bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5095: CGH+SNP microarrays for copy-neutral aberration detection in cancer research
Authors: Costa Paula , Curry Bo , Peter Brian , Anderson Paige , Sampas Nick , Ashutosh Ashutosh , Vadapalli Arjun , Ijpma Arne , Ruvolo Michael , Lucas Anne , LeProust Emily , Srinivasan Maithreyan , Ghosh Jayati , Fulmer-Smentek Stephanie , Witte Anniek De
Published: August 2012

Advances in cancer research have greatly benefited from high resolution copy number (CN) measurements provided by oligo array Comparative Genomic Hybridization (aCGH). The addition of single nucleotide polymorphism (SNP) measurements to CGH microarrays enables the detection of copy-neutral loss of heterozygosity (cnLOH) events and allelic imbalances arising from genomic instability during cancer development and progression. Nonetheless, cancer studies face challenges associated with genomes’ aneuploidy, polyclonality and mosaicism due to the admixtures of tumor and normal cells. In the quest to decipher tumor complexity, the power and sensitivity of Agilent's CGH+SNP platform has been expanded with new computational methods capable of determining clonal fraction, total CN and allele-specific CN in aneuploid samples. Genomic DNA from hematology-oncology samples, cell lines and a genotyped control sample was digested with AluI and RsaI restriction endonucleases to allow for SNP profiling at the enzymes’ restriction sites. Experimental and reference samples were differentially labeled and hybridized to the Agilent Cancer CGH+SNP microarray, containing ∼20K cancer associated CGH probes, ∼100K backbone probes and ∼60K SNP probes. The data were analyzed using algorithms with extended capability to determine clonal fraction, total and allele-specific CN of the aberrant clone. As expected, significant diversity was found in different chronic lymphocytic leukemia (CLL) tumors. In one CLL sample a small homozygous deletion and cnLOH were identified on chromosome 13. In another CLL case a trisomy of the entire chromosome 12 was observed, together with a small hemizygous deletion and cnLOH on chromosome 18. An Acute Lymphoblastic Leukemia (ALL) tumor sample was found to harbor an amplification on chromosome 6, as well as two small amplifications on chromosome X. To assess the ability of detecting low level mosaicism, we conducted a mixed sample experiment whereby a sample with a known aberration was mixed at known ratios with a matched sample not containing the aberration. The computed clonal fraction, total and allele-specific CN matched the expected values. We have shown that the new algorithms developed for cancer sample analysis determined the genotypes, total copy numbers and clonal fractions in aneuploid samples highly mixed with normal cell populations.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5095. doi:1538-7445.AM2012-5095
Abstract 5096: Targeted genome sequencing of 184 cancer-related genes for molecular targeted therapy in gastric cancer
Authors: Kim Hwang-Phill , Cho Geuna , Shin Jong-Yeon , Lee Won-Cheol , Yoon Young-Kwang , Han Sae-Won , Kim Jong-Il , Seo Jeong-Sun , Kim Tae-You
Published: August 2012

Cancer is a result of complex genetic alterations including mutations, copy number alterations, and gene fusions. Recent advance in sequencing technology enables comprehensive genome-wide study of the complex genetic alterations. Better understanding of the genetic alteration of cancer will ultimately lead to better treatment options and improved survival of the patients. We have established a set of cancer-related genes comprised of 184 genes for targeted sequencing using next generation sequencing technology. Targeted genome sequencing of the coding regions in the gene set was performed using the Illumina Genome Analyzer IIx. We analyzed the gene set in 13 gastric cancer cell lines. Average coverage depth was 140X. A total of 237 somatic sequence variations were found in 111 genes. The frequently mutated genes were ATM in 7, MSH6 in 6, TP53 in 5, and KRAS in 4 gastic cancer cells. Using the coverage data, we could also analyze copy number variations. Copy number gain (>2.0 fold) was found in 5 genes, whereas copy number loss (<2.0 fold) was found in 17 genes in gastric cancer cell lines. Moreover, the association between the genetic alterations and in vitro sensitivity to various targeted agents is being analyzed. The analyses are aimed at identifying sensitive/resistant mechanisms of the targeted agents and identification of novel therapeutic target. Validation and detailed analysis is currently underway. Targeted genome sequencing of cancer-related genes using NGS can be an accurate and efficient method for studying mutations and copy number variations with a single analysis. Moreover, investigational therapies with their molecular characteristics will reveal biomarkers that can be used to guide clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5096. doi:1538-7445.AM2012-5096
Abstract 5097: Genome-wide analysis of DNA copy number aberrations in gastric cancer
Authors: Tsukamoto Yoshiyuki , Noguchi Tsuyoshi , Uchida Tomohisa , Nakada Chisato , Aizawa Tetsuya , Hijiya Naoki , Matsuura Keiko , Moriyama Masatsugu
Published: August 2012

Genomic copy number aberrations (CNAs) are believed to play a major role in the development and progression of human cancers. Although many CNAs have been reported in gastric cancer, their relevance to gastric carcinogenesis are poorly understood. In this study, we investigated CNAs of 30 advanced gastric cancers (AGC) and 20 gastric carcinoma in situ (CIS) by using array comparative genomic hybridization (array CGH). By comparing the frequencies of CNAs in CISs with those in AGCs, we identified several CNAs that were more frequent in AGCs than CISs. Among them, gain of 20q13 was detected in almost all cases of AGC (29/30). We further analyzed the expression of genes located on 20q13 and found that 8 genes were upregulated more than 2-fold on average compared with normal tissues, suggesting that these genes may play an important role in the progression of gastric cancer. Taken together, our data may provide useful information about the involvment of genomic CNAs in gastric carcinogenesis. References: 1. Tsukamoto et al, J Pathol. 2008;216(4):471-82. 2. Uchida et al, J Pathol. 2010;221(1):96-105.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5097. doi:1538-7445.AM2012-5097
Abstract 5107: Unravelling the complexity of individual cancer genomes
Authors: Menghi Francesca , Woo XingYi , Gong Min , Rouam Sigrid L. , Bertrand Denis , Lee Wah H. , Wang Wendy , Rama Nona , Stronach Euan A. , Karuturi Krishna , Gabra Hani , Agarwal Roshan , Liu Edison T.
Published: August 2012

Ovarian cancer affects ∼ 204,000 women each year, and it is one of the leading causes of cancer-related death among women world-wide, as most patients present with advanced stage (III/IV) tumours, and only 40% survive 5 years after diagnosis. Improvement in the clinical management of ovarian patients is likely to derive from a better understanding of the molecular aberrations which initiate and maintain tumour growth as well as from the discovery of novel drug-able targets for the development of personalised targeted therapies. In this study, we comprehensively characterised the cancer genome of a patient diagnosed with grade III serous ovarian carcinoma, using a combination of massive-parallel sequencing technologies, including long distance DNA Paired-End-Tag (DNA-PET) sequencing, RNA-sequencing and exome-capture sequencing. This approach ensures deep coverage (from 50-180X) of critical mutational elements in a cancer genome. By comparing the genomic abnormalities identified in the tumour sample with those found in its normal counterpart (peripheral blood lymphocytes), we were able to compile a catalogue of all of the somatic events which occurred during oncogenesis and to define the overall complexity of this specific cancer genome. Interestingly, chromosomal arm loss appeared to be the predominant feature of this tumour, with more than 30% of the haploid genome being affected by a decrease in copy number. This associated with a prevalence of inter-chromosomal rearrangements, suggesting chromosomal translocations as a preferred consequence of genomic instability. Here we describe some examples of somatic translocation events causing the loss of tumour suppressor genes. Using this genotype as a possible model of ovarian carcinoma evolution, we aim to explain a subset of ovarian cancers displaying a similar chromosomal profile by applying an analysis of copy number variations, and to define common mechanisms of cancer gene loss.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5107. doi:1538-7445.AM2012-5107
Abstract 5118: A functional genetic approach in patient-derived glioblastoma stem cells reveals pre-mRNA splicing components to be cancer-lethal gene targets
Authors: Hubert Christopher G. , Ding Yu , Toledo Chad , Paddison Patrick J. , Olson James M. , Berndt Jason
Published: August 2012

Glioblastoma multiforme (GBM) is the most aggressive and common form of brain cancer in adults and is among the deadliest cancers with a median survival period of 12-14 months. This poor prognosis despite aggressive therapy underscores the need for novel therapeutic targets specifically required by GBM cells. Many GBM are thought to arise from a neural stem cell (NSC) origin and, consistent with this premise, tumor-initiating GBM stem cells (GSCs) isolated from patients retain the NSC-like phenotype and molecular profile of primary tumors. Importantly, unlike serum-cultured cell lines, GSCs retain the developmental potential and specific genetic mutations acquired as each patient's tumor progressed from its cell of origin. We hypothesized that these genetic alterations driving GBM growth might also give rise to unique molecular vulnerabilities within the cancer cells. To identify such novel gene targets required for GBM cell growth, but which are dispensable to normal cells, we performed genome-scale RNAi screens in multiple patient-derived GSC isolates and simultaneously counter-screened against primary untransformed human NSCs. From these results, we identified and validated the existence of GBM-lethal genes that, when inhibited, render patient GSCs sensitive to cellular stresses arising within these transformed cells. From these targets, we show that GSCs have an increased requirement for the expression and function of multiple specific members of the pre-mRNA splicing machinery. Notably, loss of specific key splicing proteins resulted in cell cycle arrest and subsequent cell death only in GSCs, identifying the spliceosome as a specific molecular vulnerability in GBM. New treatment strategies for this disease are urgently needed. The identification of spliceosomal proteins as essential for the growth and maintenance of GSCs both adds to our understanding of glioblastoma biology and suggests novel targets for therapeutic intervention.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5118. doi:1538-7445.AM2012-5118
Abstract 5131: Identification of the genes that chemosensitize hepatocellular carcinoma cells to interferon-α and 5-fluorouracil therapy
Authors: Shiota Goshi , Sakabe Tomohiko , Nagano Hiroaki , Tsuchiya Hiroyuki
Published: August 2012

The increasing numbers of patients with hepatocellular carcinoma (HCC), which is characterized by an extremely poor prognosis, are causing serious problems worldwide. To improve their prognosis, the development of a more effective therapy is required. Combination therapy with interferon-α (IFN-α) and 5-fluorouracil (5-FU) was reported to provide benefits to patients with advanced HCC, although its efficacy is still unsatisfactory. In this study, taking advantage of functional screening, three chemosensitizing genes for this therapy against HCC cells were identified. Identification of chemosensitizing genes was performed using ribozyme-based functional screening. We constructed a plasmid DNA library expressing random ribozyme genes with ∼6.0 × 106 target recognition sequences. We performed a positive-selection strategy based on the ability of 5-FU to eliminate cells expressing a ribozyme that targets chemosensitivity-unrelated genes. The ribozyme library was transfected into HepG2 cells, which were subsequently treated with the IC50 dose of 5-FU. This cycle was repeated 10 times to fully concentrate the ribozymes targeting those genes involved in the sensitivity of 5-FU. Consequently, 100 target ribozyme recognition sequences that were recovered before screening and after 10 cycles of screening were subjected to a BLAST search to retrieve genes with homologous sequences. This analysis indicated 3 genes (PRKAG2, TGFBR2, and EXT1) as candidate genes. The chemosensitizing effects of these genes on IFN-α/5-FU as well as 5-FU treatment were confirmed by adenovirus-mediated gene transfer. In addition, the mechanisms of effects of these genes on enhancing effects on combination therapy with IFN-α/5-FU were also evaluated. Furthermore, to evaluate the relationship between their expression levels in tumor tissues and the clinical outcome of IFN-α/5-FU combination therapy in advanced HCC patients, mRNA expression was measured using real-time reverse transcription-polymerase chain reaction. As a result, PRKAG2, TGFBR2, and EXT1 were identified as novel chemosensitizing genes for the combination therapy against HCC. TGFBR2 and EXT1 induced transforming growth factor-β signal activation and endoplasmic reticulum stress, respectively. Moreover, PRKAG2 and TGFBR2 were suggested to be promising predictive markers for the clinical outcome of this therapy. In conclusion, the identified genes are promising candidates as targets to enhance the therapeutic effect of IFN-α/5-FU combination therapy in advanced HCC and could be used as predictive markers for response to this therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5131. doi:1538-7445.AM2012-5131
Abstract 5135: Functional genomic shRNA screens reveal that the canonical Wnt pathway protects lung cancer cells from EGFR inhibition
Authors: Casas Matias
Published: August 2012

Lung cancers are the leading cause of cancer deaths globally. In particular, non-small cell lung cancer (NSCLC) is a devastating disease, and is rarely curable. Hyperactivation of mitogenic signaling cascades is a hallmark of cancer development, including for lung cancers. Targeted therapies inhibiting specific receptor tyrosine kinases have shown clinical promise, but rarely produce complete responses and are not curative, suggesting the existence of escape mechanisms promoting cell survival. Using a loss-of-function, whole genome shRNA screen, we identified various components and mediators of the canonical Wnt pathway that contribute to the maintenance of NSCLC cells during inhibition of the epidermal growth factor receptor (EGFR). Modulation of tankyrases, casein kinases and different components of the Wnt pathway with shRNAs or small molecules, in vitro and in vivo, significantly increased the efficacy of EGFR inhibitors, thus revealing a critical role for the canonical Wnt pathway in maintaining lung cancer cells during EGFR inhibition. Targeting the Wnt/β-catenin pathway together with EGFR inhibition in the clinic could lead to more durable remissions for non-small cell lung cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5135. doi:1538-7445.AM2012-5135
Abstract 5138: Development of a high-throughput screen for breast cancer metastatic potential
Authors: Harris Janelle L. , Khanna Kum Kum
Published: August 2012

Metastasis is a frequent cause of cancer-associated morbidity and mortality. Identification of genetic signatures or expression profiles which are associated with metastasis typically involves microarray analysis or genomic sequencing of isolated metastases, either in a clinical setting or animal models. In order to explore the contribution of individual genes to metastasis, we have developed an in vitro culture method for preferential expansion of breast cancer cells with high metastatic potential. Growing breast cancer cells in low growth factor, anchorage independent conditions maintains cells with low metastatic potential in a dormant or non-proliferative state while cells with more aggressive characteristics rapidly proliferate to form ascini. Low-throughput manual testing of this system is currently being carried out using two ‘dormant’ cell lines (PMC 42 LA and MCF-7) and one aggressive metastatic line (MDA MB 231) and ectopic expression of a subset of genes which we anticipate could promote metastasis (PTK2, MYLK, ROCK1, S100A4, Rad51 and ID1). In the high-content genetic screens, genes which cause a significant change in the number or size of ascini will be identified and validated in culture manually as well as in mouse models of breast cancer metastasis. The genetic data derived from these screens will identify genes which promote metastasis, yielding mechanistic information about the progress of metastatic disease. It will also provide convenient platform for testing therapeutic strategies to maintain micrometastases in a dormant state and prevent the onset of clinically relevant metastatic disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5138. doi:1538-7445.AM2012-5138
Abstract 5143: ErbB2 activation up-regulates glutaminase 1 expression via NF-κB pathway
Authors: Qie Shuo , Chu Clarissa , Li Weihua , Wang Chenguang , Reginato Mauricio , Sang Nianli
Published: August 2012

Active glutamine utilization is essential for cell proliferation in many tumors, for it provides critical carbon and nitrogen sources. Glutaminolysis represents the first and rate-limiting step of glutamine utilization and is catalyzed by glutaminase. Previous studies have shown that c-Myc regulates glutaminolysis by increasing glutaminase expression in tumor cells. However, it remains unclear whether other oncogenic signaling pathways promote glutaminolysis. Breast cancer is the second most common cause of death for women in the United States and ErbB2 activation is one of the major causes of breast cancer. Using MCF10A and MCF10A-derived NeuT cells, we studied the effect of ErbB2 activation on glutaminase expression, and found that ErbB2 activation increased glutaminase 1 expression at both mRNA and protein levels. Knockdown of ErbB2 decreased glutaminase 1 expression in several human ErbB2-positive cell lines. Consistently, blocking ErbB2 signaling pathway by trastuzumab repressed glutaminase 1 expression. We further showed that in these cells, ErbB2-mediated up-regulation of glutaminase 1 was independent of c-Myc expression. In addition, we found that activation of PI3K/Akt or MAPK pathway was not sufficient to up-regulate glutaminase 1 expression. Instead, inhibition of NF-κB down-regulated glutaminase 1 expression whereas stimulation of NF-κB induced glutaminase 1 expression, suggesting a PI3K/Akt-independent activation of NF-κB signaling pathway up-regulates glutaminase 1. Finally, inhibition of glutaminase activity significantly decreased human breast cancer cell proliferation. Our data indicate that ErbB2 activation promotes glutaminase 1 expression via NF-κB in breast cancer cells, identifying another oncogenic signaling pathway which stimulates glutamine utilization. These findings may facilitate the identification of novel targets for cancer therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5143. doi:1538-7445.AM2012-5143
Abstract 5156: Hyperglycemia enhances IGF-I responses and alters the AMPK activation profile that protects against metformin-induced growth inhibition in pancreatic cancer cells
Authors: Karnevi Emelie , Said Katarzyna , Andersson Roland , Rosendahl Ann H.
Published: August 2012

Epidemiological studies have shown direct associations between type 2 diabetes (T2D) and obesity, both conditions associated with hyperglycemia and hyperinsulinemia, and the risk of pancreatic cancer. Up to 80% of pancreatic cancer patients are diagnosed with either impaired glucose tolerance or new-onset T2D one or two years prior to their cancer diagnosis. Recent population studies indicate that the incidence of pancreatic cancer is reduced among diabetics taking metformin, an insulin-sensitizing and glucose lowering drug. We investigated the effects of exposure of pancreatic cancer cells to high glucose levels on their growth and response to metformin. We found that metformin significantly reduced the proliferation of pancreatic cancer cells under normal glucose conditions, but that hyperglycemia protected against the metformin-induced growth inhibition. The antiproliferative actions of metformin were associated with an activation of AMP-activated protein kinase AMPK(T172) together with a dose-dependent suppression of the insulin/insulin-like growth factor (IGF) signaling mediators IRS-1 and phosphorylated AKT. Furthermore, exposure to metformin during normal glucose led to increased apoptosis as measured by poly(ADP-ribose) polymerase (PARP) cleavage. In contrast, exposure to high glucose promoted a more robust IGF-I response and AKT activation, that stimulated AMPK(S485) phosphorylation and suppressed AMPK(T172) phosphorylation resulting in reduced antiproliferative and apoptotic effects by metformin. In conclusion, our results indicate that metformin has direct antitumor activities in pancreatic cancer cells involving AMPK(T172) activation and modulation of the insulin/IGF signaling pathways. However, hyperglycemic conditions enhance the insulin/IGF-I responses resulting in an altered AMPK activation profile that prevent metformin from fully switching of the growth promoting signals in pancreatic cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5156. doi:1538-7445.AM2012-5156
Abstract 5159: Investigation of local vitamin D metabolism in breast cancer using an ex vivo tissue explant system
Authors: Suetani Rachel J. , Ho Kristen , Jindal Shalini , Neilsen Paul M. , Gill Grantley , Morris Howard A. , Callen David F.
Published: August 2012

Vitamin D is an essential molecule obtained either by UV irradiation of the skin, or ingested through diet or supplementation. Sequential hydroxylations of vitamin D are required to generate the inactive precursor 25-hydroxyvitamin D (25D), and the active metabolite 1,25-dihydroxyvitamin D (1,25D) which activates the vitamin D receptor to alter transcription of target genes. In addition to its role in calcium homeostasis, vitamin D has been shown to have anti-proliferative, apoptotic and immunomodulating effects. Furthermore, many epidemiological studies have shown a link between a low vitamin D status and an increased risk of breast cancer. However, the mechanistic basis of this link remains unknown. We have utilized a breast explant system whereby fresh tumour and matched non-malignant breast tissues are obtained from women undergoing mastectomies. These tissues are placed on gelatin sponges in tissue culture media supplemented with either 25D or 1,25D. Tissues are then collected and analysed for alterations in gene expression and biological response. This system allows for the analysis of dynamic changes in viable tissue in response to exogenously applied compounds, and can easily be used to examine other compounds of interest. Explanted non-malignant and tumour tissue both displayed an increase in expression of the vitamin D metabolizing enzyme CYP24A1 when directly treated with 1,25D. In response to the precursor molecule 25D, non-malignant breast tissue showed a similar pattern of enzyme induction, indicating an intact and competent vitamin D metabolic pathway in these tissues. Metabolism in breast tumour tissue however was somewhat disrupted, with some cases showing little potential in converting 25D to the active metabolite 1,25D. Proliferation markers have also been examined in the non-malignant and tumour breast samples. Interestingly, some tumour specimens did not show any change in proliferation despite favourable changes in gene expression, indicating further dysregulation of vitamin D metabolism in these samples. These data will have implications for any future clinical studies of vitamin D supplementation to complement current chemopreventative therapies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5159. doi:1538-7445.AM2012-5159
Abstract 5164: Using metabolomic flux to uncover new targets for the modulation of breast cancer metastasis
Authors: Van Wassenhove Lauren D. , Wynn Michelle L. , Ashrafzadeh Sepideh , Merajver Sofia D.
Published: August 2012

Inflammatory breast cancer (IBC) is the most deadly breast cancer because of its ability to rapidly metastasize, often before the primary lesion is detected. To better understand what drives this highly aggressive form of cancer, we studied central carbon metabolism in IBC. The Warburg effect, which is characterized by high rates of glucose uptake and glycolysis even under aerobic conditions, occurs in most cancer cells and plays an important role in the tumorigenic capacity of breast cancer. Specifically, we examined metabolic changes that occur when breast cancer cells switch from a proliferating phenotype to a highly motile phenotype. In primary tumor formation rapid proliferation is crucial, while in metastasis motility to new sites is critical. We hypothesize that alterations in the regulation of metabolic pathways leads to the direction of energy from proliferation to motility, and that this change is important in the progression of breast cancer towards metastases. Understanding if a coincident metabolic shift occurs when malignant cells switch from a proliferative to a more aggressive motile form may help identify new therapeutics for highly aggressive breast cancers. For our studies, we are using “normal-like” MCF-10A breast cells, which proliferate quickly and move slower than cancer cells, and two highly metastatic breast cancer cell lines (MDA-MB-231 and the IBC-derived line SUM 149), which have the ability to move quickly and proliferate relatively slowly. Using these cell lines as a model, we have examined metabolic changes as cancer progresses. We are conducting targeted metabolomic studies to determine relative concentrations of metabolites in the glycolysis, tricarboxylic acid (TCA) cycle, and mevalonate pathway. In addition, we are using 13C-labeled pyruvate and glucose to measure the transient and steady state flux through these metabolic pathways. Our preliminary results indicate interesting and unexpected changes in both the transient and steady state metabolic fluxes across the different cell lines. In addition, the metabolic flux through the TCA cycle is very active in the metastatic cancer cells despite an increase in glycolysis. In agreement with flux studies of other cancer cell lines, our results suggest that the cancer cells are likely using intermediates from the TCA cycle to generate nucleotides and fatty acids needed for replication. The results from this work, once validated, can be used to identify new therapeutic metabolic targets to modulate breast cancer metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5164. doi:1538-7445.AM2012-5164
Abstract 5166: Tumor adiponectin signaling underlies the association between obesity and lethal outcome in prostate cancer
Authors: Martin Neil E. , Gerke Travis , Loda Massimo , Stampfer Meir , Finn Stephen , Rider Jennifer , Mucci Lorelei A.
Published: August 2012

Background: Obesity is associated with poor outcomes in men with prostate cancer. The biologic drivers of this relationship are poorly understood. Adiponectin is produced by adipose tissue and has been shown to regulate multiple pathways including inflammation, angiogenisis and insulin sensitivity. Low circulating levels are associated both with obesity and with lethal prostate cancer. Here we investigate whether adiponectin signaling has a differential effect on cancer outcomes in the tumors of obese compared to healthy weight men. Methods: The study cohort consisted of men diagnosed with prostate cancer from the Physicians’ Health Study (1982-2004) and the Health Professionals Follow Up Study (1986-2004) for whom archival tumor specimens were available. Follow-up data for development of metastases and cancer-specific mortality (defined as lethal prostate cancer) was available through 2010. Gene set analyses was undertaken for 116 men using gene expression profiling data from cDNA-mediated annealing, selection, extension, and ligation (DASL) assay of 6000 genes. Genes differentially expressed between lethal (n=30 lethal events) and indolent disease among the obese and healthy weight men were explored. Immunohistochemistry for adiponectin receptor 2 (ADIPOR2) protein expression in tumors and adjacent normal was performed on tissue microarrays representing tumor tissue from 1017 patients (n=98 lethal events). Anthropometric measures were obtained from validated questionnaires. Results: Gene set analysis identified distinct gene sets that were enriched in the comparison of lethal to indolent among the obese and healthy weight men. In the obese men, five gene sets were enriched in the lethal cases with a false discovery rate <0.1. The second most enriched set associated with lethality was the KEGG adiponectin signaling pathway. We found that protein expression of ADIPOR2 in prostate tumors, but not normal prostate, was higher among obese compared to healthy weight men. Moreover, high expression of ADIPOR2 protein expression was associated with a 3-fold greater risk of lethal outcomes. Conclusion: Tumor adiponectin signaling appears to be key pathway underlying lethal outcomes in obese men with prostate cancer. This work supports the hypothesis that there are unique drivers of poor outcome among obese men with prostate cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5166. doi:1538-7445.AM2012-5166
Abstract 5171: Restoration of fibronectin matrix assembly impedes invasion and reduces tumor-stromal cell affinity in prostate cancer
Authors: Jia Dongxuan , Entersz Ildiko , Butler Christine , Vaca Connan , Foty Ramsey A.
Published: August 2012

Cell-cell and cell-ECM interactions are important regulators of cohesion, a property previously demonstrated to mediate malignant invasion. Intermixing of the epithelial and stromal compartments is often associated with increased invasive capacity and poor prognosis. The studies reported here propose a novel role for α5β1 integrin, the principle mediator of fibronectin matrix assembly (FNMA), as an invasion suppressor in prostate cancer cells. Employing a combination of biophysical and cell biological techniques we explore the relationship between cohesion, invasion, FNMA and tumor-stromal interactions, using a well-characterized prostate cancer model. We show that cohesion is inversely proportional to invasive capacity. We also show that the most invasive cells are deficient in FNMA and express reduced levels of α5β1 integrin. We generated cells over-expressing either wild-type α5 integrin or a non-functional α5 in which the cytoplasmic domain was replaced with that of α2. We show that only wild-type α5 was able to increase cohesion and reduce the invasiveness of aggressive cells. In order to further explore the influence of FNMA on tumor-stromal cell interaction we employed a liquid miscibility model in which interactions between tumor and stroma are rooted in the same thermodynamic principles that govern the mixing of simple fluids. In this model, whether tumor and stromal cells segregate or intermix depends on the ratio of the strength of self-cohesion to cross-adhesion; a high ratio corresponds with segregation, whereas a lower ratio denotes intermixing. We hypothesized that segregation between tumor and stroma can be promoted by increasing the strength of tumor cell cohesion. To test this hypothesis, we first showed that aggressive prostate cancer cells intermix with prostate fibroblasts. We then treated the cancer cells with MEK inhibitors (MEKi) and showed that treatment restored FNMA, leading to increased cell-cell cohesion. When mixed with prostate stromal cells, MEKi-treated cells segregated, suggesting that increasing cohesion between tumor cells is sufficient to reduce their affinity for stromal cells. These data suggest that by increasing tumor cell cohesion through a fibronectin matrix mediated adhesion mechanism, it may be possible to effectively reduce affinity of tumor cells for stroma, thus inhibiting invasion.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5171. doi:1538-7445.AM2012-5171
Abstract 5172: The role of vimentin in FAK-mediated cell adhesion and metastasis
Authors: Havel Lauren S. , Marcus Adam
Published: August 2012

Lung cancer is the leading cause of cancer death in the United States. Specifcally, non-small cell lung cancer (NSCLC) is responsible for about 80% of lung cancer deaths and has a 5-year survival rate of only 15-25%. This poor survival statistic results from diagnosis at advanced stages of the disease when metastasis of the primary tumor has already occurred; therefore, identifying the molecular mechanisms guiding metastasis is crucial for advances in treatment. The intermediate filament vimentin is overexpressed in metastatic cancers and its expression correlates with increased invasive capacity and reduced patient survival. A population of vimentin filaments localizes to focal adhesions, which attach to the extracellular matrix and are required for cell motility. Focal adhesions are composed of a complex of proteins, including focal adhesion kinase (FAK), which is overexpressed in NSCLC. Since evidence suggests that both FAK and vimentin co-localize at focal adhesions, we hypothesize that vimentin acts through FAK to regulate cell adhesion and thereby cell migration. Our results show that vimentin depletion leads to decreased cell adhesion and migration, with decreased FAK activation and expression. We show that vimentin filaments directly enter total FAK and pFAK- Y397 sites at the cellular leading edge of motile lung cancer cells. Using immunoprecipitation, we show vimentin interacts with several members of the focal adhesion complex, suggesting that vimentin can regulate focal adhesion activity. To address the mechanism by which this may occur, we hypothesize that the phosphorylation state of vimentin, which is important for cell migration, is crucial for proper functioning at focal adhesions. This is the focus of our future work with the goal of identifying potential new therapeutic targets to block metastasis and improve lung cancer patients’ prognosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5172. doi:1538-7445.AM2012-5172
Abstract 5175: Discovering a novel role of claudin-7 as a basolateral protein maintaining cell-matrix interactions in human lung cancer cells and in mouse intestines
Authors: Lu Zhe , Ding Lei , Lu Qun , Chen Yan-Hua
Published: August 2012

The ability to invade host tissues and metastasize is the major cause of cancer-related death. During tumor invasion, metastasizing cells disrupt normal cell-matrix adhesion and acquire an invasive phenotype. Claudins are adhesion proteins localized at tight junctions (TJs). Claudin-7 is a unique TJ membrane protein in that it has a stronger basolateral membrane distribution than that of apical TJs in epithelial cells. To study the basolateral function of claudin-7, claudin-7 gene silencing experiments were carried out in a lung cancer cell line using the lentivirus shRNA approach. We found that claudin-7 knockdown (KD) cells showed disrupted cell-matrix interactions. Consequently, when claudin-7 KD cells were plated on the uncoated glass surface, they were unable to attach to the glass and died the day after plating. In contrast, control cells adhered well and grew normally. Using immunofluorescent microscopy and biochemistry methods, we found that claudin-7 co-localized and co-immunoprecipitated with integrin β1 at the basolateral compartment. Knockdown of claudin-7 by lentivirus claudin-7 shRNA downregulated integrin β1 at both mRNA and protein levels. This cell-matrix defect can be rescued by transfecting claudin-7 or integrin β1 back to the claudin-7 KD cells. Suppression of claudin-7 also upregulated matrix metalloproteinase-3 (MMP-3) at both mRNA and protein levels and downregulated several collagens, such as type V α1, type XV α1, and type XVI α1, at the mRNA level. We also generated claudin-7-deficient (Cldn7−/−) mice using targeted-gene deletion method. Cldn7−/− mice displayed severe intestinal defects including attenuated villi, mucosal ulcerations and epithelial cell sloughing. Using immunofluorescent microscopy and biochemistry methods, we found that claudin-7 co-localized and co-immunoprecipitated with integrin α2 in Cldn7+/+ mouse intestines where claudin-7 was highly expressed. Deletion of claudin-7 reduced integrin α2 expression and altered its localization. Electron microscopy revealed significant intercellular gaps below TJs and the cell-matrix loosening in Cldn7−/− intestines. Thus, our current study highlights a novel non-TJ function of claudin-7 in maintaining epithelial cell-matrix interactions by interacting with integrins.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5175. doi:1538-7445.AM2012-5175
Abstract 5176: Imaging αv integrin-GFP during tumor blood vessel interaction in the chick CAM
Authors: Tome Yasunori , Sugimoto Naotoshi , Yano Shuya , Momiyama Masashi , Maehara Hiroki , Tsuchiya Hiroyuki , Tomita Katsuro , Kanaya Fuminori , Hoffman Robert M.
Published: August 2012

In this study, we describe imaging of αv integrin-GFP in cancer cells interacting with blood vessels. Osteosarcoma cells (143B) (5×106) expressing αv integrin-GFP were injected in the 10-day old chick chorioallontoic membrane (CAM). In vivo imaging with the Olympus OV-100, visualized tumor-induced angiogenesis one week after transplantation. Osteosarcoma cells expressing αv integrin-GFP were aligned along the tumor-induced blood vessels. High resolution ex vivo imaging with the Olympus FV1000 confocal microscope showed narrow capillary vessels surrounded by osteosarcoma cells which strongly expressing αv integrin-GFP compared to those cells far from vessels. This result imply that expressed of osteosarcoma αv integrin is related to the interaction of the osteosarcoma cells and blood vessels. The ability to image αv integrin during interaction of cancer cells and blood vessels will enable the further understanding of its role in this process.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5176. doi:1538-7445.AM2012-5176
Abstract 5177: Molecular targeting of αvβ3 integrin by echistatin inhibits tumor progression and metastasis in osteosarcoma
Authors: Tome Yasunori , Kimura Hiroaki , Maehara Hiroki , Sugimoto Naotoshi , Tomita Katsuro , Tsuchiya Hiroyuki , Kanaya Fuminori , Bouvet Michael , Hoffman Robert M.
Published: August 2012

Expression of αvβ3 integrin is associated with tumor progression and metastasis in several types of cancer, including metastatic osteosarcoma. We tested echistatin, an antagonist of αvβ3 integrin, as a molecular targeting drug, in an imageable orthotopic mouse model of human metastatic osteosarcoma. αvβ3 integrin expression was measured in a series of human osteosarcoma cell lines (143B parental and LM1-LM4) with increasing metastatic potential, using RT-PCR and immunoprecipitation. The cell lines expressed GFP in the nucleus and RFP in the cytoplasm and therefore could be imaged down to the subcellular level in vitro and in vivo. αvβ3 integrin expression correlated with the metastatic potential of the cells, with the LM4 cells showing the highest expression and highest metastatic capability, 6-fold and 18-fold, respectively, compared to parental 143B cells. In vitro cell proliferation of LM4 was inhibited by echistatin (p<0.01). In vitro migration, invasion, and adhesion of LM4 were also inhibited by echistatin (p<0.01). Tumor-induced angiogenesis by LM4 in the chick CAM model was also inhibited by echistatin (p<0.05). Mice treated with an echistatin-doxorubicin conjugate had significantly smaller primary tumors and longer survival compared to control untreated tumor-bearing mice (p<0.05 and p<0.01, respectively). Treatment with echistatin resulted in decreased experimental pulmonary metastases, imaged at the cellular level, in a nude mouse model compared to the untreated control (p<0.01). The results of this study indicate that αvβ3 integrin plays an essential role in osteosarcoma metastasis and that echistatin has potential to target αvβ3 integrin and inhibit tumor progression and metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5177. doi:1538-7445.AM2012-5177
Abstract 5178: Evaluation of lumican expression profiles in prostate cancer
Authors: Yoshimura Kazuhiro , De Velasco Marco A. , Hatanaka Yuji , Yamamoto Yutaka , Kodama Mitsumasa , Tanaka Motoyoshi , Shimizu Nobutaka , Nozawa Masahiro , Yoshikawa Kazuhiro , Nishio Kazuto , Uemura Hirotsugu
Published: August 2012

The transcription factor, Stat3, has been implicated in the growth and progression of several cancer types including lung skin, liver, ovarian, kidney, colon cancer and prostate. Stat3 has also been shown to induce the metastatic behavior of prostate cancer cells in vitro and in vivo. Activation of Stat3 occurs by the binding of various cytokines to its receptors leading to the activation of the JAK/Stat3 signaling pathway. Of these cytokines, interleukin-6 (IL-6) has been implicated in regulating growth of various malignant tumors and activated IL-6 has been shown to be elevated in the sera from patients with metastatic prostate cancer. In the present study we examined the IL6-induced Stat3 transcriptional activation in the development and progression of prostate cancer and its possible role as a target for the treatment of prostate cancer using the prostate-specific conditional targeting PTEN knockout mouse model. Homozygous PTEN-mutants develop prostate cancer in a stage-specific manner and have the ability to grow and develop castration-resistant prostate cancer after total androgen ablation by castration. ELISA assays revealed increased expression of soluble IL-6 receptor (IL-6R) in tissue lysates from prostate tumors in non-castrated and castrated mice (77.0 ± 6.17 vs. 23.75 ± 5.17 ng/g of tissue, p<0.001, respectively) at 32 weeks of age. Immunohistochemical analysis revealed correlated expression of with Stat3 transcriptional activation, proliferation and androgen receptor in tumors from castrated mice. To determine the role of IL-6R induced transcriptional activation of Stat3, non-castrated and castrated tumor bearing mice were treated with an anti-mouse IL-6R antibody (MR-16). Mice treated with MR-16 showed downregulation of phosphorylated Stat3, however, treatment failed to inhibit tumor development and cellular proliferation. However, it was discovered that therapeutic doses of MR16 treatment resulted in increased phosphorylation of Erk1/2. Our data suggests that inhibition of IL-6R-induced Stat3 activation alone is insufficient to suppress tumor growth; however, further studies need to be conducted to evaluate the efficacy of horizontal combination strategies targeting Stat3 and MAP kinase signaling cascades

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5178. doi:1538-7445.AM2012-5178