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Transcriptional regulation of prostate cancer metabolism
EACR24 Poster Sessions / European Journal of Cancer 61, Suppl. 1 (2016) S9–S218 ligand-independent signalling of Notch may be a new strategy to specifically interfere with Notch mutants. No conflict of interest. 755 HER2 positivity is high in breast cancer of young women: Experience of a single institute in Turkey E. Celikmakas1 , Y. Karslioglu2 , I. Hacibey1 , S. Kilic3 , M. Ozturk4 , M. Safali2 , O. Onguru2 . 1 Gulhane Military Medical Academy, School of Medicine, Ankara, Turkey, 2 Gulhane Military Medical Academy, Department of Pathology, Ankara, Turkey, 3 Gulhane Military Medical Academy, Department of Public Health, Ankara, Turkey, 4 Gulhane Military Medical Academy, Department of Oncology, Ankara, Turkey Background: Breast cancer can be devastating issue in young women with some different clinical, epidemiological and genetic characteristics compared to the ones in older patients. The incidence of breast cancer show geographical differences. In this study, we systematically analyzed the clinicopathologic features of breast cancer in young women from a single tertiary institute of Turkey. Methods: We retrospectively collected information of 91 women diagnosed with breast cancer, aged <40 years. All of the patients were diagnosed, operated and treated at Gulhane Military Medical Academy. The data included the following variables: age, family history, pregnancy at the time of diagnosis, histologic type, histologic grade, tumor size, lymph node status, distant metastasis, HER2 and hormonal receptor expression and stage. Results: Mean age of patients was 33.2 (19−39). 28.6% (26/91) of patients were 30. A positive family history of breast cancer was present in 10.9% (10/91). Majority (94.5%) of tumors were invasive ductal carcinoma, NOS type and most (62.6%) of them were histologically grade III. Mean tumor diameter was 2.9 (0.2−6.5). Of all cancers, 34.4% were T1, 59.0% T2, 6.6% T3 and T4. 78.5% of cases presented with lymph node metastasis. Thirteen of the patients (14.3%, 13/91) had distant metastasis at presentation. Bone and liver was the most frequent site of metastasis. Four (4.4%) tumors were presented in pregnancy. At the time of diagnosis, contralateral breast cancer was detected in five cases (5.5%). Hormone receptor was positive in 49.2%. Her2 was positive in 38.1%. Of the cases, 12.7% were triple negative breast cancer (TNBC). The frequency of TNBC and Her2 positivity at very young age (30) was not different to the other subjects in our series. Conclusion: Breast cancers in young patients typically present as large palpable tumor with frequent nodal metastasis. Histologically, most of the tumors tend to be high grade. Her2 positivity is approximately two times more frequent compared to overall incidence of Her2 overexpression (15−20%) in all breast cancer cases. No conflict of interest.
Monday 11 July 2016 Poster Session
Signalling Pathways II 756 Transcriptional regulation of prostate cancer metabolism L. Valcarcel1 , V. Torrano1 , A.R. Cortazar1 , X. Liu2 , J. Urosevic3 , M. Castillo4 , G. Morciano5 , M. Graupera6 , P. Pandya7 , M. Unda-Urzaiz8 , N. Schultz9 , A. Aransay10 , V. Sanz-Moreno11 , R. Barrio12 , G. Velasco13 , P. Pinton5 , C. Cordon-Cardo4 , R.R. Gomis3 , J. Locasale2 , A. Carracedo1 . 1 CICbioGUNE, Proteomics, Derio, Spain, 2 Cornell University, Division of Nutritional Sciences, Ithaca, NY, USA, 3 Institute for Research in Biomedicine IRB, MetLab − Oncology Programme, Barcelona, Spain, 4 Icahn School of Medicine at Mount Sinai, Pathology, Manhattan, USA, 5 University of Ferrara, Department of Morphology- Surgery and Experimental Medicine Section of General ` de Bellvitge Pathology, Ferrara, Italy, 6 Institut d’Investigacio´ Biomedica IDIBELL, Vascular Signalling Laboratory, Barcelona, Spain, 7 New Hunt’s House- Guy’s Campus, Tumour Plasticity Team- Randall Division of Cell and Molecular Biophysic, London, United Kingdom, 8 Basurto University Hospital, Department of Urology, Bilbao, Spain, 9 Memorial Sloan-Kettering Cancer Center, Computational Biology, New York, USA, 10 CICbioGUNE, Genome Analysis Platform, Derio, Spain, 11 King’s College London- New Hunt’s House- Guy’s Campus, Tumour Plasticity Team- Randall Division of Cell and Molecular Biophysics, London, United Kingdom, 12 CICbioGUNE, Functional Genomics, Derio, Spain, 13 Complutense University and Instituto de Investigaciones Sanitarias San Carlos IdISSC, Department of Biochemistry and Molecular Biology I- School of Biology, Madrid, Spain Background: The current view of cellular transformation and cancer progression supports the notion that cancer cells must undergo a metabolic switch in order to survive and progress in a hostile environment. The cancer metabolism field emerged with the observation of a variety of cancer genes (oncogenes and tumour suppressors) regulating cellular and organism metabolism, in turn linking metabolic disorders to cancer pathogenesis.
S171
Metabolic reprogramming in cancer is the integration of a wide array of metabolic routed that need to be coordinatedly altered. Despite the extensive information about the metabolic changes occurring in cancer, we still do not understand which genetic cues are responsible for the coordination of such complex program. Master co-regulators of metabolism orchestrate the modulation of multiple metabolic pathways through transcriptional programs, and hence constitute a probabilistically parsimonious mechanism for general metabolic rewiring. Therefore, the driving force of this study is to define the molecular cues that are at the core of the complex metabolic rewiring occurring in prostate cancer. Methods: We have approached the study of prostate cancer (PCa) metabolism starting from extensive bioinformatics analysis of large prostate cancer datasets followed by genetic mouse models of prostate cancer, cellular analysis and integrative transcriptomics with state-of-the-art untargeted metabolomics, stable isotope tracing and biochemical assays. Results: We have applied meta-analysis constrains that ensure the selection of relevant candidates contributing to the global metabolic switch, based on two premises: i) they are altered in a significant proportion of prostate cancer studies and ii) they are associated to aggressiveness of the disease. 9 metabolic co-regulators were found altered in the pilot database, while only 3 complied with premise 1. From these 3 co-regulators, only one was associated with aggressiveness. After this medium throughput screen, we validated the relevance of our metabolic regulator through the use of Genetically Engineered Mouse Models of prostate cancer, and surrogate cellular systems. Furthermore, we have used cellular systems to recapitulate the features observed in GEMMs and defined the transcriptional and metabolic means that contribute to prostate cancer progression. Conclusions: In this study we demonstrate that a global metabolic rewiring driven by a anabolic/catabolic switch and the unfavoured use of mitochondrial oxidative programs (i) can be elicited through the alteration of master transcriptional metabolic program, and (ii) opposes the progression and dissemination of prostate cancer. No conflict of interest. 758 PREX1 integrates G protein-coupled receptor and phosphoinositide 3-kinase signaling to promote glioblastoma invasion A. Gont1,2 , M. Daneshmand1,3 , J. Woulfe1,2,3 , I. Lorimer1,2 . 1 Ottawa Hospital Research Institute, Cancer Theraputics, Ottawa, Canada, 2 University of Ottawa, Department of Biochemistry- Microbiology and Immunology, Ottawa, Canada, 3 University of Ottawa, Department of Pathology and Laboratory Medicine, Ottawa, Canada Introduction: A defining feature of the brain cancer glioblastoma is its highly invasive nature. The Rac subclass of Rho GTPases has been shown to promote invasive behaviour in glioblastoma. PREX1 was originally identified as a Rac GEF specifically responsive to the phosphoinositide 3-kinase (PI-3-Kinase) pathway second messenger phosphatidylinositol 3,4,5trisphosphate (PIP3). Further research showed that PREX1 is synergistically activated by binding of both PIP3 and G-protein couple receptor bg subunits. PREX1 expression and function have not been investigated in glioblastoma. Methods and Results: We found that PREX1, although absent in the U87MG glioblastoma cell line, is readily detected in glioblastoma cells isolated from patients under serum-free conditions. PREX1 expression was detected both when these primary glioblastoma cells were grown in cell culture and when they were grown as xenografts in immunocompromised mice. The level of PREX1 expression varied between cultures isolated from different patients. Some of this variation appears to be related to the molecular subtype of glioblastoma, as analysis of the TCGA database showed that PREX1 is generally expressed at higher levels in the classical subtype. Analysis of a tissue microarray containing samples from thirty five glioblastoma patients showed that PREX1 was expressed in most cases, with a subset of patients showing high expression. PREX1 expression was low or absent in normal brain tissue. Knockdown of PREX1 with siRNA duplexes decreased cell invasion through coated transwell chambers and decreased motility in time-lapse videomicroscopy experiments in multiple primary glioblastoma cell cultures, while exhibiting no effects on cell growth. To investigate the upstream activators of PREX1, gallein, a G protein-coupled receptor bg subunit inhibitor, and BKM120, a PI-3-kinase a/b inhibitor were used. Individually, gallein and BKM120 inhibited glioblastoma cell culture motility. Combination treatment did not have any additional effects, suggesting that both PI-3-kinase and G proteincoupled receptor signalling are required for optimal cell migration. Glioblastoma cell motility was also inhibited by high concentrations of haloperidol, suggesting that dopamine, serotonin or adrenergic receptor activity may contribute to PREX1 activation in glioblastoma. Conclusions: Overall, PREX1 is overexpressed in many glioblastomas, particularly in the classical subtype. PREX1 is also expressed in glioblastoma cells isolated from patients, where it integrates signals from both PI-3-kinase pathway signalling and G protein-coupled receptor signaling to drive invasion. The PI-3-kinase pathway is activated by mutation in most glioblastomas; our
Monday 11 July 2016 Poster Session
Signalling Pathways II 756 Transcriptional regulation of prostate cancer metabolism L. Valcarcel1 , V. Torrano1 , A.R. Cortazar1 , X. Liu2 , J. Urosevic3 , M. Castillo4 , G. Morciano5 , M. Graupera6 , P. Pandya7 , M. Unda-Urzaiz8 , N. Schultz9 , A. Aransay10 , V. Sanz-Moreno11 , R. Barrio12 , G. Velasco13 , P. Pinton5 , C. Cordon-Cardo4 , R.R. Gomis3 , J. Locasale2 , A. Carracedo1 . 1 CICbioGUNE, Proteomics, Derio, Spain, 2 Cornell University, Division of Nutritional Sciences, Ithaca, NY, USA, 3 Institute for Research in Biomedicine IRB, MetLab − Oncology Programme, Barcelona, Spain, 4 Icahn School of Medicine at Mount Sinai, Pathology, Manhattan, USA, 5 University of Ferrara, Department of Morphology- Surgery and Experimental Medicine Section of General ` de Bellvitge Pathology, Ferrara, Italy, 6 Institut d’Investigacio´ Biomedica IDIBELL, Vascular Signalling Laboratory, Barcelona, Spain, 7 New Hunt’s House- Guy’s Campus, Tumour Plasticity Team- Randall Division of Cell and Molecular Biophysic, London, United Kingdom, 8 Basurto University Hospital, Department of Urology, Bilbao, Spain, 9 Memorial Sloan-Kettering Cancer Center, Computational Biology, New York, USA, 10 CICbioGUNE, Genome Analysis Platform, Derio, Spain, 11 King’s College London- New Hunt’s House- Guy’s Campus, Tumour Plasticity Team- Randall Division of Cell and Molecular Biophysics, London, United Kingdom, 12 CICbioGUNE, Functional Genomics, Derio, Spain, 13 Complutense University and Instituto de Investigaciones Sanitarias San Carlos IdISSC, Department of Biochemistry and Molecular Biology I- School of Biology, Madrid, Spain Background: The current view of cellular transformation and cancer progression supports the notion that cancer cells must undergo a metabolic switch in order to survive and progress in a hostile environment. The cancer metabolism field emerged with the observation of a variety of cancer genes (oncogenes and tumour suppressors) regulating cellular and organism metabolism, in turn linking metabolic disorders to cancer pathogenesis.
S171
Metabolic reprogramming in cancer is the integration of a wide array of metabolic routed that need to be coordinatedly altered. Despite the extensive information about the metabolic changes occurring in cancer, we still do not understand which genetic cues are responsible for the coordination of such complex program. Master co-regulators of metabolism orchestrate the modulation of multiple metabolic pathways through transcriptional programs, and hence constitute a probabilistically parsimonious mechanism for general metabolic rewiring. Therefore, the driving force of this study is to define the molecular cues that are at the core of the complex metabolic rewiring occurring in prostate cancer. Methods: We have approached the study of prostate cancer (PCa) metabolism starting from extensive bioinformatics analysis of large prostate cancer datasets followed by genetic mouse models of prostate cancer, cellular analysis and integrative transcriptomics with state-of-the-art untargeted metabolomics, stable isotope tracing and biochemical assays. Results: We have applied meta-analysis constrains that ensure the selection of relevant candidates contributing to the global metabolic switch, based on two premises: i) they are altered in a significant proportion of prostate cancer studies and ii) they are associated to aggressiveness of the disease. 9 metabolic co-regulators were found altered in the pilot database, while only 3 complied with premise 1. From these 3 co-regulators, only one was associated with aggressiveness. After this medium throughput screen, we validated the relevance of our metabolic regulator through the use of Genetically Engineered Mouse Models of prostate cancer, and surrogate cellular systems. Furthermore, we have used cellular systems to recapitulate the features observed in GEMMs and defined the transcriptional and metabolic means that contribute to prostate cancer progression. Conclusions: In this study we demonstrate that a global metabolic rewiring driven by a anabolic/catabolic switch and the unfavoured use of mitochondrial oxidative programs (i) can be elicited through the alteration of master transcriptional metabolic program, and (ii) opposes the progression and dissemination of prostate cancer. No conflict of interest. 758 PREX1 integrates G protein-coupled receptor and phosphoinositide 3-kinase signaling to promote glioblastoma invasion A. Gont1,2 , M. Daneshmand1,3 , J. Woulfe1,2,3 , I. Lorimer1,2 . 1 Ottawa Hospital Research Institute, Cancer Theraputics, Ottawa, Canada, 2 University of Ottawa, Department of Biochemistry- Microbiology and Immunology, Ottawa, Canada, 3 University of Ottawa, Department of Pathology and Laboratory Medicine, Ottawa, Canada Introduction: A defining feature of the brain cancer glioblastoma is its highly invasive nature. The Rac subclass of Rho GTPases has been shown to promote invasive behaviour in glioblastoma. PREX1 was originally identified as a Rac GEF specifically responsive to the phosphoinositide 3-kinase (PI-3-Kinase) pathway second messenger phosphatidylinositol 3,4,5trisphosphate (PIP3). Further research showed that PREX1 is synergistically activated by binding of both PIP3 and G-protein couple receptor bg subunits. PREX1 expression and function have not been investigated in glioblastoma. Methods and Results: We found that PREX1, although absent in the U87MG glioblastoma cell line, is readily detected in glioblastoma cells isolated from patients under serum-free conditions. PREX1 expression was detected both when these primary glioblastoma cells were grown in cell culture and when they were grown as xenografts in immunocompromised mice. The level of PREX1 expression varied between cultures isolated from different patients. Some of this variation appears to be related to the molecular subtype of glioblastoma, as analysis of the TCGA database showed that PREX1 is generally expressed at higher levels in the classical subtype. Analysis of a tissue microarray containing samples from thirty five glioblastoma patients showed that PREX1 was expressed in most cases, with a subset of patients showing high expression. PREX1 expression was low or absent in normal brain tissue. Knockdown of PREX1 with siRNA duplexes decreased cell invasion through coated transwell chambers and decreased motility in time-lapse videomicroscopy experiments in multiple primary glioblastoma cell cultures, while exhibiting no effects on cell growth. To investigate the upstream activators of PREX1, gallein, a G protein-coupled receptor bg subunit inhibitor, and BKM120, a PI-3-kinase a/b inhibitor were used. Individually, gallein and BKM120 inhibited glioblastoma cell culture motility. Combination treatment did not have any additional effects, suggesting that both PI-3-kinase and G proteincoupled receptor signalling are required for optimal cell migration. Glioblastoma cell motility was also inhibited by high concentrations of haloperidol, suggesting that dopamine, serotonin or adrenergic receptor activity may contribute to PREX1 activation in glioblastoma. Conclusions: Overall, PREX1 is overexpressed in many glioblastomas, particularly in the classical subtype. PREX1 is also expressed in glioblastoma cells isolated from patients, where it integrates signals from both PI-3-kinase pathway signalling and G protein-coupled receptor signaling to drive invasion. The PI-3-kinase pathway is activated by mutation in most glioblastomas; our