bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 805: Subcellular distribution of RAGE affects its functions in melanoma growth and progression
Authors: Reith Maike , Gebhardt Wolf-Henning , Tarnanidis Kathrin , Wagner Nikolaus B. , Ikenberg Kristian , Kehrel Coretta , Utikal Jochen , Gebhardt Christoffer
Published: August 2015

The receptor for advanced glycation end-products (RAGE) is known to act as a central driver of tumorigenesis by sustaining a chronic inflammatory tumor microenvironment. Until to date, RAGE has been exclusively described as a cell surface receptor being activated upon engagement with its various extra-cellular ligands, e.g. S100B, S100A8/A9, HMGB1, and others. This study aimed at elucidating the functional role of RAGE and its isoforms depending on their subcellular distribution in the context of melanoma development, growth and progression. Therefore, various in vitro models using melanoma cells or melanocytes and melanoma mouse models as well as tissue-microarrays representing human specimens of malignant melanoma and benign nevi were applied.

The expression analyses revealed an overexpression of RAGE in melanoma cells compared to melanocytes/nevocytes. Moreover, RAGE protein was found to be localized primarily in the nucleus of melanocytes/nevocytes whereas a predominant cell surface/cytoplasmic localization of RAGE is observed in melanoma cells. Nuclear translocation of RAGE depends on the nuclear transport machinery and site-directed mutagenesis of predicted DNA binding sites within the RAGE protein was applied in order to study its functional role in the nucleus. Furthermore, knockdown of RAGE indicated its central function in apoptosis induction.

In conclusion, RAGE and its isoforms are overexpressed and aberrantly localized in malignant melanoma cells compared to melanocytes/nevocytes. Our data point towards a novel tumor-protective function of RAGE in melanoma development depending on its nuclear localization.

Citation Format: Maike Reith, Wolf-Henning Gebhardt, Kathrin Tarnanidis, Nikolaus B. Wagner, Kristian Ikenberg, Coretta Kehrel, Jochen Utikal, Christoffer Gebhardt. Subcellular distribution of RAGE affects its functions in melanoma growth and progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 805. doi:10.1158/1538-7445.AM2015-805
Abstract 812: Processing of core protein by signal peptide peptidase participates in propagation and pathogenesis of hepatitis C virus
Authors: Okamoto Toru , Aizawa Sayaka , Kouwaki Takahisa , Suzuki Tatsuya , Fukuhara Takasuke , Moriishi Kohji , Koike Kazuhiko , Matsuura Yoshiharu
Published: August 2015

Hepatitis C virus (HCV) is a major causative agent of chronic liver diseases including steatosis, cirrhosis and hepatocellular carcinoma (HCC). The HCV core protein is cleaved off from a precursor polyprotein by a signal peptidase and then further processed by signal peptide peptidase (SPP) within the transmembrane region. However, the biological significance of the processing of HCV core protein by SPP remains largely unknown. Transgenic mice expressing HCV core protein exhibit insulin-resistance, steatosis and finally develop HCC, suggesting that core protein plays crucial roles in pathogenesis of HCV. In this study we have established SPP-knockout (KO) mice and cell lines to examine the roles of SPP on the propagation and pathogenesis of HCV. SPP KO mice were embryonic lethal at around E13.5, therefore we prepared SPP-KO mouse embryonic fibroblast cells (MEFs). In SPP-KO cells, HCV core protein was quickly degraded by proteasome and propagation of HCV was severely impaired. Treatment of cells with LY-411575, an inhibitor for SPP, also induced degradation of HCV core protein and suppressed the production of infectious HCV particles. PA28γ or E6AP, which had been reported to degrade core protein, were not involved in SPP-mediated degradation, suggesting the other mechanisms were involved in SPP-induced core protein degradation. Furthermore, oral administration of LY-411575 into HCV core transgenic mice reduced expression of core protein and cured insulin-resistance. The deletion of one allele of SPP gene in core transgenic mice led to impair the expression of core protein and cure insulin-resistance. These results suggest that SPP is a novel drug target for chronic hepatitis C through suppression of viral propagation and liver failures.

Citation Format: Toru Okamoto, Sayaka Aizawa, Takahisa Kouwaki, Tatsuya Suzuki, Takasuke Fukuhara, Kohji Moriishi, Kazuhiko Koike, Yoshiharu Matsuura. Processing of core protein by signal peptide peptidase participates in propagation and pathogenesis of hepatitis C virus. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 812. doi:10.1158/1538-7445.AM2015-812
Abstract 815: DTX1 is an epigenetically regulated tumor suppressor gene discovered by integrative analysis of epigenetic and transcriptional alterations in HNSCC
Authors: Gaykalova Daria A. , Zizkova Veronika , Tiscareno Ilse , Wei Yingying , Vatapalli Rajita , Hennessey Patrick T. , Ahn Julie , Danilova Ludmila V. , Khan Zubair , Bishop Justin A. , Koch Wayne M. , Westra William H. , Ochs Michael F. , Califano Joseph A.
Published: August 2015

Half a million new cases of Head and Neck Squamous Cell Carcinoma (HNSCC) are diagnosed annually worldwide. Given the advanced stage at diagnosis, 5 year survival after initial diagnosis is only 50% for HNSCC patients despite the current therapeutic strategies.

Recently, high throughput technologies have accelerated the genome-wide characterization of HNSCC, but given the high sensitivity of this methodology, these data require careful evaluation. To improve the detection of cancer-related changes in HNSCC we applied outlier statistics to high throughput DNA methylation and gene expression data, and identified 76 top-scoring candidate proto-oncogenes and tumor suppressor genes with significantly changed expression in tumors compared to normal tissues. We identified epigenetically regulated candidates with potential role in HNSCC by selecting genes with strong negative correlation between gene expression and DNA methylation of their promoters. Differential expression and methylation of selected candidates were confirmed with assays on an independent HNSCC cohort and on public domain high throughput data on TCGA-HNSCC cohort (The Cancer Genome Atlas). We further performed functional evaluation of a lead candidate - DTX1 and demonstrated that DTX1 is a negative regulator of cell migration. We confirmed that epigenetic-dependent downregulation of DTX1 enhanced invasiveness of head and neck cancer cells.

Outlier analysis integrated with high throughput expression and methylation can be used to identify an epigenetically- regulated tumor suppressor genes, including DTX1.

Citation Format: Daria A. Gaykalova, Veronika Zizkova, Ilse Tiscareno, Yingying Wei, Rajita Vatapalli, Patrick T. Hennessey, Julie Ahn, Ludmila V. Danilova, Zubair Khan, Justin A. Bishop, Wayne M. Koch, William H. Westra, Michael F. Ochs, Joseph A. Califano. DTX1 is an epigenetically regulated tumor suppressor gene discovered by integrative analysis of epigenetic and transcriptional alterations in HNSCC. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 815. doi:10.1158/1538-7445.AM2015-815
Abstract 816: Persistent effects of radiation on intestinal stem cells: implications for colorectal carcinogenesis
Authors: Datta Kamal , Suman Shubhankar , Kumar Santosh , Fornace Albert J.
Published: August 2015

Adult stem cells, similar to somatic cells, are also subject to a similar array of insults and are therefore presumed to accrue radiation damage. Unlike somatic cells, however, lesions arising in the stem cell compartment are propagated both to daughter stem cells and to downstream lineages through the processes of self-renewal and differentiation. The impact of damage accrued in individual stem cells can thus have major ramifications for all levels of the developmental hierarchy leading to cancer initiation and promotion in human as well as in animal models of human colorectal cancer (CRC) such as in APCMin/+. Differentiation of intestinal stem cell (ISC) and ordered migration of differentiated cells along crypt-villus axis is essential for maintenance of intestinal homeostasis and provides a model system to study stem cell alterations after radiation. Furthermore, considering that tumors in animal models of CRC almost exclusively develop in the small intestine and increased intestinal tumor frequency was radiation quality dependent, we investigated persistent effects of radiation quality on ISC as well as on differentiated intestinal epithelial cells in mice. Mice (6 to 8 week; C57BL/6J) were exposed to γ (0.5 and 2 Gy) and 56-Fe (0.5 and 1.6 Gy) radiation, samples were collected 7 d and 2 m after radiation exposure, and carcinogenic precursor events such as persistent oxidative stress and DNA damage were assessed. Higher persistently increased oxidative stress and oxidative DNA damage was observed after 56-Fe relative to γ radiation. However, increased oxidative stress did not increase cell death implying that the oxidative stress was sub-lethal. Autophagy, a self-cannibalistic process of cell involved in removing damaged cell and cell constituents, was consistently downregulated in a radiation quality dependent manner even 2 m after exposure suggesting persistence of damage-bearing cells. We believe radiation-induced autophagy downregulation and sub-lethal oxidative stress is working in tandem to trigger proliferative signaling such as PI3K/Akt and mTOR in intestinal cells. Taken together our results provide insight into radiation quality dependent persistent cancer-related molecular events, which could have altered phenotypic behavior of ISC such as proliferation, differentiation, and migration.

Note: This abstract was not presented at the meeting.

Citation Format: Kamal Datta, Shubhankar Suman, Santosh Kumar, Albert J. Fornace. Persistent effects of radiation on intestinal stem cells: implications for colorectal carcinogenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 816. doi:10.1158/1538-7445.AM2015-816
Abstract 817: Lipid, glucose and global oxidation in relation to prostate cancer development
Authors: Wu Tianying
Published: August 2015

Oxidative stress has been implicated in the development of prostate cancer (PCa); however, no study has comprehensively examined the associations between lipid, glucose and global oxidation with the development of prostate cancer. Using a prospective study design, we collected blood and urine samples among participants who are free of cancer in Ohio area. These participants were followed up 14 years. We selected 24 men who developed PCa during this period and matched them with 24 healthy controls by age and race among these participants (who did not develop prostate cancer) during this period. We measured the following biomarkers in these 48 samples (24 PCa cases and 24 health controls). Biomarkers included urine F2-isoprostanes (a marker of lipid oxidation), plasma advanced glycation end products (AGEs; a marker of advanced glycation and glucose oxidation), and plasma fluorescent oxidation products (FlOPs; markers of global oxidation). We have found that only AGEs were significantly higher in PCa cases than in health controls (182 vs. 152 μg/ml, P < 0.05). In the conditional logistic regression model, one standard deviation higher levels of AGEs were associated with increased risk of incident PCa (Relative risk = 1.79, 95% confidence interval = 1.00-3.21). In the model, one standard deviation higher levels of AGEs accounted for ∼8% variance of prostate cancer liability. Our results suggest that glycation and glucose oxidation but not lipid and global oxidation is an important etiology pathway for prostate cancer development. Lipid and global oxidation do not occur in parallel for the development of prostate cancer. If our results are confirmed by other studies, it may suggest a new pathway for prostate cancer prediction and treatment.

Note: This abstract was not presented at the meeting.

Citation Format: Tianying Wu. Lipid, glucose and global oxidation in relation to prostate cancer development. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 817. doi:10.1158/1538-7445.AM2015-817
Abstract 819: Fatty acid synthase, cycloxygenase-2 (COX-2) and osteoprotegerin (OPG) expression in invasive breast cancer: Implications in carcinogenesis
Authors: Goswami Sudeshna , Sharma-Walia Neelam
Published: August 2015

Breast cancer, the leading cause of death among women with an onset frequency of one in eight, is the most common type of cancer among women. However, despite the advancement in therapy, the mortality rate in breast cancer patients still remains high. The role of various different genes, directly or indirectly, regulating these processes have tremendously added to our knowledge of the complex process of proliferation, which has remained the most important prognostic factor so far. In our previous study, we established Osteoprotegerin (OPG)'s role in aneuploidy, cell proliferation and angiogenesis in breast cancer. We demonstrated that OPG is expressed and secreted at very high levels from the highly invasive breast cancer cell lines SUM149PT and SUM1315MO2 as compared to human normal mammary epithelial cells (HMEC). Here, we demonstrated by mass spectrometry that OPG pulls down fatty acid synthase (FASN), a key enzyme of the fatty acid biosynthetic pathway in breast cancer cells. We observed large number of lipid droplets in SUM149PT and SUM1315MO2 cells in comparison to HMEC by electron microscopy. Human breast cancer tissue samples showed high expression of FASN. Treatment with FASN inhibitor C75 decreased the number of lipid droplets/cell, altered expression of genes involved in cell proliferation, cell signaling (Akt/PI3K, ERK, GSK3β) apoptosis (caspase-3 and caspapse-9) and cell cycle. Treatment with C75 lowered the inflammatory pathway enzyme cyclooxygenase-2 (COX-2) expression and reduced secretion of its inflammatory metabolite prosataglandin E2 (PGE2). We reasoned that there might be crosstalk between OPG, FASN, and COX-2, that sustains the inflammatory pathways, and drives the progression of breast cancer. Here, we identified cis-acting elements involved in the transcriptional regulation of COX-2 by recombinant human OPG. Collectively, our study indicates the crosstalk between FASN, COX-2 and OPG. Combinatorial drug treatment of these main players of carcinogenesis can be used as a potential therapeutic target to treat highly invasive breast cancer.

Citation Format: Sudeshna Goswami, Neelam Sharma-Walia. Fatty acid synthase, cycloxygenase-2 (COX-2) and osteoprotegerin (OPG) expression in invasive breast cancer: Implications in carcinogenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 819. doi:10.1158/1538-7445.AM2015-819
Abstract 821: Human mammary tumor virus(HMTV) is deeply involved in breast cancer and metastasis
Authors: Holland James F. , Melana Stella , Jaffer Shabnam , Etkind Polly , Nartey Teiko , Lee Tina , Pogo Beatriz G.T.
Published: August 2015

Human Mammary Tumor Virus (HMTV), a betaretrovirus 90-95% homologous to Mouse Mammary Tumor Virus (MMTV), present in 40% of American women's breast cancers, but not in the normal tissue from the same breast indicates an infection (Cancer Res 1995, 55:5173-79). Absence of murine intracisternal particles and murine cyclo-oxygenase 2 by PCR and nanoString excludes contemporaneous mouse DNA, and thus MMTV contamination (Proc AACR 2014 #1595). We have detected the entire 9.9 kb proviral sequence. Proviral DNA is visible in nuclei by FISH, and budding virus was observed by EM. HMTV in cells is present in the milk of 8% of unselected American women, but in 21% if they had had a prior biopsy based upon clinical or radiologic suspicion of a mass. (Infectious Agents and Cancer 2014, 9:20).

HMTV has been found in cells of 30 of 32 (94%) metastatic breast cancer effusions (MSSM cells). HMTV protein and virion RNA were detected in MSSM cells but not in normal human mammary epithelium. (J Virol Methods 2010,163:157-61). HMTV isolated from MSSM cells is infectious for B and T lymphocytes, MCF10A andF and human mammary epithelial cells (HMEC). Infection led to proviral insertion sites with different flanking sequences. Epithelial-mesenchymal transition was observed with upregulation of vimentin and downregulation of E-cadherin. Restriction factors including the cytidine deaminase APOBEC G and F, TRIM 5, 21 and 25 and Tetherin were also increased after in vitro HMTV infection.

Epidemiologically, breast cancer is less frequent in 5 countries of Central and East and Asia (28-43/100,000/y. World Cancer Report 2014) where HMTV positivity in breast cancers ranges from 0 to 20%, compared to 7 countries of Western Europe and its former colonies in North and South America and Australia (47-92/100,000/y op.cit.) where 30 to 60% are positive. This correlates with indigenous mouse species, Mus musclus and M.castaneus in the East, and M. domesticus with much more HMTV in its genome in the West.

HMTV positive lymphomas have been reported in patients with HMTV positive breast cancers (Clin Cancer Res 2004 10:56565-64). The parallel with MMTV behavior in the mouse with respect to breast cancer and lymphomas is striking. These composite data support a significant role of HMTV in human breast cancer and its metastatic process.

Citation Format: James F. Holland, Stella Melana, Shabnam Jaffer, Polly Etkind, Teiko Nartey, Tina Lee, Beatriz G.T. Pogo. Human mammary tumor virus(HMTV) is deeply involved in breast cancer and metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 821. doi:10.1158/1538-7445.AM2015-821
Abstract 822: Genetically engineered mouse models of catastrophic pancreatic ductal adenocarcinoma
Authors: Yamaguchi Takashi , Ikehara Sanae K. , Nakanishi Hayao , Ikehara Yuzuru
Published: August 2015

Carcinogenesis of pancreatic ductal adenocarcinoma (PDAC) has been well studied using genetically engineered mouse (GEM) models with abnormalities in activation of both RAS and E2 factor (E2F)-which is associated with the Rb-E2F pathway, and dysfunction of p53-which is associated with the transformed 3T3 cell double minute 2 (MDM2)-p53 pathway. Catastrophic progression of the malignancy is one of unique manifestation, which has not been recapitulated in the GEMs to study underlined mechanisms, while we have recently reported a new GEM developing PDAC to die in 3 weeks after birth due to the catastrophic progression [Yamaguchi T. et al, J Pathol. 2014; 234(2):228-138]. The GEM had the specific expression of SV40 tsA58 large T antigen (tsTAg) to inhibit p53 and Rb family molecules (Rb1, Rbl1 and Rbl2) on pancreas with the expression of oncogenic Kras (KrasG12D) by Pdx1 promoter-mediated Cre-loxP recombination.

In this study, we sought to determine if the induced tsTAg on pancreatic epithelial cells with constitutive KrasG12D expression are enough to develop PDAC and catastrophic manifestation, by the generation of a new GEM based on Cre-loxP recombination and Tet-On system. In the GEM, KrasG12D constitutively expresses on pancreatic epithelial cells after the Cre-loxP recombination under the control Pdx1 promoter, while tsTAg expression can be regulated by Tet-On system after the recombination by Pdx1 promoter. Any of the mice did not developed PDAC within 6 month, but all of the mice developed PDAC by Dox administration in within 1 week and died by the progression of PDAC within 3 weeks after the start.The rapid progression and development of PDAC appeared to be associated with the stronger tsTAg expression depending on reverse tetracycline trans-activator.

These results demonstrated that KrasG12D driven PDAC carcinogenesis was promoted by the presence of tsTAg expression, resulting in production of PDAC cells with aggressive growth. In conclusion, our established new mouse model can contribute to increase knowledge about catastrophic PDAC.

Citation Format: Takashi Yamaguchi, Sanae K. Ikehara, Hayao Nakanishi, Yuzuru Ikehara. Genetically engineered mouse models of catastrophic pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 822. doi:10.1158/1538-7445.AM2015-822
Abstract 823: Gene expression profiling for oncogenic Kras mutation in mice and human colorectal cancer
Authors: Niitsu Hiroaki , Hinoi Takao , Kawaguchi Yasuo , Sentani Kazuhiro , Oue Naohide , Sotomaru Yusuke , Adachi Tomohiro , Saito Yasufumi , Miguchi Masashi , Kochi Masatoshi , Shimomura Manabu , Yasui Wataru , Ohdan Hideki
Published: August 2015

Purpose: Oncogenic KRAS mutations are found in 40-50% of human colorectal cancers. However, specific therapeutics has not been yet available. We aimed to search new therapeutic molecular targets or biomarkers in KRAS mutated colorectal cancers.

Experimental design: We generated Apcflox/flox; CDX2P9.5-G22Cre and LSL-KrasG12D; Apcflox/flox; CDX2P9.5-G22Cre mice, that had been considered as sporadic colon cancer mouse model with Kras wild and mutant, respectively. We observed the carcinogenesis at the age of 3-4 weeks by necropsy, and harvested the tumors for gene expression profiling. We compared the gene expression by Microarray (GeneChip, Affymetrix) between each 3 tumors from these mice strains. Genes with >5 fold changes between the two groups were selected as candidate genes, and subsequently narrow these down by bibliographic search for further analyses in human colorectal cancers.

Result: In macroscopic findings, extensive polyps were generated in cecum and proximal colon, but not in other site of gastrointestinal tract. Tumors occurred in the both strains were considered as well differentiated adenocarcinomas in hematoxylin-eosin staining. It was confirmed that tumors generated in Apcflox/flox; CDX2P9.5-G22Cre mice had Cre-targeted Apc 580D alleles and wild-type Kras, and that tumors generated in LSL-KrasG12D; Apcflox/flox; CDX2P9.5-G22Cre mice had both Cre-targeted Apc 580D alleles and activated oncogenic KrasG12D. In the subsequent gene expression profiling, we identified 31 genes with >5 fold change in Microarray analyses. Of these, we focused on some genes that showed lower expression in Kras mutant tumor than in Kras wild type tumor. We confirmed the lower expressions of these genes in murine tumors by quantitative RT-PCR, and the lower expression in early-staged human colorectal cancers with oncogenic KRAS mutant by immunohistochemistry staining. To analyze the functions, the retroviral vectors that lead overexpression of these genes were constructed and infected to SW480 and RKO CRC cell lines. Proliferation and migration is being analyzed using these cell lines.

Conclusion: We identified novel genes associated with oncogenic KRAS mutation, using the analyses of gene expression in colon cancer mouse model.

Citation Format: Hiroaki Niitsu, Takao Hinoi, Yasuo Kawaguchi, Kazuhiro Sentani, Naohide Oue, Yusuke Sotomaru, Tomohiro Adachi, Yasufumi Saito, Masashi Miguchi, Masatoshi Kochi, Manabu Shimomura, Wataru Yasui, Hideki Ohdan. Gene expression profiling for oncogenic Kras mutation in mice and human colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 823. doi:10.1158/1538-7445.AM2015-823
Abstract 828: Methylation status of HPV16 E2-binding sites identifies subtypes of HPV-associated oropharyngeal squamous cell carcinomas
Authors: Speel Ernst-Jan M. , Reuschenbach Miriam , Huebbers Christian U. , Prigge Elena-Sophie , Bermejo Justo L. , Kalteis Simon , Preuss Simon , Kolligs Jutta , Olthof Nadine C. , Kremer Bernd , Wagner Steffen , Klussmann Jens P. , Vinokurova Svetlana , von Knebel Doeberitz Magnus
Published: August 2015

Background: The viral E2 protein is a transcriptional repressor of the HPV oncogenes E6/E7 and loss of E2 function is considered a key step in carcinogenesis. Integration of HPV into the host genome may disrupt the E2 gene. Furthermore, methylation of CpG dinucleotides in E2 binding sites (E2BSs) in the HPV upstream regulatory region (URR) may interfere with transcriptional repression of E6 and E7 by the E2 protein. We hypothesized that CpG methylation of the E2BS is found in a proportion of HPV16-associated oropharyngeal squamous cell cancers (OPSCC) in association with E2 gene integrity and viral integration.

Methods: Methylation of 10 CpGs within the URR, encompassing E2BSs 1, 2, 3 and 4, was quantitatively analyzed by bisulfite pyrosequencing in 57 HPV16-associated OPSCC. E2 status was analyzed by gene amplification and quantitative real-time RT-PCR. Viral integration was determined by integration-specific PCR methods.

Results: Three subgroups with differential E2BSs3 and 4 methylation were identified: a) complete methylation (>80%) associated with presence of integrated HPV genomes with intact E2 gene, b) intermediate methylation levels (20-80%) with predominantly episomal HPV genomes with intact E2, and c) no methylation (<20%) with integrated HPV with disrupted E2. Patients with complete and without methylation had poor 5-year overall survival compared to patients with intermediate methylation (stage-adjusted hazard ratio 3.8 (95%CI 1.3-11.2) and 3.1 (95%CI 1.14-8.28); p = 0.03).

Conclusions: E2BSs3 and 4 methylation in oropharyngeal cancers with episomal HPV and integrated HPV with intact E2 gene might explain deregulated viral oncogene expression in the presence of E2. The methylation level appears to be of prognostic significance for patients with HPV-associated OPSCC.

Citation Format: Ernst-Jan M. Speel, Miriam Reuschenbach, Christian U. Huebbers, Elena-Sophie Prigge, Justo L. Bermejo, Simon Kalteis, Simon Preuss, Jutta Kolligs, Nadine C. Olthof, Bernd Kremer, Steffen Wagner, Jens P. Klussmann, Svetlana Vinokurova, Magnus von Knebel Doeberitz. Methylation status of HPV16 E2-binding sites identifies subtypes of HPV-associated oropharyngeal squamous cell carcinomas. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 828. doi:10.1158/1538-7445.AM2015-828
Abstract 833: Persistent alterations of gene expression profiling of human peripheral blood mononuclear cells from smokers
Authors: Weng Daniel Y. , Chen Jinguo , Taslim Cenny , Hsu Ping-Ching , Marian Catalin , David Sean P. , Loffredo Christopher A. , Shields Peter G.
Published: August 2015

The number of validated biomarkers of tobacco smoke exposure is limited, and none exist for tobacco-related cancer. Additional biomarkers for smoke effects on cellular systems in vivo are needed to improve early detection of lung cancer, and to assist the Food and Drug Administration in regulating exposures to tobacco products. We assessed the effects of smoking on the gene expression in vitro and in vivo using human cell cultures and blood from a cross-sectional study. We profiled global transcriptional changes in cultured smokers’ peripheral blood mononuclear cells (PBMCs) treated with cigarette smoke condensate (CSC) in vitro and from well-characterized smokers’ blood (n = 36). ANOVA and linear regression with adjustment for covariates were used for statistical analysis. CSC in vitro altered the expression of 1178 genes (177 genes with > 1.5 fold-change) at P < 0.05. Phase I and II metabolizing enzyme genes were up-regulated, cytokines and chemokines were down-regulated. Nine putative oncogenes and 5 tumor suppressor genes were dysregulated. In vivo, PBMCs of heavy and light smokers differed for 614 genes (29 with > 1.5 fold-change) at P < 0.05 (309 remaining significant after adjustment for age, race and gender). Seventy eight genes had a significant linear correlation (P < 0.01) with cigarettes per day and 28 genes were linearly correlated with cumulative pack-year smoking. Forty-one genes were persistently altered both in vitro and in vivo, 22 having the same expression pattern reported for non-small cell lung cancer. Our data provides evidence that persistent alterations of gene expression in vitro and in vivo may relate to carcinogenic effects of cigarette smoke, and the identified genes may serve as potential biomarkers for cancer. The use of an in vitro model to corroborate results from human studies provides direct evidence for a cause and effect.

Citation Format: Daniel Y. Weng, Jinguo Chen, Cenny Taslim, Ping-Ching Hsu, Catalin Marian, Sean P. David, Christopher A. Loffredo, Peter G. Shields. Persistent alterations of gene expression profiling of human peripheral blood mononuclear cells from smokers. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 833. doi:10.1158/1538-7445.AM2015-833
Abstract 839: Effects of occupational exposure to carbon black on peripheral white blood cell counts
Authors: Dai Yufei , Zhang Rong , Niu Yong , Duan Huawei , Meng Tao , Ye Meng , Shen Meili , Bin Ping , Yu Shanfa , Vermeulen Roel , Rothman Nathaniel , Lan Qing , Zheng Yuxin
Published: August 2015

Carbon black (CB) is a material produced by the combustion of hydrocarbon gases or liquids and widely used in various industrial settings. The International Agency for Research on Cancer has classified CB as possibly carcinogenic to humans. Previous studies suggest that CB might induce lung inflammation and injury and it is well known that inflammation can play an important role in tumor development. However, the relationship between CB exposure and peripheral white blood cell counts is still not clear. To address this issue, we conducted a cross-sectional molecular epidemiology study of 106 CB-exposed male workers responsible for packing CB in a CB manufacturing facility that produced very high purity CB and 112 unexposed male control workers from the same geographic region who were frequency-matched to the exposed workers by age. The average number of years worked in the factory for the exposed workers was 12.5 years. Peripheral blood samples were collected and a complete blood cell count (CBC) and lymphocyte subset analysis were conducted. There was a borderline significant increase in eosinophil count, and a significant decrease in total lymphocyte and NK cell counts, among the exposed vs. control workers. Among non-smokers (35 exposed and 29 controls), CB exposure was associated with a borderline significant increase in total white blood cell and neutrophil counts, a significant increase in eosinophil counts, and a significant decrease in total lymphocyte, CD3+ T cell, CD8+ T cell and NK cell counts. There were no significant differences between exposed and control workers for any white blood cell measure among smokers. Also, there was some evidence of an interaction between carbon black exposure and smoking status for neutrophil, total lymphocyte, CD3+ T cell, and CD8+ T cell counts. Our results suggest that CB exposure may be associated with altered levels of cells that play an important role in the inflammatory process among non-smokers. Also, there was evidence for an interaction between CB exposure and tobacco use. These findings may provide new insights into particulate-associated disease.

Citation Format: Yufei Dai, Rong Zhang, Yong Niu, Huawei Duan, Tao Meng, Meng Ye, Meili Shen, Ping Bin, Shanfa Yu, Roel Vermeulen, Nathaniel Rothman, Qing Lan, Yuxin Zheng. Effects of occupational exposure to carbon black on peripheral white blood cell counts. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 839. doi:10.1158/1538-7445.AM2015-839
Abstract 840: Occupational exposure to trichloroethylene and LINE-1 methylation
Authors: Hu Wei , Zhang Luoping , Kim Christopher , Tang Xiaojiang , Kim4 Sungkyoon , Bassig Bryan , Seow Wei-Jie , Shen Min , Qiu Chuangyi , Ge Yechen , Reiss Boris , Purdue Mark , Moore Lee , Li Laiyu , Yue Fei , Huang Hanlin , Smith Martyn T. , Vermeulen Roel , Rothman Nathaniel , Lan Qing
Published: August 2015

Trichloroethylene (TCE) is a volatile chlorinated organic compound that is commonly used in industrial settings as a degreaser. Epidemiologic studies suggest that TCE exposure is associated with an increased risk of kidney cancer and may also be associated with non-Hodgkin lymphoma. However, the mechanisms for these associations are still not clear. To evaluate the relationship between occupational exposure to trichloroethylene (TCE) and LINE-1 methylation, a measure of global methylation, we conducted a cross-sectional molecular epidemiology study of 79 workers exposed to TCE and 81 comparable unexposed controls in Guangdong, China. Personal air exposure measurements and urine samples were collected prior to phlebotomy. The urine TCE metabolite, trichloroacetic acid (TCA), was measured by liquid chromatography electrospray ionization tandem mass spectrometry in urine samples and LINE-1 methylation was assessed by pyrosequencing DNA isolated from peripheral blood. LINE-1 methylation levels [median (range) in percent] were 78.40 (75.80-81.20) in controls, 77.80 (76.10-81.00) in lower exposed, and 77.50 (74.20-80.20) in higher exposed workers based on median urine TCA level (p trend = 0.012). These results suggest that TCE may be associated with a decrease in global methylation measured by the LINE-1 assay. Gene-specific methylation is currently being conducted to follow up these findings.

Citation Format: Wei Hu, Luoping Zhang, Christopher Kim, Xiaojiang Tang, Sungkyoon Kim4, Bryan Bassig, Wei-Jie Seow, Min Shen, Chuangyi Qiu, Yechen Ge, Boris Reiss, Mark Purdue, Lee Moore, Laiyu Li, Fei Yue, Hanlin Huang, Martyn T. Smith, Roel Vermeulen, Nathaniel Rothman, Qing Lan. Occupational exposure to trichloroethylene and LINE-1 methylation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 840. doi:10.1158/1538-7445.AM2015-840
Abstract 841: SNP-FEMS: a method to identify DNA binding proteins interacting with enhancer elements
Authors: Mendoza-Fandino Gustavo A. , Woods Nicholas , Baskin Rebekah , Gjyshi Anxhela , Monteiro Alvaro N.
Published: August 2015

An important goal of human genetics is to understand the etiology of genetic traits. In the past few years, genome-wide association studies (GWAS) have identified thousands of chromosomal loci associated with hundreds of traits. GWAS provide a list of variants (single nucleotide polymorphisms, SNP) that are statistically associated with a phenotype, but do not offer any direct evidence about the biological mechanism. Most GWAS cancer susceptibility loci are located in non-coding regions suggesting that they function by modifying tracsriptional regulatory elements, such as enhancers and promoters of target genes. Thus, methods to help identify transcription factors that interact with regulatory regions containing risk associated variants can reveal biological mechanisms. We developed a method, SNP-FEMS (FPLC, EMSA, Mass Spec) that combines liquid chromatography (FPLC), electrophoretic mobility shift assays (EMSAs), and mass spectrometry to identify transcription factors interacting with putative regulatory regions at the 8q24 ovarian cancer risk locus. Human embryonic kidney 293FT nuclear extract was subjected to Sepharose 6 10/300 GL size fractionation and all the fractions were screened for allele-specific binding proteins by EMSA using a probe containing a genome-wide significant SNP. Fractions with positive signal were combined, concentrated and further fractionated with an ion exchange column MonoQ 5/10 GL to further reduce the complexity of the fractions. Fractions were screened by EMSA with the same probe, and the positive fractions were combined, concentrated, subjected to PAGE, in-gel trypsin digestion, and analyzed by Mass Spectrometry to identify proteins in the fractions. Bioinformatics analysis predicted different transcription factors binding to the putative regulatory region and the only transcription factor that was also present in our experimental dataset was the Cyclin AMP-dependent transcription factor ATF-1. This data suggests that ATF1 is implicated in ovarian cancer susceptibility and demonstrate the utility of the SNP-FEMS method to identify regulatory factors involved in ovarian cancer risk. This technique is applicable to any predisposition loci mediated by changes in regulatory elements.

Citation Format: Gustavo A. Mendoza-Fandino, Nicholas Woods, Rebekah Baskin, Anxhela Gjyshi, Alvaro N. Monteiro. SNP-FEMS: a method to identify DNA binding proteins interacting with enhancer elements. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 841. doi:10.1158/1538-7445.AM2015-841
Abstract 842: Two new loci and gene sets related to sex determination and cancer progression are associated with susceptibility to testicular germ cell tumor
Authors: Grotmol Tom , Kristiansen Wenche , Karlsson Robert , Rounge Trine B. , Whitington Thomas , Andreassen Bettina K. , Magnusson Patrik KE , Adami Hans-Olov , Turnbull Clare , Haugen Trine B. , Wiklund Fredrik
Published: August 2015

Genome-wide association (GWA) studies have reported 19 distinct susceptibility loci for testicular germ cell tumor (TGCT). The aim of the present study was to identify additional loci and gene sets associated with susceptibility to TGCT. A GWA study for TGCT was performed by genotyping 610,240 single nucleotide polymorphisms (SNPs) in 1,326 cases and 6,687 controls from Sweden and Norway. We put forward 34 SNPs from 17 novel regions and 11 SNPs previously reported, for replication in 710 case-parent triads and 289 cases and 290 controls. Predefined biological pathways and processes, in addition to a custom-built sex determination gene set, were subject to enrichment analyses using Meta-Analysis Gene Set Enrichment of Variant Associations (M) and Improved Gene Set Enrichment Analysis for Genome-wide Association Study (I). In the combined meta-analysis, we observed genome-wide significant association for rs7501939 on chromosome 17q12 (OR = 1.29, 95% CI = 1.19-1.40, P = 1.1 × 10-9) and rs2195987 on chromosome 19p12 (OR = 1.31, 95% CI: 1.19-1.45, P = 3.2 × 10-8). The marker rs7501939 on chromosome 17q12 is located in an intron of the HNF1B gene, encoding a member of the homeodomain-containing superfamily of transcription factors. The sex determination gene set (FDRM < 0.001, FDRI < 0.001) and pathways related to NF-κB, glycerophospholipid and ether lipid metabolism, as well as cancer and apoptosis, was associated with TGCT (FDR < 0.1). In addition to revealing two new TGCT susceptibility loci, our results support the notion that genes governing normal germ cell development in utero are implicated in the development of TGCT.

Citation Format: Tom Grotmol, Wenche Kristiansen, Robert Karlsson, Trine B. Rounge, Thomas Whitington, Bettina K. Andreassen, Patrik KE Magnusson, Hans-Olov Adami, Clare Turnbull, Trine B. Haugen, Fredrik Wiklund. Two new loci and gene sets related to sex determination and cancer progression are associated with susceptibility to testicular germ cell tumor. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 842. doi:10.1158/1538-7445.AM2015-842
Abstract 849: The changing role of GDF15 (growth/differentiation factor 15) during prostate carcinogenesis
Authors: Rybicki Benjamin A. , Chitale Dhananjay , Gupta Nilesh , Jackson LaToya , Wheeler Travis , Trudeau Sheri , Jankowski Michelle , Bobbittt Kevin , Rundle Andrew , Tang Delian
Published: August 2015

GDF15 (growth/differentiation factor 15) is a divergent member of the TGFβ superfamily of cytokines and is highly expressed in prostate tumors, but its role in prostate carcinogenesis and utility as a prognostic biomarker is unclear. We studied 58 prostate cancer cases (35 Whites and 23 African Americans) that underwent surgery as their primary treatment and were then subsequently followed for biochemical recurrence (BCR). In these cases, which also had a benign prostate biopsy at least one year before their prostate cancer diagnosis, we quantified GDF15 expression in their pre-malignant benign prostate, prostate tumor and tumor adjacent benign prostate tissue. GDF15 expression levels were analyzed for association with high grade prostate cancer and BCR. Twenty-six percent of cases had high grade cancer defined as Gleason score 8 or higher or a Gleason score of 7 and a primary Gleason of 4. During follow-up, 11 cases (19%) experienced BCR (80 percent of men without BCR had at least 2 years of follow-up). GDF15 expression was not associated high grade cancer in any prostate tissue types. GDF15 expression was associated with BCR, but the direction of the association was dependent upon the type of prostate tissue expression measured. Evidence of GDF15 expression in pre-malignant benign prostate was associated with an almost 9-fold increased risk of BCR (Hazard Ratio (HR) = 8.90; 95% confidence interval (CI) = 1.09-72.49), even after adjusting for race, tumor grade and stage. When we stratified men into low and high expression groups, we also found a significant gradient of risk with increased GDF15 expression (p = 0.03). The opposite was found for GDF15 expression in tumor tissue, where men with low GDF15 expression had a decreased risk of BCR (HR = 0.30; 95% CI = 0.06- 1.59), and men with high GDF15 expression were at an even lower risk (HR = 0.10; 95% CI = 0.02-0.61). Expression of GDF15 in tumor adjacent benign prostate tissue was not associated with BCR. The role GDF15 plays in prostate carcinogenesis is complex and may change during the course of tumor development. Our results suggest GDF15 exerts a pro-tumorgenic effect very early in prostate carcinogenesis, but later after the tumor becomes clinically apparent, GDF15 may act more in an anti-tumorgenic manner.

Citation Format: Benjamin A. Rybicki, Dhananjay Chitale, Nilesh Gupta, LaToya Jackson, Travis Wheeler, Sheri Trudeau, Michelle Jankowski, Kevin Bobbittt, Andrew Rundle, Delian Tang. The changing role of GDF15 (growth/differentiation factor 15) during prostate carcinogenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 849. doi:10.1158/1538-7445.AM2015-849
Abstract 851: Characteristics of adult men who are willing to give HPV vaccine to their daughters in the United States: a national survey
Authors: Rogers Jessica , Idowu Kolapo A. , Coker Rahmatalai , Segunmaru Hamidat , Daremipouran Mohammad , Burnside Clinton , Williams Carla D. , Laiyemo Adeyinka O.
Published: August 2015

Background: The uptake of Human Papillomavirus (HPV) vaccination needs improvement. However, efforts to improve uptake of HPV vaccine is often concentrated on women. Although men can influence the uptake of the vaccine by their children, the opinion of adult men in the United States on HPV vaccination has not been well studied.

Aim: To evaluate the willingness of adult men to vaccinate their daughters for HPV

Methods: Health Information National Trends Survey (HINTS) 2007 was used to capture the opinion of 1,927 men (weighted population size = 72,633,670) who answered questions on their willingness to let their daughters receive HPV vaccine. We compared the characteristics of those who were willing to let their daughters receive HPV vaccine with those who were not. We used survey weights in all analyses, and variance estimations were performed using Taylor series linearization to account for the complex survey design. We used logistic regression models to calculate odds ratios (OR) and 95% confidence intervals (CI).

Results: A total of 1,554 (80.7%) men were willing to let their daughters receive HPV vaccine while 373 (19.3%) would not let their daughters receive the vaccine. The most common reasons for disagreeing with vaccination were: not knowing enough about it (27.7%); vaccine safety concerns (16.5%); and “my child is not sexually active” (13.7%). Of note, less than 1% felt the vaccine will promote promiscuity. Demographic (age, race, marital status, education status, and health insurance) and lifestyle characteristics (smoking and body mass index) did not predict men who would let their daughters get HPV vaccine.

Conclusion: A high percentage of men are willing to let their daughters receive HPV vaccine suggesting that all men should be engaged in the conversation to improve HPV vaccine uptake in the United States. Primary care physicians should make it a priority to discuss the HPV vaccine with men and women more frequently to enlighten the population and allay their fears about the safety of the vaccine.

Citation Format: Jessica Rogers, Kolapo A. Idowu, Rahmatalai Coker, Hamidat Segunmaru, Mohammad Daremipouran, Clinton Burnside, Carla D. Williams, Adeyinka O. Laiyemo. Characteristics of adult men who are willing to give HPV vaccine to their daughters in the United States: a national survey. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 851. doi:10.1158/1538-7445.AM2015-851