bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 94: DrugPath: A web database for academic investigators to identify drugs that match oncology molecular targets
Authors: Shah Eric D. , Zhao Peng , Ghent Matthew V. , Arceci Robert J. , Buckley Jonathan D. , Reaman Gregory H. , Sorensen Poul H. , Triche Timothy J. , Reynolds C Patrick
Published: February 2011

High-throughput genomics can identify oncology molecular targets, but translation into novel therapeutics requires access to relevant drugs. Pharmaceutical pipeline databases, such as MedTRACK (www.medtrack.com), can identify such drugs, but these databases are geared for commercial operations and present significant cost barriers to non-profit academic research. We have developed the DrugPath database (www.drugpath.org) as a comprehensive, free-of-charge resource for academic investigators. DrugPath can identify drugs within the cancer pipeline that may act against molecular targets of interest. DrugPath searchable data include names and mechanisms of drugs in the oncology pipeline (organized by clinical trial phase and route of administration), drug sponsor, and also tested combinations with other drugs. Drugs and drug sponsors are initially identified from ClinicalTrials.gov, biotech company websites, and other online resources; specific pipeline information is extracted by DrugPath personnel according to written protocols. PERL scripts to automate a portion of the data mining are being developed, although manual verification will remain to ensure data accuracy and relevance. The website is authored in XHTML and PHP, and content is stored in a MySQL database. We compared the extent of the DrugPath database to that of MedTRACK. DrugPath covers more than 475 companies, or 50% of 938 companies with oncology therapeutics in development as identified by MedTRACK. A search of DrugPath for inhibitors of VEGF, a well-established target, yielded 58 lead drug candidates (17% of 332 drugs identified by MedTRACK) from 45 total companies (47% of 95 companies identified by MedTRACK). A search for Hsp90 inhibitors yielded 15 companies (78% of 19 in MedTRACK) and 17 lead drug candidates (30% of 56 in MedTRACK), and a search for aurora kinase inhibitors yielded 17 companies (94% of 18 in MedTRACK) and 17 lead drug candidates (70% of 24 in MedTRACK). Ongoing efforts to populate the DrugPath database are focused on identifying drug development programs for emerging drug targets rather than expanding existing coverage of drug candidates against well-known targets. To support the end-goal of DrugPath–to facilitate obtaining drugs for academic research–DrugPath contains contact information (when available) for each company to assist with the process of developing material transfer agreements (MTAs). This database should be useful to academic investigators studying particular molecular targets and also to clinical investigators seeking to develop novel clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 94.
Abstract 97: In silico computational modeling and virtual screening of novel noscapinoid library as potent microtubule interacting anticancer drugs
Authors: Naik Pradeep K. , Aneja Ritu , Vangapandu Surya , Santoshi Seneha , Joshi Harish C.
Published: February 2011

Tubulin remains a persistent and promising target for anti-cancer therapy. This is well validated by the already approved clinically used drug classes such as taxoids and vincas. We have previously discovered a new class of anti-microtubule (MT) drugs, noscapinoids. The lead compound, noscapine, is an orally available anti-cough plant alkaloid that binds tubulin and makes MTs less dynamic. Using computational algorithms such as Glide Extra Precision, Quantum Polarized Ligand Docking (QPLD), Molecular Mechanics Generalized Born Surface Area (MM-GB/SA), EmBrace and Solvation Generalized Born-Linear Interaction Energy (SGB-LIE), we have generated molecular models of noscapine-tubulin complex. Furthermore, based upon structure-function analyses of noscapine, we have identified a key residue on the isoquinoline ring of noscapine that can be modified without losing its biological activity. We report a library of novel derivatives, based upon combinatorial in silico approaches that can be chemically synthesized and tested in vitro and in vivo in animal models of human tumor xenografts in immune compromised mice. The prototype analogs synthesized so far have non-toxic to normally dividing healthy tissues but pro-apoptotic in a wide variety of cancer cells in vitro and in vivo. Some of these prototypes have shown already that these approaches can generate compounds more effective than the lead compound. Unlike currently available clinical anticancer MT-drugs, noscapine and its derivatives are orally available, and display no toxicities such as myelosuppression and immunosuppression, peripheral neuropathies, or pathology of gastrointestinal tract. Moreover, noscapine readily crosses the blood brain barrier. The in silico libraries generated here promise to be even more effective. Additionally, because the mechanisms by which different anti-MT drugs induce apoptosis can vary widely in both, the same cell lineage as well as in distinct lineages, there is a tremendous opportunity for a better therapeutic outcome by combining noscapinoids with already approved MT-drugs in a cancer type specific manner.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 97.
Abstract 976: Consumption of vitamins and calcium reduces breast cancer risk by their regulation of the DNA repair capacity
Authors: Vergne Yeidyly , Matta Jaime L. , Morales Luisa , Vargas Wanda , Alvarez-Garriga Carolina , Bayona Manuel
Published: February 2011

Background- Breast cancer (BC) is the most common cancer in women with over one million new cases diagnosed annually worldwide. The capacity of the DNA to repair itself (DRC) is a complex biological process involving over 200 proteins. This process involves at least five pathways and is critical in maintaining genomic stability. The objective of this study was to examine the association of DRC and BC risk in terms of consumption of vitamin and calcium supplements by means of a large scale case-control study.

Methods.- This was an incident-case case-control study design involving Puerto Rican women. The selection of potential predictors under study was based on previous published research in BC including information on vitamins, calcium, and variables that could provide an estimate of BC risk including age, BMI, family history, gynecological history, hormonal and environmental factors. The host reactivation assay with a luciferase reporter gene was used to measure the DRC in the lymphocytes from all participants. Cases were compared to controls, regarding diet supplement intake, DRC, and other selected covariates. The crude and multiple logistic regression adjusted Odds Ratio (OR) were used as measures of association and the 95% confidence interval of the OR was utilized to asses the precision of this estimate.

Results.- A total of 268 breast cancer cases and 457 controls were included in this analysis. Statistically significant associations were found between BC and ageing, low DRC levels, family history of BC, and no breastfeeding. Vitamins and calcium intake were found to be protective reducing 30% and 40% the odds of having BC respectively. Calcium reduced considerably its protective effect becoming negligibly and not statistically significant when DRC was included in the logistic regression model. This suggests that DRC explains this association. In contrast, vitamin's intake did not show an important change in the association with BC when adjusting for DRC. Calcium and vitamins’ intake were strongly associated with higher levels of DRC.

Discussion.- Vitamins and calcium intake are protective for breast cancer and are associated with higher DRC levels. Vitamins’ intake is an independent protective factor for BC while the protective effect of calcium may be explained by an increased DRC. DRC can be used to monitor the protective effect of calcium in terms of breast cancer risk. This study is supported by grants from the NCI Center to Reduce Health Disparities and NIH-MBRS Program grant #: S06 GM008239-23.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 976.
Abstract 99: The caBIG® life sciences distribution: Integrative tools to support translational research
Authors: Heiskanen Mervi A. , Klemm Juli , Nagarajan Rakesh , Sharma Mukesh , Talele Chandrakant , Goldstein Stephen , Piepenbring Chris , Jacob Shine , Tavela Eric , Marple John P. , Fore Ian , Andrews Timothy J. , Nguyen Ngoc , Floratos Aris , Basu Anand , Derr Leslie
Published: February 2011

The cancer Biomedical Informatics Grid® (caBIG®) is a collaborative network designed to accelerate the translation of discoveries from research to clinical care. This extensible informatics platform integrates diverse data types and supports interoperable software tools in the areas of clinical sciences, biospecimen management, imaging, and basic research. The Life Sciences Distribution (LSD) product bundle brings together a range of tools that support translational research, all connected via grid network technology. Tools included in the bundle support management and annotation of microarray data (caArray), biospecimens (caTissue Suite), imaging data (NBIA), clinical data (CTODS) and genome-wide association studies (caGWAS). LSD also includes integrative tools that allow scientist to search data across different data repositories connected to the grid, and analyze and integrate these data across different data types. geWorkbench supports integrative analysis and visualization of expression data, sequences, pathways and protein structures. cancer Bench-to-Bedside (caB2B) allows end-users to search biospecimens, images and array data across the grid and makes it possible to execute queries such as: “Are there any gene expression microarray data available from patients with Stage III lung cancer and are there corresponding in vivo images available for the affected patients?” Such a query would potentially span information federated across several instances of caArray, caTissue, and NBIA around the world. caIntegrator2 allows researchers to set up study specific custom web portals that bring together heterogeneous clinical, microarray and in vivo imaging data. This tool provides a graphical user interface to allow a study author to “point” to data of interest in systems on the grid and to then bring that data (or pointers to it, in the case of images) into the data warehouse. Once this information is in the caIntegrator2 environment, end user scientists can then run advanced queries, perform correlative outcomes analysis using Kaplan-Meier survival plots, and access analysis and visualization tools on and off the grid. All tools in the LSD are open source and community members are encouraged to participate and contribute. More information on the LSD suite of products, including installation packages, user and installation guides, and links to exemplar installations can be found at https://cabig.nci.nih.gov/adopt/LSD/

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 99.
Abstract 992: PR55alpha, a regulatory subunit of PP2A, specifically regulates PP2A-mediated beta-catenin dephosphorylation
Authors: Zhang Wen , Chen Xi , Yu Tianxin , jia jianhang , Liu Chunming
Published: February 2011

BACKGROUND: Wnt/beta-catenin signaling plays essential roles in development and tumorigenesis. A central question in Wnt signaling is the regulation of beta-catenin phosphorylation and degradation. Our previous work found that beta-catenin is sequentially phosphorylated by CKIalpha and GSK3, which creates binding site for beta-Trcp, leading to degradation via the ubiquitination/proteasome machinery. Mutations in beta-catenin or APC genes that prevent beta-catenin phosphorylation or ubiquitination/degradation lead ultimately to cancer. In addition to the involvement of kinases, protein phosphatases, such as PP1, PP2A and PP2C, are also implicated in Wnt/beta-catenin regulation. However, which phosphatase dephosphorylates beta-catenin in vivo and how the specificity of beta-catenin dephosphorylation is regulated are not clear. In this study, we show that PP2A regulates beta-catenin phosphorylation and degradation in vivo. PP2A consists of a catalytic subunit (PP2Ac), a structure subunit (PR65/A) and variable regulatory B subunit (PR/B, PR/B’, PR/B”, or PR/B’’’). The substrate specificity of PP2A is thought to be determined by its B subunit. We further identified that PR55alpha is the regulatory B subunit that controls PP2A-mediated beta-catenin dephosphorylation.

RESULTS: Toward the goal of understanding the mechanism of beta-catenin phosphorylation, we carried out siRNA screening targeting several major phosphatases, in which we found that PP2A dephosphorylates beta-catenin. We demonstrate that PP2A is required for Wnt/beta-catenin signaling in Drosophila. Moreover, by siRNA screening, we have identified PR55alpha as the regulatory subunit of PP2A that controls beta-catenin phosphorylation and degradation. Mechanistically, we found that PR55alpha directly interacts with beta-catenin and regulates PP2A-mediated beta-catenin dephosphorylation in Wnt signaling. PR55alpha but not the catalytic subunit, PP2Ac, directly interacts with beta-catenin. RNAi knockdown of PR55alpha elevates beta-catenin phosphorylation and decreases Wnt signaling, whereas overexpressing PR55alpha enhances Wnt signaling. Taken together, our results suggest that PR55alpha specifically regulates PP2A-mediated beta-catenin dephosphorylation and plays an essential role in Wnt signaling.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 992.
Abstract LB-10: Defective nucleolar localization and dominant interfering properties of a parafibromin L95P missense mutant causing the hyperparathyroidism-jaw tumor syndrome
Authors: Jacob Leelamma P. , Zhang Jian Hua , Dagur Pradeep K. , Simonds William F.
Published: February 2011

The hyperparathyroidism-jaw tumor syndrome (HPT-JT) is a familial cancer syndrome that results from germline loss-of-function mutation of HRPT2/CDC73, a putative tumor suppressor gene that encodes parafibromin, a component of the transcriptional regulatory PAF1 complex with homology to the yeast protein Cdc73p. The vast majority of HRPT2/CDC73 germline mutations identified have been truncation or frameshift mutations, and loss-of-function due to missense mutation is rare. We report here a kindred with HPT-JT due to a germline L95P missense mutation in parafibromin. The mutant parafibromin was studied in vitro to understand the basis of its presumed loss-of-function. When transfected in cultured cells the L95P mutant was expressed to a lower level than wild-type parafibromin, a difference that was not overcome by inhibition of the proteasome degradation pathway. The L95P mutant parafibromin retained the ability to assemble with endogenous PAF1 complex components as evidenced by co-immunoprecipitation. Analysis of subcellular localization showed that the L95P mutant was markedly deficient in nucleolar localization compared to the wild-type, an impairment likely resulting from disruption of a nucleolar localization signal immediately upstream of the L95P mutation. Transfection of the L95P parafibromin mutant, but not the wild type, increased HEK 293 cell survival and stimulated cell-cycle progression in NIH-3T3 cells, properties that appear to result from dominant interference with endogenous parafibromin signaling pathways. The simultaneous loss of nucleolar localization and acquisition of a growth stimulatory phenotype with the L95P mutation raise the possibility that parafibromin must interact with targets in the nucleolus to fully execute its tumor suppressor functions.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-10.
Abstract LB-102: The p21Cip1/Waf1 cyclin-dependent kinase inhibitor is required for the activation of the FA-BRCA pathway
Authors: Rego Meghan A. , Mauro Maurizio , Harney Julie A. , Shen Mae , Kolling Frederick W. , Howlett Niall G.
Published: February 2011

Fanconi anemia (FA) is a rare recessive disorder characterized in part by pronounced cancer susceptibility. The FA proteins, and the protein products of the major breast cancer susceptibility genes BRCA1 and BRCA2, function cooperatively in the FA-BRCA pathway, to repair damaged DNA and to prevent cellular transformation. A major step in the activation of the FA-BRCA pathway is the mono-ubiquitination of the FANCD2 and FANCI proteins, targeting these proteins to discrete chromatin-associated nuclear foci. Importantly, the regulation of FANCD2 (and FANCI) mono-ubiquitination remains poorly understood. To gain further insight into this important post-translational modification step, here we have examined the roles of p53 tumor suppressor as well its downstream target p21Cip1/Waf1 in the regulation of the mono-ubiquitination of FANCD2.

In this study we have used the isogenic HCT116 p53+/+, p53−/−, p21+/+, and p21−/− cell lines, to determine that p21, and not p53, plays a key role in the regulation of the mono-ubiquitination of FANCD2 following exposure to DNA damaging agents. For example, robust FANCD2 mono-ubiquitination is observed in HCT116 p21+/+ cells 2 h following exposure to 20 J/m2 UV irradiation, while this effect is markedly attenuated in p21−/− cells. Using immunofluorescence microscopy we also demonstrate that p21 is necessary for the efficient recruitment of FANCD2 to nuclear foci following exposure to UV irradiation. Furthermore, preliminary protein-protein interaction analyses in COS-7 cells demonstrate that FLAG-p21 and 6xHis/V5-FANCD2 physically interact.

Interestingly, despite a failure to efficiently activate the mono-ubiquitination of FANCD2 following exposure to DNA damaging agents, p21−/− cells are not hypersensitive to the cytotoxic effects of the DNA crosslinking agent mitomycin C (MMC). However, elevated levels of distinct chromosome aberrations, including telomere associations and dicentric chromosomes, are observed in p21−/− cells compared with p21+/+ cells following exposure to MMC. Taken together, our results demonstrate an important upstream role for the p21 cyclin-dependent-kinase inhibitor in the activation of the FA-BRCA pathway. A greater understanding of the regulation of this important tumor suppressor pathway will lead to improved diagnostic and therapeutic approaches to FA and enhance our understanding of the elevated cancer susceptibility of FA patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-102.
Abstract LB-113: PKC-iota induces uncontrolled glioblastoma cell survival by modulating proapoptotic function of Bad
Authors: Desai Shraddha , Acevedo-Duncan Mildred
Published: February 2011

Brain tumors are a major challenge since they destroy the most important part of the human body. The aggressive and infiltrative behavior is a hallmark of glioblastoma multiforme which makes it highly incurable. PKC-iota is reported to be highly expressed in transformed phenotype of human glioma, benign and malignant meningiomas (Cell Prolif. 41:122-35, 2008) but the signaling mechanism by which PKC-iota induces glioma cell survival remains elusive. Hence, in the current study, we investigated whether PKC-iota phosphorylates and de-activates Bad, a pro-apoptotic BH3-only molecule of the Bcl-2 family. Our results demonstrated that PKC-iota co-localizes as well as directly associates and phosphorylates Bad at Ser-112, Ser-136 and Ser-155. In addition, recombinant PKC-iota induced direct phosphorylation of exogenous as well as endogenous Bad at all three serine residues as shown by in-vitro kinase assay. Recombinant PKC-zeta (is 70% homologous to PKC-iota) and PKC-delta (known to regulate the activity of Bad in rat cardiac myocytes; Mol Cells. 24:224-231, 2007) in contrast phosphorylated Bad only at Ser-112 residue in in-vitro kinase assays. Reduction of PKC-iota upon RNA interference exhibited a corresponding reduction in Bad phosphorylation at these three sites. Conversely, inhibition of PKC-zeta and delta by their corresponding siRNA did not lead to reduction of Bad phosphorylation. Additionally, disruption of Bad/Bcl-XL heterodimerization was not observed in either of the RNA silenced samples. Thus, our data strongly supports the hypothesis that PKC-iota may function as a Bad kinase directly inhibiting its pro-apoptotic function and impeding the normal homeostasis of glioma cells thereby enhancing prolonged survival of glioma cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-113.
Abstract LB-117: A novel tween-80 free, water soluble formulation of docetaxel for cancer treatment
Authors: Tan Lily , Qiao Hongqun
Published: February 2011

Docetaxel is a semi-synthetic anticancer drug in the taxane family. It is a potent treatment for a wide range of tumors. The approved indications include breast cancer, NSCL cancer, ovarian cancer, and prostate cancer. The current docetaxel application, Taxotere, uses tween-80 and ethanol as solvents and has shown undesirable adverse effects, including hypersensitivity from the active as well as inactive ingredients.

Captisol (sulfobutylether cyclodextrin) is a modified cyclodextrin that has been used to improve drugs’ water solubility for parental use, such as in Pfizer's Vfend IV and Geodone IM.

In this paper, we introduce ML-061, an improved water soluble formulation of lyophilized docetaxel with captisol using patented technology. While the efficacy of ML-061 is similar to taxotere at the equivalent dose, a vastly better safety profile has been shown for the new formulation. ML-061 showed no detectable hemolytic activities in a study in Beagle dogs compared to the placebo group, no hypersensitivity effect on blood pressure and serum complement activation while taxotere did.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-117.
Abstract LB-121: Serrated colorectal adenocarcinoma: Specific copy number alterations
Authors: Tuppurainen Karoliina , Li Yilong , Kaasinen Eevi , Tuomisto Anne , Aaltonen Lauri A. , Karhu Auli , Makinen Markus J.
Published: February 2011

The serrated pathway in the colorectal cancer (CRC) development shows characteristic morphological and genetic differences as compared with the traditional Vogelstein's adenoma-carcinoma sequence. According to current estimations, about 15-20% of colorectal cancers originate from serrated polyps. Activating mutations of BRAF and KRAS genes and microsatellite instability are common features in serrated polyps and cancers. To identify the driver genes behind the serrated colorectal pathway we analyzed gene copy number variations (CNVs) in 19 serrated colorectal carcinomas.

DNA was extracted from fresh-frozen tissues according to standard procedures. By using the Illumina HumanCNV-370 platform we analyzed the samples for somatic copy number alterations. Raw-data were normalized using the built-in Illumina BeadStudio normalization method. To segment the LRRs (tumor vs. normal logarithmic intensity ratios) we used CBS (circular binary segmentation) and BAFsegmentation (B allele frequency) and the driver genomic alterations were searched using GISTIC algorithm. To determine genetic changes specific for serrated adenocarcinomas, the recurrent genomic variations detected were compared with those previously reported in colorectal carcinomas.

The recurrent genomic alterations in serrated carcinomas overlap substantially with the previously reported alterations seen in colorectal carcinomas; losses in 1p, 4p, 4q, 5q, 8p, 17p, 18p and 18q, and the gains in 7p, 7q, 8q, 13q and 20q. However, regions that are frequently altered specifically in serrated carcinomas were also found. For example, the gains of 1q, 5p, 6p and 6q are not commonly found in primary tumors in colorectal cancer, but only in liver metastases. The frequent losses specific for serrated carcinoma occur in 3p, 6q, 9p, 11q, 16p, 22q, Xp and Xq. The highest deletion peaks observed in the current sample set are those located at 9p21.3.

In summary, our whole-genome screening of serrated colorectal adenocarcinomas with SNP arrays revealed several copy number alterations previously reported in colorectal cancers. Moreover, we detected several novel copy number alterations, which were specific for the serrated adenocarcinoma. The upcoming sequence analysis of genes within regions of copy number alterations may allow detection of mutations of genes linked to serrated adenocarcinoma pathogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-121.
Abstract LB-141: Multiple protein secretion profiling analysis of cancer cells at single cell and small cell colony level with integrated barcode chip
Authors: Shin Young Shik , Hwang Kiwook , Fan Rong , Heath James R.
Published: February 2011

Pathophysiology of a complex disease such as cancer requires capability to analyze highly heterogeneous cellular events at single cell level, which can measure large numbers of molecular events from individual cells to be analyzed in statistically meaningful manner. Here we report on an approach that integrates microfluidic cell handling and in situ protein secretion profiling to assess the functional heterogeneity of single cells, with extensions to small cell colonies. We measured a dozen proteins secreted from cells for the most aggressive type of primary brain tumor, glioblastoma multiforme (GBM). We observed functional phenotypes in terms of secreted proteins with profound cellular heterogeneity but still in a statistically meaningful manner. The unique features that we confirmed from single cell analysis can present additional useful information to the conventional bulk analysis. Combining physical status of the system such as cell-cell distance and the protein secretion profiles enables to study tumor microenvironment. We further demonstrate the potential application of this platform to the clinic by analyzing solid tumor cells derived from a GBM patient. This platform is inexpensive, requires minute amounts of cells and yields large volume of molecular information, showing great potential for clinical assessment of cellular characteristics in human disease lesions.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-141.
Abstract LB-148: Depletion of myeloid derived suppressor cells in pancreatric cancer results in decreased tumor growth
Authors: Mitchem Jonathan , Porembka Matthew , Hawkins William G. , Goedegebuure Simon Peter , Linehan David C.
Published: February 2011

Introduction: Pancreatic adenocarcinoma (PA) is a highly aggressive and chemo-resistant malignancy with a median survival of only 6 months. Myeloid derived suppressor cells (MDSC) have been implicated in the immune suppression and growth of several malignancies. MDSC are highly prevalent in the tumor microenvironment, and represent a potential therapeutic target. Here, we assess the tumor-promoting ability of MDSC in a model of PA and explore MDSC targeting strategies using ZA, a drug known to inhibit MDSC.

Methods: C57BL/6 mice were inoculated in the right flank with 100,000 or 50,000 of the syngeneic PA line, Pan02. Tumor volume was determined by standard calculation using caliper measurements. Mice were treated with Gemcitabine 20mg/kg Qwk IP and/or ZA 1ug QOD SQ once tumors were palpable. Analysis for MDSC accumulation was determined in the tumor, spleen and bone marrow of mice at sacrifice using flow cytometry for standard markers of MDSC (CD45, CD11b, Gr1, Ly6G, and Ly6C). MDSC prevalence is expressed as a percentage of total leukocytes (CD45+). Statistical analysis was performed using non-parametric student's t-test.

Results: MDSC are up regulated in the spleen and bone marrow of mice with flank injected Pan02 compared to non-tumor bearing mice. MDSC accumulate over time and increase in number until 4 weeks post-inoculation (CD45 + 11b+Gr1+, Bone Marrow: 26.06% 42.04%, spleen 1.07% 7.18%, p=0.0021). Treatment of tumor bearing mice with ZA decreases tumor associated MDSC (CD45+11b+Gr1+: 50.36 29.38; CD45+11b+Ly6G+: 49.45 30.47) and tumor volume (2.45 1.71 p=0.034). ZA also works in concert with Gemcitabine to decrease MDSC (Gr1 +: 50.36 39.38; Ly6G+: 49.45 37.28;) and tumor volume (2.45 1.02, p=0.0011).

Conclusion: MDSC play an integral role in the development of PA. Depletion of MDSC by ZA results in significantly smaller tumors and can be added to the standard chemotherapeutic regimen (Gemcitabine) successfully. The addition of ZA to Gemcitabine represents an attractive and innovative regimen to improve the treatment of this deadly malignancy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-148.
Abstract LB-15: Oncogenic BRAF mutation, IGFBP7 silencing and evasion of senescence in colorectal tumorigenesis
Authors: Spring Kevin J. , Miranda Mariska , Fernando Winnie , Togashi Kazutomo , Young Graeme P. , Leggett Barbara A.
Published: February 2011

Serrated polyps are now recognized as the precursor lesion from which sporadic BRAF (V600E) mutant/CIMP-H colorectal cancers (CRC) develop, but it appears likely BRAF mutation alone is not enough to drive progression to carcinoma. CIMP is generally localized to the proximal colon and may cause the epigenetic silencing of key genes such as IGFBP7 that may act in concert with BRAF mutation to promote neoplasia. We are conducting a detailed molecular analysis of the mechanism of serrated neoplasia involving IGFBP7 and evasion of oncogene-induced senescence (OIS). Normally senescence functions as a defense mechanism against tumorigenesis in precursor lesions.

We have analysed 44 CRCs and 217 colorectal polyps including 45 advanced serrated polyps (sessile serrated adenomas, serrated adenomas and mixed polyps) and 109 simple serrated polyps (goblet cell rich hyperplastic polyps and microvesicular hyperplastic polyps) for BRAF, K-ras and CIMP relative to clinicopathologic variables. In addition a MethyLight assay was developed to determine IGFBP7 methylation patterns in serrated polyps and cancers. For serrated polyps, both BRAF mutation (75.5%) and CIMP (46.6%) was most frequent in advanced serrated polyps (ASPs). Overall the frequency of CIMP was significantly associated with proximal location (P<0.0001) and larger size (P<0.005). When serrated polyps were analyzed separately, histological type and location showed strong associations, with CIMP frequency differing significantly in proximal vs distal BRAF mutant ASPs and GC/MV-HPs. We find that BRAF mutation is highly associated with both SSAs and MV-HPs throughout the colorectum, but only in proximal SSAs is CIMP highly associated with BRAF mutation (70.8%). For CRCs we found IGFBP7 methylation frequently associated with CIMP and BRAF mutation (87%). For serrated polyps, IGFBP7 methylation was significantly associated with ASPs (P<0.0002), larger size (P<0.0002) and proximal location (P<0.0004). In advanced serrated polyps, which harbour both BRAF mutation and exhibit CIMP, methylation of IGFBP7 was observed 65% of cases. We are currently determining p53 expression levels, Ki67 and the senescence markers p19ARF and Dec1 in different serrated polyp types. This investigation will provide evidence for the involvement of IGFBP7 loss, by DNA hypermethylation, in the evasion of senescence and resulting in the progression of BRAF mutant/CIMP-H carcinoma of the colon.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-15.
Abstract LB-153: PDE4D as a new prostate cancer drug target
Authors: Hammer Kimberly DP , Marker Paul C.
Published: February 2011

We have been conducting a two-species screen for prostate cancer oncogenes that are suitable for use as new prostate cancer drug targets. The initial screen utilized transposon-based somatic mutagenesis in mice to induce prostate tumors and molecularly tag candidate oncogenes. This approach identified several candidate prostate cancer genes including known oncogenes such as Nras and Braf as well as new candidate oncogenes including Pde4d. A secondary screen used human prostate tissue microarrays (TMAs) to validate the relevance of new candidate oncogenes for understanding human prostate cancer. Automated quantitative analysis of PDE4D protein expression on the human TMAs confirmed its over-expression in human prostate cancer patients. The Pde4d mRNA is alternatively spliced to encode a family of related PDE4D enzymes. These enzymes share a common catalytic domain that gives all of the enzymes cAMP-specific 3′,5′-cyclic phosphodiesterase activity. Protein domains outside the catalytic domain allow for differential protein regulation and sub-cellular localization of different PDE4D isoforms. Several small molecules have been identified that are highly selective inhibitors of PDE4D activity including cilomilast and NVP-ABE171. In vitro testing of these small molecules has shown that they inhibit the growth of some but not all commonly used prostate cancer cell lines. Growth-inhibition was associated with down-regulation of the MEK-ERK and Shh signaling pathways. Current studies are investigating the molecular mechanisms associated with differential sensitivity/resistance to PDE4D inhibition as well as the potential use of PDE4D inhibitors as anti-prostate cancer drugs in vivo.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-153.
Abstract LB-154: Loss of PTEN cooperates with activated RAS in melanoma genesis and progression
Authors: Nogueira Cristina , Kim Kwan Hyun , Paraiso Kim T. , Dannenberg Jan-Hermen , Bosenberg Marcus , Chin Lynda , Kim Minjung
Published: February 2011

MAPK and AKT pathways are frequently co-activated in melanoma through overexpression of receptor tyrosine kinases, mutations in their signaling surrogates, such as RAS and BRAF, or loss of negative regulators such as PTEN. Since RAS is a known positive upstream regulator of PI3-K, it has been suggested that the loss of PTEN and the activation of RAS are redundant events in melanoma pathogenesis (Tsao et al., 2000). Here, in genetically engineered mouse models of cutaneous melanomas, we addressed the genetic interactions of activated HRAS and PTEN inactivation in melanoma genesis and progression in vivo. We showed that loss of Pten cooperates with HRAS activation in Ink4a/Arf−/− background and increased penetrance of melanoma development and emergence of metastasis. Correspondingly, Pten loss increases invasion and migration of melanoma cells and non-transformed melanocytes, which correlates with E-cadherin down-regulation. Together, this study demonstrates a synergistic interaction between Pten inactivation and RAS activation in melanoma development and progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-154.
Abstract LB-160: ITRI-260, a promising candidate for treatment of acute myeloid leukemia
Authors: Liu Chih-Peng , Liu Hsingjin Eugene , Lee On , Chen Chih-Hung , Huang Jui-Wen , Chen Ting-Shou , Ko Ching-Huai , Wang Ling-Mei , Chen Yen-Chun , Tseng Hsiang-Wen , Shih Ying-Chu , Chiou Shu-Jiau , Tsai Yuan-Jang , Hwang Chrong-Shiong
Published: February 2011

Acute myeloid leukemia (AML) is a highly malignant hematopoietic tumor characterized by an abnormal proliferation of myeloid progenitor cells, decreased rate of self-destruction and an arrest in cell differentiation. The internal tandem duplication mutations of FMS-like tyrosine kinase-3 (FLT3/ITD mutations) are common in AML and linked to poor prognosis. ITRI-260 generated from designed by computer-assisted rationale drug design and in silico prioritized approaches, was proved to be a potent cell growth inhibitor against FLT3/ITD mutated AML cells through FLT3 and c-Kit inhibition. The in vivo efficacy of ITRI-260 via oral route was demonstrated in MV4-11 xenograft mice model. Besides, two more orthotopic murine models were used to assess the in vivo therapeutic efficacy of ITRI-260. The first one is MV4-11 cell orthotopically engraft into NOD/SCID mice bone marrow and the second one is primary AML cells from AML patient engraft into NOD/SCID mice bone marrow. The results show that ITII-260 can decrease FLT3/ITD mutant leukemia blasts in peripheral blood and bone marrow in both animal models. Finally, ITRI-260 prolongs the survival in both bone marrow engraft mice models. These findings strongly suggest clinical benefits of ITRI-260 in patients with FLT3/ITD AML. Currently we are planning the bridged projects to push ITRI-260 toward IND status.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-160.
Abstract LB-164: Aurora Kinase A as a target for sensitizing glioma cells to radiation treatment
Authors: Patel Disha , Gordon Nicolaus , Shirai Katsyuki , Chakravarti Arnab
Published: February 2011

Purpose: Eukaryotic cells have a complex cell cycle process that is regulated by many factors. Tumors arise when cell cycle progression is deregulated. Mitosis is a well coordinated event and occurs only when proper signals allow the cell cycle machinery to proceed beyond the G2/M checkpoint. Aurora kinase A (AurKA) is a mitotic regulator involved in bipolar-spindle assembly and chromosome segregation, and also recruits other proteins and cofactors that are important for mitotic events. AurKA is upregulated in many cancers and has been shown to cause aneuploidy and induce oncogenic transformation. AurKA overexpression allows cells to surpass the G2/M checkpoint and enter mitosis prematurely, which leads to chromosomal abnormalities and genomic instability.

Experimental Design: Here we are investigating AurKA in established (ATCC) and primary glioblastoma (GBM) cell lines. Assays measuring radiosensitivity were conducted using wild-type and AurKA knockdown GBM cells. These include the clonogenic survival assay, MTS proliferation assay, cell-cycle analysis, and Annexin V apoptosis assay. Protein profiles were used to investigate pro-apoptotic, anti-apoptotic, and other downstream and upstream signaling pathways after radiation treatment.

Results and Discussion: Irradiation induces AurKA expression in both our established and primary GBM cell lines. Next we silenced AurKA using lentiviral mediated shRNA transduction. AurKA knockdown cells exhibit greater radiosensitivity when compared to cells transduced with a mock vector. AurKA and p53 interact at multiple levels. Preclinical data with AurKA inhibitors suggest p53-negative tumors may be more sensitive to AurKA inhibitors than p53-positive tumors. To investigate which subset of GBMs show greater response to AurKA silencing, we will conduct experiments using our panel of isogenic GBM cells lines. Our hypothesis is that AurKA knockdown cells will show reduced proliferation and increased apoptosis, thereby leading to cell death. The outcome of this preclinical study would be to evaluate AurKA as a potential therapeutic target for the treatment of GBM.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-164.
Abstract LB-178: Characterization of estrogen responses in breast cancer cell lines highlights ERα as an LRH-1 regulated gene
Authors: Thiruchelvam Paul T.R , Hua Hui , Lai Chun Fui , Photiou Andy , Kyle Fiona , Coombes Charles , Buluwela Laki , Ali Simak
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-178.
Abstract LB-179: Enhanced resistance to tamoxifen by the c-ABL proto-oncogene in breast cancer
Authors: Zhao Hua Jun , Ou-Yang Fu , Chen I-Fen , Hou Ming-Feng , Yuan Shyng-Shiou F. , Chang Hsueh-Ling , Lee Yi-Chen , Plattner Rina , Waltz Susan , Ho Shuk-Mei , Sims Jonathan , Wang Shao-Chun
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-179.
Abstract LB-180: Prevention of NNK-induced lung cancer in mice by anti-estrogen therapy
Authors: Siegfried Jill M. , Rothstein Mary E. , Lenzner Diana , Gubish Christopher T. , Land Stephanie R. , Stabile Laura P.
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-180.
Abstract LB-194: Restoration of miR-31 function in already-established metastases elicits an anti-metastatic therapeutic response in vivo.
Authors: Valastyan Scott , Chang Amelia , Benaich Nathan , Reinhardt Ferenc , Weinberg Robert A.
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-194.
Abstract LB-201: Nuclear localization of Src, androgen receptor and PSA in prostate cancer
Authors: Saxena Parmita , Wang Tao , Adams Dave S. , Gioeli Daniel , FitzGerald Thomas J. , Languino Lucia R.
Published: February 2011

Src signaling plays an important role in prostate cancer (PrCa) progression. Dasatinib, an oral src family kinase inhibitor, is being tested in patients with PrCa in several Phase II and III clinical trials. It has been shown that src interacts with androgen receptor (AR) and enhances AR transactivation. Although it has been shown that src promotes AR activity, the underlying pathway has not been defined.

To characterize the src-AR pathway, we first analyzed the localization of src, AR, and Prostate Specific Antigen (PSA, an AR target gene). Using sub-cellular fractionation and immunofluorescence, p-src and src were found in the nucleus, apart from their normal cytoplasmic localization, in androgen-dependent LNCaP cells upon androgen stimulation or deprivation conditions. Also, their localization was not affected by androgen stimulation. Similar to src and p-src, AR as well as pSer81-AR (an AR site indirectly phosphorylated by src) were found in the nucleus as well as in the cytoplasm. Unexpectedly, we found PSA localization in the nucleus upon androgen stimulation in LNCaP and C4-2B cells as well as in the nucleus of C4-2B cells upon androgen deprivation.

We further studied the effect of src on AR activity by transfection of dominant negative src (SrcK298M) in LNCaP and androgen-independent C4-2B cells. Transfection with SrcK298M did not affect PSA expression in LNCaP cells whereas in C4-2B cells SrcK298M transfection inhibited PSA expression.

These data show that src is required for AR activity and, consequently, PSA expression in androgen-independent prostate cancer cells, but not in androgen-dependent cells.

In conclusion, these data suggest that the nuclear co-localization of p-src, AR and PSA might allow macromolecular interactions which can further enhance AR transactivation and promote disease progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-201.
Abstract LB-208: Radiation-induced liver fibrosis is mitigated by gene therapy against transforming growth factor-ß signaling in the rat
Authors: Shi-suo Du , Zhao-Chong Zeng , Min Qiang , Zheng-Yu Zhang , Zhong-shan Liu
Published: February 2011

Purpose: We investigated whether blockage the Transforming growth factor (TGF)-ß signaling ameliorated established liver radiation-induced liver fibrosis (RILF).

Materials and Methods: A replication-defective adenoviral vector expressing the extracellular portion of mouse TßRII and the Fc portion of human immunoglobulin IgG fusion protein(AdT RIIFc) was produced. The entire liver was exposed to 30 Gy irradiation to generate a RILF model (RILFM). Then, RILFM animals were treated with AdT RIIFc (1× 1011 PFU, TßRII), Control virus (1×1011 PFU, AdGFP), or saline (SALIN). Delayed radiation liver injury was assessed using histology, morphometry, and immunohistochemistry. Chronic oxidative stress damage, hepatic stellate cell (HSC) activation, and hepatocyte regeneration were also analyzed.

Results: Blocking TGF-ß signaling with AdT RIIFc successfully mitigated established radiation induced liver fibrosis (RILF). Compared with RILFM or AdGFP rats, AdT RIIFc-treated mice exhibited less oxidative stress damage, less HSC activation, preserved liver function.

Conclusions: Modulation of TGF-ß with AdT RIIFc is feasible and ameliorates established radiation-induced liver fibrosis. These data confirm the putative role of TGF-ß in delayed radiation-induced liver damage.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-208.