bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2301: Myeloid cells promote fibroblast invasion and lead mammary adenocarcinoma cell invasion.
Authors: Shaw Aubie K. , Novitskiy Sergey , Coussens Lisa M. , Moses Harold L.
Published: August 2014

Myeloid cells localize to invasive edges of solid tumors where they promote invasive tumor growth, intravasation into vasculature and metastasis of malignant cells1,2. Fibroblasts, on the other hand, lead collective cell migration3 of carcinoma cells via fostering extracellular matrix (ECM) remodeling and generating tracks of collagen fibers that act as “highways” for cell migration4,5. To identify soluble molecules mediating these juxtacrine communication mechanisms between myeloid cells and fibroblasts in potentiating metastasis, we conducted pilot studies with subsets of spleen- or tumor-derived myeloid cells, tumor-derived fibroblasts, and mammary adenocarcinoma cells. Whereas spleen-derived CD11b+Ly6ChiLy6G− monocytes promote fibroblast migration ex vivo, two populations of tumor-derived myeloid cells, e.g., CD11b+Gr1+ and CD11b+Gr1− cells possessed this capability. In ex vivo migration assays, fibroblast migration was dependent on myeloid cell-derived factors, e.g., CXCL11, CXCL15, FGF2, IGF-1 and Shh, as well as fibroblast Akt and ERK1/2 phosphorylation. Inhibition of fibroblast MEK reduced myeloid cell-induced fibroblast migration, whereas FGF-RI inhibition completely abolished this capability. These studies indicate a role for myeloid cell-fibroblast communication in fostering migration (and potentially activation) of fibroblasts that lead collective cell migration properties of mammary carcinoma cells. Ongoing studies are identifying specific subpopulations of tumor-associated myeloid cells possessing these activities, as well as the role of FGF receptor signaling pathways in facilitating metastasis.

1.

Wyckoff, J.B., et al. Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. Cancer Res 67, 2649-2656 (2007).

2.

Qian, B.Z. & Pollard, J.W. Macrophage diversity enhances tumor progression and metastasis. Cell 141, 39-51 (2010).

3.

Friedl, P., Locker, J., Sahai, E. & Segall, J.E. Classifying collective cancer cell invasion. Nat Cell Biol 14, 777-783 (2012).

4.

Khalil, A.A. & Friedl, P. Determinants of leader cells in collective cell migration. Integr Biol (Camb) 2, 568-574 (2010).

5.

Gritsenko, P.G., Ilina, O. & Friedl, P. Interstitial guidance of cancer invasion. J Pathol 226, 185-199 (2012).

The authors acknowledge generous support from the NIH/NCI, a Dept of Defense (DoD) Breast Cancer Research Program Postdoctoral Fellowship, a DoD Era of Hope Scholar Expansion Award, Susan G. Komen Fndt, and the Breast Cancer Research Fndt.

Citation Format: Aubie K. Shaw, Sergey Novitskiy, Lisa M. Coussens, Harold L. Moses. Myeloid cells promote fibroblast invasion and lead mammary adenocarcinoma cell invasion. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2301. doi:10.1158/1538-7445.AM2013-2301
Abstract 2410: Circulating tumor cells to guide the choice between chemotherapy and hormone therapy as first line treatment for metastatic breast cancer patients: the STIC CTC METABREAST trial.
Authors: Pierga Jean-Yves , Baffert Sandrine , Hajage David , Brain Etienne , Armanet Sebastien , Simondi Cécile , Mignot Laurent , Asselain Bernard , Tresca Patricia , Bidard François-Clément
Published: August 2014

Background

Baseline CTC count already demonstrated its very good performance as an independent prognostic marker in metastatic breast cancer (MBC), before the start of any treatment (Cristofanilli, NEJM 2004; Pierga Ann Oncol 2011). Multivariate analyses performed in both a pooled analysis (Liu, ASCO 2011) and in the IC 2006-04 study showed that the other independent prognostic factors were the performance status and hormone-receptor (HR) status. Oppositely, the other criteria that are frequently used to choose between hormone therapy and chemotherapy for the treatment of first line hormone receptor-positive MBC patients (e.g. metastatic sites, metastasis-free interval…) were not independent prognostic factors. On the basis that CTC count may be a better criterion for this important choice than the currently used empiric criteria, we designed a large pivotal phase III trial.

Methods

N=996 HR+ M+ breast cancer patients will be randomized between the clinician choice and CTC count-driven choice. Main inclusion criteria include hormone receptor-positive MBC patients with no previous treatment for the metastatic disease, and who can receive either hormone therapy or chemotherapy as first line treatment. In the CTC arm, patients with ≥5 CTC/7.5ml will receive chemotherapy whereas patients with <5 CTC/7.5ml will receive endocrine therapy as first line treatment. As every patient will receive a treatment, this pivotal trial has been designed to show a non-inferiority of the CTC arm for PFS (primary clinical endpoint) and a superiority of the CTC arm for the medico-economics study (co-primary endpoint). The STIC CTC METABREAST trial has been funded by the French Ministry of Health (STIC program) and Veridex. The promoter is the Institut Curie (Paris).

Results

This trial began in February 2012 and more than 15 French cancer centers participate to this study. The accrual should be completed in 2 to 3 years.

Conclusion

This large pivotal phase III trial is intended to demonstrate the clinical relevance of baseline CTC count.

Citation Format: Jean-Yves Pierga, Sandrine Baffert, David Hajage, Etienne Brain, Sebastien Armanet, Cécile Simondi, Laurent Mignot, Bernard Asselain, Patricia Tresca, François-Clément Bidard. Circulating tumor cells to guide the choice between chemotherapy and hormone therapy as first line treatment for metastatic breast cancer patients: the STIC CTC METABREAST trial. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2410. doi:10.1158/1538-7445.AM2013-2410
Abstract 2552: A genome-wide association study of prostate cancer in West African men.
Authors: Cook Michael B. , Wang Zhaoming , Yeboah Edward D. , Adjei Andrew A. , Tettey Yao , Biritwum Richard B. , Tay Evelyn , Truelove Ann , Niwa Shelley , Chu Lisa , Yeager Meredith , Hutchinson Amy , Yu Kai , Haiman Christopher A. ,   African American Prostate Cancer GWAS Consortium, Robert N. Hoover, Hsing Ann , Chanock Stephen J.
Published: August 2014

Age-adjusted mortality rates for prostate cancer are higher for Africa compared with North America or Western Europe. In addition, African American men are noted to have higher age-adjusted incidence rates of this malignancy than European American men. Coupled with the fact that West Africa is the principal ancestral region of African-American men has led to the hypothesis that there may exist distinct ancestral genetic profiles which mediate prostate cancer risk. In addition, advantages of conducting a genome-wide association study (GWAS) of prostate cancer in African men include a more discrete linkage disequilibrium (LD) structure, a higher number of private single nucleotide polymorphisms (SNPs), the predominance of symptomatic disease, and assessment of unique exposures. The Ghana Prostate Study was conducted collaboratively involving the US National Cancer Institute (NCI) and the University of Ghana during 2006-2012. The NCI Cancer Genomics Research Laboratory genotyped 494 prostate cancer cases and 498 population controls using the Illumina HumanOmni5-Quad BeadChip. Associations were assessed using multivariate logistic regression adjusted for age and genetic ancestry. We sought to validate the 30 most promising SNP associations with prostate cancer through the African American Prostate Cancer GWAS Consortium. A novel locus at 10p14 for prostate cancer risk was the strongest signal detected, and the 8 SNPs at this locus were in LD. This locus is located 360 kb 5’ of GATA3 and the 8 SNPs reside within an intron of LincRNA gene RP11-543F8.2. Analysis of African 1000 Genomes Project data did not indicate LD between 10p14 SNPs and splice or exonic SNPs of this gene, while HaploReg found no significant enrichment of enhancer elements. None of the most promising 30 SNPs replicated in the African American Prostate Cancer GWAS Consortium. This may be due to chance or differences in population genetics, environment, and/or proportion of symptomatic disease. Further genetic studies of prostate cancer in African men are needed to validate the 10p14 susceptibility locus.

Citation Format: Michael B. Cook, Zhaoming Wang, Edward D. Yeboah, Andrew A. Adjei, Yao Tettey, Richard B. Biritwum, Evelyn Tay, Ann Truelove, Shelley Niwa, Lisa Chu, Meredith Yeager, Amy Hutchinson, Kai Yu, Christopher A. Haiman, African American Prostate Cancer GWAS Consortium, Robert N. Hoover, Ann Hsing, Stephen J. Chanock. A genome-wide association study of prostate cancer in West African men. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2552. doi:10.1158/1538-7445.AM2013-2552
Abstract 2794: Ex vivo chemoresponse assay using patient-derived tumor xenografts in combination with high content imaging platform as models for oncology drug development.
Authors: Garg Mansi , Vo Martin , Ricono Jill , Broudy Thomas , Mirsaidi Cyrus , Praveen Kesavan Nair
Published: August 2014

Current strategies for developing new anticancer drugs rely heavily on preclinical testing in cancer cell lines and their derived in vivo cell line xenograft models. The value of a predictive assay is heavily dependent on the extent to which it can recapitulate the biology of disease. It has been reported that heterogeneity of cell populations within the tumor microenvironment has significant correlation with the efficacy of any treatment regimen. Cell line models fail to recapitulate the heterogeneous nature of tumor. The purpose of this study was to investigate whether the use of patient-derived primary tumor cells (PTC) could be expanded as xenografts for in vitro chemosensitivity assays to predict clinical outcomes in anticancer drug testing. We have developed preclinical cancer models using PTCs grown in 3D cultures that better mimic in vivo conditions. One of the disadvantages of using patient derived cells is the limited number of cells available from biopsy samples. We utilized patient-derived xenografts (PDX) as a strategy to expand the human tumor cells in mice, while taking efforts to retain the actual tumor biology of the patient. We used high content imaging to evaluate colony segmentation, morphological features, proliferative and apoptotic endpoints to determine efficacy of oncology drug candidates. We conducted a retrospective study using PTCs from solid tumor indications including endometrial and lung to evaluate the correlation between various models and usefulness in predicting the outcome of a specific drug candidate. Our results indicate that PDX models in combination with 3D PTC models have predictive value and enable secondary interrogation beyond single agent anti-proliferative activity to better profile the antitumor activity of targeted agents. Improved preclinical models are required to advance our understanding of the molecular aberrations that underpin cancer and are critical for developing and deploying targeted therapies that will improve patients’ lives.

Citation Format: Mansi Garg, Martin Vo, Jill Ricono, Thomas Broudy, Cyrus Mirsaidi, Kesavan Nair Praveen. Ex vivo chemoresponse assay using patient-derived tumor xenografts in combination with high content imaging platform as models for oncology drug development. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2794. doi:10.1158/1538-7445.AM2013-2794
Abstract 2811: A prostate stroma-derived profile is predictive of early relapse and reflects potential mechanisms of aggressive disease.
Authors: Jia Zhenyu , Rahmatpanah Farahnaz , Chen Xin , McClelland Michael , Mercola Dan
Published: August 2014

The microenvironment of prostate cancer plays a major role in tumor initiation, growth, and metastatic spread. Using RNA expression data from fresh frozen human prostatectomies for which the clinical outcome was followed for over 5 years, we have developed a general approach to identify gene expression changes of the tumor-adjacent stroma that distinguishes aggressive prostate cancer cases from indolent cases. Highly significant gene associations were defined by correlating expression with Disease-free Survival (DFS) and also, separately, by comparing RNA expression between cases that remained disease free for at least 4 years following surgery with those cases that exhibited relapse within 1 year of surgery. A total of 193 highly significant genes were identified, of which 15 were common to the two approaches. These 15 genes were used to develop a PAM (Prediction Analysis for Microarrays)-based algorithm to classify new cases as indolent or aggressive and to provide a probability for each classification. The algorithm was tested on stroma-enriched samples from 47 new cases with known outcome with an accuracy of 87% [1]. A potential functional basis for some the genes associated with aggressive disease was suggested by the observation that p53 expression is decreased in aggressive disease whereas p53-target genes and cell cycle inhibitors including p21, GADD45A, and GADD45B are increased, which is a pattern others have noted as suggestive of senescence [2]. Increased IGFBP6, Tubulin alpha 1a, and Lamin A/C further suggest a senescence-associated secretory phenotype [3]. A potential secretory factor of known significance in the growth of prostate cancer (IGF-1/Somatomedin-C) has elevated RNA levels in the stroma of the same cases. These features argue that an accurate and general prediction of aggressiveness may be derived from the tumor microenvironment and may be composed of genes that have functional roles in bringing about aggressive disease.

[1] Jia et al. 2012 PLoS ONE, 7(8):e41371

[2] Liebermann and Hoffman 2008 Journal of Molecular Signaling, 3:15

[3] Coppe et al. 2008 PLoS Biology, 6(12):e301

Citation Format: Zhenyu Jia, Farahnaz Rahmatpanah, Xin Chen, Michael McClelland, Dan Mercola. A prostate stroma-derived profile is predictive of early relapse and reflects potential mechanisms of aggressive disease. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2811. doi:10.1158/1538-7445.AM2013-2811

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2825: Using SHG to study breast cancer tumor progression.
Authors: Burke Kathleen A. , Brown Edward
Published: August 2014

The majority of cancer mortality is a result of metastatic events, making it increasingly important to uncover prognostic markers that can predict the metastatic potential of a primary tumor to aid in determining the best course of patient treatment. Stromal extracellular matrix restructuring increases efficiency of cell migration away from the primary tumor into surrounding tissues and vasculature, thereby enhancing metastatic potential. We have determined that the ratio of the forward propagating to backward propagating Second Harmonic Generation (SHG) signals from type I collagen can be used as a quantitative method of differentiating healthy and tumor breast tissue, and tracking stromal changes throughout tumor progression. Tumors were graded and staged according to standard pathological qualifications, incorporating morphological changes in the tissue as well as extent of tumor growth and invasion. As the tumor cells initiate development in the ducts of the breast tissue the F/B ratio of the fibrillar collagen does not change. Upon progression from an in situ to an invasive carcinoma the F/B ratio of ductal tumors significantly decreases. Taking a closer look at this phenomenon, to examine whether the invasion through the basement membrane into the surrounding tissue causes an immediate change in the F/B ratio, tumors of varying grade and stage were analyzed showing significant trends throughout development. Increased tumor grade was associated with a gradual decrease in the F/B ratio, whereas size of the tumor had no significant effect on the F/B but all sizes were significantly lower than healthy tissue. Interestingly, there was a significant increase in the F/B ratio associated with the furthest progression of lymph-node involvement. Invasive Lobular Carcinoma analysis revealed a similar decrease in F/B, but was not affected by lymph node involvement. On the other hand F/B of these tumors was significantly affected by an interaction with the chest or skin. These results provide insight into the different development processes undergone by two related breast carcinomas, which could differ due to their alternative methods of invasion and stromal components. F/B ratio measurements can be used alongside pathological analysis to provide information about the metastatic potential of the primary tumor, as demonstrated by the relationship between F/B ratio of IDC and lymph node metastasis. Enhanced scientific understanding of stromal development could be applied to the development of improved biomarkers and metastatic treatment methods.

Citation Format: Kathleen A. Burke, Edward Brown. Using SHG to study breast cancer tumor progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2825. doi:10.1158/1538-7445.AM2013-2825
Abstract 2844: ATATESTM activated the same patient's immune system to eliminate hundreds of melanoma tumors for cure of cancer.
Authors: Qian Yong
Published: August 2014

A 26 year old male melanoma cancer patient had over 1000 solid tumors originated from a laser treatment of a mold on his back. Mutated cancer cells metastasized to his lymph nodes first then spread to the skin all around the body. He had symptoms of vomitting, poor apetide, loosing weight and lack of strength for several months before seeking BioMedicure's help. ATATESTMproducts, with antigenic material information from millions of tumor cells preserving in spheres, were made out of his melanoma tumors for immunotherapy over one month. His symptoms have been reversed after a couple of months’ ATATESTM immunotherapy. Over 95% of his melanoma tumors have been under control and disappearing over time. Multiple death pathways elicited by ATATESTM have been observed and investigated. The detailed mechanisms of action for ATATESTM will be presented.

Citation Format: Yong Qian. ATATESTM activated the same patient's immune system to eliminate hundreds of melanoma tumors for cure of cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2844. doi:10.1158/1538-7445.AM2013-2844

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 288: TGF-beta induced epithelial-to-mesenchymal transition is attenuated when the MNK/eIF4E pathway is functionally impaired.
Authors: del Rincon Sonia V. , Huor Bonnie , Ngan Elaine , Petruccelli Luca , Siegel Peter , Miller Wilson H.
Published: August 2014

The epithelial-mesenchymal-like transition (EMT) is a process enabling epithelial cells to gain the motile characteristics of mesenchymal cells, in a manner resembling metastasis. TGF-beta, via well-defined transcriptional mechanisms, is considered a master regulator of EMT. However, the idea that TGF-beta can regulate the translational machinery to drive EMT remains largely unexplored. The eukaryotic translation initiation factor eIF4E is known to be overexpressed in breast cancer, has been linked to increased invasiveness, and is a promising target for the treatment of breast cancer. Our hypothesis is that phosphorylation of eIF4E stimulated by TGF-beta is required for inducing EMT and metastasis in breast cancer. Our novel preliminary data show that TGF-beta can stimulate eIF4E phosphorylation as normal mammary epithelial cells become mesenchymal. Silencing of eIF4E attenuates molecular and behavioral changes associated with EMT. Moreover, decreasing eIF4E levels can impair TGF-beta induced migration and invasion of ErbB2-expressing breast cancer cells. In keeping with a role of phosphorylated eIF4E in driving the metastatic phenotype, we show that chemically and genetically inhibiting the eIF4E kinase MNK1 attenuates TGF-beta-stimulated EMT. We hypothesized that the expression of master regulators of EMT could be restricted when the eIF4E/MNK pathway is functionally impaired, which prompted us to look at the expression of Twist and Snail in TGF-beta treated eIF4E- and MNK- silenced cells. Our data shows that TGF-beta-induced Snail protein expression, but not mRNA level, is repressed when the MNK/eIF4E pathway is functionally impaired. Our results indicate that MNK and eIF4E are essential for EMT and suggest that therapeutic inhibition of MNK/eIF4E pathway may be a useful strategy for the control of tumor invasion and metastasis.

Citation Format: Sonia V. del Rincon, Bonnie Huor, Elaine Ngan, Luca Petruccelli, Peter Siegel, Wilson H. Miller, Jr. TGF-beta induced epithelial-to-mesenchymal transition is attenuated when the MNK/eIF4E pathway is functionally impaired. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 288. doi:10.1158/1538-7445.AM2013-288
Abstract 2880: A prospective study of 51 serum immune markers and risk of non-Hodgkin lymphoma.
Authors: Purdue Mark P. , Hofmann Jonathan N. , Kemp Troy J. , Chaturvedi Anil K. , Lan Qing , Park Ju-Hyun , Pfeiffer Ruth M. , Hildesheim Allan , Pinto Ligia A. , Rothman Nathaniel
Published: August 2014

Background: Although severe immune dysregulation is an established risk factor for non-Hodgkin lymphoma (NHL), the etiologic relevance of subclinical immunologic effects is unclear. Few prospective studies have investigated the relationship between circulating immune markers and NHL, and those conducted to date have been limited in sample size, the number of evaluated analytes, or in their ability to examine markers of risk many years preceding disease diagnosis. To better understand this question, we conducted a nested case-control study investigating pre-diagnostic serum levels of 51 immune markers within the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial.

Methods: Fifty-one immune markers (including cytokines, chemokines, extracellular matrix proteins, growth factors and soluble products of immune activation) were measured in baseline serum from 301 NHL cases diagnosed 5+ years after blood collection (median 8.0 years, range 5.0-13.9) and 301 individually matched controls using Luminex bead-based assay panels. Odds ratios (ORs) and 95% confidence intervals (CIs) relating marker levels with NHL risk were computed using conditional logistic regression modeling.

Results: We observed strong associations with NHL for elevated levels of several analytes: a pro-inflammatory marker, soluble tumor necrosis factor receptor 2 (sTNFRII; fourth quartile vs. first: OR 3.4, 95% CI 2.0-5.8; Ptrend = 5.9 x 10−6); a regulator of B-cell traffic, B-cell attracting chemokine 1 (BCA-1; 2.7, 1.7-4.2; Ptrend = 1.1 x 10−5); a regulator of angiogenesis, soluble vascular endothelial growth factor receptor 2 (sVEGFR2; 2.3, CI 1.4-3.9; Ptrend = 0.0005); and the pro-inflammation and apoptotic cytokine TNF-related apoptosis inducing ligand (TRAIL; 1.6, 1.0-2.5; Ptrend = 0.02). These associations remained significant in analyses of cases diagnosed 8-13 years after blood collection and, with the exception of TRAIL, following Bonferroni correction and simultaneous model adjustment.

Conclusions: Our findings suggest that elevated levels of sTNFRII, BCA-1, sVEGFR2, and, possibly, TRAIL may reflect biologic processes contributing to lymphomagenesis. Additional research is needed to replicate these findings, identify cell sources of these markers, and elucidate the underlying biologic mechanisms.

Citation Format: Mark P. Purdue, Jonathan N. Hofmann, Troy J. Kemp, Anil K. Chaturvedi, Qing Lan, Ju-Hyun Park, Ruth M. Pfeiffer, Allan Hildesheim, Ligia A. Pinto, Nathaniel Rothman. A prospective study of 51 serum immune markers and risk of non-Hodgkin lymphoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2880. doi:10.1158/1538-7445.AM2013-2880
Abstract 2895: Phylogenetic models of tumor progression from fluorescence in situ hybridization (FISH) data on many single cells of a solid tumor.
Authors: Chowdhury Salim A. , Schäffer Alejandro A. , Shackney Stanley E. , Wangsa Darawalee , Heselmeyer-Haddad Kerstin , Ried Thomas , Schwartz Russell
Published: August 2014

Purpose: Characterizing the common pathways through which tumors progress is critical to understanding the molecular basis of cancer and developing effective treatments. Algorithms for phylogenetics, i.e., evolutionary tree building, can be used to infer progression pathways of single tumors when there is widespread intra-tumor heterogeneity from cell to cell. We describe computational methods to compute likely evolutionary histories from single-cell copy number data and apply the methods to single-cell gene copy number data derived from paired primary and metastatic samples of cervical cancers.

Materials and methods: The inputs to our methods are lists of cell types for each patient sample, where a cell type is an array of copy numbers of gene probes measured by FISH. In the cervical cancer data set, copy numbers of four genes (LAMP3, PROX1, PRKAA1 and CCND1) were measured on up to 250 cells each of paired primary and metastatic samples from 16 patients. An example of a cell type is (4, 1, 3, 2). Our algorithms derive maximum parsimony phylogenetic trees explaining how all observed cell types might have evolved from a common diploid ancestor (2,2,2,2), seeking to explain the observed cells with as few copy number changes as possible across the tree. Due to the computational difficulty, we apply a heuristic algorithm to infer an approximately optimal tree by identifying likely unobserved ancestral cell states through a variant of the median joining method and then find the most parsimonious tree connecting these states. Our software also includes a method of computing consensus trees that facilitate the comparison of the tree models for each primary/metastasis sample pair.

Results and Conclusion: Our methods allow us to build models of evolution of single tumors at the cellular level that provide insights into possible progression pathways and varying selective pressures in primary and metastatic sites. Analysis of tree geometry shows significant variability in evolutionary patterns between primary and metastatic sites consistent with more specific selective pressures in the metastases. Analysis of specific patterns of gene gain or loss in each environment shows a complicated portrait of tumor evolution with substantial variability from patient to patient.

Citation Format: Salim A. Chowdhury, Alejandro A. Schäffer, Stanley E. Shackney, Darawalee Wangsa, Kerstin Heselmeyer-Haddad, Thomas Ried, Russell Schwartz. Phylogenetic models of tumor progression from fluorescence in situ hybridization (FISH) data on many single cells of a solid tumor. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2895. doi:10.1158/1538-7445.AM2013-2895
Abstract 2923: Glucose-regulated protein 78(GRP78) mediated the therapeutic efficacy of epirubicin via the downregulation of intracellular oxidative stress in colon cancer cells.
Authors: Chang Yu-Jia , Chen Ching-Hsein , Wei Po-Li
Published: August 2014

Background: The 78-kDa glucose-regulated protein (GRP78) is induced in the cancer microenvironment and can be considered as a novel predictor of responsiveness to chemotherapy in many cancers. However, there is little information regarding whether the expression level of GRP78 affects epirubicin treatment in colon cancer.

Methods: The expression levels of GRP78 were determined in HT-29 and DLD-1 colon cancer cells. GRP78 expression was suppressed in DLD-1 cells using a short interfering RNA (siRNA) technique. The anti-cancer effects of epirubicin were examined using a MTT assay, TUNEL assay, and cell cycle analysis. The status of intracellular reactive oxygen species (ROS) was determined.

Results: In this study, we found thateEpirubicin resistance reduced in cells with suppressed GRP78, in contrast to the parental cells, as determined by survival assays. Cell cycle analysis and TUNEL assays revealed that the apoptotic cell population was increased in the GRP78 knockdown cells treated with epirubicin compared to the scrambled control cells. We found that intracellular reactive oxygen species were higher in GRP78 knockdown DLD-1 colon cancer cells compared with scrambled control cells. Treatment with epirubicin in GRP78-knockdown DLD-1 cells enhanced apoptosis and was associated with decreased production of intracellular ROS.

Conclusions: In summary, our results indicate that GRP78 expression levels were highly correlated with the therapeutic efficacy to epirubicin which was associated with decreased production of intracellular ROS.

Citation Format: Yu-Jia Chang, Ching-Hsein Chen, Po-Li Wei. Glucose-regulated protein 78(GRP78) mediated the therapeutic efficacy of epirubicin via the downregulation of intracellular oxidative stress in colon cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2923. doi:10.1158/1538-7445.AM2013-2923
Abstract 2927: Breast tumor kinase (Brk/PTK6) mediates hypoxia/HIF-1α-associated breast cancer progression.
Authors: Anderson Tarah Regan , Hubbard Gregory , Daniel Andrea R. , Peacock Danielle , Wenger Roland H. , Schörg Alexandra , Hoogewijs David , Vande Woude George , Seagroves Tiffany N. , Lange Carol A.
Published: August 2014

Triple negative (basal type) breast cancers are more aggressive and difficult to treat relative to HER2/ER/PR positive (luminal type) breast cancers; these tumors frequently express abundant Met receptors and contain high constitutive levels of hypoxia inducible factor alpha (HIF-1α), a principal mediator of cellular stress that predicts cancer relapse and increased risk of metastasis. Breast tumor kinase (Brk/PTK6; Sik in mice) is a soluble tyrosine kinase that is not found in normal breast tissue, but is aberrantly expressed in up to 86% of all breast cancers. We recently showed that Brk is a critical downstream effector of Met signaling and required for HGF-induced cell migration. Herein, we show that Brk and HIF-1α are co-expressed in human triple negative cell lines and tumors. Brk mRNA and protein levels are strongly upregulated during conditions that induce cellular stress in vitro, including low glucose, H2O2, and hypoxia. Chromatin-immunoprecipitation (ChIP) assays revealed that Brk is a direct early transcriptional target of HIF-1α; HIF knockdown demonstrated that Brk expression is dependent upon both HIF-1α and HIF-2α. Notably, Brk expression rescued the growth of MDA-MB-231 xenografts knocked-down for both HIF1α and HIF2α in vivo. Finally, we crossed WAP-Brk (FVB) transgenic mice into the MMTV-METmut model (FVB mice engineered to express MMTV-driven constitutively active mutant Met receptors); the addition of the human wt Brk transgene decreased survival in this model. Notably, mouse mammary tumors that arose in MMTV-METmut mice expressed abundant Sik, the mouse homolog of Brk; high Sik expression conferred markedly decreased latency to tumor formation independently of the presence of the Brk transgene. These results identify HIF-1α as a novel regulator of Brk mRNA expression and suggest that Brk is a key mediator of HIF-1α-induced breast cancer progression. Targeting Brk expression or activity (downstream of the HGF/Met receptor signaling pathway) may provide an effective means to block the progression of deadly triple-negative breast cancers.

Citation Format: Tarah Regan Anderson, Gregory Hubbard, Andrea R. Daniel, Danielle Peacock, Roland H. Wenger, Alexandra Schörg, David Hoogewijs, George Vande Woude, Tiffany N. Seagroves, Carol A. Lange. Breast tumor kinase (Brk/PTK6) mediates hypoxia/HIF-1α-associated breast cancer progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2927. doi:10.1158/1538-7445.AM2013-2927
Abstract 2931: Synergy between secreted factors promotes a hyper-secretory phenotype which may fuel ovarian cancer progression.
Authors: Davalos Albert R. , Schaum Nicholas , Campisi Judith
Published: August 2014

The challenge in ovarian cancer research is twofold: 1) identification of early biomarkers of the disease and 2) uncovering targetable molecules to treat malignancy while minimizing toxicity. Mutations in the BRCA1/BRCA2 gene locus account for approximately 15% of patients with ovarian cancer, but several other risk factors are known, including increasing age and obesity. We recently uncovered that a nuclear protein (HMGB1) becomes secreted when cells undergo cellular senescence and we detect increased levels in plasma from elderly compared to young individuals. Significantly, we detect abundant extranuclear HMGB1 staining in tissue from patients with serous ovarian carcinoma, suggesting the presence of senescence cells in the tumor.

We find elevated levels of circulating HMGB1 in patients diagnosed with serous carcinoma. Tissue microarray data analysis correlates with reduced survival in ovarian cancer patients staining with higher levels of HMGB1.

Like aging, obesity is a risk factor for ovarian cancer but how remains poorly understood. We found that plasma from mice on high fat diets contained higher levels of HMGB1 compared to mice on normal diets. To extend this observation, we found that culturing ovarian cancer derived cells with saturated fatty acids promoted significantly increased secretion of HMGB1, IL-6 and MMP-3. Moreover, senescent cells which exhibit a secretory phenotype became “hyper-secretory” when cultured with saturated fatty acids. Recent data revealed that the redox state of HMGB1 establishes the protein's function. Secreted HMGB1 synergizes with other secreted factors to stimulate secretion of inflammatory molecules. Indeed we found cells cultured with fatty acid released cf DNA (cell free DNA) which can complex with secreted HMGB1 to promote cytokine secretion. Immunocytochemsitry revealed that tissue from serous ovarian cancer patients exhibited strong FABP-4 expression, a marker for adipoctyes but also shown to be secreted in obese individuals. Taken together these results lead to the possibility that multiple cells types synergize to create a permissive microenvironment to foster ovarian cancer development and progression.

Citation Format: Albert R. Davalos, Nicholas Schaum, Judith Campisi. Synergy between secreted factors promotes a hyper-secretory phenotype which may fuel ovarian cancer progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2931. doi:10.1158/1538-7445.AM2013-2931
Abstract 2946: AY4, an agonistic anti-death receptor 4 monoclonal antibody, induces death receptor 4-mediated apoptotic cell death in anaplastic thyroid cancer cells via downregulation of Bcl-xL with reactive oxygen species generation.
Authors: Lee Bok-Soon , Cha Hyun-Young , Shin Yoo Seob , Kang Sung Un , Hwang Hye Sook , Kang Sam , Chang Jae Won , Kim Yong- Sung , Kim Chul-Ho
Published: August 2014

Background: Anaplastic thyroid carcinoma (ATC) is an aggressive human tumor with a median survival of 6 months. We previously developed an agonistic anti-death receptor 4 monoclonal antibody, AY4, and demonstrated the anti-tumor effects of AY4 in head and neck cancer cells. Presently, we show that ATC cells are sensitive to AY4 and that the sensitivity correlates with reduced expression level of Bcl-xL and reactive oxygen species (ROS) generation.

Methods: The effects of AY4 in Thyroid cell lines (C-643, U-HTH7, HTH83 and SW1736) was studied on apoptosis using MTT assy, Annexin V assay and Western blot analysis and was examed on the production of reactive oxygen species with DCFDA staining. HTH83 and C-643 cells were transfected with Bcl-xL siRNA using Lipofectamine 2000.

Results: AY4 induced cell death of C-643, U-HTH 7, HTH83, and SW1736 cells. To elucidate the role of ROS generation in AY4-induced apoptosis of ATC cells, U-HTH 7 and SW1736 cells were pretreated with an anti-oxidant (N-acetyl-cysteine, NAC) followed by AY4 treatment. The cell death was blocked by NAC. AY4-induced cell death was accompanied by the down-regulation of the anti-apoptotic protein, Bcl-xL. To examine the link between the apoptotic response and Bcl-xL protein expression, U-HTH 7 cells were transfected with Bcl-xL plasmid. The consequence of the over-expression of Bcl-xL appeared to decrease AY4-mediated cell death by blocking ROS generation in U-HTH 7 cells. In contrast, Bcl-xL knock-down using small interfering RNA of Bcl-xL enhanced AY4 sensitivity in HTH83 and C-643 cells, and rendered the cells sensitive to AY4-induced cell death..

Conclusion: The results support the conclusion that the expression level of Bcl-xL is important in the AY4-induced apoptosis of ATC cells through ROS generation. AY4 may be a promising tool for ATC therapy.

Keywords: Death receptor 4, agonistic antibody, apoptosis, ROS, anaplastic thyroid carcinoma (ATC)

Citation Format: Bok-Soon Lee, Hyun-Young Cha, Yoo Seob Shin, Sung Un Kang, Hye Sook Hwang, Sam Kang, Jae Won Chang, Yong- Sung Kim, Chul-Ho Kim. AY4, an agonistic anti-death receptor 4 monoclonal antibody, induces death receptor 4-mediated apoptotic cell death in anaplastic thyroid cancer cells via downregulation of Bcl-xL with reactive oxygen species generation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2946. doi:10.1158/1538-7445.AM2013-2946
Abstract 2969: A Novel pathway for regulation of ANXA2 expression in lung cancer cell metastasis by SETDB1/SMAD3 repressor complex.
Authors: Tzeng Tsai-Yu , Hsu Ming-Ta
Published: August 2014

Aberrant histone methylation contributes to tumor development and progression by histone methyltransferase. SETDB1 is an H3K9 methyltransferase that contributes to the epigenetic silencing of target genes in cancer cells, but the role of SETDB1 in the progression of tumorigenesis remains largely unknown. We show here that H3K9Me2 and SETDB1 are down-regulated in metastatic lung cancer cell lines. Immunohistochemistry examination of the lung carcinoma tissue microarray revealed that expression of SETDB1 is significant decreased in metastasis status. Ectopic expression of SETDB1 in highly invasive lung cancer cells suppressed cancer cell migration, invasion and filopodia formation. In contrast, knockdown of endogenous SETDB1 in non-invasive lung cancer cells by RNA interference increases cell migration and invasion in vitro. Mechanistic investigations suggested that SETDB1 is specifically down-regulated by miR-29b in metastatic lung cancer cells. Ectopic miR-29b expression in non-invasive lung cancer cells increases cell invasion ability. Furthermore, we also establish a causal role for the SETDB1/SMAD3 complex in driving metastasis and identify ANXA2 as a critical target in this process among numerous target genes. These findings suggest a novel pathway regulated by SETDB1/SMAD3 complex in lung cancer metastasis.

Citation Format: Tsai-Yu Tzeng, Ming-Ta Hsu. A Novel pathway for regulation of ANXA2 expression in lung cancer cell metastasis by SETDB1/SMAD3 repressor complex. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2969. doi:10.1158/1538-7445.AM2013-2969
Abstract 297: Loss of giant obscurins expression in breast epithelium disrupts epithelial junctions and promotes cell motility and invasion.
Authors: Shriver Marey , Stroka Kimberly , Konstantopoulos Konstantinos , Kontrogianni-Konstantopoulos Aikaterini
Published: August 2014

Obscurin A (∼720kDa) and B (∼870kDa) are giant multidomain proteins, encoded by the single OBSCN gene, originally shown to play important roles in the structural organization and contractile activity of striated muscles. Early studies have indicated that the OBSCN gene is highly mutated in different types of cancers. In light of this observation, our laboratory set forth to examine the expression profile and roles of giant obscurins in normal and cancer breast tissue. Obscurins are readily expressed in breast epithelial cells, but their expression is dramatically diminished in breast cancer cells. Down-regulation of obscurin A and B using small hairpin RNAs (shRNAs) in non-tumorigenic MCF10A breast epithelial cells resulted in decreased protein expression of β-catenin and E-cadherin, which are major components of adherens junctions and have been heavily implicated in the formation and metastasis of breast tumors. Specifically, β-catenin exhibited a preferential nuclear accumulation accompanied by a concomitant loss from cell-cell junctions. Additionally, loss of obscurins led to alterations in the expression levels and localization of multiple proteins associated with epithelial to mesenchymal transition (EMT). Obscurin-deficient MCF10A cells showed significantly increased motility in 2-Dimensional (2-D) substrata and confined spaces, and invasion through a matrigel coated chamber. Consistent with this, actin filaments re-distributed to extending filopodia-like protrusions where they exhibited increased dynamics. Taken together, our findings indicate that loss of giant obscurins from breast epithelium results in disruption of cell contacts and acquisition of a mesenchymal phenotype that leads to enhanced migration and invasiveness.

Citation Format: Marey Shriver, Kimberly Stroka, Konstantinos Konstantopoulos, Aikaterini Kontrogianni-Konstantopoulos. Loss of giant obscurins expression in breast epithelium disrupts epithelial junctions and promotes cell motility and invasion. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 297. doi:10.1158/1538-7445.AM2013-297
Abstract 2977: Malignant transformation associated with repetitive Ras/Rac1 activation, increased production of superoxide anion, and global DNA methylation alterations.
Authors: Molognoni Fernanda , Melo Fabiana M. , Tainah Camila , Jasiulionis Miriam G.
Published: August 2014

Driver mutations in N-Ras and BRAF, and more recently in Rac1, were described in melanomas. Using a malignant transformation model of murine melanocytes developed in our laboratory, in which melanoma cell lines were obtained after submitting the melanocyte lineage melan-a to sequential cycles of anchorage blockade, we identified dysfunction in both N-Ras and Rac1 during stressful condition. Our group has already showed increased global DNA methylation as well increased DNA methyltransferase 1 (Dnmt1) protein level few hours after anchorage blockade. This increase seems to be related with elevated amounts of superoxide anion. Previous works have showed that both Ras may regulate superoxide level as Ras activation may affect DNA methylation. Therefore, we aimed to study the relation among Ras/MEK/ERK signaling pathway, oxidative stress and DNA methylation during melan-a anchorage blockade. We showed that Ras, Rac1 and ERK are activated few hours after melan-a anchorage blockade and inhibiting this activation abrogates superoxide increase. Furthermore, global DNA methylation levels decrease during anchorage blockade after treating cells with superoxide scavenger or Ras inhibitor (FTI). Dnmt1 protein level is also decreased after treating melan-a cells during anchorage blockade with FTI or ERK inhibitor (U0126). Ras/MEK/ERK pathway seems to be regulated by and regulates superoxide production during melan-a anchorage restriction. In this way, superoxide anion depletion impairs Ras and ERK activation and FTI or U0126 treatment decreases superoxide during adhesion impediment. This aberrant signaling seems to have a significant impact in melanoma genesis since the malignant transformation was drastically compromised when melan-a melanocytes were pre-treated with superoxide scavenger before each anchorage impediment cycle. Elucidating the relation among oxidative stress, epigenetic changes and melanocyte malignant transformation may provide novel avenues to develop alternative therapeutic approaches.

Citation Format: Fernanda Molognoni, Fabiana M. Melo, Camila Tainah, Miriam G. Jasiulionis. Malignant transformation associated with repetitive Ras/Rac1 activation, increased production of superoxide anion, and global DNA methylation alterations. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2977. doi:10.1158/1538-7445.AM2013-2977