bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract C116: The KidsCancerKinome: Validation of drug targets for high risk childhood cancers
Authors: Caron Huib N. , Westerhout Ellen M. , Molenaar Jan J. , Boer den Monique L. , Clifford Steve C. , Delattre Olivier , Geoerger Birgit , Lanvers Claudia , Pietsch Torsten , Scotlandi Katia , Sera Massimo , Shipley Janet , Vassal Gilles , Versteeg Rogier , Verschuur Arnauld 
Published: June 2012

Abstract
The aim of KCK is the pre-clinical selection and validation of drug targets and corresponding targeted drugs for 6 high risk childhood cancers. The KCK consortium focuses on six aggressive childhood tumors, killing ∼2000 children in Europe annually, i.e Ewing sarcoma, osteosarcoma, rhabdomyosarcoma, neuroblastoma, medulloblastoma and ALL. The KidsCancerKinome (KCK) project is funded by the EU in the FP6 programme and consists of 9 tumor biology labs in 4 European countries.
The experimental approach encompasses the following points:target presence analyses (mRNA and protein expression of the human kinome)molecular validation of kinase tumor dependency (RNAi)kinase mutation analysisin vitro drug efficacy testingin vivo proof-of-principle of drug efficacy
The KCK consortium has generated gene expression profiles (Affy U133plus2 arrays) of &gt;500 clinical tumor samples form those six tumor types. We have performed extensive analyses of mRNA expression of human kinases. Tissue microarrays with &gt;800 tumor samples have been analysed for various kinases and for phosphorylation status of downstream proteins. Examples of interesting expression patterns of the human kinome will be presented. Detailed analyses for the first 5 kinases for which targeted drugs are available, i.e. PI3K, IGF1R, AURKA+B, and CDK2 will be presented. Lentiviral shRNA mediated knockdown of kinase protein expression has been used in cell line panels for each tumor type to select kinases with tumor dependency as validated drug targets.
Many novel kinase inhibitors are under development for adult oncology and KCK will test their in vitro activity against the tumor-driving kinases identified in this program. We are currently testing small molecule inhibitors for the first 5 kinases. For those kinases that have no small molecule inhibitors, a novel generation of siRNA based nucleic acid drugs (LNAs), produced by the Santaris company, will be applied and tested in vitro. Successful small molecule inhibitors and LNAs will be taken further to in in in vivo validation in established xenograft models of the six childhood tumor types. Pharmacokinetic studies of these drugs will finally prepare them for evaluation in future clinical studies in childhood cancer patients.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C116.
Abstract C136: An analysis of BCAR3-mediated regulation of p130Cas localization and focal adhesion formation in breast cancer cells
Authors: Borre Pierre Vanden , Near Richard , Lerner Adam 
Published: June 2012

Abstract
BCAR3 and p130Cas, two focal adhesion proteins that bind to one another and that were previously identified in a screen for genes that confer anti-estrogen resistance in ER+ breast cancer cells, were analyzed in breast cancer cell lines to determine the effect of BCAR3 expression on cell morphology and protein localization including the Src-mediated redistribution of p130Cas.
The effect of altered BCAR3 expression and role of BCAR3 and p130Cas association was studied in MCF7 cells stably over-expressing wildtype BCAR3 or BCAR3 containing an R743A missense mutation that prevents p130Cas association. Protein localization and cellular morphology were examined by immunofluorescence and Western analyses of cells were conducted to observe p130Cas post-translational modifications and the potential involvement of the TGFβ and β-catenin signaling pathways.
Stable over-expression of BCAR3 in MCF7 epithelial cells induces the loss of cohesive association between cells and the formation of membrane projections as well as large focal adhesions that contain abundant p130Cas. This BCAR3-mediated increase in p130Cas localization to focal adhesions is independent of the association of BCAR3 and p130Cas. Src, which is known to phosphorylate p130Cas, also induces augmented localization of p130Cas and this effect is also further enhanced by BCAR3 expression. Once again, physical interaction between BCAR3 and p130Cas is not necessary for the ability of BCAR3 to increase Src-mediated redistribution of p130Cas to focal adhesions. In spite of inducing a loss of epithelial morphology in MCF7 cells, BCAR3 over-expression did not induce SMAD2 phosphorylation or nuclear translocation of β-catenin, suggesting that neither the TGFβ or β-catenin signaling pathways are activated in response to BCAR3 over-expression in epithelial breast cancer cells. Consistent with these results, shRNA-mediated reduction of BCAR3 expression in MDA-231 mesenchymal cells ablates basal p130Cas phosphorylation, alters p130Cas and focal adhesion localization and induces an epithelial-like growth pattern with attenuated stress fiber formation and decreased tensin expression. The inhibition of stress fiber formation in MDA-231 cells prevents p130Cas phosphorylation. BCAR3 expression regulates the morphology and cytoskeleton of epithelial and mesenchymal breast cancer cells by affecting focal adhesion formation, p130Cas localization and the cytoskeletal response to Src signaling. The effect of BCAR3 appears not to involve either the TGFβ or β-catenin signaling pathways and is largely independent of BCAR3's direct physical interaction with p130Cas. Our current studies are seeking to examine the role of BCAR3's SH2 domain and serine-proline-rich region in the regulation of focal adhesion biology.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C136.
Abstract C149: Exon array analyses across the NCI-60 reveals potential regulation of TOP1 by transcription pausing at guanosine quartets in the first intron
Authors: Reinhold William C. , Mergny Jean-Louis , Liu Hongfang , Ryan Michael , Pfister Thomas D. , Kinders Robert , Parchment Ralph , Doroshow James , Weinstein John N. , Pommier Yves 
Published: June 2012

Abstract
Because Topoisomerase I (TOP1) is critical for the relaxation of DNA supercoils and because it is the target for anticancer activity of camptothecins, we assessed TOP1 transcript levels in the 60 cell line panel (the NCI-60) of the National Cancer Institute's anticancer drug screen. TOP1 expression levels varied over a 5.7-fold range across the NCI-60. HCT116 colon and MCF-7 breast cancer cells were the highest expressers; SK-MEL-28 melanoma and HS578T breast carcinoma cells were the lowest. TOP1 mRNA expression was highly correlated with Top1 protein levels, indicating that TOP1 transcripts could be conveniently used to monitor Top1 protein levels and activity in tissues. Assessment of the TOP1 locus by array comparative genomic hybridization across the NCI-60 showed copy numbers ranging from 1.71 to 4.13 and a statistically significant correlation with TOP1 transcript levels (p&lt;0.01). Further analyses of TOP1 expression on an exon-specific basis revealed that exon 1 expression appeared to be generally higher, and less variable than the other exons, suggesting some form of transcriptional pausing regulation between exons one and two. Accordingly, we found the presence of multiple evolutionarilly-conserved potential G-quadruplex-forming sequences in the first TOP1 intron. Physico-chemical tests for actual quadruplex formation by several of those sequences yielded quadruplex formation for two of them and duplex formation for one. The observations reported here suggest the hypothesis that there is a conserved negative transcription regulator within intron 1 of the TOP1 gene, and the regulator is a quadruplex-prone region.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C149.
Abstract C156: Molecular targeting CK2 by siRNA or antisense delivered by tenfibgen ligand nanocapsules modulates NF-kappaB, TP53 and the malignant phenotype in head and neck cancer
Authors: Chen Zhong , Brown Matthew S. , Diallo Oumou T. , Hu Michael , Ehsanian Reza , Yang Xinping , Arun Pattatheyil , Lu Hai , Korman Vicci , Unger Cretchen , Van Waes Carter 
Published: June 2012

Abstract
We previously reported that protein kinase CK2 contributes to aberrant NF-kappaB activation, but the molecular mechanisms and antitumor effects of targeting CK2 have not been thoroughly investigated in head and neck squamous cell carcinoma (HNSCC). In the present study, we showed elevated CK2alpha, alpha' and beta mRNA and protein expression in a panel of HNSCC lines and tissues. siRNA specifically knocked down CK2 subunits and differentially inhibited IKKbeta mediated IkappaBalpha degradation, p65 synthesis and phosphorylation at serine 536 and 529, p65 and p50 binding, and NF-kappaB reporter activities. CK2alpha knockdown also upregulated two genes promoting apoptosis, TP53 and TAp63, which together with inhibition of NF-kappaB downregulated genes promoting growth, survival and adhesion, such as CyclinD1, BCL-XL, ITGA3, and ITGB4. Knockdown of CK2alpha' or beta also exhibited differential effects on gene expression. Consequently, CK2alpha siRNA most significantly inhibited proliferation, cell cycle, and migration, while inducing apoptosis and cisplatin sensitivity in vitro. Furthermore, delivery of antisense targeting CK2alpha and alpha' using tenfibgen (fibrinogen fragment of tenascin) coated nanocapsules significantly suppressed tumor growth in HNSCC xenograft models. Consistent with CK2 inhibition, antisense-treated tumor tissue specimens showed decreased expression of CK2alpha and alpha', NF-kappaB p65, Cyclin D1, Bcl-XL and Bcl2 proteins, decreased NF-kappaB p65 phosphorylation at serine 536 and 529, as well as increased TP53 and p63 by immunohistochemistry. Our study revealed the novel role of CK2 subunits in co-regulating NF-kappaB and TP53/p63 expression and signaling, as well as downstream gene expression, which result in anti-tumor effects in vitro and in vivo. Targeting the CK2 subunits may be an effective antitumor strategy with the ability to enhance the efficacy of cisplatin chemotherapy.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C156.
Abstract C157: Identification of a novel DNA topoisomerase II inhibitor by proteomic profiling
Authors: Kawatani Makoto , Takayama Hiroshi , Muroi Makoto , Kimura Shinya , Maekawa Taira , Osada Hiroyuki 
Published: June 2012

Abstract
Background: BNS-22 is a chemically synthesized derivative of the plant natural product GUT-70 isolated from the stem bark of Calophyllum brasiliense. BNS-22 possesses antiproliferative activity on human cancer cells, but its mechanism of action is unknown. We recently developed a proteome-based profiling system to predict molecular targets and mechanisms of action of bioactive small molecules. Here we report on the identification of a molecular target for BNS-22 by using the proteomic profiling approach.
Materials and Methods: HeLa cells were treated with BNS-22 and 19 well-characterized small molecules including signal transduction inhibitors and anticancer drugs for 18 h, and the cell lysates were subjected to 2-dimensional fluorescence differential gel electrophoresis (2D-DIGE). Then, we extracted 314 spots detected in all analysis gels, and performed cluster analysis based on the expression levels of those spots.
Results: Proteomic profiling analysis indicated that BNS-22 was classified into the same cluster as ICRF-193, a DNA topoisomerase II (TOP2) catalytic inhibitor. This profiling analysis also distinguished clearly the action between ICRF-193 and VP-16 (etoposide), a TOP2 poison. These results suggested that BNS-22 acted as a TOP2 catalytic inhibitor and not a TOP2 poison in cells. BNS-22 inhibited the DNA decatenation activity of human TOP2 in vitro, with an IC50 value of 2.4 M. BNS-22, like ICRF-193, induced the abnormal mitotic figure characterized by the unusually-distorted spindle and the failure of chromosome alignment and segregation, as assessed by immunofluorescence staining with anti- -tubulin antibody, anti- -tubulin antibody, and DAPI. Flow cytometric analysis showed that BNS-22-treated cells were accumulated at G2/M phase after 24 h in a dose-dependent manner. In the time course experiment, peaks corresponding to 8C were observed after 36 h in cells treated with 10 M BNS-22, indicating that BNS-22 as well as ICRF-193 induced polyploidy. BNS-22, unlike VP-16, did not cause DNA double-strand breaks, as measured by the phosphorylation of histone H2AX on Ser 139 (-H2AX), and furthermore antagonized VP-16-mediated DNA damage.
Conclusion: These results strongly suggest that BNS-22 targets TOP2 and acts as its catalytic inhibitor in cells.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C157.
Abstract C163: Gene expression signature-based Connectivity Map analysis revealed vorinostat as a novel therapeutic target agent in human gastric cancer
Authors: Claerhout Sofie A. , Lim Jae Yun , Choi Woonyoung , Kim Sang-Bae , Park Yun-Yong , Kim KyoungHyun , Mills Gordon B. , Hong Waun Ki , Lee Ju-Seog , Cho Jae Yong 
Published: June 2012

Abstract
Gene-expression profiling has been applied in specific settings such as to elucidate the mechanisms of biological pathways, to classify subtypes of a disease, and to predict cancer prognosis. Furthermore, the Connectivity Map using gene-expression signature can categorize the biological responses to a large number of diverse small molecule therapeutics.
To find new agents targeting gastric cancer, we analyzed global human gastric cancer gene expression profile using the Connectivity Map. From 3360 gastric cancer specific genes, it was revealed that the candidate compounds for targeting gastric cancer were histone deacetylase inhibitors (HDACi) such as vorinostat. We validated the therapeutic efficacy of vorinostat in gastric cancer cell lines. Vorinostat induced both apoptosis and autophagy in gastric cancer cell lines, but showed a different pattern in each cell line. Pharmacological or genetic inhibition of autophagy respectively by chloroquine and a siRNA against Beclin 1 increased the therapeutic efficacy of vorinostat, indicating a mechanism of drug resistance. Further transcriptome analysis will make it possible to find a predictive biomarker gene signature for vorinostat and as such can help in the future to better understand the action mechanisms of vorinostat. In conclusion, we showed that systems analysis using Connectivity Map with a human gastric cancer gene signature can be novel approach to find targeting agents for gastric cancer.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C163.
Abstract C178: Enhanced suppression of tumor growth by concomitant treatment of human lung cancer cells with SAHA and arsenic trioxide
Authors: Chien Chia-Wen , Yao Ju-Hsin , Lee Te-Chang 
Published: June 2012

Abstract
The efficacy of arsenic trioxide (ATO) against acute promyelocytic leukemia (APL) and relapsed APL has been well-documented. ATO may cause DNA damage through the generation of reactive oxygen intermediates. Histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), modulates gene or protein expression via histone dependent or independent pathway that may result in chromatin decondensation, cell cycle arrest, differentiation, and apoptosis. The objective of this study is to investigate whether ATO and SAHA could synergistically enhanced cell killing to cancer cells. The present studies showed that combined treatment with ATO and SAHA results in synergistic cytotoxicity in non-small cell lung carcinoma H1299 cells. By analyzing the annexin V positive cells, apoptotic cell death is significantly enhanced in combined treatment. By aid of Comet assay, we demonstrated that ATO induces DNA breaks in H1299 cells, but not SAHA. However, co-treatment with SAHA and ATO significantly increases ATO-induced DNA damage. At the doses used, ATO does not interfere with the cell cycle progression, whereas SAHA induces the expression of p21WAF1/CIP1 and leads to G1 arrest. Our present results suggest that SAHA may result in G1 arrest, which becomes more susceptible to ATO for causing DNA damage.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C178.
Abstract C186: A glycoengineered anti-CD19 antibody (MEDI-551) is effective as monotherapy or in combination treatment in mouse models of B-cell malignancies
Authors: Ward Beth K. , Mittereder N. , Kuta E. , Sims G. , Damschroeder M. , Dall'Acqua W. , Bowen M. A. , Wu H. , Tedder T. F. , Kiener P. A. , Jallal B. , Coyle T. , Herbst R. 
Published: June 2012

Abstract
Human cluster of differentiation (CD) antigen 19 is a B cell-specific surface antigen that is expressed by early pre-B cells and throughout B cell differentiation until it is down-regulated during terminal differentiation of plasma cells. Expression of CD19 is maintained following malignant transformation and therefore CD19 is expressed on the majority of B cell-derived leukemias and lymphomas.
MEDI-551 is a humanized immunoglobulin G1 kappa (IgG1) monoclonal antibody (MAb) directed against CD19. The Fc portion of MEDI-551 does not contain a fucose sugar moiety due to its expression in BioWa's Potelligent® cells. This afucosylated form of MEDI-551 exhibits enhanced CD16 binding and antibody-dependent cellular cytotoxicity (ADCC). Rituximab, another B-cell targeted therapy, is directed against CD20 and is widely used for the treatment of B cell malignancies. However, a subset of patients is unresponsive to rituximab therapy and the majority of patients will relapse following treatment. We have previously shown that the glycoengineered, ADCC-enhanced anti-CD19 MAb is effective in in vitro and in vivo models of B cell leukemia and lymphoma. The purpose of the present study was to explore the anti-tumor activity of the anti-CD19 MAb in SCID mouse models of human lymphoma as single agent and in combination with rituximab. In several models the combination of anti-CD19 with anti-CD20 resulted in prolonged suppression of tumor growth with no untoward adverse events observed. The results suggest that combination therapy targeting both CD19 and CD20 could be an effective approach in the clinic for the treatment of B cell lymphomas.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C186.
Abstract C21: Evaluation of an 8-plex cytokine immunoassay on the AVANTRA®, a novel automated biomarker workstation
Authors: Maimonis Peter 
Published: June 2012

Abstract
Decision Biomarkers, Inc. (DBI) has developed a novel, automated, immunoassay platform to simultaneously evaluate multiple biomarkers in a single reaction vessel. Biomarker panels are used for many research and clinical indications. We have generated data from a multiplex assay measuring a panel of 8 cytokines: IL-1β, IL-2, IL-5, IL-6, Il-8, IL-10, IL-12, and TNFα. The heart of the system is a microarray of the biomarker specific monoclonal antibodies spotted onto a glass slide which has been coated with an ultra-thin, two-dimensional film of nitrocellulose. This microarray is assembled in a plastic biochip (MAX BIOCHIP™): a self-contained device with a series of micro fluidic channels, valves, sensors, and reagent chambers containing dried, biotinylated detection antibodies, streptavidin-conjugated Cy3, and wash buffer. The biochip is processed in the AVANTRA® biomarker workstation following injection of 200 µl of sample (100 µl of plasma diluted in 100 µl of PBS). The instrument initiates, in sequence, the flow of sample, detection antibody, label, and buffer, across the array. The end result is in a fluorescent labeled complex that is then imaged by an internal CCD camera. Spot intensity is correlated with analyte concentration by instrument software. “Master” curves” electronically stored in the instrument are used to calculate analyte concentrations through a 2-point calibration scheme. Each cytokine assay has a 4-log dynamic range and individual analytical sensitivities ranging between 2.7 and 20 pg/ml. Both the intra- and interassay precision results yielded mean CV's consistently &lt;10%
Reproducibility was demonstrated by overlapping curves following replicate standard evaluation. The unique surface chemistry, marker specific reagents, and enhanced biochip micro fluidics and instrument automation, all combine to produce a robust and easy to use multiplex immunoassay. Currently in development are a 10-plex angiogenesis panel and a 12-plex cardiac panel that will soon be available. Additionally, assays for numerous other biomarker panels are slated for future development. Custom assay development is also available.
In summary, DBI has developed a sensitive and precise, multiplex immunoassay for 8 cytokines that can run on its new, automated platform, the AVANTRA® biomarker workstation.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C20.
Abstract C207: Checkpoint abrogation and potentiation of cytotoxic chemotherapeutics with a novel checkpoint kinase 1 inhibitor
Authors: Massey Andrew J. , Stokes Stephen , Browne Helen , Borgognoni Jennifer , Foloppe Nicolas , Scrace Simon , Fiumana Andrea , Bedford Simon , Webb Paul , Ray Stuart , Jordan Allan , Drysdale Martin J. , Wood Mike 
Published: June 2012

Abstract
Conventional chemotherapeutic agents such as gemcitabine, cisplatin or irinotecan induce DNA damage and activate cell cycle checkpoints. P53 defective tumors lack a functional G1 checkpoint and rely heavily on the S and G2 checkpoints, and the effector kinase Chk1, for protection against this DNA damage. Inhibiting Chk1 potentiates the anti-tumor effects of these cytotoxic chemotherapeutic agents. Targeting Chk1 is a potential therapeutic opportunity for potentiating the anti-tumor efficacy of DNA damaging cytotoxic chemotherapeutic drugs without increasing their toxicity to normal cells. Elaboration of a designed kinase directed fragment core utilizing X-ray structure guided design lead to the identification of a potent pyridone series of Chk1 inhibitors. Further profiling identified V158411 as the lead candidate. X-ray crystallography identified V158411 as being bound to the ATPase site in the kinase domain of Chk1. V158411 potently inhibited Chk1 and Chk2 with IC50s of 4.4 and 4.5nM respectively. The addition of V158411 to gemcitabine or camptothecin treated cells abrogated the cell cycle checkpoints induced by these agents resulting in the expected modulation of cell cycle proteins and increased apoptosis. V158411 potentiated the cytotoxicity of gemcitabine, cisplatin, SN38 and camptothecin in a variety of p53 deficient but not proficient human tumor cell lines in vitro. V158411 could be formulated in a simple aqueous form suitable for i.v. dosing. In nude mice, V158411 was well tolerated as a single agent (MTD &gt;100mg/kg) and in combination with irinotecan. Intravenous administration to rats and mice resulted in low plasma clearances (20mL/min/kg) and long half-lives (2.9–3.7h). In tumor bearing animals, V158411 was detected at high concentrations in the tumor (tumor:plasma AUC ratio of 4.7) with a long tumor elimination half life of 22 hours. No pharmacologically relevant in vivo drug-drug interaction with irinotecan was identified. V158411 potentiated the anti-tumor activity of gemcitabine and irinotecan in a variety of human tumor xenograft models without additional systemic toxicity. These results demonstrate the potential of combining V158411 with standard of care chemotherapeutic agents to potentiate the therapeutic efficacy of these agents without increasing their toxicity to normal cells. Based on this data, the clinical development of V158411 is currently being actively pursued.
Citation Information: Mol Cancer Ther 2009;8(12 Suppl):C207.