bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract B3: HSP27 is required for invasion and metastasis triggered by hepatocyte growth factor
Authors: Pavan Simona , Musiani Daniele , Torchiaro Erica , Migliardi Giorgia , Erriquez Jessica , Olivero Martina , Renzo Maria Flavia Di
Published: August 2014

The Hepatocyte Growth Factor (HGF) also known as Scatter Factor, activates cancer cell invasion and metastasis. We show that in ovarian cancer cells HGF induced the phosphorylation of the small heat shock protein of 27KDa (HSP27) by activating the p38MAPK. HSP27 is increased in many cancers at advanced stage including ovarian cancer and associated with cancer resistance to therapy and poor patients' survival. The phosphorylation of HSP27 regulates both its chaperone activity and its control of cytoskeletal stability. We show that HSP27 was necessary for the motility in vitro and the remodeling of actin filaments induced by HGF. In vivo, HSP27 silencing impaired the ability of the highly metastatic, HGF-secreting ovarian cancer cells to give rise to spontaneous metastases. This was due to defective motility across the vessel wall and reduced growth. Indeed, HSP27 silencing impaired both the ability of ovarian cancer cells to form experimental haematogenous metastases and to grow as subcutaneous xenografts. Moreover, HSP27 suppression resulted in the sensitization of xenografts to low doses of the chemotherapeutic Paclitaxel, likely because HSP27 protected microtubules from bundling caused by the drug. Altogether, these data show that the HSP27 is required for the pro-invasive and pro-metastatic activity of HGF and suggest that HSP27 might be not only a marker of progression of ovarian cancer, but also a suitable target for therapy. Grant sponsors: This work has been supported by grants to M.F.D.: 2012 IG grant and 2010 Special Program Molecular Clinical Oncology 5xMille of the Italian Association of Cancer Research (AIRC), Project n° 9970, and Grant of the CARIPLO Foundation.

Citation Format: Simona Pavan, Daniele Musiani, Erica Torchiaro, Giorgia Migliardi, Jessica Erriquez, Martina Olivero, Maria Flavia Di Renzo. HSP27 is required for invasion and metastasis triggered by hepatocyte growth factor. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B3.
Abstract B48: Targeting chemo-resistance in CCNE1-amplified ovarian cancer
Authors: Taylor-Harding Barbie , Agadjanian Hasmik , Aspuria Paul Joseph , Mizuno Takako , Cheon Dong-Joo , Orsulic Sandra , Karlan Beth , Walsh Christine , Wiedemeyer Wolf Ruprecht
Published: August 2014

The putative oncogene CCNE1 (cyclin E1) is frequently amplified in human high-grade serous ovarian cancer. CCNE1-amplified cancers tend to be wildtype for BRCA1 and BRCA2 and are associated with shorter survival and resistance to platinum-based chemotherapy. Cyclin E1 is an activating cofactor for cyclin-dependent kinases (CDK), which stimulate cell cycle progression through phosphorylation of the retinoblastoma (RB) protein and subsequent induction of E2F transcriptional activity (cyclin E1-RB-E2F signaling). Since BRCA1 and BRCA2 are known target genes of E2F transcription factors, we hypothesize that cyclin E1 signaling actively contributes to high DNA repair capacity and chemo-resistance in a BRCA1/2-dependent manner. We predict CCNE1-amplified ovarian cancers to exhibit an “anti-BRCAness” phenotype. In two independent gene expression datasets, generated by TCGA and The Women's Cancer Program at Cedars-Sinai, we found that CCNE1-amplified ovarian cancers maintain higher levels of BRCA1 than CCNE1-wildtype tumors. Moreover, we have employed genetic and pharmacological targeting approaches in order to assess if inhibition of cyclin E1-dependent signaling can induce BRCAness and restore chemo-sensitivity. Here, we show that pharmacological CDK inhibitors (CDKi) effectively shut down E2F-mediated transcription, resulting in downregulation of BRCA1 and BRCA2 in CDKi-treated cells. Long-term exposure of ovarian cancer cell lines to CDKi selected for cells with reduced dependency on cyclin E1, and genomic profiling revealed de novo DNA copy number changes which compensate for loss of cyclin E1 function. Importantly, CDKi-resistant subclones retained lower levels of BRCA1 and were significantly more sensitive to cisplatin than parental cell lines. Collectively, our results suggest that cyclin E1 signaling is required for BRCA1 expression and chemo-resistance. Currently available CDKi such as Dinaciclib, which is in a phase 3 clinical trial, may be useful to specifically sensitize CCNE1-amplified ovarian cancers to cisplatin.

Citation Format: Barbie Taylor-Harding, Hasmik Agadjanian, Paul Joseph Aspuria, Takako Mizuno, Dong-Joo Cheon, Sandra Orsulic, Beth Karlan, Christine Walsh, Wolf Ruprecht Wiedemeyer. Targeting chemo-resistance in CCNE1-amplified ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B48.
Abstract B78: Osteopontin-c splicing isoform is a key molecule in ovarian cancer progression
Authors: Tilli Tatiana M. , Mello Kivvi D. , Franco Vanessa , Robbs Bruno , Wanderley João Luiz , Rass Fabrício , Viola João P B , Weber Georg , Castronovo Vincent , Bellahcène Akeila , Gimba Etel R P , Gimba Etel
Published: August 2014

Introduction: Osteopontin (OPN) is a glyphosphoprotein overexpressed in ovarian carcinoma (OC) and is involved in tumorigenesis and metastasis. Alternative splicing of OPN primary transcript generates 3 isoforms, named as OPNa, OPNb, and OPNc. Objective: This study aimed to characterize the expression profile and functional role of each OPN splicing isoform (OPN-SI) in OC. Methodology: The expression patterns of OPN-SI and a series of genes involved in key cancer pathways were analyzed by real-time PCR. In vitro and in vivo functional assays were performed using the ovarian cancer cell line OvCar-3, stably overexpressing the three OPN-SI and conditoned medium secreted by these cells. Results: OPNc, but not OPNa and OPNb, was specifically expressed on OC samples. OPNc splice variant significantly activated OvCar-3 cell proliferation, migration, invasion and anchorage independent cells growth, besides inducing tumor formation in nude mice. We also found that these features are mainly mediated by PI3K/Akt signaling pathway. By using a real time PCR Cancer Gene Array, we found that OvCar-3 OPNc-overexpressing cells are also able to modulate the expression of 34 genes involved in key cancer pathways. Notably, tumors formed by OPNc-overexpressing cells present high expression levels of typical angiogenic markers, such as VEGF-A, VEGFR-2 and CD34. Based on these data, we also investigated the molecular mechanisms by which OPNc stimulates angiogenic processes. Our data showed that OPNc overexpression activates VEGF-A expression and secretion, besides upregulating the expression of c-Fos, c-Jun and phospho-c-Jun. OPNc role on activating the phosphorylation of c-Jun is mediated by the integrin receptor (αvβ3), in an RGD-dependent manner. In addition, OPNc-conditioned medium is able to induce HUVEC endothelial cell proliferation, migration and adhesion. Conclusion: In summary, our data demonstrate that OPNc is able to activate sereral aspects of OC progression, including angiogenesis, mainly by the PI3K/Akt pathway, indicating that this splice variant could be a putative target for further studies aiming to investigate this molecule for new therapeutic approaches for OC.

Citation Format: Tatiana M. Tilli, Kivvi D. Mello, Vanessa Franco, Bruno Robbs, João Luiz Wanderley, Fabrício Rass, João P B Viola, Georg Weber, Vincent Castronovo, Akeila Bellahcène, Etel R P Gimba, Etel Gimba. Osteopontin-c splicing isoform is a key molecule in ovarian cancer progression. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B78.
Abstract IA10: Comprehensive proteomic analyses of TCGA ovarian cancer specimens: Update from CPTAC
Authors: Rodland Karin D.
Published: August 2014

The Cancer Genome Atlas project included ovarian serous cystadenocarcinoma as one of the first four tumor types examined, and TCGA has currently completed comprehensive genomic analyses of over 500 HGSOC tumor specimens, including complete exome sequencing, SNP and methylation analysis, as well as mRNA and miRNA profiling. The Clinical Proteomics Tumor Analysis Consortium (CPTAC) has collaborated with TCGA to provide equally comprehensive proteomic analysis of the same tumors analyzed by TCGA, including quantitative global proteomics, phosphoproteomics, and glycoproteomics. The objective of this study is to further understanding of the execution of genome-level aberrations at the protein level, and to identify functional changes associated with clinical outcome that may provide new insights into the mechanisms of disease progression, factors contributing to chemoresistance, and potentially the identification of new therapeutic targets. Initial analysis of a subset of the TCGA ovarian tumors at Pacific Northwest National Laboratory and Johns Hopkins Medical Institutes has provided preliminary evidence of pathway-level dysregulation associated with platinum resistance and poor clinical outcome.

Citation Format: Karin D. Rodland. Comprehensive proteomic analyses of TCGA ovarian cancer specimens: Update from CPTAC. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr IA10.
Abstract IA21: Panel Discussion: Survivorship Issues
Authors: Levine Douglas A.
Published: August 2014

This one-of-a-kind discussion panel with a mix of both scientists and survivors, will help to bridge the gap between researchers and patients. The main goal is for scientists to appreciate issues of important priority to patients and understand where the ultimate fruits of their labors reside. Scientists will gain a better understanding of the complex issues surrounding first in human studies and the balance of efficacy and toxicity that face patients and doctors on a daily basis. Survivors and clinicians will appreciate the complex environment of drug development in the laboratory and how to take a lead compound through to clinical testing from a human trials perspective. We will discuss issues faced when trying to overcome barriers in improving ovarian cancer outcome and survival. Financial aspects of clinical trials and correlative science continually plague decision-making processes. In addition, how scientists can help advocates in bringing effective drugs into market even when the market share may be small. Scientists will have the opportunity to hear patient stories of diagnosis and treatment as well as specific survivorship issues and healthcare needs of survivors. The balance between efficacy and quality of life will also be discussed.

Citation Format: Douglas A. Levine. Panel Discussion: Survivorship Issues. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr IA21.
Abstract IA22: Causes and consequences of cancer-associated thrombocytosis
Authors: Sood Anil K. , Stone Rebecca L. , Afshar-Kharghan Vahid
Published: August 2014

Platelets represent one of the largest storage pools of angiogenic and oncogenic growth factors in the human body. The observation that thrombocytosis (platelet count >450,000/μL) occurs in patients with solid malignancies was made over 100 years ago. However, mechanisms of paraneoplastic thrombocytosis and the role that platelets play in abetting cancer growth are unclear. We have used clinical data coupled with sophisticated mouse models to identify the mechanisms and biological implications of paraneoplastic thrombocytosis. Thrombocytosis was significantly associated with advanced disease and shortened survival. Plasma levels of thrombopoietin and interleukin-6 were significantly elevated in patients who had thrombocytosis as compared with those who did not. In mouse models, increased hepatic thrombopoietin synthesis in response to tumor-derived interleukin-6 was an underlying mechanism of paraneoplastic thrombocytosis. Tumor-derived interleukin-6 and hepatic thrombopoietin were also linked to thrombocytosis in patients. Silencing thrombopoietin and interleukin-6 abrogated thrombocytosis in tumor-bearing mice. Anti–interleukin-6 antibody treatment significantly reduced platelet counts in tumor-bearing mice and in patients with epithelial ovarian cancer. In addition, neutralizing interleukin-6 significantly enhanced the therapeutic efficacy of paclitaxel in mouse models of epithelial ovarian cancer. The use of an antiplatelet antibody to halve platelet counts in tumor-bearing mice significantly reduced tumor growth and angiogenesis. Biologically, platelets were detected within the tumor microenvironment and affected tumor growth and response to chemotherapeutic agents. These findings support the existence of a paracrine circuit wherein increased production of thrombopoietic cytokines in tumor and host tissue leads to paraneoplastic thrombocytosis, which fuels tumor growth. Blocking the stimulatory effects of platelets may have implications for new therapeutic approaches.

Citation Format: Anil K. Sood, Rebecca L. Stone, Vahid Afshar-Kharghan. Causes and consequences of cancer-associated thrombocytosis. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr IA22.
Abstract IA17: Exploiting the synthetic lethal properties of selective PI3K-β inhibition in PTEN deficient cells with GSK2636771
Authors: Greshock Joel
Published: August 2014

The phosphoinositide 3-kinase (PI3K) pathway is among the most commonly mutated pathways in human cancer. The majority of first-generation PI3K inhibitors currently in clinical trials inhibit multiple PI3K isoforms. The PIK3CB gene encodes p110β isoform, which has been reported to be required for pathway activation in phosphatase and tensin homolog (PTEN) deficient cancer cells. The observed dependence of PTEN-deficient tumors on p110β provides an opportunity for identification of a novel isoform-selective therapeutic with a synthetic lethal effect. During an extensive knowledge-based lead identification and optimization effort, we identified substituted benzimidazole GSK2636771 as a potent, orally bioavailable, PI3Kβ-selective inhibitor. The observed enzymatic potency for PI3Kβ is approximately 1000 and 10 fold higher than the potency against PI3Kα and PI3δ respectively. Pre-clinical data demonstrate the synthetic lethal relationship between PTEN deficiencies and PI3K-β inhibition. PTEN-deficient cells dosed with GSK2636771 show concentration dependent decreases in cellular levels of phosphorylated AKT (pAKT), and PRAS40 both in-vitro and in-vivo. A 3-day proliferation assay performed on a large panel of cell lines (n=80) grown in-vitro demonstrates selective growth inhibition in PTEN-deficient cells at lower concentrations of GSK2636771. In-vivo studies in PTEN-deficient prostate cancer models also show dose dependent tumor growth delay. PTEN is commonly mutated, deleted, or epigenetically silenced in a wide range of solid tumors. Loss of PTEN protein is particularly frequent in prostate (50%), endometrial (57%), and triple negative breast cancers (30%). Prospective selection of patients with this protein loss serves as a development path through the course of study P3B115717 (NCT01458067), a Phase I/IIa first-time-in-human study with GSK2363771. This study is currently in dose-escalation.β

Citation Format: Joel Greshock. Exploiting the synthetic lethal properties of selective PI3K-β inhibition in PTEN deficient cells with GSK2636771. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Synthetic Lethal Approaches to Cancer Vulnerabilities; May 17-20, 2013; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(5 Suppl):Abstract nr IA17.
Abstract B82: Olanzapine for cachexia in patients with advanced cancer.
Authors: Abdelrahim Maen , Naing Aung , Dalal Shalini , Wheler Jennifer , Hess Kenneth , Fu Siqing , Hong David S. , Janku Filip , Falchook Gerald S. , Ilustre Alyson , Ouyang Fengying , Kurzrock Razelle
Published: August 2014

Background: This study assessed the safety, tolerability as well as effectiveness of olanzapine as a treatment for cachexia in patients with advanced cancer.

Methods: Patients with advanced cancers were recruited into this exploratory, open-label, study of olanzapine for cachexia starting at 2.5 milligram (mg) once-daily oral dosing. All patients received anti-neoplastic treatments for their cancers in addition to our study drug olanzapine. Serum samples from patients were collected at baseline, and after weeks 1, 2, 4, and 8 for analysis of levels of leptin, growth hormone, ghrelin, and interleukin-6 (IL-6).

Results: Thirty-nine patients received olanzapine. Of the 39 participants, 31 were evaluable for weight change. Six were excluded because they had developed ascites, and two were excluded because of incomplete body weight data. Toxicities related olanzapine are Grade II somnolence (n=1), grade II pancreatitis (n=1), grade II extrapyramidal symptoms (n=1) and grade II nausea/ vomiting (n=1). No dose-limiting toxicities were reported. The recommended dose was defined as 20 mg daily by mouth once daily. Samples from twenty-nine patient were eligible for analysis of serum cytokine levels. Mean values of leptin levels before treatment and on treatment were 3153 pg/ml vs 2488 pg/ml (p=0.085); for growth hormone the values were 2961 pg/ml vs 3758 pg/ml (p=0.33); for interleukin-6, the values were 27.5 pg/ml vs 40.9 pg/ml (p=0069) and for ghrelin they were 0.18 ng/ml vs 0.19 ng/ml p=0.52. The mean change in slope in weight from prior to and during the study is 0.24 with 95% CI = (-0.08, 0.56) and p = 0.13. Associations between changes in different cytokines and weight were inconclusive.

Conclusion: Though some patients’ serum cytokines levels increased or decreased, these changes did not correlate with change in body weight. Treatment with Olanzapine had a modest effect in changing the trajectory of weight loss.

Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B82.

Citation Format: Maen Abdelrahim, Aung Naing, Shalini Dalal, Jennifer Wheler, Kenneth Hess, Siqing Fu, David S. Hong, Filip Janku, Gerald S. Falchook, Alyson Ilustre, Fengying Ouyang, Razelle Kurzrock. Olanzapine for cachexia in patients with advanced cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B82.
Abstract 4067: Pegylated arginine deiminase (ADI-PEG20) as a potential novel therapy for argininosuccinate synthetase-deficient acute myeloid leukemia
Authors: Szlosarek Peter W. , Luong Fiona , Clear Andrew , Taussig David , Joel Simon , Calaminici Maria , Debernardi Silvana , Fitzgibbon Jude , Bomalaski John S. , Frankel Arthur E. , Bonnet Dominique
Published: August 2014

Acute myeloid leukemia (AML) is the most common leukemia in adults and the second most common leukemia in children, accounting for a significant share of health care costs with ten thousand cases diagnosed per annum in the US alone. Enzyme-based therapy in the form of asparaginase has revolutionized the treatment of acute lymphoblastic leukemia and there is increasing interest in exploiting analogous tractable metabolic defects in AML. Interestingly, arginine auxotrophic tumors due to deficiency of the rate-limiting enzyme for arginine production, argininosuccinate synthetase (ASS1) are susceptible to arginine-degrading enzymes. Here, we studied whether ASS1 negativity would predict for the efficacy of pegylated arginine deiminase (ADI-PEG20) using AML cell lines and primary AML samples. A lack of ASS1 protein was identified in three of seven leukemic cell lines (K562, Kasumi and KG-1) and in all nine samples from patients with cytogenetically normal and abnormal karotype AML. Methylation of the ASS1 promoter correlated with reduced levels of ASS1 mRNA and absence of ASS1 expression. Bone marrow trephines from patients with AML revealed absence of ASS1 protein in 87% (46/53) of samples by immunohistochemistry, indicating that ASS1 expression may be a biomarker of response to ADI-PEG20 in vivo. Increased levels of ASS1 mRNA and detectable ASS1 protein expression were noted in acute promyelocytic leukemia with the translocation t(15;17). Significantly, ADI-PEG20 reduced the viability of ASS1-negative AML lines whereas the ASS1-positive control lines, Fujioka and U937, were resistant to drug-induced arginine deprivation. Recently, we have identified primary ASS1-negative AML samples with good engraftment in NOD/SCID mice and are proceeding to test the efficacy of ADI-PEG20 using this primary AML xenograft model. Based on our preliminary data and the potential efficacy with low toxicity of arginine deprivation in humans, a phase II trial of ADI-PEG20 is planned in patients with relapsed AML.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4067. doi:10.1158/1538-7445.AM2011-4067
Abstract 2377: MCT1 and MCT4 in NSCLC: Overexpression of MCT1 in tumor and stroma is an independent prognostic marker for NSCLC survival.
Authors: Eilertsen Marte , Andersen Sigve , Al-Saad Samer , Kiselev Yury , Donnem Tom , Stenvold Helge , Al-Shibli Khalid , Richardsen Elin , Busund Lill-Tove , Bremnes Roy Martin
Published: August 2014

Background: As non-small cell lung cancer (NSCLC) is a leading cause of cancer deaths, new prognostic and predictive markers are highly warranted. Monocarboxylate transporters (MCTs) 1 and 4 are transmembrane transporters of lactate and puruvate and are central in cancer cell metabolism. The biological and prognostic importance of MCT1 and MCT4 is not thoroughly studied in NSCLC. Herein, we aimed to explore the prognostic impact on disease specific survival (DSS) in both tumor and stromal cells from NSCLC-patients.

Methods: Tissue micro arrays (TMAs) were constructed, representing both epithelial tumor tissue and stromal tissue from 335 unselected patients diagnosed with NSCLC I-IIIA. Immunohistochemistry was used to evaluate the expression of MCT1 and MCT4.

Results: MCT1 and MCT4 expression was located to the cell membrane. In univariate analyses, patients with high MCT1 expression in tumors had a favorable disease-specific survival (DSS) (p=0.021). While high expression of MCT1 in stroma was associated with poor DSS (p=0.003). High expression of MCT4 in tumor was a negative prognostic factor (p=0.027), while high expression of MCT4 in stroma showed a trend toward being a positive prognostic factor (p=0.11). In the multivariate analysis, MCT1 in tumor was independently and significantly associated with a favorable DSS (HR = 1.80, CI 95% 1.23-2.64, p=0.003). MCT1 in stroma was independently and significantly associated with a poor DSS (HR = 1.7, CI 95% 1.1-2.7, p=0.016). MCT4 in tumor had no significant prognostic impact in the multivariate analysis.

Conclusions: High expression of MCT1 in tumor and stroma is an independent prognostic marker for NSCLC survival, and is therefore a candidate marker for prognostic stratification in NSCLC. These results also highlight the differential, but possible cooperative energy metabolism in tumor and stroma for NSCLC-patients.

Citation Format: Marte Eilertsen, Sigve Andersen, Samer Al-Saad, Yury Kiselev, Tom Donnem, Helge Stenvold, Khalid Al-Shibli, Elin Richardsen, Lill-Tove Busund, Roy Martin Bremnes. MCT1 and MCT4 in NSCLC: Overexpression of MCT1 in tumor and stroma is an independent prognostic marker for NSCLC survival. [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 2377. doi:10.1158/1538-7445.AM2013-2377
Abstract 299: Targeting epithelial-mesenchymal transition in breast cancer cells using Honokiol, a natural phenolic compound.
Authors: Avtanski Dimiter B. , Nagalingam Arumugam , Kupusamy Panjamurthi , Saxena Neeraj K. , Sharma Dipali
Published: August 2014

Introduction: The epithelial-mesenchymal transition (EMT) presents a critical step in the acquisition of metastatic state. Tumor cells undergoing EMT not only exhibit increased migration and invasion potential but also acquire increased resistance to chemotherapy and radiation therapy. Hence, EMT is an attractive target for therapeutic interventions directed against tumor metastasis. Honokiol (HNK) is a natural phenolic compound isolated from an extract of seed cones from Magnolia grandiflora. Recent studies from our lab show that honokiol inhibits breast carcinogenesis. The present study is designed to systematically elucidate if nontoxic lower doses of honokiol can be used to specifically inhibit EMT in breast cancer cells.

Results: Our studies show that HNK inhibits EMT in human breast cancer cells. Exposure of estrogen-independent and estrogen-responsive breast cancer cells to nontoxic lower doses of HNK results in significant downregulation of mesenchymal marker proteins (Fibronectin, Vimentin) along with simultaneous upregulation of epithelial markers (E-cadherin, Cytokeratin-18). HNK also inhibits expression and nuclear translocation of transcriptional modulators of EMT-related genes (Snail, ZEB1/2 and Twist). Experimental EMT induced by exposure to TGFβ and TNFα in a spontaneously immortalized nontumorigenic human mammary epithelial cell line (MCF-10A) was also completely reversed by HNK treatment as evidenced by morphological changes (inhibition of fibroblast-like appearance, decreased pseudopodia and actin reorganization to membrane-bound location in TGFβ and TNFα treated MCF10A cells) as well as molecular changes (downregulation of Fibronectin, Vimentin, Snail, Zeb1/2 and upregulation of E-cadherin and Cytokeratin-18). Mechanistically, HNK inhibits MTA1-Wnt1 axis leading to stabilization and nuclear translocation of β-catenin to inhibit EMT. Analysis of breast tumors treated with honokiol show modulation of EMT markers and inhibition of key molecules of MTA1-Wnt1-β-catenin axis corroborating our in vitro findings.

Conclusions: Taken together, these data provide the first in vitro and in vivo evidence of the potential of HNK as a novel, non-toxic and non-endocrine therapeutic strategy for breast carcinoma. MTA1, Wnt1 and β-catenin appear to be novel biomarkers associated with HNK treatment.

Citation Format: Dimiter B. Avtanski, Arumugam Nagalingam, Panjamurthi Kupusamy, Neeraj K. Saxena, Dipali Sharma. Targeting epithelial-mesenchymal transition in breast cancer cells using Honokiol, a natural phenolic compound. [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 299. doi:10.1158/1538-7445.AM2013-299
Abstract 329: A mouse model of epithelial ovarian cancer with defined oncogenic drivers.
Authors: Agadjanian Hasmik , Cheon Dong-Joo , Diaz Elena , Laury Anna , Miller Carl , Karlan Beth , Orsulic Sandra , Walsh Christine
Published: August 2014

Epithelial ovarian cancers (EOC) represent the gynecologic malignancy with the highest mortality rate. The Cancer Genome Atlas demonstrated a high degree of genetic heterogeneity among the papillary serous (PS) tumors, the most lethal of the EOC subtypes. Aside from almost universal mutation in TP53, there was a low prevalence of recurrent mutations in other genes, presenting a challenge to the development of targeted therapeutics against this aggressive tumor type. We are developing a mouse model of EOC by introducing defined genetic alterations to mouse ovarian surface epithelial (MOSE) cells. We had a particular interest in developing a cyclin E amplified tumor model, as CCNE1 amplification is found in 20% of human EOC tumors, making this one of the most prevalent PS EOC subtypes. We infected MOSE cells from p53-/- mice with different combinations of oncogenes encoded in retroviral vectors including CCNE1, myc and H-RAS. MOSE cells were infected with one or two oncogenes, passaged and injected into the peritoneal cavity of nude mice. Myc-HRAS caused the rapid development of hemorrhagic ascites and intraperitoneal carcinomatosis in 5 of 5 mice by day 18. HRAS alone and HRAS-CCNE1 both caused intraperitoneal and omental tumors to develop with clear ascites by day 35 in 5 of 5 mice. Myc caused bloating due to the development of frank hemoperitoneum in all 5 mice between days 101 and 150. Only 4 of the 5 mice had small volume intraperitoneal tumors. Myc-CCNE1 similarly caused bloating due to ascites in 2 mice and hemoperitoneum in 2 mice. Three of these four mice had small volume intraperitoneal tumors. One mouse has not developed any phenotype by 150 days. CCNE1 alone has not caused tumor formation in any of 5 mice by day 150. Histology of the Myc-HRAS, HRAS, HRAS-CCNE1, and Myc tumors were all consistent with high-grade undifferentiated carcinoma. Tumor cells generated in the Myc-HRAS, HRAS and HRAS-CCNE1 experiments were capable of causing intraperitoneal tumor development in immunocompetent C57BL/6 mice by day 17, 29 and 54 respectively. Taken together, our data demonstrate a reproducible mouse model for the development of high-grade epithelial ovarian carcinomas with defined genetic alterations that are capable of study in both immunodeficient and immunocompetent mice. Tumor phenotypes varied based on the combination of activated oncogenes.

Citation Format: Hasmik Agadjanian, Dong-Joo Cheon, Elena Diaz, Anna Laury, Carl Miller, Beth Karlan, Sandra Orsulic, Christine Walsh. A mouse model of epithelial ovarian cancer with defined oncogenic drivers. [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 329. doi:10.1158/1538-7445.AM2013-329
Abstract 3504: Plasma growth differentiating factor-15 (GDF-15) and other inflammatory markers are associated with weight loss and poor prognosis in cancer patients.
Authors: Lorena Lerner , Hayes Teresa , Tao Nianjun , Krieger Brian , Feng Bin , Nicoletti Richard , Bai Ailin , Weng Zhigang , Liu Qing , Chiu Maria Isabel , Gyuris Jeno , Garcia Jose M.
Published: August 2014

Background: Cachexia is associated with increased inflammatory markers and decreased survival in cancer. Also, elevated GDF-15 has been associated with poor prognosis in several cancer types but its role in cachexia is not well-understood.

Methods: We measured body weight change, appetite, plasma GDF-15, and other inflammatory markers in 62 males with cancer-cachexia (CC), 72 non-cachectic cancer subjects (CNC) and 64 non-cancer controls (Co) matched by age, gender and pre-illness body weight. In a subset of patients we also measured grip strength (HGS), appendicular lean body mass (aLBM), ECOG and KPS.

Results: GDF-15, IL-6 and IL-8 were increased in CC vs. other groups. Activin and G-CSF were significantly upregulated in CC vs. Co. A subset analyses showed that GDF-15, Activin A and IL-8 were increased in CC vs. CNC in lung cancer patients and that GDF-15, IL-6 and IL-8 were increased in CC patients treated with platinum-based chemotherapy.

GDF15, IL-6 and IL-8 levels significantly correlated with 6-month weight loss and with IL-6, IL-1ra, IL-2, IL-4, IL-9, IL-10, IFN, MCP-10, MIP-1a, MIP-1b, TNF-a, VGEF and activin in cancer patients. Analysis in a subset of patients showed that CC had lower grip strength, aLBM, and fat mass; and that ECOG and KPS were lower in CC and CNC compared to controls. GDF-15 and IL-8 correlated negatively with aLBM, HGS and fat mass. Activin correlated negatively with aLBM.

Survival analysis showed that GDF-15 and IL-8 predicted survival adjusting for stage and weight change (Cox regression p<0.01, HR 3).

Conclusion: GDF-15 and other inflammatory markers are associated with weight loss, decreased muscle mass and strength and poor survival in cancer patients. GDF-15 may serve as a prognostic indicator in cancer patients and be a novel therapeutic target for cancer cachexia.

Citation Format: Lerner Lorena, Teresa Hayes, Nianjun Tao, Brian Krieger, Bin Feng, Richard Nicoletti, Ailin Bai, Zhigang Weng, Qing Liu, Maria Isabel Chiu, Jeno Gyuris, Jose M. Garcia. Plasma growth differentiating factor-15 (GDF-15) and other inflammatory markers are associated with weight loss and poor prognosis in cancer patients. [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 3504. doi:10.1158/1538-7445.AM2013-3504
Abstract 366: Downregulation of THBS1 is a critical step in glioblastoma angiogenesis.
Authors: Brooks Michael D. , Jackson Erin , Piwnica-Worms David , Mitra Robi D. , Rubin Joshua B.
Published: August 2014

One of the hallmarks of Glioblastoma multiforme (GBM) is its abundant vasculature. These blood vessels support the tumor in a number of ways, including providing a niche for glioblastoma cancer stem cells. GBM tumors drive the formation of their vasculature and associated germinal niches through reciprocal inductive interactions between GBM cells and endothelial cells. Targeting the primary drivers of angiogenesis, VEGF and bFGF, in anti-angiogenic cancer treatments has value but is not curative. We hypothesize that identification of additional angiogenic molecules and pathways will provide new avenues for treatment. We used microarrays to perform global gene expression profiling to identify genes that are increased or decreased in expression as a consequence of functional interactions between GBM and brain microvascular endothelial cells. We identified several regulators of angiogenesis whose expression is modulated by this cell-cell interaction, but were particularly interested in the downregulation of Thrombospondin-1 (THBS1) and its associated receptors. THBS1 is a powerful anti-angiogenic protein and has previously been implicated in cancer, but its precise mechanism in glioblastoma angiogenesis is not yet known. A number of in-vitro experiments validated the importance of THBS1 in the interaction between GBM and endothelial cells. In-vivo mouse xenograft experiments showed that overexpression of THBS1 strongly inhibited tumor formation. We believe these results reveal the importance of factors other than VEGF in glioblastoma angiogenesis and provide a rationale to evaluate combinatorial targeted anti-angiogenic therapy for GBM.

Citation Format: Michael D. Brooks, Erin Jackson, David Piwnica-Worms, Robi D. Mitra, Joshua B. Rubin. Downregulation of THBS1 is a critical step in glioblastoma angiogenesis. [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 366. doi:10.1158/1538-7445.AM2013-366
Abstract 3806: Progressive biallelic loss of TP53 is associated with progression of pleuropulmonary blastoma initiated by germline loss and somatic mutation of DICER1.
Authors: Pugh Trevor J. , Yu Weiying , Yang Jiandong , Ambrogio Lauren , Carter Scott L. , Kiezun Adam , McKenna Aaron , Giannikopoulos Petros , Getz Gad , Messinger Yoav , Meyerson Matthew , Hill D. Ashley
Published: August 2014

Pleuropulmonary blastoma (PPB) is a rare, aggressive childhood cancer arising from lung mesenchymal cells. In early stages, these cells reside in the walls of epithelial-lined cysts and may remain dormant or progress to thickened cysts and then highly malignant masses over several years. Familial germline loss-of-function DICER1 variants have been described, although carriers may not be affected. Therefore, we sought to uncover additional genetic events underlying tumor development through whole exome sequencing of 15 tumor/normal pairs. In addition to known loss-of-function germline variants found throughout DICER1, this analysis identified somatic missense mutations clustered in the DICER1 RNase IIIb domain in nearly every case. While the somatic mutations impact novel amino acid residues, expression analysis demonstrated depletion of 5p-derived miRNAs similar to that described in other DICER1-mutant tumors. While mutation rates were relatively high compared to other pediatric solid tumors, most mutations, except those in DICER1 and TP53, appeared to be subclonal. Early stage tumors (PPB Type II) had predominantly single-copy deletion of TP53 whereas late stage tumors (PPB Type III) had secondary TP53 mutations. Several late stage tumors also had activating mutations in RAS pathway members, including NRAS and BRAF. Therefore, compound DICER1 loss appears to be initiating and synergistic with incremental loss of TP53 and acquisition of additional driver mutations. We subsequently characterized this molecular pattern using deep sequencing and immunohistochemistry in an extension cohort of 45 additional tumors. This step-wise series of germline and somatic alterations illustrates a genetic mechanism consistent with the progressive nature of pleuropulmonary blastoma.

Citation Format: Trevor J. Pugh, Weiying Yu, Jiandong Yang, Lauren Ambrogio, Scott L. Carter, Adam Kiezun, Aaron McKenna, Petros Giannikopoulos, Gad Getz, Yoav Messinger, Matthew Meyerson, D. Ashley Hill. Progressive biallelic loss of TP53 is associated with progression of pleuropulmonary blastoma initiated by germline loss and somatic mutation of DICER1. [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 3806. doi:10.1158/1538-7445.AM2013-3806
Abstract 3893: Endocan is upregulated on tumor vessels in invasive bladder cancer and mediates VEGF-A-induced angiogenesis.
Authors: Roudnicky Filip , Poyet Cedric , Wild Peter J. , Krampitz Sarah , Negrini Fabrizia , Huggenberger Reto , Rogler Anja , Stöhr Robert , Hartmann Arndt , Provenzano Maurizio , Otto Vivianne I. , Detmar Michael
Published: August 2014

Tumor-associated blood vessels differ from normal vessels at the morphological and molecular level. Proteins that are only present on tumor vessels may serve as biomarkers and as therapeutic targets for inhibition of angiogenesis in cancer. To identify factors contributed by the tumor-associated endothelium, we performed immuno-laser capture microdissection (i-LCM) of blood vascular endothelial cells in conjunction with transcriptional profiling from surgically harvested cancer and normal matched bladder tissue of five patients with invasive bladder cancer. Comparing the transcriptional profiles of blood vascular endothelium from human invasive bladder cancer and from normal bladder tissue, we found that endocan (endothelial cell-specific molecule-1) was highly elevated on tumor vessels and associated with the filopodia of angiogenic endothelial tip cells in invasive bladder cancer. Notably, endocan expression on tumor vessels strongly correlated with the tumor stage and invasiveness and predicted a shorter recurrence-free survival time in non-invasive bladder cancers. Furthermore, both endocan and VEGF-A levels were significantly higher in plasma of patients with invasive bladder cancer (n=53) compared to healthy individuals (n=60). Using cultured blood vascular endothelial cells and an in vivo transgenic mouse model we found that endocan expression was strongly upregulated by VEGF-A activation of VEGFR-2. RNA interference-mediated knockdown of endocan in cultured blood vascular endothelial cells revealed that endocan was required for the promotion of cell migration and tube formation by VEGF-A. Furthermore, we found that endocan potentiates VEGFR-2 phosphorylation in response to VEGF-A. Therefore, blocking the interaction of endocan with either VEGFR-2 or VEGF-A might represent a promising approach for inhibiting tumor angiogenesis. Endocan might also serve as a novel biomarker for monitoring disease progression and the efficacy of VEGF-A-targeting therapies in patients with bladder cancer.

Citation Format: Filip Roudnicky, Cedric Poyet, Peter J. Wild, Sarah Krampitz, Fabrizia Negrini, Reto Huggenberger, Anja Rogler, Robert Stöhr, Arndt Hartmann, Maurizio Provenzano, Vivianne I. Otto, Michael Detmar. Endocan is upregulated on tumor vessels in invasive bladder cancer and mediates VEGF-A-induced angiogenesis. [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 3893. doi:10.1158/1538-7445.AM2013-3893
Abstract 4238: BROMOscan - a high throughput, quantitative ligand binding platform identifies best-in-class bromodomain inhibitors from a screen of mature compounds targeting other protein classes.
Authors: Quinn Elizabeth , Wodicka Lisa , Ciceri Pietro , Pallares Gabriel , Pickle Elyssa , Torrey Adam , Floyd Mark , Hunt Jeremy , Treiber Daniel
Published: August 2014

Post-translationally appended acetyllysine marks on histone tails are key regulatory features of the epigenetic code. Bromodomains are “readers” of this specific lysine acetylation code, playing an important role in chromatin remodeling and regulation of gene expression. Bromodomains have emerged as an important new druggable target class in small-molecule inhibitor drug discovery, and several bromodomain-containing proteins have been associated with disease. There are 57 bromodomains contained in 41 different proteins; however, few small molecule bromodomain inhibitors have been reported. One primary factor limiting the discovery of new inhibitors is the absence of a comprehensive biochemical bromodomain screening platform. Here we describe the application of DiscoveRx Corporation's proven ligand binding assay technology (KINOMEscan) to the development of quantitative ligand binding assays for human bromodomains (BROMOscan). We have developed a carefully validated assay panel that covers >30 percent of the human bromodomain family, and this panel is suitable for HTS, selectivity profiling, and quantitative affinity (Kd) assessment. We have used this panel internally to identify novel bromodomain inhibitors and, remarkably, have demonstrated that known, mature inhibitors thought to be selective for targets from other protein families have best in class affinity for bromodomains as well. These data shall be presented, as will a description of the BROMOscan panel replete with extensive assay validation data.

Citation Format: Elizabeth Quinn, Lisa Wodicka, Pietro Ciceri, Gabriel Pallares, Elyssa Pickle, Adam Torrey, Mark Floyd, Jeremy Hunt, Daniel Treiber. BROMOscan - a high throughput, quantitative ligand binding platform identifies best-in-class bromodomain inhibitors from a screen of mature compounds targeting other protein classes. [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 4238. doi:10.1158/1538-7445.AM2013-4238
Abstract 5066: Matriptase mediated signaling in breast cancer.
Authors: Zoratti Gina L. , Bergum Christopher , Boerner Julie L. , List Karin
Published: August 2014

Historically proteases have been associated with tumor progression and metastasis through degradation of the basement membrane. In recent years these enzymes have also been shown to play key roles in the activation of growth factors and cytokines, thereby activating pro-oncogenic signaling pathways. Matriptase is an epithelia-specific pericellular protease which has received considerable attention recently spurred by its consistent dysregulation in human epithelial tumors including breast cancer. Several institutions are currently developing small and large molecule inhibitors of matriptase, however thus far no studies have been published focusing on the role of matriptase in breast cancer growth and progression. Presently we do not know whether matriptase plays a causal or promotional role in breast carcinogenesis and therefore its potential as an anti-cancer drug target is still unexplored. In the following study we will perform both functional and mechanistic studies of matriptase in breast cancer using parallel, complementary in-vitro and in-vivo genetic “loss-of-function” strategies. Matriptase efficiently cleaves pro-hepatocyte growth factor (pro-HGF) into active HGF, the cognate ligand of the proto-oncogene tyrosine kinase receptor c-Met, and thereby elicits pro-tumorigenic downstream signaling events. Both c-Met and matriptase are expressed on the surface of mammary epithelial cells and breast carcinoma cells, whereas the c-Met ligand and matriptase proteolytic target pro-HGF is secreted by stromal fibroblasts thus defining a paracrine growth factor activation and signaling mechanism. Our goal with this project is to determine whether matriptase is a novel target which could provide an opportunity for developing improved breast cancer intervention drugs in the future.

Citation Format: Gina L. Zoratti, Christopher Bergum, Julie L. Boerner, Karin List. Matriptase mediated signaling in breast 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 5066. doi:10.1158/1538-7445.AM2013-5066