 AML1 mutated cases (43%) had a normal karyotype (p=0.001). Three of the AML1 mutated cases (21%) also had FLT3 mutations. One had an FLT3-LM, one an FLT3-TKD mutation, and one case both LM and TKD mutations. Clinical follow up data were available for 33 patients (22 AML1 non- mutated, 11 AML1 mutated). The median OS and EFS of the AML1 non-mutated versus the mutated group was 276 days versus 63 days (p = 0.0679) and 276 vs. 63 days (p=0.0630) respectively. Thus the AML1 mutated cases tend to have a worse clinical outcome. When other AML subtypes were screened for AML1 mutations, i.e. M1 (n=26), M2 (n=21) and M4 (n=3), only 1 additional AML1 mutation was detected, confirming the highest prevalence of AML1 mutations in M0. In conclusion, 1) we established a new assay to screen for AML1 mutations. 2) We confirmed the high incidence of AML1 gene mutations in AML M0, both in cases with normal and aberrant karyotype. 3) For the first time we demonstrated that AML1 mutations define an unfavorable subentity in AML M0.
A New Polycythemia Syndrome: Congenital Polycythemia with High Erythropoietin and Propensity for Malignant Hypertension Due to Paraganglioma.
Authors: Guan Yongli , Wu John K. , Jastaniah Wasil , Moss Larry G. , Digman Colleen , Mostacci Salvatore , Prchal Jaroslav F. , Prchal Josef T. 
Published: April 2020

Abstract
We report three patients, two females and one male, with congenital polycythemia and high erythropoietin. All were extensively investigated in childhood; no known polycythemic conditions were diagnosed nor were any tumors known to be associated with polycythemia uncovered. At the ages of 14, 16, and 32 years the patients developed malignant hypertension secondary to multiple paragangliomas. In addition, one of patients has also developed a malignant pancreatic carcinoid tumor that expressed somatostatin. After the surgical resection of paragangliomas, hypertension subsided, but polycythemia persisted in all three subjects.
Comprehensive screening for germ-line mutations of von Hippel-Lindau (VHL) gene as well as genes known to be causing familial paragangliomas - catalytic succinate dehydrogenase subunits B (SDHB), and catalytic succinate dehydrogenase subunit D (SDHD), has not revealed any mutations. In one of these patients studied so far there was no abnormality of the any of the three hypoxia control associated proline hydroxylase genes expressions found. Unlike the erythroid progenitors of Chuvash Polycythemia, these patients had normal sensitivity of erythroid progenitors to erythropoietin. The search for somatic mutations of VHL, SDHB, and SDHD genes, as well as quantitative analysis of the expressions of hypoxia inducible factor 1 alpha subunit (HIF-1a), vascular endothelial growth factor (VEGF), VHL, SDHB, and SDHD mRNAs by real-time PCR is ongoing.
In summary, we report a new polycythemic syndrome of congenital polycythemia with high erythropoietin and the normal sensitivity of erythroid progenitors to erythropoietin, which is not associated with VHL mutation. This is in contrast to our previous experience with about 200 patients, homozygous for Chuvash Polycythemia mutation or compound heterozygous for Chuvash Polycythemia R200W and other VHL mutations, who have not developed paraganglioma or any VHL syndrome tumors. This new polycythemic syndrome shares some clinical features with both von Hippel Lindau syndrome and with the Chuvash polycythemia. While its molecular defect is yet to be elucidated, it is a distinct clinical and genetic entity.
Analysis of Common Single Nucleotide Polymorphisms in MDR1 Gene in Patients with Multiple Myeloma.
Authors: Jamroziak Krzysztof , Balcerczak Ewa , Smolewski Piotr , Piaskowski Sylwester , Cebula Barbara , Panczyk Mariusz , Janus Agnieszka , Mirowski Marek , Robak Tadeusz 
Published: April 2020

Abstract
P-glycoprotein (P-gp) encoded by MDR1 gene is a xenobiotic transporter involved in protection of normal tissues against environmental toxicants and multi-drug resistance of cancer cells. Recently, a number of authors, including our group, have indicated that phenomenon of single nucleotide polymorphism (SNP) in MDR1 may influence susceptibility or prognosis in various diseases as well as pharmacokinetics of P-gp substrates. Although environmental chemicals are suspected carcinogens in multiple myeloma (MM) and P-gp transported drugs are used in the treatment of this malignancy, little data is known on the significance of MDR1 SNPs in MM. In this study, three common MDR1 SNPs: C1236T, G2677T/A and C3435T were assessed in 89 MM patients using automated sequencing and RFLP method. Moreover, in the control group of 215 healthy individuals MDR1 C3435T SNP was genotyped in all subjects and P-gp expression and activity were examined in a proportion of subjects. The haplotypes were inferred using Partition-Ligation algorithm showing highest frequency of the haplotype with wild-type alleles at all sites studied (1236C-2677G-3435C, 30%), followed by haplotype with all mutant alleles (1236T-2677T-3435T; 27%) and 1236C-2677G-3435T haplotype (12%). Carriers of a mutant allele at MDR1 1236 site showed a trend to longer overall survival as compared to wild-type homozygotes MDR1 1236CC (p=0.08), while no correlations with survival were observed regarding other polymorphisms. As compared to the control group, the MDR1 C3435T SNP allele frequency did not differ in MM patients, however the carriers of at least one T-allele (CT and TT genotypes) were significantly more frequent in the MM cohort (79% vs. 65%, p=0.02). The flow cytometry results of anti-P-gp MRK16 antibody staining and functional rhodamin123/PSC833 assay performed on peripheral blood mononuclear cells, CD19+ B-lymphocytes and CD3+ T-lymphocytes were similar among healthy individuals having MDR1 3435CC, CT and TT genotypes. In conclusion, we observed that MDR1 gene SNPs may exert some effect in genetic predisposition and prognosis in MM, which should be further examined in larger cohorts of uniformly treated patients.