bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5602: Identification of ethnic specific differences in breast cancer and normal breast tissue
Authors: Baumbach Lisa L. , Gomez Carmen , Yan Jim , Halsey Tom , Ahearn Mary Ellen , Jorda Merce , Pegram Mark
Published: June 2012

Disparities in breast cancer stage of presentation and survival rates exist in patients of different ethnicities. These differences are undoubtedly a result of a combination of factors, including socio-economic, lifestyle, tumor characteristics and inherent factors, such as genetic composition. Our group is analyzing the genetic contributions to these disparities, with the goal to increase understanding of the underlying biology, leading ultimately to individualized, ethnic-specific diagnostic and therapeutic approaches. Here we report our results of a recently completed study focusing on gene expression profiling in a multi-ethnic cohort of triple negative breast cancer patients. We analyzed breast cancer and self-matched normal tissue samples from 10 African-American (AA), 10 Hispanic (His), and 10 non-Hispanic white (Caucasian) patients from south Florida. Study samples were cut from FFPE (Formalin Fixed Paraffin-embedded tissue) blocks marked by pathology as normal vs. tumor tissue, and sent to Almac Diagnostics for RNA isolation, cDNA preparation, and hybridization of tumor/normal cDNAs to a breast cancer focused gene expression array (Breast Cancer DSA Research Tool). From the Breast Cancer DSA arrays data, a two-way ANOVA (disease state and ethnicity) was used to identify transcripts with a p-value less than 0.01. Data QC indicated that samples clustered well with respect to ethnicity and adjacent normal vs. tumor tissue. We have identified ethnic-specific expression patterns in the matched normal and tumor samples. Initial pathway analysis using MetaCore Program shows that a number of genes related to the DNA repair pathway are differentially expressed across the ethnicities. In a set of ten DNA repair/cell cycle genes, the direction of change (increased or decreased expression) was the same for all three ethnic groups, however, the level of change differed greatly between ethnic groups. Fold change in this set of ten genes ranged from -6.54 to +5.53 with all being greater than 2 fold change in at least one ethnic group. Additional pathway analysis and validation of these results is ongoing. In follow up to this study, we have initiated parallel analysis in normal tissue samples (reduction mammoplasty) samples from AA and Cau non-cancer patients. Combined analysis of tumor and normal expression data will help to better understand the possible significance of gene expression differences in breast tissue between ethnic groups. These studies have important implications for addressing BC health disparities, as well as tailored approaches to prediction, prevention and treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5602. doi:10.1158/1538-7445.AM2011-5602
Abstract 5612: Identification of shared regions of genetic susceptibility to breast cancer in Arab and Jewish women with a family history of consanguinity
Authors: Gornick Michele C. , Shakour Sana , Li Jun , Rennert Gad , Gruber Stephen B.
Published: June 2012

Background: Breast cancer is the most commonly diagnosed cancer in Israel, and a leading cause of death among women regardless of their ethnic origin. However, younger ages at diagnosis, larger primary tumor size, and lower 5-year survival rate have been reported among Arab women compared with the Jewish women. Arabs have a high rate of parental consanguinity, ranging from 25% to 60% in different countries in the Middle East. Consanguinity leads to a higher probability of sharing alleles identical-by-descent and increased risks for autosomal recessive disorders.

Methods: Genome-wide analysis of germline DNA from 32 breast cancer patients and 26 first degree relatives was performed using the Affymetrix Human SNP Array 6.0 or the Illumina CytoSNP12 genotyping platform. Probands were screened for mutations in BRCA1 and BRCA2 to exclude common forms of inherited susceptibility. Genotype data were analyzed using PLINK to screen for runs of homozygosity of at least 500 kilobases.

Results: Long runs of homozygosity were observed in five samples, including two consanguineous Arab and 3 Ashkenazi Jewish probands. The largest region of homozygosity was on chromosome 10q23.1-25.3 which harbors several important candidate genes, including the tumor suppressor gene PTEN.

Conclusion: Preliminary analysis of breast cancer patients from consanguineous families of Arab or Jewish descent provide suggestive evidence of recessive susceptibility to breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5612. doi:10.1158/1538-7445.AM2011-5612
Abstract 5616: Assessment of differential DNA methylation by genotype in ovarian cancer cases and controls
Authors: Cicek Mine S. , Armasu Sebastian M. , Kalli Kimberly R. , Fridley Brooke L. , Sellers Thomas A. , Tsai Ya-Yu , Cunningham Julie M. , Goode Ellen L.
Published: June 2012

Ovarian carcinoma is a significant cause of cancer mortality in women in the United States. Increasing evidence has demonstrated that differential methylation status in lymphocyte DNA may be linked to risk of carcinogenesis. We are investigating the differential DNA methylation in germline DNA obtained from pre-treatment ovarian cancer cases (n=256) and individually-age matched controls (n=274) using the Illumina Infinium Human Methylation 27K BeadChip. We are first studying methylation and the common moderate-risk alleles for ovarian cancer on chromosomes 2q31, 3q25, 8q24, 9p22, 17q21, and 19p13. Within each locus, we correlated single nucleotide polymorphism (SNP) genotypes with methylation values at relevant nearby CpG sites. Preliminary results are available on the first set of 84 cases and 90 controls. At 2q31, cases with TG or TT genotype at rs2072590 were hypomethylated at HOXD1 (a homeobox transcription factor) compared to cases with GG genotype (mean beta TG/TT=0.08, GG=0.09, p=0.034); and at 17q21, cases with AG or GG genotype at rs9303542 were hypomethylated at SKAP1 (a T-cell adaptor protein) compared to cases with AA genotype (mean beta AG/GG=0.14, AA=0.16, p=0.045). At these two loci, similar trends were seen among controls. Logistic regression adjusted for age and methylation chip was then used to assess the association between SNP genotype and ovarian cancer risk, with and without inclusion of the relevant methylation value. At HOXD1 rs2072590, the per-allele ovarian cancer risk of 1.34 attenuated to 1.25 with inclusion of the HOXD1 methylation value as a covariate, suggesting that the disease association may be mediated in part through methylation. Though based on preliminary data, results are consistent with a role for inherited variation on germline methylation at HOXD1 and SKAP1. Analysis of methylation on additional genotyped cases and controls is underway. Finally, because, trans effects may also exist, we are assessing relationships between genotype and methylation at other loci throughout the genome. Such correlative molecular studies have the potential to help elucidate mechanisms underlying associations from genome-wide association studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5616. doi:10.1158/1538-7445.AM2011-5616
Abstract 5618: Polymorphisms in DNA repair genes are associated with endometrial cancer risk among Japanese women
Authors: Hosono Satoyo , Matsuo Keitaro , Ito Hidemi , Hirose Kaoru , Watanabe Miki , Nakanishi Toru , Tajima Kazuo , Tanaka Hideo
Published: June 2012

Common polymorphisms in DNA repair genes may affect an individual's capacity to repair damaged DNA. Although many former studies have reported an association between several types of cancers and DNA repair polymorphism, the association with endometrial cancer (EC) risk has not been investigated fully, especially in Asian population. To examine the association between DNA repair polymorphisms and postmenopausal EC risk, we conducted a case-control study with 91 EC cases (59 endometrioid adenocarcinoma and 32 other types of carcinoma) and 261 non-cancer female controls from database of Hospital based Epidemiologic Research Program at Aichi Cancer Center. We examined 16 coding SNPs (single nucleotide polymorphism) of 12 DNA repair genes (OGG1, MUTYH, XRCC1, APE1, ADPRT, XPA, XPC, XPD, ERCC1, RAD23B, XPG, ERCC6) by TaqMan Methods. We surveyed lifestyle/environmental information by self-administered questionnaire. Impact of each SNPs on EC risk was evaluated by multivariate unconditional logistic regression models. Further, we examined the gene-gene interaction.

We observed significant association between EC risk and rs1799782 (C>T, Arg194Trp) of XRCC1 [CT: Odds ratio (OR) = 1.74, 95% confidence interval (95% CI) = 0.99-3.04, TT: OR = 2.76, 95% CI = 1.23-6.16; trend p = 0.007]; rs25487 (G>A, Arg399Gln) of XRCC1 [GA: OR = 0.66, 95% CI = 0.39-1.12, AA: OR = 0.15, 95% CI = 0.02-1.16; trend p = 0.019]; rs1136410 (T>C, Val762Ala) of ADPRT [TC: OR = 1.49, 95% CI = 0.84-2.64, CC: OR = 1.92, 95% CI = 0.88-4.18; trend p = 0.076]; and rs17655 (G>C, Asp1104His) of XPG [GC: OR = 0.68, 95% CI = 0.39-1.21, CC: OR = 0.54, 95% CI = 0.26-1.12; trend p = 0.082]. Although both XRCC1 and ADPRT involve in base excision repair, gene-gene interaction of these polymorphisms were not significant; the interaction p between rs1799782 and rs25487 was 0.687; that between rs25487 and rs1136410 was 0.328.

In conclusion, we found significant association between EC risk among postmenopausal Japanese women and DNA repair polymorphisms. Gene-gene interactions between XRCC1 and ADPRT were not significant. These results suggests that DNA repair polymorphisms might be associated with etiology of EC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5618. doi:10.1158/1538-7445.AM2011-5618
Abstract 5619: Exploration of pleiotropic effects of cancer GWAS hits with risk of endometrial cancer in the PAGE study
Authors: Peters Ulrike , Fesinmeyer Megan , Carty Cara , Kocarnik Jonathan , Lin Yi , Haessler Jeffrey , Kooperberg Charles ,   PAGE Consortium
Published: June 2012

Introduction: Endometrial cancer is the most common gynecological cancer in developed countries. While so far no genome-wide association study (GWAS) has been published for endometrial cancer this approach has been very successful in identifying susceptibility loci for various cancer sites. Interestingly, these GWAS have discovered susceptibility loci that are associated with multiple cancer sites, such as in the 8q24 or TERT-CLPTM1L locus. To comprehensively explore such pleiotropic effects for endometrial cancer we evaluated whether GWAS hits for different cancer sites are associated with risk of endometrial cancer.

Methods: We included 965 endometrial cancer cases and 5759 controls aged 50-79 years from the Women's Health Initiative, which were genotyped as part of the Population Architecture using Genomics and Epidemiology (PAGE) study. We selected 175 single nucleotide polymorphisms (SNPs) previously shown to be associated with different cancer sites mostly in GWAS. We coded SNPs using additive genetic models and used logistic regression to estimate per allele odds ratios (OR) and 95% confidence intervals (CI). Race/ethnicity-specific models were adjusted for age and post-menopausal hormone use.

Results: Among whites the two most independent significant SNPs were rs7501939/17q12 [odds ratio (OR)=0.82; 95% confidential interval (CI)=0.74-0.92; p-value=3.4e-4], and rs4324715/5q31 (OR=0.85; 95%CI=0.76-0.94; p-value=1.5e-3). Interestingly, these two variants have been related to cancers of the male reproductive system (prostate and testicular cancer, respectively) in prior GWAS. When adjusting for multiple comparisons (p=2.86e-4) only one variant (rs7501939) remained marginally significant. The number of other ancestral groups, such as African American or Asian (n=72) was too small to allow separate analysis.

Conclusion: This first analysis suggests that susceptibility loci of the male reproductive system may be associated with endometrial cancer. The next step in this analysis will be to incorporate data from the other PAGE studies to improve the power, explore effects across different ethnicities and to expand our analysis to evaluate gene-environment interactions, particularly looking at hormone related factors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5619. doi:10.1158/1538-7445.AM2011-5619
Abstract 562: Multi-Kinase Inhibitor: AFPep inhibits the growth and invasion of U87 glioblastoma cells through inhibition of FAK, MAPK, and PKC
Authors: Mesfin Fassil B. , Riccio Alexander , Andersen Thomas T. , Bennett James
Published: June 2012

INTRODUCTION: Glioblastomas are the most frequent tumors of the central nervous system. Patients with a diagnosis of glioblastomas have poor prognosis, and new therapeutic agents are in demand for management of this disease. Here, we report on a novel multi-protein kinase inhibitor, AFPep, with regard to its effects on the growth, invasion, and localization in U87 cells.

METHODS: Proliferation/Invasion Assay: Cells were seeded into wells of plastic tissue culture plates and treated with various concentrations of AFPep. Crystal violet method was used to determine cell numbers. The Matrigel chemoinvasion assay was used to determine the effect of AFPep on U87 cell invasion. During Xenograft growth assay, U87 cells were solidified in fibrin clot and implanted under the kidney capsule of severe combined immunodeficient (SCID) mice. Various doses of AFPep were administrated i.p and tumor growth was monitored during survival laparotomy. Localization assay was performed by utilizing fluorescent-labeled AFPep. Multi-immunoblotting Assay was performed using Kinexus, the KinetworksTM methodology to screen for protein kinase expression profile in GBM before and after AFPep treatment.

RESULTS: AFPep inhibited the growth of U87 cells in culture with it is maximal effect at 10−7 M. Higher concentrations of AFPep (10−6M to 10−4M) inhibited the invasion of U87 cells in a dose dependent manner. Treatment of SCID mice bearing U87 xenografts with 4 mg/Kg/day AFPep resulted in a 50% inhibition of tumor growth. Further, AFPep-Bodipy appeared to be localized in mitochondria and interferes with phosphorylation of FAK, MAPK, and c-kit.

CONCLUSION: AFPep has an inhibitory effect on U87 cells. All studies to date have shown no toxicity. As such AFPep is expected to be tolerated and may be a new agent for the treatment of malignant glioblastomas

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 562. doi:10.1158/1538-7445.AM2011-562
Abstract 569: Shear stress radiatiosensitizes colon cancer cells by downregulating FAK via integrin β1
Authors: Luo Chi-Wen , Wu Chia-Ching , Ch'ang Hui-Ju Mandy
Published: June 2012

Introduction: Interstitial fluid flow plays important role in delivering anticancer agents to tumor tissue. Recent studies suggested that anti-angiogenic or vasculature disrupting agents may enhance the therapeutic effect of radiotherapy by regulating tumor blood flow and interstitial fluid pressure. Although the importance of interstitial fluid flow in modulating tumor microenvironment has been studied, the molecular mechanisms of shear flow on tumor cell survival and sensitivity to therapeutic agents have not been much explored. We would like to investigate the interaction of shear stress with irradiation on tumor cells.

Materials and methods: Human T84 and SW480 colon adenocarcinoma cells were used in this study. Cells were seeded onto glass slides pre-coated with fibronectin, and then subjected to 0-12 dyne/cm2 laminar shear stress in parallel plate flow chamber (PPFC) system for 0-24 hours. Cells were then exposed to 0-8Gy irradiation (IR) by a RS2000 X-ray irradiator. MTT and clonogenic assays were used to study cell survival. Tumor cell cycle and apoptosis were evaluated by flow cytometry and Western blot. Candidate signal molecules were measured by real time PCR and immunoblots with or without pretreatment with cyclohexamide or MG132.

Results: MTT and clonogenic assays revealed a synergistic effect of 24 hour- shear stress and IR on tumor cytotoxicity. Shear stress downregulates FAK, phosphor-FAK, AKT and phosphor-AKT expression but not ILK and GSK3B through integrin β1. Decreased protein expression was mainly through proteosomal degradation but not transcriptional suppression. By enhancing G2/M phase cell accumulation, shear stress increases tumor apoptosis after IR. The above phenomenon was not observed in IR pretreated with 3 hour-shear stress.

Conclusion: The above observation suggests that shear stress may enhance colon cancer cell radiation sensitivity by inducing G2/M arrest, and FAK protein degradation via integrin β1 signaling pathway

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 569. doi:10.1158/1538-7445.AM2011-569
Abstract 570: Radiation resistance of orthotopic glioblastoma xenografts is mediated by increasing CD133+ cell fraction, mobilizing G0 cells into G1 phase, and activating self-renewal and mesenchymal genes
Authors: Liu Zhi-gang , Zhao Xiu-mei , Baxter Patricia , Tao Ya-Lan , Su Jack , Adesina Adekunle , Chintagumpala Murali , Blaney Susan , Xia Yun-Fei , Li Xiao-Nan
Published: June 2012

Pediatric glioblastoma multiforme (pGBM) is a highly aggressive brain tumor with poor prognosis. Resistance to ionizing radiation, which is one of the major therapeutic regimes, is believed to play a critical role in tumor recurrence and ultimate treatment failure. However, little is known about the cellular and molecular mechanisms of radioresistance in pGBM, particularly the in vivo responses of tumor cells toward clinically relevant fractionated radiotherapy. To gain a better understanding of the radioresistance mechanisms, we administered fractionated radiation (2Gy per day × 5 times) to SCID mice bearing pre-established orthotopic xenograft tumors derived from two independent fresh patient tumors. Examination of the xenograft tumors immediately at the end of radiation treatment showed that the fractionated radiotherapy significantly increased the abundance of CD133+ tumor cells, and drove quiescent G0 phase cells (both CD133+ and CD133-) into G1 phase in both models. Immunohistochemical examination of key regulators of stem cell self-renewal revealed elevation of BMI-1 and SOX2 gene expression in vivo in the xenograft tumors. Additionally, we found that radiotherapy promoted mesenchymal transition, as the intensity and distribution of fibronectin, a mesenchymal marker, was remarkably enhanced. Further analysis of the recurrent xenograft tumors ∼ 5 weeks post-fractionated radiation showed that the increased CD133+ fraction, the elevated expressions of BMI-1 and SOX2 as well as fibronection were maintained. In conclusion, our data demonstrated, for the first time, that failure of radiotherapy is mediated by activated pathways related to self-renewal of cancer stem cells and mesenchymal transition. Since this study was conducted in vivo in clinically relevant animal models in a setting that is the closest to human brain tumors, our findings provided strong preclinical evidence to suggest that new stratigies are needed to inhibit these radiation triggered-pathways to improved therapeutic efficacy in pediatric GBM patient.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 570. doi:10.1158/1538-7445.AM2011-570
Abstract 576: Gadd45a sensitizes medulloblastoma cells to irradiation and modulates the cellular switch from migratory to stationary cell phase by downregulating MMP-9
Authors: Asuthkar Swapna , Nalla Arun Kumar , Dinh Dzung H. , Gujrati Meena , Mohanam Sanjeeva , Rao Jasti S.
Published: June 2012

Gadd45a protein is induced in medulloblastoma cell lines by treatment with ionizing radiation (IR), which requires p53 stabilization. The role of Gadd45a in IR-induced G2-M arrest of medulloblastoma cells is demonstrated by its increased binding to Cdc2, thereby inactivating Cdc2 kinase activity. Knockdown of Gadd45a alleviates G2-M blockage and results in decreased binding of Gadd45a to Cdc2. Further, the anti-tumorigenic role of Gadd45a is mediated by the negative regulation of β-catenin and its nuclear translocation, which might decrease the β-catenin/LEF-1-mediated transactivation of cell invasion proteins, such as MMP-9, examined in the present study. Overexpression of Gadd45a protein with IR treatment resulted in decreased nuclear levels of β-catenin, increased cytoplasmic levels of p-β-catenin, and increased distribution of β-catenin on membrane E and N-cadherins. The IR-treated control cells showed contrary results with demonstrated loss of E-cadherin, gain of N-cadherin, and inhibition of β-catenin binding to membrane cadherins as well as increased nuclear translocation of β-catenin. Zymography studies demonstrate the negative regulation of IR-induced MMP-9 activity by Gadd45a overexpression. Wound healing and Matrigel invasion assays confirmed the anti-tumorigenic and anti-metastatic role of Gadd45a in medulloblastoma cell lines. Furthermore, siRNA-mediated knockdown of MMP-9 results in prominent expression levels of Gadd45a protein in animal tumors. Thus, we confirm that overexpression of Gadd45a in combination with ionizing radiation has therapeutic implications by preventing recurrence of MMP-9-mediated cancer cell invasion and effectively containing cells in a static phase by maintaining CCA/CCI.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 576. doi:10.1158/1538-7445.AM2011-576
Abstract 578: Roles of mitochondrial protein kinases in targeted cancer therapy
Authors: Guo Tiannan , Kon Oi Lian , Sze Siu Kwan
Published: June 2012

Targeted cancer therapy is based on the curious dependence that some cancer cells have on a single oncogenic molecule, typically a protein kinase (the phenomenon of “oncogene addiction”). However, the molecular mechanisms of such dependence are incompletely understood. We have adopted a quantitative proteomic approach to investigate the phenomenon that MET-overexpressing cancer cells are paradoxically more sensitive to MET inhibition than cells with basal MET expression. Unexpectedly, we found that mitochondria are a direct target of MET kinase inhibition, in addition to inhibition of plasma membrane MET. Effects on activated MET in the mitochondria of cancer cells that are sensitive to MET inhibition could reflect a novel and critical non-canonical mechanism that contributes to the efficacy of MET-targeted therapeutics. We hypothesize that protein kinases which aberrantly translocate to mitochondria modulate mitochondrial proteins and functions. Here we have employed advanced LC-MS/MS-based stable isotope labeling with amino acids in cell culture (SILAC) quantitative phosphoproteomic approaches to identify, quantify and functionally annotate phosphorylated mitochondrial proteins of various cancer cell lines, including those that are sensitive or resistant to targeted agents. We identified several phosphorylated protein kinases e.g. MET, EGFR, HER2 and HER3, that were translocated to mitochondria of cancer cells. In addition to documenting the first kinome and phosphoproteome of mitochondria in cancer cells, we present evidence showing that the aberrant presence of protein kinases in mitochondria predicts sensitivity to specific kinase inhibitors. In conclusion, our data reveal a novel mitochondrial mechanism of oncogene addiction, and suggest that mitochondrial protein kinases might play decisive roles in molecularly-directed therapies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 578. doi:10.1158/1538-7445.AM2011-578
Abstract 58: Understanding target biology using protein interactomes
Authors: Mohan Amrita , Bhagwat Shripad V. , Epstein David M. , Miglarese Mark , Pachter Jonathan A.
Published: June 2012

The use of systematic genome-wide physical protein-protein interaction detection methods has led to the generation of large networks of putatively interconnected human proteins including a number of therapeutically relevant disease-associated proteins. These networks have intrinsic value for understanding the pathophysiology of proteins, identification of tractable drug targets within a network, and hypothesis generation with regard to potential pharmacodynamic and patient selection biomarkers for a drug target of interest.

Here, we present a novel approach to utilize a high-confidence human focal adhesion kinase (FAK) protein interaction network to better understand FAK target biology. FAK is a well-studied signaling protein known to directly control cellular motility via mechanosensing. As a large 125 kDa protein with both protein tyrosine kinase and scaffolding functions, FAK is particularly rich in its potential to interact with numerous proteins through SH2 and other interaction domains. By forming a diverse array of target interactions that regulate cytoskeletal structural changes, cellular adhesion, cell migration and cellular proliferation, FAK is believed to be a key regulator of several critical processes in tumorigenesis. It has also been established that human tumors often exhibit FAK gene amplification with resulting increased FAK expression which correlates with cancer progression and poor prognosis. Although the plausible roles of FAK have been well-elaborated, there are limited data available on the precise interplay of biological entities controlling the FAK signaling pathway. In this study, we built a prototypical model representing cross-talk between multiple proteins believed to be involved in the FAK signaling pathway. In doing so, we describe a step by step procedure to assemble a FAK interactome using integrative data mining procedures with publicly available interaction databases such as MINT, BioGRID, DIP and HPRD. Next, we filter this data set by incorporating confidence scores and lines of evidence available corresponding to each interaction to finally derive a high-confidence FAK interactome.

We propose that this approach to generate high-confidence protein interactomes could also be applied to other proteins of interest to facilitate a systems biology approach to elucidate additional biological pathways influencing tumorigenesis, metastasis and other disease states amenable to rational drug discovery.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 58. doi:10.1158/1538-7445.AM2011-58
Abstract 583: Inecalcitol, a 14-epi-analogue of 1,25D3, induces growth inhibition through apoptosis and cell cycle arrest in a squamous cell carcinoma model system
Authors: Ma Yingyu , Yu Wei-Dong , Kong Rui-Xian , Delansorne Remi , Trump Donald L. , Johnson Candace S.
Published: June 2012

Epidemiological and experimental studies have demonstrated that 1,25D3, the active vitamin D metabolite, has broad spectrum anti-tumor activity. However, hypercalcemia may limit its clinical application when administrated inappropriately. An orally available 14-epi-analogue of 1,25D3, inecalcitol (19-nor-14-epi-23-yne-1,25-(OH)2D3, TX522), has shown good clinical tolerance at high and frequent exposure without side effects and hypercalcemia. Therefore, we examined the antitumor activity of inecalcitol in a squamous cell carcinoma (SCC) model system. Inecalcitol at 10 or 100 nM readily induced vitamin D receptor expression in SCC cells. Inecalcitol suppressed SCC cell proliferation in a dose dependent manner as shown by MTT assays. SCC cells underwent G0/G1 cell cycle arrest following inecalcitol treatment, which was accompanied by the induction of p27 protein as shown by immunoblot analysis. Inecalcitol markedly induced apoptosis in SCC cells as examined by annexin V/7AAD staining and DNA fragmentation ELISA assays. The cleavages of caspases 8, 9, and 3 and PARP were observed following inecalcitol treatment in SCC cells. In addition, migration assay revealed that inecalcitol inhibited the motility of SCC cells. Inecalcitol also suppressed the invasion of SCC cells as assessed by Matrigel-based invasion assay. To have a better understanding of the effects of inecalcitol, 1,25D3-resistant variant of SCC (SCC-DR) were employed. The above findings were not observed in SCC-DR cells, indicating the critical role of inecalcitol. Our studies suggest that inecalcitol strongly suppress SCC growth through the promotion of apoptosis and cell cycle arrest. Inecalcitol also inhibits SCC migration and invasion. Thus, inecalcitol may be a promising 1,25D3 analogue for cancer treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 583. doi:10.1158/1538-7445.AM2011-583
Abstract 600: The HSV-2 oncolytic virus ΔPK induces multiple death and inflammatory programs associated with inhibition of melanoma tumor growth
Authors: Colunga Aric , Li Biquan , Aurelian Laure
Published: June 2012

Metastatic melanoma is a highly aggressive and drug resistant cancer. Oncolytic virotherapy is a novel therapeutic strategy designed to circumvent cancer resistance through virus replication-induced cancer cell lysis. Unfortunately, its clinical efficacy is modest, apparently related to poor virus replication within the tumors and the failure to stimulate inflammatory/immunotherapeutic pathways. The growth compromised herpes simplex virus type 2 (HSV-2) mutant ΔPK causes metastatic melanoma cell death through activation of multiple non-redundant death pathways. Activation of calpain and caspases-3 and -7 provide the bulk of the death programs, as evidenced by the finding of 80% protection in cells treated with the combination of calpain (PD150606) and pancaspase (zVAD-fmk) inhibitors. In addition, ΔPK upregulates the autophagy proteins Beclin-1 and LC-II and the autophagy inhibitor 3-methyladenine (3-MA) reduces ΔPK-induced cell death by 15-17%. Cell death is further decreased (to a total of 28-34%) by treatment with 3-MA together with the JNK inhibitor SP600125, indicating that JNK signaling and autophagy contribute independently to ΔPK-induced cell death. Inflammatory-related pathways contribute to ΔPK-induced melanoma cell death, as evidenced by caspase-1 activation, interleukin (IL)-1β release into the conditioned medium and upregulation of pro-apoptotic/stress signaling molecules and pro-inflammatory cytokines and chemokines identified by microarray analysis. These include BAD, BIK, BOK, NLRC4, CIDEB, IL-1β, IL-6, IL-8, IL-12α, IL-12β, LTα, CXCL-10, c-JUN and caspases-4, -5, -6, -8, and -10. Tumor burden is significantly reduced by intratumoral injection of ΔPK and xenografts generated from a fresh clinical isolate, were completely eliminated for a period of 1 year following treatment. Growth inhibition is associated with activation of calpain and caspases-3, -7 and -1 and expression of the inflammatory cytokine TNF-α. Collectively, the data indicate that ΔPK eliminates tumor burden through the simultaneous activation of multiple non-redundant death programs and pro-inflammatory signaling, providing a distinct and promising virotherapeutic approach.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 600. doi:10.1158/1538-7445.AM2011-600
Abstract 602: Silencing BRG1 in human cancers
Authors: Reisman David R. , Gramling Sarah , Wright Tracy
Published: June 2012

BRG1 is a novel tumor suppressor that serves as the major catalytic subunit of the SWI/SNF complex. Because this complex is central to the function of a number of transcription factors and key cellular proteins and it is required for a plethora of signal transduction pathways, its loss can have devastating effects on cellular function. We know that BRG1 is not only lost in lung cancer but also in a variety of tumor types: between 10-40%. Hence, the loss of this protein plays an important role in cancer development and progression. In cell lines, it has been well document that BRG1 is mutated or altered in the vast majority of cell lines, but in primary lung cancer mutations are not found. Our analysis has led us to believe that BRG1 is epigenetically silenced like its homolog BRM. Hence, it may be possible to restore its expression in cancer cells. The loss of BRG1 impacts the sensitivity to Cisplatin by making cells more sensitive to the DNA damaging effects of this agent. Similarly, this loss also impacts sensitivity to EGFR inhibitors by making cells more resistant. Hence, the loss of BRG1 can help to explain the reciprocal sensitivity to these drugs seen in cancers. Hence, the BRG1 protein can play the role of a biomarker for chemotherapy sensitivity as well as being a potential target for therapy. Further research will reveal how this gene can best be integrated into clinical practice.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 602. doi:10.1158/1538-7445.AM2011-602
Abstract 604: Nek2 is a promising target for triple negative breast cancer treatment strategies
Authors: Lee Jaehyung , Gollahon Lauren
Published: June 2012

Although the anticancer drugs paclitaxel and doxorubicin are commonly used to treat many types of solid tumors, their effectiveness is highly variable due to tumor cell resistance. Therefore, it is important to find mechanisms that can be targeted to increase the sensitivity of cancer cells to antitumor agents such as paclitaxel and doxorubicin. NIMA related kinase 2 (Nek2), a serine/threonine kinase is emerging as an important oncogene because of its regulatory role in mammalian cell mitosis. Thus, regulation of the Nek2 expression levels may prove important as a target for cancer treatment. In this study, we investigated whether drug sensitivity was increased in the triple negative breast cancer cell lines MDA-MB-231 and MDA-MB-468 by using antisense oligonucleotides (ASOs) and siRNA against Nek2. MDA-MB-231 and MDA-MB-468 breast cancer cells transfected with Nek2 siRNA or ASO were exposed to various concentrations of paclitaxel and doxorubicin. Cell viability, cell cycle distribution, and apoptosis were evaluated. We observed that drug susceptibility in these transfected cells was dramatically increased compared with either agent alone. Interestingly, the cell proliferation of the Nek2 ASO transfected breast cancer cells was much lower than the Nek2 siRNA transfected breast cancer cells. Our FACS data showed paclitaxel treated cells transfected with ASOs arrested in G2/M phase as expected due to Nek2 regulatory function in centrosome duplication. We observed that ASOs and siRNA against Nek2 work with paclitaxel and by arresting cells at G2/M and progressing the cells to apoptosis. Our results suggest that these drugs in combination with Nek2 siRNA or ASO treatment greatly improve the sensitivity of cancer cells. Thus, these results provide a possible new strategy for the treatment of breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 604. doi:10.1158/1538-7445.AM2011-604
Abstract 610: Regression of spontaneous non-small cell lung cancers in P53 mutant transgenic mice following exposure to PRIMA-1
Authors: Duan Wenrui , Gao Li , Vereb Gregory , Wu Xin , Otterson Gregory A. , Villalona-Calero Miguel A.
Published: June 2012

P53 mutations are common in lung cancer, thus restoration of p53 function in tumor cells have the potential to be an important approach for lung cancer therapy. The small molecule PRIMA-1 (p53-dependent reactivation and induction of massive apoptosis) has been shown to induce apoptosis in human tumor cells containing mutant p53 by restoration of tumor suppressor function of p53. This is believed to be mediated by a change in the conformation of mutated p53 protein, restoring DNA binding and activation of p53 target genes.

We have developed a lung tumor model in transgenic mice in which the human mutant p53(273H) is expressed in a lung specific manner, under the control of the surfactant protein C promoter. These mice developed lung adenocarcinomas at an age range of 12-15 months. To evaluate feasibility and preliminary activity of PRIMA-1 on the murine spontaneous lung tumor, we treated SPC-p53-273H lung tumor bearing mice via intraperitoneal injection (i.p) with PRIMA-1 at a dose of 100 mg/kg in 0.2 ml PBS every other day for two weeks. Lung tumor bearing mice were identified with a micro-computed tomography (micro-CT) system and pre- and post-treatment tumor volumes were estimated based on the micro-CT image.

Five mice received the PRIMA-1, and a control group of 4 lung tumor mice received PBS every other day for two weeks. A repeat CT scan was obtained the week after the last treatment. Results of post-treatment with pre-treatment CT volume comparisons are detailed below:

These preliminary results indicate activity of PRIMA-1 in lung tumor bearing P53 mutant transgenics, and suggest the potential feasibility of using p53 restoring activity small molecules to treat lung cancer. Expansion of the sample size, as well as mechanistic and dosing studies is ongoing in order to strengthen the rationale to eventually pursue this strategy in clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 610. doi:10.1158/1538-7445.AM2011-610
Abstract 613: Depletion of Mdm2 and Mdm4 by adenovirus: Molecular mechanism and anticancer therapeutic implication
Authors: Liao Daiqing , Zhao Lisa Y. , Yang Heng
Published: June 2012

Mdm2 and Mdm4 are two paralogous proteins. They contain a conserved p53-binding domain in the N-terminal region and a RING-finger domain in the C-terminal part. Both proteins physically bind p53 and prevent p53 activation. Biochemically, Mdm2 acts as an E3 ubiquitin ligase of p53 and promotes ubiquitin-dependent p53 degradation. Mdm4 does not appear to have intrinsic E3 ubiquitin ligase activity and may negatively control p53 function through suppressing p53-mediated transactivation of gene expression. Mdm2 and Mdm4 also form heterodimer, which suggests that Mdm2 and Mdm4 might cooperatively control p53 activity. In vivo, both Mdm2 and Mdm4 have an essential but non-redundant function in suppressing p53 activity to enable embryonic development. Mdm2 and Mdm4 are overexpressed in several types of human cancer such as breast cancer and osteosarcoma. In transgenic mouse models, overexpression of Mdm2 or Mdm4 leads to tumorigenesis. Thus, both Mdm2 and Mdm4 are bona fide oncogenes. Several small molecules that disrupt the interaction between p53 and Mdm2 and Mdm4 have been shown to induce p53 activation, cell cycle arrest and apoptosis. These compounds also cause tumor repression in preclinical animal models. Thus, they have clinical potentials for treating various types of human cancer. Biological agents that specifically target Mdm2 and Mdm4 have not been described. We found that adenovirus infection of diverse type of cancer cells results in dramatic downregulation of both Mdm2 and Mdm4. Adenovirus E1B-55K binds specifically both Mdm2 and Mdm4 in cells and in vitro, demonstrating specific interactions between E1B-55K and Mdm2 or Mdm4. Cotransfection experiments indicated that E1B 55K alone was sufficient to deplete Mdm2 and Mdm4. E1B-55K is known to form a functional E3 ubiquitin ligase consisting of Cul2 or Cul5. Thus, it is likely that E1B-55K targets Mdm2 and Mdm4 for ubiquitin-mediated proteosomal degradation. Based on these observations, we hypothesize that adenovirus is effective for treating cancers with overexpression of Mdm2 or Mdm4. We are currently testing whether adenoviral therapy is efficacious on treating such cancers in cell culture and animal models. (This work was supported by Bankhead-Coley Cancer Research Program of the Florida Department of Health.)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 613. doi:10.1158/1538-7445.AM2011-613
Abstract 614: Efficacy of metformin in two mouse models of Li-Fraumeni syndrome
Authors: Kunnumakkara Ajaikumar B. , Memmott Regan M. , Maier Colleen R. , Kawabata Shigeru , Quinn Brendan , Gills Joell , Dallos Matthew , Hollander Christine M. , Dennis Phillip A.
Published: June 2012

Li-Fraumeni Syndrome (LFS) is a rare autosomal hereditary disorder that is commonly associated with germline mutations in the TP53 gene. People with LFS are at high risk to develop a wide range of malignancies including breast cancer, brain tumors, acute leukemias, soft tissue sarcomas, bone sarcomas, and adrenal cortical carcinomas. Although no therapies are approved to treat or prevent LFS, preclinical studies have shown that the anti-diabetic drug metformin is preferentially cytotoxic to p53 null cells in the absence of glucose. This led us to hypothesize that metformin has therapeutic potential in LFS. To confirm the p53 dependence of metformin in vitro, isogenic human and murine cancer cell lines were treated with metformin in the absence or presence of glucose. Metformin induced greater toxicity in p53-deficient cells in a dose-dependent manner. Metformin was then tested in two mouse models that recapitulate the increased tumorigenesis and cancer-related mortality in LFS (p53 mutant mice (p53R172H/+) and p53 null mice (p53-/-)). When administered at 5 mg/ml in drinking water, metformin reached steady state plasma concentrations similar to those in diabetic humans. Biomarker studies showed that oral administration of metformin activated AMPK and inhibited the mTOR pathway in liver tissues. To determine the effects of metformin on overall survival, two sets of studies were performed. For each model, treatment with metformin was begun either late in life (for p53R172H/+, 12 mo; for p53-/-, 4 mo) or early in life (for p53R172H/+, 4 mo; for p53-/-, 1 mo). Mice were treated until death. Long-term oral administration of metformin did not cause weight loss or other discernible toxicity. In the late treatment study, metformin significantly prolonged median survival (for p53R172H/+ mice, 20.6 mo vs. 15.8 mo; p=0.0004)(for p53-/- mice, 9.1 mo vs. 5.3 mo; p=0.0006). Likewise, early treatment with metformin also prolonged median survival (for p53R172H/+ mice, median survival 20.5 compared to 13 months; p=0.0025). The early study with p53-/- mice is ongoing, but median survival is undefined in metformin group, compared to 3.6 mo in control group, p=0.0387). These studies show that metformin is well tolerated and highly effective when given early or late in the life span of two mouse models of LFS. Given that metformin is an inexpensive oral drug that is FDA approved, a pilot clinical trial in LFS is planned.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 614. doi:10.1158/1538-7445.AM2011-614
Abstract 623: CLU inhibition using OGX-011 is a new adjuvant therapeutic strategy for HSP90 inhibition in prostate cancer
Authors: Lamoureux Francois , Yin Min-Jean , Zoubeidi Amina , Gleave Martin E.
Published: June 2012

Introduction and Objective: Prostate cancer (PCa) responds initially to anti-androgen therapies, however, progression to castration resistant disease (CRPC) frequently occurs. Several small molecule inhibitors of HSP90 show promise in CRPC and other cancers. However, most HSP90 inhibitors (17-AAG or PF-04928473 and its prodrug PF-04929113) trigger the elevation of HSPs (HSP90, 70, 27 and clusterin), which lead to tumor cell survival and treatment resistance. We hypothesized that targeting clusterin (CLU) using siRNA or the antisense drug, OGX-011, may enhance HSP90 inhibitors-induced cell death in PCa.

Methods: Inducible and constitutive CLU and other HSP mRNA and protein levels were measured by real-time RT-PCR and immunoblot assays. The combination of OGX-011 with PF-04928473 or 17-AAG was evaluated in vitro on LNCaP and PC3 cells growth and apoptosis. The HSP90 inhibitor PF-04929113 was evaluated in combination with OGX-011 in vivo in athymic mice bearing castration resistant LNCaP xenografts, while the combination of OGX-011 with 17-AAG was evaluated in PC3 xenografts.

Results: In prostate tumor cell lines, PF-04928473 and 17-AAG induced expression of HSPs in a dose and time dependent manner, and especially CLU at RNA and protein level, by increasing HSF-1 nuclear translocation and transcription activity. In vitro, OGX-011 synergistically enhanced the activity of HSP90 inhibitors on cell growth and apoptosis with increased sub-G1 fraction and PARP cleavage. These biologic events were accompanied by decreased expression of HSPs, Akt, AR and PSA, and HSF-1 transcriptional activity. In vivo, OGX-011, administered 3 times a week (IP, 15mg/kg), potentiated the effect of orally administered HSP90 inhibitors (PF-04929113: 25mg/kg; 17-AAG: 50mg/kg), significantly inhibiting tumor growth by 80% and prolonging survival in PC3 and castrate resistant LNCaP xenograft model compared to the HSP90 inhibitors alone.

Conclusions: These results indicate that HSP90 inhibitor-mediated induction of CLU expression can be attenuated by OGX-011, with synergistic effects on delaying progression of CRPC.

This work was supported by the Canadian Institutes of Health Research fellowship and Association pour la Recherche sur le Cancer, France.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 623. doi:10.1158/1538-7445.AM2011-623