bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-12: Inhibition of the proliferation and invasion of hepatocellular carcinoma cells by lipocalin 2 through blockade of JNK and PI3K/Akt signaling
Authors: Lee Eun-Kyoung , Kim Hyun-Jun , Lee Kong-Ju , Lee Ho-Jeong , Kim Dae-Ghon , Hong Soon-Won , Yoon Yeup , Kim Jang-Seong
Published: February 2011

Lipocalin 2 (Lcn2) has long been regarded to induce cellular proliferation because expression of Lcn2 has been associated in a variety of proliferative cells. Consistent with this, it has been demonstrated in the present study that Lcn2 was significantly elevated in human HCC (hepatocellular carcinoma) tissues compared with the non-tumor liver tissues. However, the roles of Lcn2 in hepatocarcinogenesis are far from clear. To investigate the effects of Lcn2 expression on the hepatocarcinogenesis, Chang liver and SK-Hep1, the HCC cell lines, were genetically manipulated to express Lcn2 and their effects on the proliferation and invasion of HCC cells were analyzed. Ectopic expression of Lcn2 in HCC cells significantly inhibited the growth of HCC cells in vitro and in vivo. In addition, its expression reduced the invasive potential of cells and inhibited the expression of matrix metalloproteinase 2 (MMP-2). Lcn2 may exert its function partly through the inhibition of the c-Jun N-terminal kinase (JNK) and phosphatidyl inositol 3′-kinase (PI3K)/Akt signaling pathways in HCC cells, since selective inhibition of those pathways using pharmacological inhibitors significantly inhibited the proliferation, invasion and MMP-2 expression, whereas Lcn2 expression suppressed the JNK and PI3K/Akt pathways. In conclusion, these results clearly indicate that Lcn2 may play a protective role in the progression of HCCs by suppressing the proliferation and invasion of HCC cells, although clinical significance of the present findings remains to be evaluated.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-12.
Abstract LB-126: A pooled shRNA screen for erlotinib sensitizers in Kras-mutant non-small cell lung cancer
Authors: Cordes Shaun , Dompe Nick , Lee James , Li Li , Rivers Celina , Tang Zhijun , Luk Wilman , Bennett Greg , Shames David , Modrusan Zora , Davis David P.
Published: February 2011

Erlotinib is a small-molecule inhibitor of the epithelial growth factor receptor (EGFR) that provides a survival benefit in patients with chemoresistant non-small cell lung cancer (NSCLC). In the second and third line setting, patients whose tumors harbor activating mutations in Kras showed less benefit than patients with tumors harboring wild type Kras.

To identify genes whose inhibition sensitizes Kras-mutant NSCLC cells to erlotinib, we performed a whole-genome pooled-shRNA screen and identified 337 shRNAs that were selectively depleted in erlotinib-treated cells. We subsequently validated 148 of the top 200 hits in an siRNA-based viability assay and selected 37 hits for further analysis. We are currently characterizing these hits in a panel of ∼10 additional NSCLC lines to identify targets with broad cell line activity that synergize with erlotinib to induce apoptosis. Our current understanding of the relationships between our top hits and erlotinib sensitivity will be discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-126.
Abstract LB-129: CAFET algorithm reveals WNT/PCP specific signature in squamous cell carcinoma of lung cancer
Authors: Hu Yue
Published: February 2011

We analyzed the global gene expression patterns of 138 lung cancer samples containing Squamous Cell Carcinoma (SCC) and Adenocarcinoma (AC). Gene set enrichment analysis revealed developmental and cell cycle functional groups were specifically overexpressed in SCC samples. To refine and focus our analysis on specific signaling pathways, we developed a complementary algorithm called Coverage Analysis with Fisher Exact Test (CAFET). In our analysis, the WNT signaling pathway had the most significant enrichment of overexpressed genes in a cluster of predominantly SCC. Most interestingly, the differential expression pattern in this cluster had a strong tendency to inhibit the WNT canonical pathway while enhancing the WNT/PCP pathway. The non-canonical WNT/PCP pathway is obligatory for cell migration during embryogenesis, and it has also been proposed to be involved in cancer metastasis. While the WNT canonical pathway has been reported to be important in metastasis of AC in lung cancer, our results suggest SCC of lung makes use of a different branch of the WNT pathway for survival and development. This WNT/PCP signature for lung SCC was also confirmed in several independent sample sets. It is also present in some cell lines derived from lung SCC. To find potential clinical biomarkers, we identified 174 genes present in blood and highly expressed in samples carrying this WNT/PCP signature. These results demonstrate that the new algorithm CAFET is a promising approach for mining large gene expression data sets to identify pathways with differential activity, and could be particularly useful in exploring cancer expression datasets.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-129.
Abstract LB-161: Colospheres: 3D ex vivo microtumors with more aggressive phenotype than original human colon cancer tissues
Authors: Weiswald Louis-Bastien , Massonnet Gérald , Richon Sophie , Guinebretiere Jean-Marc , Bellet Dominique , Bieche Ivan , Dangles-Marie Virginie
Published: February 2011

The ex vivo colospheres are a newly characterised three-dimensional colon cancer multicellular model, derived from mechanically dissociated human colon cancer tissue (Br J Cancer 2009, 101:473-82). We have previously demonstrated that this short term culture model is exclusively formed by cancer cells and associated with tumor aggressiveness.

To further investigate the potential interest of colospheres as micrometastasis or cancer stem cell model, we used 3 human colon cancer xenografts directly established from 3 patient colon adenocarcinoma in Nude mice (Cancer Res 2007, 67:398-407). These xenograft tissues are able to give rise to numerous colospheres in only 3 days after mechanical dissociation. This approach allows working with reproducible biological tumor material, in which all human part is known to be cancer cells.

Xenograft tissues and derived colospheres were collected for gene expression study. Expression of 78 genes, involved in stemness, proliferation, epithelial-to-mesenchymal transition and metastasis, has been studied using real-time RT-PCR. All the probes used are human specific and do not cross with mouse genome. Consequently, the comparison of gene expression profiles corresponds to the comparison of gene expression in colosphere-forming cells versus that in the cancer cell counterpart from xenograft tissue.

Gene expression clustering analysis clearly showed that for the 3 colon adenocarcinoma, colospheres matched with their parent xenografts. This first point is important because it demonstrated 1) the lack of ex vivo culture artefact (which would have led to classification into 2 groups: all xenografts opposed to all colospheres); 2) the relevance of colospheres for mimicking in vivo tumor cells. Nevertheless, in all 3 pairs xenograft/colospheres, 3 genes were found overexpressed: ALDH1, KLF4 and PLAUR. This overexpression has to be put in line with high tumorigenicity of these colospheres when injected into mice (Br J Cancer 2009, 101:473-82). Gene expression increase will be confirmed at protein level. To gain also information about location of the cells expressing ALDH1, KLF4 and PLAUR, we developed an immunostaining protocol for confocal microscopy and in situ protein detection (BMC Cancer, in revision).

In conclusion, these preliminary data underline the interest of colospheres as ex vivo microtumor model with cancer initiating-cell functions.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-161.
Abstract LB-175: Tumor regression in patients with metastatic synovial sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1
Authors: Paul Robbins F. , Morgan Richard A. , Feldman Steven A. , Yang James C. , Dudley Mark E. , Li Yong F. , El-Gamil Mona , Schwarz Susan L. , Rosenberg Steven A.
Published: February 2011

Discussant: To be announced.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-175.
Abstract LB-183: The predictive value of CYP2D6 genotypes for plasma levels of active tamoxifen metabolites in early breast cancer
Authors: Muerdter Thomas E. , Schroth Werner , Bacchus Liza , Simon Wolfgang , Eichelbaum Michel , Schwab Matthias , Brauch Hiltrud B.
Published: February 2011

Discussion

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-183.
Abstract LB-190: RAF inhibitors transactivate RAF dimers and ERK signaling in a wild-type BRAF-dependent manner
Authors: Poulikakos Poulikos I. , Zhang Chao , Joseph Eric W. , Persaud Yogindra , Pratilas Christine A. , Bollag Gideon , Solit David B. , Shokat Kevan M. , Rosen Neal
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-190.
Abstract LB-196: Treatment with anti-CD137 mAbs causes intense accumulations of liver T cells without selective antitumor immunotherapeutic effects
Authors: Dubrot Juan , Palazon Asis , Milheiro Francisca , Martinez-Forero Ivan , Melero Ignacio
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-196.
Abstract LB-219: Elevated expression of cellular proliferative index MIB-1 in post-Chornobyl papillary thyroid carcinoma with coexisting thyroiditis
Authors: Dinets Andrii , Hulchiy Mykola , Sofiadis Anastasios , Höög Anders , Larsson Catharina , Zedenius Jan
Published: February 2011

Introduction: Most patients with papillary thyroid cancer (PTC) have an excellent prognosis. Ki-67 is marker of proliferation, and proliferative index more than 5% as calculated with the MIB-1 antibody indicates a worse prognosis in PTC. This study was performed in order to investigate and evaluate expression of MIB-1 in a cohort of adult PTC patients who were exposed in their childhood to radioactivity from the Chornobyl accident in 1986.

Materials and Methods: 70 patients (60 females and 10 males, mean age 30,5 (19-39)) operated on for PTC during 2004-2008 were identified for the study. Two groups were established according to diagnosis. Group 1 consisted of patients with PTC only (n=53). Group 2 consisted of patients with PTC and co-existing chronic lymphatic thyroiditis (n= 17). Proliferative activity was measured by immunohistochemistry using monoclonal MIB-1 antibody against nuclear factor Ki-67. A minimum of 2000 tumor cells were calculated and the percentage of positive cells (MIB-1 index) was compared with clinical data.

Results: MIB-1 index was higher in Group 2 (mean 3,65) compared with Group 1 (mean 1,97) (p=0,005). No other clinical features could be correlated with the MIB-1 index. 26% of patients in Group 1 had lymph node metastases, and 23% in Group 2. The mean tumor size was slightly higher in Group 1 (2 cm) compared with Group 2 (1,75 cm). Age at diagnosis (30,5) and age at the Chornobyl accident (11) were identical between the groups.

Conclusions: The clinical features of this cohort of adult PTC patients exposed to the radioactive fallout from the Chornobyl accident seem no different to other PTC cohorts. However, a rather large proportion of patients showed signs of coexisting thyroiditis. Whether the elevated MIB-1 index in this group of patients reflects a worse prognosis is yet not possible to say due to the short follow-up time. Still, a closer follow-up may be justified.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-219.
Abstract LB-225: EGFR and EGFRvIII interact with PUMA to inhibit mitochondrial translocalization of PUMA and PUMA-mediated apoptosis independent of EGFR kinase activity
Authors: Zhu Hu , Cao Xinyu , Keir Steve , Ali-Osman Francis , Lo Hui-Wen
Published: February 2011

Glioblastoma is the most common and most intractable brain malignancy in adults. Patients with glioblastoma have dismal prognosis with a median survival of 12-14 months. This unfortunate poor prognosis is, in part, due to the aggressive nature of these tumors and the lack of effective therapy. EGFR and its constitutively activated variant EGFRvIII are linked to glioblastoma resistance to therapy, the mechanisms underlying this association, however, are still unclear. We report here that in glioblastoma cell lines, xenografts and primary specimens, EGFR and EGFRvIII paradoxically co-express with p53-upregulated modulator of apoptosis (PUMA), a proapoptotic member of the Bcl-2 family of proteins primarily located on the mitochondria. Both EGFR and EGFRvIII bind to PUMA constitutively and under apoptotic stress, and subsequently sequester PUMA in the cytoplasm. The EGFR-PUMA interaction is independent of EGFR activation and is sustained under EGFR inhibition. A Bcl-2/Bcl-xL inhibitor that mimics PUMA activity sensitizes EGFR- and EGFRvIII-expressing glioblastoma cells to EGFR kinase inhibitor, Iressa. Collectively, we uncovered a novel kinase-independent function of EGFR and EGFRvIII that leads to drug resistance of glioblastoma and potentially other tumor types.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-225.
Abstract LB-227: Receptor tyrosine kinase phosphorylation: Simultaneous detection of 10 kinases upon inhibitor treatment
Authors: Henzler Tanja , Pohl Stefanie , Schneiderhan-Mara Nicole , Rimmele Stefanie , Livengood April , Kovelman Robert , Herget Thomas
Published: February 2011

Receptor tyrosine kinases have been shown to be major pharmaceutical targets involved in disease states such as diabetes and cancer. Target driven therapeutics like small molecules or monoclonal antibodies are already in clinical use and have tremendous influence on new cancer therapies. In most cases either ligand binding to the extracellular domain followed by receptor dimerization or ATP binding to the intracellular kinase domain is inhibited. However, it is still challenging to gain specificity of those inhibitors for certain receptor tyrosine kinase targets.

Bead-based assay technologies, such as the xMAP® platform, are ideal tools for the multiplex analysis of 5-20 proteins. Low sample volumes, simple assay protocols, flexibility and robustness are characteristics of bead-based assays that make them well suited for the immunological detection of bio marker and cell signaling proteins.

In this presentation we will show results for the simultaneous detection of the ten most relevant receptor tyrosine kinases. The expression profile of more than 50 cell lines as well as breast tumor samples were evaluated with this powerful multiplex tool in a very robust and convenient way. Inhibitor profiling of small molecules under physiological conditions on ten receptor tyrosine kinases will be demonstrated by the detection of their phospho-Tyrosine level. These specificity profiles will help to improve the drug development process and to obtain potentially better drugs in future.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-227.
Abstract LB-234: Bcl-2, a novel target of Bmi-1 (the stem cell associated factor), induced Wnt signaling: Implications on prostate cancer
Authors: Saleem Mohammad , Siddiqui Hifzur R. , Tarapore Rohinton S. , Kohl Amanda M.
Published: February 2011

In the current report we provide a novel finding about the regulation of Bcl-2 expression in prostate cancer (CaP) cells. Bcl-2 is known to be regulated by Sonic Hedgehog (SHH) signaling. Here, we provide evidence that human CaP cells exhibit SHH-independent Bcl-2 activation which is regulated by Bmi-1-induced Wnt signaling. Bmi-1 is a member of polycomb group gene family and is reported to be involved in stemness and self-renewal capability of cancer stem cells. We investigated the role of Bmi-1 in human CaP development and delineate its mechanism of action. On the basis of immunohistochemical analysis of prostatic specimens of 125 human CaP patients, we show that Bmi-1 protein levels are highly elevated in patients with advanced disease. To understand the mechanism of action of Bmi-1, we employed two-prong strategy. Firstly, Bmi-1 was knocked down in CaP cells (LNCaP, DU145 and PC-3) by employing siRNA technique. Bmi-1-silenced CaP cells exhibited decreased proliferative and clonogenic potential. Secondly, Bmi-1 was over-expressed in CaP cells by transfecting Bmi-1 overexpressing plasmid (pbabe-Bmi-1) in CaP cells. Bmi-1-overexpressing cells exhibited increased clonogenicity and rate of proliferation. We performed a micro-array analysis of Bmi-1-silenced cells and found that silencing of Bmi-1 caused a decrease in the Cyclin D1 (Wnt signaling target) and Bcl-2 (SHH signaling target), however an increase in p16 was observed. These studies were validated by performing immunoblot analysis. We generated a hypothesis that the Bmi-1 regulates the expression of Cyclin D1 and Bcl-2 by interacting with Wnt /SHH signaling in CaP cells. We show that despite blocking SHH signaling (by treating CaP cells with cyclopamine, a SHH signaling inhibitor), Bmi-1 induces the Bcl-2 expression in CaP cells suggesting the involvement of SHH-independent pathway in the regulation of Bcl-2 activation. We identified Wnt signaling as the pathway that regulates Bcl-2 independent of SHH signaling in CaP cells. We provide evidence that Bcl-2 promoter region of Bcl-2 gene has binding sites for Tcf transcriptional factor (Wnt signaling component). We found that Bmi-1 induces Wnt signaling in CaP cells. Taken together, these studies provide deep-insight into the mechanism of CaP cell proliferation (even after chemotherapy such as cyclopamine treatment). We suggest that Bmi-1 could be developed as a novel molecular and therapeutic target for human CaP treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-234.
Abstract LB-237: Interaction between myopodin and integrin linked kinase (ILK) induces tumor suppression
Authors: Yu Yanping , Wang Hui , Luo Jian-Hua
Published: February 2011

Myopodin, a tumor suppressor, is frequently deleted in invasive prostate cancers. Forced expression of myopodin in prostate cancer cells reduced tumor invasiveness and slowed cell motility. In our recent yeast two hybrid system analysis, we found that the N- terminal 326 amino acids of myopodin interacts with integrin linked kinase (ILK). This result was validated by co-immunoprecipitation of myopodin or ILK in cells expressing both proteins. In addition, the binding assays between ILK and myopodin were analyzed in vitro on a series of GST-myopodin deletion mutants with ILK. A binding motif of 78 amino acids in the N- terminus of myopodin was determined to be critical for ILK binding. Phosphorylation analyses through ILK knockdown or mutant assays in vivo and in vitro indicate that phosphorylation of myopodin is ILK dependent. Phosphorylation of myopodin by ILK leads suppression of cell motility and cell growth. Mutant myopodin that does not interact with ILK is not phosphorylated and is defective on suppressing cell motility and cell growth. These findings elucidate a novel signaling pathway of ILK-myopodin that leads to activation of a tumor suppressor.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-237.
Abstract LB-263: Study of Hox gene expression in normal and malignant colonic tissues implicates Hox A4 and Hox D10 in the stem cell-origin of colon cancer
Authors: Bhatlekar Seema
Published: February 2011

Our goal was to identify transcriptional regulatory mechanisms that maintain stem cell (SC) population size in normal colonic epithelium because dysregulation, following acquisition of mutations, leads to tumor SC overpopulation, tumor initiation, and colon cancer. We used microarray analysis to identify a unique gene-expression pattern for a SC-enriched population (the bottom third of the normal human crypt, where colonic SC reside). Two-color microarray analysis (18,500 genes) was used to analyze gene expression in isolated crypt subsections (top [T], middle [M], and bottom [B] thirds). About 25% of known genes were expressed (showed hybridization to RNA from colonic epithelium). Several RNA species (n=287) were preferentially expressed in one of the three crypt subsections (T, M, or B). Six of the known genes up regulated in the bottom third of the crypt mapped to the P13 kinase pathway and two were Hox genes (Hox A4 and D10). Immunohistochemistry (IHC) was then used to analyze expression of Hox A4 and Hox D10 in human colonic tissues. IHC showed that both Hox A4 and D10 are expressed in cells residing at the bottom of the normal crypt and are co-expressed with aldehyde dehydrogenase 1 (ALDH1), a SC marker in these cells. Moreover, IHC showed that Hox A4 and D10 are overexpressed in tumors compared with normal colonic epithelium. These data represent the first gene-expression profiles for sub-regions of the human colonic crypt, especially the SC-rich crypt base, which we now know has a unique gene-expression signature. Co-expression of Hox A4 and D10 with ALDH1 in normal colonocytes suggests that these Hox genes have a role in regulation of the crypt SC population size. Overexpression of Hox A4 and D10 in colon cancer suggests that these Hox genes, when dysregulated, contribute to SC overpopulation and colon tumorigenesis. Identification of the Hox gene-based mechanisms that are involved in the regulation of normal crypt SC population size and in dysregulation of neoplastic SC populations could lead to identification of SC-based targets and ways to develop new, more effective treatments for advanced colon cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-263.
Abstract LB-271: The biological role of the transcription factor Kaiso in breast tumorigenesis
Authors: Weerawardane Sonali , Daniel Juliet M.
Published: February 2011

Introduction: The canonical Wnt signalling pathway regulates various cellular processes (e.g. proliferation, differentiation, apoptosis) and leads to tumorigenesis when aberrantly activated. -catenin, the key downstream effector in Wnt signalling, translocates to the nucleus upon Wnt pathway activation, and activates target genes involved in tumorigenesis. The transcription factor Kaiso is a novel BTB/POZ (Broad complex, Tramtrak, Bric à brac/Pox virus and zinc finger) zinc finger (POZ-ZF) protein that specifically interacts with the cell adhesion cofactor p120ctn. Recent studies from our lab and others have revealed that Kaiso negatively regulates the canonical Wnt pathway by repressing activation of Wnt/ -catenin target genes. Some studies implicate Kaiso as a tumor suppressor (decreased expression in breast, ovarian and colon tumors), and others implicate Kaiso in tumor initiation (Kaiso-deficient mice are resistant to intestinal cancer). The goal of this study is to elucidate the biological role of Kaiso in breast tumorigenesis using various cellular assays.

Methods: To assess Kaiso's role in breast cancer, Kaiso was misexpressed in MCF7 (human breast adenocarcinoma) cell lines. Stable Kaiso overexpressing and depleted cell lines were generated and assessed for cell proliferation and cell migration using focus formation, wound healing and matrigel invasion assays. Western blot analysis was performed to assess the effects of Kaiso misexpression on p120 and E-cadherin as well as epithelial-mesenchymal transition markers such as N-cadherin, Snail, Twist and Vimentin. The subcellular localization of Kaiso and its putative target genes in vivo was examined using immunohistochemistry (IHC) analysis of matched pairs of human normal and tumor breast tissue.

Results: Focus formation assays revealed increased transformation by cells over-expressing Kaiso, suggesting an oncogenic role for Kaiso in cell culture. Preliminary wound healing and matrigel assays show increased cell motility and migration respectively in Kaiso over-expressing cells compared to parental control cells, thereby suggesting a role for Kaiso in the epithelial-mesenchymal transition. IHC analysis revealed predominant nuclear Kaiso expression in lobular and ductal breast carcinomas in breast tissues. These data support Kaiso's potential role as an oncogene in human breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-271.
Abstract LB-278: BRMS1 promoter methylation correlates with decreased expression of BRMS1 in non-small cell lung cancer
Authors: Nagji Alykhan S. , Liu Yuan , Stelow Edward B. , Stukenborg George J. , Jones David R.
Published: February 2011

Background: Breast cancer metastasis suppressor gene-1 (BRMS1) mRNA and protein expression are significantly decreased in non-small cell lung cancer (NSCLC) cell lines and human NSCLC when compared to normal bronchial epithelial cells and adjacent noncancerous tumor tissue, respectively. We hypothesized that one mechanism through which BRMS1 expression was decreased in NSCLC was through BRMS1 promoter methylation.

Materials and Methods: Human NSCLC BRMS1 promoter methylation was evaluated using methylation-specific PCR and bisulfite sequencing. The effect of methylation on BRMS1 mRNA and protein expression was assessed following treatment with 5-azacitidine, a demethylating agent in NSCLC cell lines. BRMS1 transcriptional activity was assessed using a human BRMS1-luc reporter gene construct cloned by our group. Student's T-Test and Chi-square tests were used to determine significance between variables of two patient populations separated based on tumor histology (squamous cell or adenocarcinoma) and to evaluate the methylation profile between the two tumor histologies. Immunohistochemical analysis for BRMS1 protein was performed on both tumor and adjacent noncancerous bronchial epithelium. A general linear model tested the association between BRMS1 promoter methylation in human NSCLC and specific clinical and histopathologic variables.

Results: The BRMS1 promoter region contains a CpG island that was hypermethylated in all NSCLC cell lines and human tumors. This strongly correlated with reduced BRMS1 promoter activity. NSCLC cell lines treated with 5-azacitidine showed a robust recovery of BRMS1 mRNA and protein levels, suggesting that promoter methylation is involved in BRMS1 transcriptional regulation. Analysis of 40 patient NSCLC samples (20 adenocarcinoma and 20 squamous cell) identified a significant increase in the amount of BRMS1 promoter methylation when comparing tumor specimen to adjacent noncancerous tissue for both tumor histologies (p<0.0001). Additionally, the BRMS1 promoter region was significantly more methylated in squamous cell compared to adenocarcinoma histology (p=0.01). Subsequent immunohistochemical analysis confirmed a correlation between BRMS1 promoter methylation and decreased bronchial epithelium BRMS1 protein expression (p=0.06). Moreover, in the squamous cell histology, pathologic tumor stage was associated with elevated BRMS1 methylation (p=0.04).

Conclusions: These studies identify BRMS1 promoter methylation as an important mechanism for silencing BRMS1 expression in NSCLC and highlight the potential clinical relevance of this epigenetic signature with respect to tumor histology and stage.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-278.
Abstract LB-284: Combination of antiandrogens and PI3-kinase pathway inhibitors in a model of androgen-dependent prostate cancer
Authors: Efferson Clay , Zhu Joe , Ware Chris , Tammam Jennifer , Angagaw Minilik , Reilly John F. , Sur Cyrille , Clark Edwin , Winter Chris , Zhang Weisheng , Majumder Pradip K.
Published: February 2011

Prostate cancer is the third most common cause of cancer-related deaths in men and is most prevalent in developed countries. The androgen signaling pathway plays a critical role in the development and maintenance of the prostate gland and is hyper-activated in prostate cancer. Additionally, gain of function mutations in the PI3 kinase pathway is associated with progression of this disease. Using the prostate-specific Pten-deleted mouse model of cancer, we show that these two pathways are distinct for tumor growth and disease progression. Depletion of androgens by either surgical or chemical castration significantly inhibits tumor growth without altering the activation of Akt and mTOR. We demonstrate that inhibition of mTOR with ridaforolimus or inhibition of Akt with MK-2206, combined with androgen deprivation (bicalutamide), results in additive anti-tumor effects in vivo when compared to single agent treatment. In this model, we also demonstrate that Akt inhibition does not block mTOR kinase activation, and that inhibiting both Akt and mTOR kinases together significantly enhances anti-tumor efficacy. Our data suggest that the combination of an anti-androgen with an mTOR inhibitor and/or Akt inhibitor may be more effective than standard therapies in treating androgen-dependent prostate cancer patients with an activated PI3 kinase pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-284.
Abstract LB-286: Use of circulating tumor cells to monitor pharmacodynamic responses of patients in early-stage clinical trials
Authors: Kinders Robert J. , Wang Lihua , Kummar Shivaani , Pfister Thomas , Parchment Ralph , Tomaszewski Joseph , Doroshow James
Published: February 2011

Circulating tumor cells (CTCs) in peripheral blood of cancer patients potentially represent a fraction of solid tumor cells available for more frequent pharmacodynamic assessment of drug action than is possible using tumor biopsy. We have developed and validated an assay that directly examines increases in the number of cells positive for the nuclear DNA-damage marker H2AX in individual CTCs of patients treated with DNA-damaging chemotherapeutic agents. The method uses AexaFluor 488-conjugated monoclonal anti- H2AX (JBW301), to stain putative CTCs collected from blood by the CellSearch AutoPrep instrument, followed by enumeration and fluorescence analysis on the CellTracks Analyzer. Patient baselining studies demonstrated that in the absence of treatment the number of CTCs per blood aliquot is relatively constant over short time periods (2 weeks off therapy). CTC measurements are currently in use supporting four NCI new agent trials, two with cytotoxic agents and two with targeted agents (aTOP1 inhibitor and a tyrosine kinase inhibitor, TKI). CTC changes in 40 patients have been monitored on therapy to date. H2AX positive cells increased in 7/14 patients on cytotoxic therapies, 3/14 were unchanged and 4/14 were unevaluable; the fraction of H2AX positive CTCs did not decrease in any patient on cytotoxic therapy, and did not change in patients treated with TKI. The increased H2AX signal in patients on cytotoxic agents appears definitive for the cycle of therapy in which it is measured. Interestingly, 10/40 patients had increased CTC numbers immediately post-therapy, and several of these patients had few or no CTC immediately prior to therapy, suggesting that the cells were mobilized or shed from the tumor as a consequence of drug exposure. Six patients had decreases in CTC number, 12 were unchanged and 12 were unevaluable due to low CTC numbers.

These data show promise for monitoring dynamic changes in nuclear biomarkers in CTCs, in addition to a CTC count alone, to rapidly assess drug pharmacodynamic activity, in clinical trials of cytotoxic agents and targeted anti-cancer therapeutics, as well as for translational research. This method may have the potential to inform the development of new anti-cancer agents and enable longitudinal monitoring of target response.

Funded by NCI Contract No. HHSN261200800001E.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-286.
Abstract LB-289: Synthetic Hsp90 inhibitor, PF-04928473 and prodrug PF-04929113, demonstrates potent antitumor activity in head and neck cancer by inhibiting multiple proliferative and prosurvival signaling pathways
Authors: Friedman Jay , Contag Stephanie , Hu Michael , Freudlsperger Christian , Kannabiran Vishnu , Arun Pattatheyil , Mitchell James , Skillings Jamey , Kan Julie , Chen Zhong , Van Waes Carter
Published: February 2011

Purpose: Heat shock protein 90 (Hsp90) is a molecular chaperone that stabilizes client substrates involved in cell proliferation, survival, and apoptosis. Cancer cells display increased dependency on Hsp90 and client oncoproteins, and Hsp90 has been identified as a druggable target. This study investigated the biological activity of a synthetic small molecule inhibitor of Hsp90, PF-04928473 and the orally available pro-drug PF-04929113 in head and neck squamous cell carcinoma (HNSCC).

Experimental Design: Hsp90 expression was examined in nine human HNSCC cell lines (UMSCC). In vitro, the effect of PF-04928473 on cell proliferation, death and cell cycle were examined by MTT assay and flow cytometry. The effects on signaling molecules and cytokine production were determined using Western blot, reporter gene, and Luminex assays. In vitro combination with radiation was examined using the clonogenic survival assay. The effect of pro-drug PF-04929113 on UMSCC-11A tumor growth and Hsp90 client proteins were examined in the murine xenograft model and by immunohistochemistry (IHC).

Results: UMSCC cell lines over-expressed Hsp90 mRNA and protein when compared with normal keratinocytes. In vitro, PF-04928473 potently inhibited UMSCC cell growth, induced cell death and modulated cell cycle, with an IC50 range of 32-73nmol/L. Combining radiation and PF-04928473 increased radiation sensitivity by a dose modifying effect of 1.4. PF-04928473 significantly modulated signaling molecules, including down-regulation of c-Met receptor, IKK, IKK, and BCLXL, as well as up-regulation of cleaved PARP, pro-apoptotic p53, PUMA, and p21 protein expression. The drug inhibited AKT, ERK, and STAT3 phosphorylation, suppressed NF- B, AP-1, BCLXL, and IL8 reporter activities, and blocked IL8, IL6, and VEGF production. In vivo, pro-drug PF-04929113 20mg/kg daily for 3-7 weeks significantly inhibited tumor growth. After treatment, tumor specimens showed increased IHC staining for p53, PUMA, and TUNEL, and decreased staining for c-Met, SRC, p-STAT3, p-AKT, p-ERK, IKK, RELB, Ki67 and CD31.

Conclusion: Synthetic Hsp90 inhibitor PF-04928473 effectively modulated proliferation, cell survival, and inflammatory and angiogeneic cytokine production in HNSCC in vitro. Pro-drug PF-04929113 inhibits tumor growth in vivo by promoting apoptosis through inhibition of several key signaling pathways. [Supported by NIDCD project ZIA-DC-000073]

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-289.
Abstract LB-290: Tranilast inhibits breast cancer stem cells
Authors: Prud'homme Gerald J. , Glinka Yelena , Toulina Anna , Subramaniam Venkateswaran , Chakrabarti Rabindranath , Jothy Serge
Published: February 2011

Cancer stem cells (CSCs) have increased resistance to anti-cancer drugs, and may be responsible for chemotherapeutic failure. CSCs can be enriched from cancer cell lines by growth with anti-cancer drugs (e.g., doxorubicin, mitoxantrone), and form mammospheres in low-adherence, serum-free cultures. We are studying tranilast, a non-toxic orally active drug in clinical use for allergic diseases in Japan, but that we found also targets cancer cells. Previously, we reported that tranilast exerts multiple anti-cancer effects and protects against breast cancer metastasis. Our recent studies show that tranilast strongly inhibits breast CSCs in several assays, at pharmacologically relevant concentrations. We enriched CSCs from breast cancer cell lines by cell sorting for ALDH-1 positive cells (ALDEFLUOR+), or by growth in mitoxantrone-containing medium to select drug-surviving CSCs. In both cases, CSCs readily formed mammospheres in vitro. Tranilast inhibited mammosphere formation,’ and dissociated formed mammospheres. It was more effective than paclitaxel or etoposide. We recently identified molecular targets of this drug. We found it has aryl hydrocarbon receptor (AHR) agonist activity and this might explain its anti-cancer effects. The AHR is a receptor for toxins such as dioxin and polycyclic aromatic hydrocarbons. It is a transcription factor that exerts a number of ligand-dependent effects, such as cell-cycle arrest, reduced RB phosphorylation, and decreased cytokine and growth factor production. Unlike many AHR ligands, tranilast has very low toxicity (LD50 > 1 gm/kg in rats), and is well tolerated by patients. In accord with a role for the AHR, breast CSCs expressed higher levels of this receptor than the main population of cancer cells, and the effects of tranilast were markedly diminished by an AHR antagonist (alpha-naphthoflavone). Moreover, we found tranilast inhibits some ATP-binding cassette (ABC) multiple drug resistance transporters, and it enhances the effects of other anti-cancer drugs. Our studies show that tranilast is a drug of low toxicity that has potential as an anti-CSC drug.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-290.
Abstract LB-300: Small molecule modulators of MET kinase for treatment of human malignancies
Authors: Wise Scott C. , Smith Bryan D. , Hood Molly M. , Kaufman Michael D. , Lu Wei-Ping , Flynn Daniel L.
Published: February 2011

MET kinase is involved in numerous cancers including lung, melanoma and gastric. MET can not only be the oncogenic driver of the cancer cell but also is a key player in the cells ability to metastasize and invade tissues distant from the primary tumor site. It has been shown that 2 out of 3 cancers involving MET leads to a poor prognosis for patient survival. Using Deciphera Pharmaceutical's approach to kinase inhibition, compounds have been designed which effectively inhibit wild type and mutant MET kinase forms. These inhibitors are selective, including sparing of close family members AXL and RON. This presentation will highlight the attributes and development status of these compounds for treatment of human malignancy. Deciphera's MET program has afforded potent inhibitors that achieve low nanomolar inhibition of wild type MET and secondary activating mutant forms. Data will be presented on DP-4693 and DP-4756.IC50MET Kinasep-MET MKN-45MKN-45 Proliferationp-MET EBC-1EBC-1 ProliferationA549 MigrationDP-46934nM13 nM44 nM10 nM3nM260 nMDP-47566nM6nM23 nM2nM5nM21 nM

DP-4693 and DP-4756 are selective, having as few as three kinases within 50 fold of MET potency. Inhibitor bound crystal structures will be shown which demonstrate the compounds mechanism of action. In addition, proliferation and MET phosphoprotein analysis performed with EBC-1 and MKN-45 cell lines have demonstrated excellent inhibitory profiles with potencies in the low nanomolar range. The compounds also inhibit HGF dependent cell motility in an A549 migration assay. In vivo evaluation of DP-4693 in an MKN-45 gastric cancer xenograft PK/PD model revealed complete target suppression at doses as low as 6 mpk. Futher studies will be presented highlighting the efficacy in both autocrine and paracrine (HGF-dependent) xenograft models.

Deciphera's MET inhibitors also show acceptable ADME properties and are orally bioavailable in rat and dog using simple vehicles. These inhibitors have also been evaluated in two week toxicology studies, exhibiting excellent tolerability up to doses of 300 mpk. Further studies are ongoing to enable Development Candidate selection for progression into clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-300.
Abstract LB-328: Granulocytic myeloid-derived suppressor cells that have suppressive function: CD15+/CD16low human granulocytes from advanced cancer patients
Authors: Choi Jahyang , Suh Beomseok , Ahn Yong-Oon , Kim Tae Min , Lee Se-Hoon , Heo Dae Seog
Published: February 2011

Objectives: Myeloid-derived suppressor cells (MDSCs) are a subpopulation of myeloid cells with suppressive function, found to be increased in various conditions such as chronic infection, trauma, and cancer. Unlike the murine MDSC defined as CD11b+/Gr-1+, there are no specific markers for human MDSC. We have investigated human granulocytes from advanced cancer patients to delineate a specific MDSC subpopulation with clinical implication.

Methods: To find changes of granulocyte markers, we collected whole blood of 53 advanced cancer patients and 20 healthy donors and analyzed them by flow cytometry. To investigate the growth suppressive activity of granulocytes, we purified them and tested growth inhibition activity by 3H thymidine assay.

Results: Through a study of 53 advanced cancer patients, we observed heterogeneity of CD15(+)CD16(+) granulocytes: CD15+/CD16lowand CD15+/CD16high. The number of CD15+/CD16low subsets is increased in advanced cancer patients, compared with healthy donors (p = 0.009). p-methoxyamphetamine (PMA) activated granulocytes (CD16low/CD66b++/CD15+) that have similar phenotype of MDSC from cancer patients suppressed both proliferation and cytotoxicity of normal T cells. T cell proliferation is highly suppressed by granulocytes isolated from advanced cancer patients with high proportion of CD15+/CD16low granulocytes among cancer patients. We also found that patients whose peripheral blood samples with lower levels of CD15+/CD16low populations had significantly longer survival times than those with higher levels of CD15+/CD16low populations. Patients with higher levels of CD15+/CD16low are associated with poor performance status (PS, p = 0.022) as well as shorter survival time (p = 0.0036). Because these phenotypic changes of granulocytes are also shown in the case of inflammation, we speculate that some factors in tumor-bearing hosts may activate granulocytes and induce their suppressive activity.

Conclusions: These data suggest that CD15+/CD16low granulocytes from cancer patients may have a role in progression of cancer by inhibiting tumor immunity. Monitoring of these cells also can be used as a marker of terminal cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-328.