bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Nuclear Factor-kappa B Pathway Genes and Their Correlation with Clinical Features in Multiple Myeloma.
Authors: Almeida Manuella S.S. , Vettore Andre L. , Felix Roberta S. , Andrade Valeria C.C. , Carvalho Fabricio , Yamamoto Mihoko , Chauffaille Maria de Lourdes L.F. , Zago Marco Antonio , Colleoni Gisele W.B. 
Published: October 2019

Abstract
NF-k B is a transcription factor that controls the expression of a variety of genes involved in different cellular processes, including apoptosis regulation, cell proliferation and expression of adhesion molecules. In multiple myeloma (MM) patients, NF-k B confers longer survival and drug resistance to tumor cells. Therefore, a better understanding of molecular mechanisms involved in NF-k B pathway can contribute to identify new therapeutic targets for this incurable disease.
Purpose: To evaluate the expression of NF-k B pathway genes in MM patients by real-time quantitative PCR and its possible correlation with disease clinical features and survival.
Patients and methods: Expression of eight genes related to NF-k B pathway (NFKB, IKB, RANK, RANKL, OPG, IL6, VCAM and ICAM) were studied in 53 bone marrow samples from newly diagnosed MM patients and in seven normal controls, using Taqman system. Genes were considered overexpressed when tumor expression level was at least four times higher than the observed in normal samples. These results were compared to clinical variables using Pearson’s Qui-square and T-Test. Overall survival (OS) was estimated using Kaplan-Meier method.
Results: 18.9% of MM cases were classified as ISS 1, 34% ISS 2 and 39.6% ISS 3. The percentages of overexpression of the eight genes were: NFKB 0%, IKB 22.6%, RANK 15.1%, RANKL 31.3%, OPG 7.5%, IL6 39.6%, VCAM 10% and ICAM 26%. We found association between IL6 expression level and the International Staging System (ISS) (p = 0.01), meaning that MM patients with high ISS scores have more chance of overexpression of IL6. We also found association between IL6 overexpression and presence of chromosome 13 deletion (p= 0.035), a well-known marker of worse prognosis in MM. Mean value of ICAM relative expression was also associated with ISS score (p = 0.02). Regarding OS, cases with IL6 overexpression present worse evolution than cases with IL6 normoexpression (p=0.04).
Conclusions: IL6 overexpression was found in 39.6% of MM cases and confirms its participation in myeloma progression, regulating growth and survival of tumor cells. Besides, IL6 overexpression was significantly associated with advanced stage disease and del(13) and had impact on MM patient’s survival. ICAM was overexpressed in 26% of cases and its level was associated with ISS scores. Therefore, ICAM seems to have some potential as therapeutic target in MM.
Imatinib Induces Apoptosis of Myeloma Cells by Inhibiting c-Kit Signaling Transduction.
Authors: Li Juan , Huang Beihui , Zhao Ying , Luo Shaokai 
Published: October 2019

Abstract
Objective To evaluate the mechanism and influence of Imatinib on myeloma cells which express c-Kit receptor.
Methods KM3 cells were treated with Imatinib for indicated time and indicated concentration and cell growth index were evaluated by XTT assay. Cell cycle were examined by flow cytometry. Apoptosis were assessed by Annexin V/PI. c-Kit and phosphate c-Kit were examined by Western blot.
Results Imatinib inhibited proliferation of KM3 cells in a concentration more than 0.25μmol/L, induced a progressive decline in S phase cell fraction and an increase in G0/G1 cells. Annexin V/PI staining and DNA ladder indicated that Imatinib had a substantial effect on apoptosis of KM3 cells in a dose-dependent manner and induced pro-caspase-3 and PARP to cleave. Imatinib treatment inhibited expression of c-Kit and provoked a decreased of IL-6 induced c-Kit phosphorylation.
Conclusion Imatinib inhibites KM3 myeloma cells proliferation and induces apoptosis of KM3 myeloma cells by inhibiting c-Kit signaling transduction.
Selective Inhibition of Glycogen Synthase Kinase-3 by Facilitators of Insulin Response Promotes Proliferation and Survival in Multiple Myeloma Cells.
Authors: Jernberg-Wiklund Helena , Stromberg Thomas , Hammarberg Anna , Georgii-Hemming Patrik , Osterborg Anders , Nilsson Kenneth 
Published: October 2019

Abstract
There is ample evidence for a central role of IGF-1R in multiple myeloma (MM) cell survival and resistance to apoptosis. Recently, we have demonstrated the use of inhibitors of the IGF-1R tyrosine kinase (RTK) activity as a promising therapeutic strategy in MM in vitro and in vivo (Stromberg et al Blood 2006,107:669–78, Menu et al Int J Cancer 2007,121:1857–61). In the present investigation, we have examined the role of glycogen synthase kinase-3 (GSK3), a target of the IGF-1R/PI 3-K/AKT pathway, in proliferation and apoptosis of MM. Unlike most kinases GSK3 is constitutively active, but may be inhibited by e.g. insulin, EGF and FGF. GSK3 is also a converging point of the Wnt and the IGF-1R/PI 3-K/AKT survival pathway. Reduction of GSK3 activity resulted from its phosphorylation at the regulatory residues Ser21 (α isoform) and Ser9 (β isoform) catalysed by AKT. GSK3 inhibitors LiCl, AR-A014418, the maleimides SB216763 and SB41528, and the highly selective aminopyrimidine derivatives CT98014 and CT99021 (Novartis, Emeryville, CA), were investigated for their effects on MM cells when growth and survival were challenged by cytotoxic stress and/or interference with different signaling proteins. Apoptosis induced by serum starvation was reduced by CT99021, an effect comparable to the survival-promoting effect of IGF-1. The selective GSK3 inhibitor also restored survival in MM cells induced by glucocorticoids and combinations increasing the susceptibility to apoptosis using PI 3-K inhibitors LY294002, the IGF-1R antagonist αIR3, the mTOR inhibitor rapamycin and the MEK inhibitor PD98059. In addition to the anti-apoptotic effect, CT99021 was also demonstrated to counteract G0/G1 accumulation induced by serum starvation, LY294002, αIR3 and rapamycin. However, accumulation of cells in the G2/M phase induced by doxorubicin and melphalan, were not counteracted by GSK3 inhibitors. Substrates of GSK3 most prominently include beta-catenin and the IκB-analog Bcl-3, both targeted for degradation upon GSK3 activation. Selective GSK inhibition restituted protein levels of beta-catenin, Bcl-3, p21 cip1/WAF1, p27kip1 and cyclin B1, D2 and E corroborating with the effects on survival and proliferation. These data demonstrate that targeting of the GSK3 using highly selective inhibitors is associated with potent anti-apoptotic effects and propagation through the cell cycle in MM.
IgG-Derived Tregitope Peptides Suppress T Cell Responses in Vitro and in Vivo
Authors: De Groot Anne S. , Moise Leonard , Su Yan , McMurry Julie A , Martin William D , Scott David W. 
Published: October 2019

Abstract
We have identified a set of putative natural T regulatory epitopes (Tregitopes) which, when co-administered with an antigen, cause the expansion of antigen-specific adaptive Tregs in vitro and in vivo. They have the following characteristics:
they bind, in most cases, with high affinity to multiple MHC class II molecules and, when co-administered with antigen, they suppress effector T cell immune responses to the antigen and up-regulate Treg associated cytokines and chemokines.
T cells responding to Tregitopes exhibit a T regulatory phenotype (CD4+/CD25hiFoxP3+). To test whether Tregitopes derived from immunoglobulin (Ig) suppress immune responses to antigen co-administered in vivo, we performed two types of experiments. In the first, we dosed three groups of HLA DR4 mice every other week for six weeks with either
a peptide antigen (pAg) alone, pAg with murine Fc, or pAg with mTregitope289, the murine homolog of the human Tregitope289.
Mice were sacrificed and spleens harvested for assay. While the mice dosed with murine Fc demonstrated a reduced IL-4 ELIspot response to pAg, remarkably, the reduction was even greater in the mice treated with Tregitope. In a second model, C57Bl/6 mice were injected with LPS-stimulated B cells that were pulsed with either
ovalbumin (OVA) alone, mTregitopes 167 and 289 or with OVA together with the two mTregitopes.
One week later, mice were challenged with OVA 323–339 peptide in adjuvant. Two weeks after challenge, draining lymph nodes were harvested and LN cells stimulated with OVA 323–339 for measurement of T-cell proliferation by thymidine incorporation and by IFN-γ secretion by ELIspot. The mice receiving B cells previously pulsed with OVA alone demonstrated a robust IFN-gamma response to OVA re-stimulation. In contrast, the mice receiving B cells previously pulsed with OVA + Tregitopes demonstrated a comparatively reduced response. When sera were assayed for anti-OVA antibodies by ELISA, antibody response to OVA also declined following treatment with B cells co-administered with Tregitope. The mechanism of suppression appears to be due to the induction of antigen-specific adaptive tolerance induction (De Groot AS et al. Activation of natural regulatory T cells by IgG Fc-derived Peptide “Tregitopes” Blood112: in press, 2008).
Identification of the Involvement of the Tyrosine Kinase C-Ros in the Pathogenesis of Chronic Myelomonocytic Leukemia (CMML)
Authors: Cilloni Daniela , Carturan Sonia , Bracco Enrico , Defilippi Ilaria , Maffè Chiara , Iacobucci Ilaria , Pautasso Marisa , Rotolo Antonia , Giugliano Emilia , Messa Emanuela , Martinelli Giovanni , Saglio Giuseppe 
Published: October 2019

Abstract
The abnormal activation of tyrosine kinases are a common finding in chronic myeloproliferative disorders. Perturbation of RTK signalling by genetic alterations results into an abnormal proliferation advantage and finally into a malignant transformation. c-Ros is an orphan RTK displaying transforming activity whose role has been established in the development of neuronal neoplasia. The aim of this study was to evaluate the involvement of c-Ros in the pathogenesis of chronic myelomonocytic leukemia (CMML) and to establish the effects of c-Ros activation. c-Ros expression was evaluated by RQ-PCR in 133 samples collected from 96 CMML patients at diagnosis (96 BM and 37 PB) and 60 healthy donors (30 PB and 30 BM). The protein amount and localization was analyzed by westen blot and immunofluorescence assay. In order to establish the effects of c-Ros activation we generated a chimeric receptor containing the extracellular domain derived from epidermal growth factor receptor (EGFR) and the transmembrane and cytoplasmic domains from c-ros (ER). The chimeric receptor was then transfected in NIH3T3 and HEK293T cells. Transfected and control cells were then stimulated with 100 nM EGF ligand and proliferation and apoptosis evaluated by incorporation of 3H timidine and MTT assay and by FACS for the detection of annexin V, respectively. We found that Ros is undetectable in healthy subjects but it is overexpressed in CMML (p<0,0001) in both BM and PB cells with a median value of 2−ΔΔCt in BM of 380 (range 10–63303) and 212 in PB (range 6–30012). WB confirmed the presence of c-Ros protein in CMML cells but not in normal controls. Immunofluorescence staining localized the protein within the cytoplasm. We found that ROS is highly expressed in CD34+ cells and monocytes from CMML patients but not in their normal counterparts. Sequence analysis revealed the absence of mutations of c-Ros promoter. SNPs analysis exclude the presence of duplications or deletions of the gene. Moreover we found that the EGF induced activation of c-Ros affects proliferation by increasing of 3.5 folds the proliferation rate as compared to cells transfected with the empty vector and stimulated with EGF under the same conditions. Furthermore cell adhesion was 4 folds decreased as compared to control. By contrast apoptosis is not affected by c-ROS activity (p=0,2). This study demonstrates that c-Ros is abnormally expressed in patients with CMML. The abnormal activation of c-Ros is responsible for loss of adhesion and increased proliferation. In conclusion, we identified a new tyrosine kinase which may be responsible for the proliferation defect typical of CMML cells and could represent a target for molecular therapies.
Protein Phosphatase 2B Inhibition Promotes the Secretion of Von Willebrand Factor from Endothelial Cells
Authors: Nolasco Leticia , Gushiken Francisca C. , Patel Vimal , Turner Nancy , Pradhan Subhashree , Dong Jing-fei , Moake Joel L. , Vijayan K. Vinod 
Published: October 2019

Abstract
Ultra-large von Willebrand factor (ULVWF) multimers secreted from the Weibel-Palade bodies (WPBs) of endothelial cells plays a crucial role in the development of thrombotic microangiopathies. Secretion of WPB contents is regulated, in part, by the phosphorylation of vesicle trafficking proteins that constitutes the endothelial exocytotic machinery. In comparison to agonist-induced, protein kinase-dependent signaling pathways that regulate the exocytosis of WPB contents, a role for protein phosphatases in regulating endothelial exocytosis is currently undefined. In this study, we show that inhibition of serine/threonine protein phosphatase 2B (PP2B) activity by cyclosporine A (CsA), tacrolimus (FK506) or a cell-permeable PP2B autoinhibitory (AI) peptide promotes the secretion of hyper-adhesive ULVWF from human umbilical vein endothelial cells (HUVECs) in the absence of any other endothelial cell-stimulating agent. PP2B inhibitor-induced secretion and anchorage of ULVWF strings from HUVECs induce the tethering of adhesive platelets. In support of a role for PP2B in VWF secretion, we demonstrated that the catalytic subunit of PP2B expressed as a GST fusion protein interacts with the vesicle trafficking protein, Munc18c. Serine phosphorylation of Munc18c, which promotes granule exocytosis in other secretory cells, was significantly increased in CsA-treated HUVECs, suggesting that this process may be involved in CsA-mediated WPB exocytosis and secretion of VWF. Furthermore, plasma VWF antigen levels were also enhanced in CsA-treated mice, and siRNA-mediated knockdown of PP2Bb in HUVECs significantly enhanced VWF secretion. These observations suggest that CsA promotes VWF release, at least in part by inhibition of PP2B activity. These observations are compatible with the clinically observed association of CsA treatment and increased plasma VWF levels in humans, as well as the association between prolonged exposure to CsA and thrombotic microangiopathy in a subset of exposed patients.
Hematopoietic Differentiation of Human Induced Pluripotent Stem Cells
Authors: Choi Kyung-Dal , Yu Junying , Smuga-Otto Kimberly , Dias Jessica , Salvagiotto Giorgia , Vodyanik Maxim , Thomson James , Slukvin Igor 
Published: October 2019

Abstract
Induced pluripotent stem cells (iPSCs) provide an unprecedented opportunity for modeling of human diseases in vitro as well as for developing novel approaches for regenerative therapy based on immunologically compatible cells. In the present study, we employed an OP9 differentiation system to characterize the hematopoietic differentiation potential of seven human iPSC lines obtained from human fetal, neonatal, and adult fibroblasts through reprogramming with POU5F1, SOX2, NANOG, and LIN28 and compared it with the differentiation potential of five human embryonic stem cell lines (hESC; H1, H7, H9, H13, and H14). Similar to hESCs, all iPSCs in coculture with OP9 generated all types of colony forming cells (CFCs) as well as CD34+ cells that can be separated into distinct subsets based on differential expression of CD43 and CD31. CD34+CD31+CD43− cells obtained from all iPSCs expressed molecules present on endothelial cells and readily formed a monolayer when placed in endothelial conditions, while hematopoietic CFC potential was restricted to CD43+ cells. iPSC-derived CD43+ cells could be separated into three major subsets based on differential expression of CD235a/CD41a and CD45: CD235a+CD41a+/− (erythro-megakaryocytic progenitors), and lin-CD34+CD43+CD45− (multipotent), and lin-CD34+CD43+CD45+ (myeloid-skewed) primitive hematopoietic cells. Both subsets of primitive hematopoietic cells expressed genes associated with myeloid and lymphoid development, although myeloid genes were upregulated in CD45+ cells, which are skewed toward myeloid differentiation. Cytogenetic analysis demonstrated that iPSCs and derived from them CD43+ cells maintained normal karyotype. In addition short tandem repeat analysis of CFCs generated from IMR90-1 cells has been performed to confirm that blood cells are in fact derived from reprogrammed IMR90 cells, and not from contaminating hESCs. While we observed some variations in the efficiency of hematopoietic differentiation between different iPSCs, the pattern of differentiation was very similar in all seven tested iPSC and five hESC lines. Using different cytokine combinations and culture conditions we were able to expand iPSC-derived myeloid progenitors and induce their differentiation toward red blood cells, neutrophils, eosinophils, macrophages, ostoeclasts, dendritic and Langerhans cells. Although several issues remain to be resolved before iPSC-derived blood cells can be administered to humans for therapeutic purposes, patient-specific iPSCs can already be used for characterization of mechanisms of blood diseases and to identify molecules that can correct affected genetic networks.
Improved Outcome by Addition of Lomustine (CCNU) to Idarubicin and Cytarabine in Elderly Patients with De Novo Acute Myeloid Leukemia. A Report from the GOELAMS Group
Authors: Pigneux Arnaud , Witz Francis , Sauvezie Mathieu , Bene Marie-Christine , Luquet Isabelle , hunault-Berger Mathilde , Recher Christian , Vey Norbert , Lioure Bruno , Himberlin Chantal , Escoffre-Barbe Martine , Berthou Christian , Lissandre Severine , Fegueux Nathalie , Cahn Jean-Yves , Jourdan Eric , Bouscary Didier , Dreyfus Francois , Reiffers Josy M. , Milpied Noel , Harousseau Jean-Luc , Ifrah Norbert 
Published: October 2019

Abstract
In elderly patients with acute myeloid leukemia (AML) treated intensively, no improvement has been shown in the last 20 years. We performed a retrospective study in 847 patients over 60 years old, prospectively enrolled in 3 trials conducted in France between 1995 and 2005, with the aim to investigate prognostic factors for complete remission (CR) achievement and survival. Induction therapy consisted in the association of Idarubicin 8mg/m2 d1-5 and Cytarabine 100mg/m2 d1-7 (Group I, 339 patients) or the same drugs with the addition of lomustine (10mg\m2 orally at day 1)(Group II, 508 patients). Consolidation therapy consisted of anthracycline and cytarabine courses at lower doses, preceded or not by a first course with intermediate dose cytarabine. The patients’ characteristics were similar between the two groups concerning sex, WBC count, ECOG, and cytogenetics, yet patients were older in Group II versus Group I (55% versus 45% over 69 years of age, p<0.0001).The CR rate was significantly higher for patients in Group II compared to Group I (67 % vs 57%, p= 0.002). The toxic death rate was not different between groups. In multivariate analysis, three good prognostic factors emerged for achieving complete remission: good or intermediate cytogenetics (p<0.0001), ECOG < 2 (p<0.0001), and adjunction of lomustine to induction chemotherapy (p=0.002). The median overall-survival was significantly improved for patients treated with lomustine (12.7± 2.2 months vs 8.7± 2.7 months, p=0.004). In multivariate analysis, five prognostic factors affected positively overall survival: adjunction of lomustine to induction chemotherapy (p < 0.0001), age < 69 years (p =0.001), ECOG < 2 (p =0.001), FAB other than AML0,6 or 7 (p = 0.004) and good or intermediate cytogenetics(p = 0.007). The median event-freesurvival was also improved for patients treated with lomustine (10.7± 2.2 months vs 7± 2.7 months, p=0.002). Event-free-survival was affected by the same prognostic factors as overall survival. We conclude that lomustine might be added in standard induction therapy as it allowed to obtain both better CR rate and survival in this retrospective study.
Myc Drives Chromosomal Gain in Acute Myeloid Leukemia
Authors: Jones Letetia , Sevcikova Sabina , Phan Vernon , Jain Sachi , Shieh Angell , Dubansky Joshua , Li Min , Maunakea Mei Lin , Zhu George , Tennant Thelma R , Lowe Scott , Le Michelle M. , Kogan Scott C. 
Published: October 2019

Abstract
Acute Myeloid Leukemia (AML) is a disease characterized by diverse genetic pathogenesis, including both balanced and unbalanced chromosomal aberrations. Much is known regarding the pathogenic effects of balanced rearrangements in AML, whereas our understanding of how unbalanced aberrations contribute to leukemia is more limited. The balanced t(15;17) chromosomal rearrangement is a nearly constant feature of acute promyeloctyic leukemia (APL), a subtype AML. The translocation fuses the promyelocytic leukemia gene (PML) to the retinoic acid receptor α gene (RARA). Trisomy 8 is the most common secondary karyotypic lesion observed in APL, and it has been speculated but not proven that the MYC gene contributes to this chromosomal gain. We previously reported that mouse chromosome 15, which contains the mouse Myc gene in a region syntenic to human chromosome 8q24, is commonly gained in the MRP8 PML-RARA mouse model of APL. We now report our work to assess the hypothesis that increased MYC cooperates with PML-RARα to accelerate disease and that gain of MYC/Myc drives +8 in humans and +15 in mice. Expressing MYC with a retroviral vector in PML-RARA bone marrow led to the rapid development of APL-like leukemias (3 months vs. 8.5 months with PML-RARA alone). Chromosome 15 was not gained in any of the leukemias, although 70% had other clonal karyotypic abnormalities. This finding suggests that when MYC is overexpressed, there is no selective pressure to gain chromosome 15, supporting our hypothesis that Myc is driving this gain. We also generated PML-RARA mice haploinsufficient for Myc to examine the effect of decreasing MYC levels. The median latency among leukemic animals was 258 days for mice with PML-RARA and two wild-type Myc alleles, whereas the latency was increased to 339 days for PML-RARA Myc haploinsufficient mice. Hence, lower MYC expression served as a check on leukemic transformation. Furthermore, the majority of the leukemias that arose in Myc haploinsufficient mice had gained wild-type Myc. These data demonstrate a selective pressure for Myc gain. Additional experiments showed that
as MYC expression increases there is a decrease in both latency and genetic complexity of leukemias that arise, that MYC and PML-RARα interact to disrupt myeloid differentiation in vivo and that although MYC cooperates with PML-RARα to cause leukemia, additional events are required for completing transformation even at high levels of MYC.
Altogether our studies of increased and decreased MYC expression in PML-RARA mice show a strong correlation between MYC dosage and leukemic transformation. Our results suggest that agents that target MYC might be useful for the treatment of AML.