bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5679: Automatic synthesis of efficient transcription inhibitors as anti-cancer agents designing sequence-specific DNA-binding molecules.
Authors: Nagase Hiroki , Koshikawa Nobuko , Watanabe Takayoshi 
Published: August 2014

Abstract
PURPOSE: Regulation of endogenous gene expression using synthetic chemicals, so called “Transcription therapy” is a unique and promising approach. Although we have reported in vitro and in vivo anti-tumor, anti-metastatic and various anti-disease efficacies of hairpin Pyrrole-Imidazole polyamide (hPIP),
PIP still needs to improve on selectivity of target DNA sequences and sensitivity to bind specific sequences. Hence, we synthesized PIPs targeting transcription-factor-binding-sites with several different structures and tested their efficacy, selectivity and sensitivity in vitro as well as in vivo model.
METHODS: We have developed an automatic system for
PIP synthesis by using peptide synthesizer of PSSM8 (Shimazu) and synthesized hPIPs or cyclic PIPs targeting either the MMP9 AP1 site or NFkB site to evaluate sensitivity and specificity. Each PIP, which has a different beta-alanine position, was tested for target DNA binding by surface plasmon resonance (SPR) using
BiaCore system. We also test cell invasion and migration in vitro and metastatic capability in an in vivo xenograft model.
RESULTS: Each hPIPs targeting AP1 or NFkB site showed significant reduction of MMP9 expression in MDA-MB-231 cells and HeLa cells and metastatic capacity in vivo. hPIP with crossed beta-alanine position showed the almost same dissociation constant as those with parallel beta-alanine position. Cyclic PIPs also showed similar dissociation constant as hPIPs and showed better sequence specificity with reasonable efficacy at least in vitro analysis.
CONCLUSION: The result suggested that various
PIPs with different target sites or architectures give distinct efficacy, selectivity and sensitivity in at least in vitro cancer cell culture system. Depending on pharmacokinetic properties of PIP the most suitable PIP can be selected for the future trial as seen in combinatorial chemistry technique.
Citation Format: Hiroki Nagase, Nobuko Koshikawa, Takayoshi Watanabe. Automatic synthesis of efficient transcription inhibitors as anti-cancer agents designing sequence-specific DNA-binding molecules. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5679. doi:10.1158/1538-7445.AM2013-5679
Abstract 5683: Blockade of glioma proliferation via allosteric inhibition of JAK2.
Authors: He Kunyan , Qi Qi , Chan Chi-Bun , Xiao Ge , Liu Xia , Tucker-Burden Carol , Wang Liya , Mao Hui , Lu Xiang , McDonald Frank E. , Luo Hongbo , Fan Qi-wen , Weiss William A. , Sun Shi-yong , Brat Daniel J. , Ye Keqiang 
Published: August 2014

Abstract
Gliomas are the most common primary brain tumors. Glioblastoma multiforme (GBM), WHO grade IV, is the most frequent and aggressive form, with a median survival of approximately 12 months following aggressive treatment with surgical resection, radiation and chemotherapy. Epidermal growth factor receptor (EGFR) is frequently amplified and mutated in human cancers including GBM. While the rationale for treating GBM patients with EGR inhibitors is strong, the efficacy of anti-EGFR small molecular inhibitors or monoclonal antibodies in treating gliomas with PTEN mutation or deletion has been modest. Here we show a novel approach to inhibiting EGFR that involves a small molecule (G5-7) that selectively blocks JAK2 phosphorylation of EGFR, inhibits cell proliferation and reduces VEGF secretion, and leads to glioma shrinkage via apoptosis and anti-angiogenesis. G5-7 inhibited EGFR and STAT3 tyrosine phosphorylation by JAK2 through binding to its N-terminal FERM domain. Interestingly, G5-7 selectively inhibited the proliferation of U87MG/EGFRvIII cells and human GBM explant neurosphere cultures but only weakly affected U87MG/PTEN cells or normal neural progenitor cultures. Moreover, oral administration of this compound markedly diminishes the brain tumor volumes in both subcutaneous and intracranial models, supporting that this compound itself and/or the metabolites might be able to penetrate the brain-blood barrier and exert the anti-proliferative actions through blocking the oncogenic pathways in vivo. Chronic treatment of mice with 50 mg/kg G5-7 exhibits no obvious toxicity in the animals. Our finding demonstrates that G5-7 is a novel allosteric JAK2 inhibitor with potent anti-tumor activity for GBM. Our report not onlyidentify a novel drug for treating the GBM, but also provide insight into the molecular mechanisms of how these small molecules impinge on the EGFR/PTEN signaling. Optimization of these novel compounds will lead to new treatments for the patients with these devastating tumors.
[K.H. and Q.Q. contributed equally to this work.]
Citation Format: Kunyan He, Qi Qi, Chi-Bun Chan, Ge Xiao, Xia Liu, Carol Tucker-Burden, Liya Wang, Hui Mao, Xiang Lu, Frank E. McDonald, Hongbo Luo, Qi-wen Fan, William A. Weiss, Shi-yong Sun, Daniel J. Brat, Keqiang Ye. Blockade of glioma proliferation via allosteric inhibition of JAK2. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5683. doi:10.1158/1538-7445.AM2013-5683
Abstract 5684: Target tubulin polymerization with novel reverse ABI analogs interacting with the colchicine binding site.
Authors: Xiao Min , Ahn Sunjoo , Chen Jianjun , Wang Jin , Miller Duane D. , Dalton James T. , Li Wei 
Published: August 2014

Abstract
Cancer remains one of the leading causes of mortality worldwide. While current therapies are effective in treating early stage cancers, their efficacy against advanced cancers, especially multidrug-resistant cancers, is limited. Our lab recently reported a set of 2-aryl-4-benzoyl-imidazoles (ABI) derivatives with potent anti-proliferative activity against melanoma and prostate cancer cell lines and xenografts. Further optimization has led to the synthesis of a new series of 4-aryl-2-benzoyl-imidazoles with the aryl group and the benzoyl group reversed as compared to the previous ABI structure. The reverse ABI (RABI) compounds were evaluated for biological activity against 8 cancer cell lines including some which were multidrug-resistant. The in vitro results showed that several RABI compounds had excellent antiproliferative properties with IC50 values in the low namomolar range, comparable to existing tubulin-targeting agents such as paclitaxel, colchicine, or vinblastine, but with the ability to effectively circumvent Pgp-mediated drug resistance. The IC50 of the most active compound 12a was 9 nM. The mechanism of action (MOA) of the RABI compounds was investigated using cell cycle analysis, tubulin polymerization assays and molecular docking studies. These studies showed that their antitumor activity was achieved through inhibition of tubulin polymerization at the colchicine binding site. Our results suggest that the novel RABI compounds may be developed as a promising antitumor agent for advanced cancers.
Citation Format: Min Xiao, Sunjoo Ahn, Jianjun Chen, Jin Wang, Duane D. Miller, James T. Dalton, Wei Li. Target tubulin polymerization with novel reverse ABI analogs interacting with the colchicine binding site. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5684. doi:10.1158/1538-7445.AM2013-5684
Abstract 57: Validation of Next Generation Sequencing technologies in comparison to current diagnostic gold standards for BRAF, EGFR and KRAS mutational analysis.
Authors: McCourt Clare , Maxwell Perry , Catherwood Mark , Salto-Tellez Manuel 
Published: August 2014

Abstract
BACKGROUND: Next Generation Sequencing (NGS) has the potential of becoming an important tool in clinical diagnosis and therapeutic decision-making in oncology owing to its enhanced sensitivity in DNA mutation detection, fast-turnaround of samples in comparison to current gold standard methods and the potential to sequence a large number of cancer-driving genes at the one time. We aim to test the diagnostic accuracy of current NGS technology in the analysis of mutations that represent current standard-of-care, and its reliability to generate concomitant information on other key genes in human oncogenesis.
METHODS: Thirteen FFPE clinical samples (8 lung adenocarcinomas, 3 colon carcinomas and 2 malignant melanomas) already genotyped for EGFR, KRAS and BRAF mutations by current standard-of-care methods (Sanger Sequencing and q-PCR), were analyzed for detection of mutations in the same three genes using two NGS platforms and an additional 43 genes with one of these platforms. The results were analyzed using closed platform-specific proprietary bioinformatics software as well as open third party applications (including, among others, the ‘Open’ CLC Genomics Workbench).
RESULTS: Our results indicate that the existing format of the NGS technology performed well in detecting the clinically relevant mutations stated above, with apparent improved sensitivity, but may not be reliable for a broader unsupervised analysis of the wider genome in its current design. They also illustrate the relevance of different bioinformatics curation approaches, allowing different degrees of stringency, for the avoidance of potential false positives in the broader choice of genes selected, when compared with the expected mutation frequencies from sources such as the COSMIC database, for the tumor types analyzed. The systems are also able to detect other mutations outside the expected ones representing the current main "standard-of-care," the significance of which are uncertain at this point.
CONCLUSIONS: Our study represents a diagnostically lead validation of the major strengths and weaknesses of this technology in its current state of development before consideration for diagnostic use.
Citation Format: Clare McCourt, Perry Maxwell, Mark Catherwood, Manuel Salto-Tellez. Validation of Next Generation Sequencing technologies in comparison to current diagnostic gold standards for BRAF, EGFR and KRAS mutational analysis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 57. doi:10.1158/1538-7445.AM2013-57
Abstract 570: The integrated stress response kinase GCN2 regulates cell survival through p21/WAF1/CIP1 in response to nutrient deprivation.
Authors: Lehman Stacey L. , Ye Jiangbin , Koumenis Constantinos 
Published: August 2014

Abstract
The tumor microenvironment is often characterized by deficiencies in nutrients, such as oxygen, glucose, and amino acids. Activation of the nutrient-sensing kinase GCN2, which constitutes one arm of the Integrated
Stress Response (ISR), is one mechanism by which tumor cells cope with and adapt to nutrient stress. GCN2 phosphorylates the alpha subunit of the eukaryotic translation initiation factor eIF2, leading to global downregulation of translation to conserve amino acids and initiation of a transcriptional program through ATF4 to promote recovery from nutrient deprivation. Loss of GCN2 results in decreased tumor cell survival in vitro under conditions of amino acid deprivation and attenuated tumor growth in vivo in a variety of implanted animal tumor models. Our recent studies have shown that amino acid deprivation induces the cell cycle inhibitor p21 in a GCN2 dependent manner. GCN2 regulates p21 at the transcriptional and translational level. Induction of p21 mRNA occurs in both eIF2α S51A and p53 -/- cells, indicating that phosphorylation of eIF2α and expression of p53 are not required for GCN2 dependent transcriptional regulation. At the translational level, GCN2 phosphorylates eIF2α to prevent translational suppression of p21 transcript variants containing a 5’ uORF overlapping the true translation start site. Expression of p21 promotes cell survival during amino acid deprivation, suggesting a mechanism by which GCN2 regulates p21 in order to coordinate cell division with nutrient availability. In agreement with a pro-survival role, we observe p21 upregulation concomitant with ISR activation in a genetically engineered mouse of soft tissue sarcoma. These findings point to a novel mode of regulation of p21/WAF1/CIP1 by nutrient stress in the tumor microenvironment that contributes to tumor progression.
Citation Format: Stacey L. Lehman, Jiangbin Ye, Constantinos Koumenis. The integrated stress response kinase GCN2 regulates cell survival through p21/WAF1/CIP1 in response to nutrient deprivation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 570. doi:10.1158/1538-7445.AM2013-570
Abstract 573: USP7 and Daxx regulate mitosis progression and taxane sensitivity by affecting stability of Aurora A kinase.
Authors: Giovinazzi Serena , Morozov Viacheslav M. , Summers Matthew K. , Reinhold William C. , Ishov Alexander M. 
Published: August 2014

Abstract
A large number of patients are resistant to taxane-based chemotherapy. Functional mitotic checkpoints are essential for taxane sensitivity. Thus, mitotic regulators are potential markers for therapy response and could be targeted for anticancer therapy. In this study we identified a novel function of Ubiquitin Specific processing Protease-7 (USP7) that interacts and cooperates with protein Daxx in regulation of mitosis and taxane resistance. Depletion of USP7 impairs mitotic progression, stabilizes cyclin B and reduces stability of the mitotic E3 ubiquitin ligase, Checkpoint with Forkhead and RING finger (CHFR). Consequently, cells with depleted of USP7 accumulate Aurora A kinase, a CHFR substrate, thus elevating multipolar mitoses. We further show that these effects are independent from the USP7 substrate p53. Thus, USP7 and Daxx are necessary to regulate proper execution of mitosis, partially via regulation of CHFR and Aurora A kinase stability.
Results from colony formation assay, in silico analysis across the NCI60 platform and in breast cancer patients suggest that USP7 levels inversely correlate with response to taxanes, pointing at the USP7 protein as a potential predictive factor for taxane response in cancer patients. In addition, we demonstrated that inhibition of Aurora A attenuates USP7-mediated taxane resistance, suggesting that combinatorial drug regimens of Taxol and Aurora A inhibitors may improve the outcome of chemotherapy response in cancer patients resistant to taxane treatment. Finally our study offers novel insights on USP7 inhibition as cancer therapy.
Citation Format: Serena Giovinazzi, Viacheslav M. Morozov, Matthew K. Summers, William C. Reinhold, Alexander M. Ishov. USP7 and Daxx regulate mitosis progression and taxane sensitivity by affecting stability of Aurora A kinase. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 573. doi:10.1158/1538-7445.AM2013-573
Abstract 574: Investigating the role of CDC25B in inhibition of cellular proliferation.
Authors: Lee Caleb C. , Manfredi James 
Published: August 2014

Abstract
CDC25B is a dual specificity phosphatase that activates cyclin-dependent kinases to trigger entry into mitosis. Overexpression of CDC25B has been associated with breast and head and neck cancers as well as higher histological tumor grade in non-small cell lung cancers. In a transgenic mouse model, overexpression of Cdc25B in the mammary glands lead to accelerated mammary epithelial proliferation resulting in hyperplasia while in vitro, co-transfection of CDC25B with Ras allows for transformation of MEF's. However, in vitro experiments using ectopic expression of CDC25B in transformed cell lines have shown an anti-proliferative effect. This may be related to inappropriate cyclin-dependent kinase activation and premature mitotic entry, leading to both p53-dependent and independent checkpoints. In addition, S phase cells overexpressing CDC25B have higher rates of apoptosis due to premature entry into mitosis. CDC25B has also been shown to cause an early progression to S phase that is associated with increased amounts of DNA damage. Our goal is to expand on these results to further clarify the anti-proliferative effects of CDC25B as well as detail the mechanism by which tumor cells bypass these effects in vitro and in vivo.
Citation Format: Caleb C. Lee, James Manfredi. Investigating the role of CDC25B in inhibition of cellular proliferation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 574. doi:10.1158/1538-7445.AM2013-574
Abstract 575: Cdk1 prevents DNA rereplication in G2/M by phosphorylating and facilitating the removal of Cdc7 from chromatin at the end of S phase.
Authors: Knockleby James , Kim Byung Ju , Lee Hoyun 
Published: August 2014

Abstract
Cancer is inextricably linked with aneuploidy. Aneuploidy can arise for a number of reasons, including the deregulation of DNA and centrosome replication. Mechanisms have evolved that are designed to ensure that DNA is replicated only once per cell cycle. The initiation of DNA replication is a key target of these mechanisms, which includes regulating Cdc6, Cdt1 and Cyclin dependent kinases (Cdks). Although the regulatory mechanisms of Cdc6 and Cdt1 are known, the role of Cdk1 in particular is less clear. One potential target for Cdk1 regulation is the Dbf4-Cdc7 kinase (DDK), which acts as a licensing factor for DNA replication. DDK performs an essential and critical step in initiating DNA replication. As such, it is a target for novel cancer therapeutic drugs currently in clinical trials. Even so, the regulatory mechanisms dictating DDK function in human cells remains poorly understood.
A site-directed mutagenesis approach was undertaken to determine the in vivo function of Cdk-dependent phosphorylation sites on the primary Cdc7 sequence. Using over-expressed Cdc7 in human cells (HEK293T), the functional significance of Cdc7 phosphorylation was determined; both the lack of phosphorylation and constitutive phosphorylation of Cdc7 was analyzed. We found that Cdc7, the catalytic subunit of the DDK, is a target of Cdk1. Cdc7 is phosphorylated in G2/M phase by Cdk1. Phosphorylation of Cdc7 removes the catalytic subunit of DDK from origins of replication and chromatin with the assistance of the nuclear export factor CRM1, which relocates Cdc7 to the microtubule cytoskeleton. Finally, the Cdk1-dependent removal of Cdc7 from chromatin prevents the cell from initiating DNA replication, indicating that Cdc7 is a target of Cdk1-dependent inhibition of DNA re-replication.
We conclude that (1) Cdk1 phosphorylates Cdc7 in vivo. (2) Cdk1 dependent phosphorylation of Cdc7 regulates chromatin binding and DNA replication and (3) pCdc7 is associated with the nuclear export factor CRM1 in G2/M phase, suggesting a mechanism that couples the down-regulation of Cdc7 function at the end of S phase through nuclear localization and chromatin association. Taken together, our data provides key insight into the functional understanding of DDK.
Citation Format: James Knockleby, Byung Ju Kim, Hoyun Lee. Cdk1 prevents DNA rereplication in G2/M by phosphorylating and facilitating the removal of Cdc7 from chromatin at the end of S phase. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 575. doi:10.1158/1538-7445.AM2013-575
Abstract 576: Transforming growth factor-beta (TGF-β) inhibits growth of human myeloid leukemia cells and downregulates the expression of CDK inhibitor, p18INK4C.
Authors: Guardia Talia , Badaloo Reshma , Serrano Valentina , Toro Alejandra , Hu Xiaotang 
Published: August 2014

Abstract
The cell cycle progression controlled by cyclin-dependent kinases (CDKs) is counterbalanced by CDK inhibitors (CKIs). The CKIs have two families: CIP/KIP and INK. The CIP/KIP members are composed of p21WAF1/CIP1, p27KIP1 and p57KIP2, which bind to and inhibit both cyclin D-CDK4/6 kinases and cyclin-E/A-CDK2. The INK members consist of p15INK4B, p16INK4A, p18INK4C and p19INK4D. The members in this family only bind to and inhibit G1CDKs (CDK4 and CDK6). We have previously reported that p27KIP1 plays both a positive and a negative role in human myeloid leukemia cells. In this study, we investigated whether p18INK4C plays any role in TGF-β-mediated cell cycle control of human myeloid leukemia cells. TGF-β significantly inhibited proliferation of MV4-11 and TF-1 cells, with upregulation of p27KIP1and downregulation of multiple CDKs and cyclins. Surprisingly, TGF-β significantly inhibited the expression of p18INK4C, a traditional G1 CDK inhibitor. The inhibitory effect of TGFβ on p18INK4C was time dependent. The time course showed that early inhibition of p18INK4C occurred at 3h after initiating TGFβ treatment in the cells with a maximal inhibition of 70% being observed at 72h incubation with TGF-β. The control cells treated with the solvent for TGFβ did not show any significant changes in the levels of p18INK4C. As a sample loading control, the cells treated with or without TGF-β expressed about the same levels of actin.TGFβ-induced p18INK4C inhibition is also dose dependent with a maximal inhibition being detected by Western blot when 30ng/ml of TGFβ was added to the culture. Low concentrations (5ng/ml or less) did not markedly affect the expression of p18INK4C. The association of p18INK4C with CDK4 was also decreased in response to TGF-β treatment detected by immunoprecipitation. Our data also showed that the two cell lines tested barely expresses p15INK4B, p16INK4A, p19INK4D, and p21WAF1/CIP1 and these inhibitors do not appear to play an important role, either positively or negatively, in the cell cycle control of the leukemia cells. Our data suggest that p18INK4C may be required for cell cycle progression in human myeloid leukemia cells tested. Whether such a positive role of p18INK4C is caused by a possible gene mutation(s) and thereby contributes to leoplasia of the leukemia cells is currently under investigation (supported by NIH-NIGMS MBRS RISE: R25 GM059244, Barry University).
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Citation Format: Talia Guardia, Reshma Badaloo, Valentina Serrano, Alejandra Toro, Xiaotang Hu. Transforming growth factor-beta (TGF-β) inhibits growth of human myeloid leukemia cells and downregulates the expression of CDK inhibitor, p18INK4C. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 576. doi:10.1158/1538-7445.AM2013-576