Cancer-related Changes Research Articles

Metabolomic Profiling of Extracellular Vesicles and Alternative Normalization Methods Reveal Enriched Metabolites and Strategies to Study Prostate Cancer-Related Changes.

Body fluids are a rich source of extracellular vesicles (EVs), which carry cargo derived from the secreting cells. So far, biomarkers for pathological conditions have been mainly searched from their protein, (mi)RNA, DNA and lipid cargo. Here, we explored the small molecule metabolites from urinary and platelet EVs relative to their matched source samples. As a proof-of-concept study of intra-EV metabolites, we compared alternative normalization methods to profile urinary EVs from prostate cancer patients before and after prostatectomy and from healthy controls.Methods: We employed targeted ultra-performance liquid chromatography-tandem mass spectrometry to profile over 100 metabolites in the isolated EVs, original urine samples and platelets. We determined the enrichment of the metabolites in the EVs and analyzed their subcellular origin, pathways and relevant enzymes or transporters through data base searches. EV- and urine-derived factors and ratios between metabolites were tested for normalization of the metabolomics data.Results: Approximately 1 x 1010 EVs were sufficient for detection of metabolite profiles from EVs. The profiles of the urinary and platelet EVs overlapped with each other and with those of the source materials, but they also contained unique metabolites. The EVs enriched a selection of cytosolic metabolites including members from the nucleotide and spermidine pathways, which linked to a number of EV-resident enzymes or transporters. Analysis of the urinary EVs from the patients indicated that the levels of glucuronate, D-ribose 5-phosphate and isobutyryl-L-carnitine were 2-26-fold lower in all pre-prostatectomy samples compared to the healthy control and post-prostatectomy samples (p < 0.05). These changes were only detected from EVs by normalization to EV-derived factors or with metabolite ratios, and not from the original urine samples.Conclusions: Our results suggest that metabolite analysis of EVs from different samples is feasible using a high-throughput platform and relatively small amount of sample material. With the knowledge about the specific enrichment of metabolites and normalization methods, EV metabolomics could be used to gain novel biomarker data not revealed by the analysis of the original EV source materials.

PUB082 Role of Isolated Flavonoid from Indigofera Tinctoria in N-Nitrosopyrrolidine Induced Biochemical Changes Related to Lung Cancer

Lung cancer is a second leading cause of deaths throughout the globe. The key cause, tobacco smoke contains a number of nitrosamines like N-nitrosopyrrolidine (NPYR) interact with cellular system and alter redox signaling pathway. Medicinal plants like Indigofera tinctoria can be a promising source in treatment. The study was demeanor to evaluate the role of isolated compound 3,6-dihydroxy-(3’,4’,7’-trimethoxyphenyl)-chromen-4-one-7-glucoside (ITC) in lung cancer related changes. Lactate dehydrogenase (LDH), Lysosomal parameters: Acid phosphatase (ACP), Type II enzyme: Alkaline phosphatase (ALP), Clara cell marker enzyme: γ-glutamyl transpeptidase (GGT), Glycoproteins: Hexose, Hexosamine and Sialic Acid and total protein content were estimated. LDH, ACP and GGT levels showed augmentation while ALP activity was alleviated in NPYR treated subjects when compared to the normal healthy subjects. Post treatment with isolated compound significantly restored the abnormal values of lung cancer biomarkers. In group of NPYR treated animals, the level of glycoproteins significantly boost up when compared to the values of animals of control group. Total protein content was found to be heightened in the mice bearing lung cancer significantly when compared to the normal healthy group of animals. The post treatment of plant PTX and isolated compound most effectively normalize these heightened amounts. These biochemical enzymatic parameters stipulated the potential of isolated compound from experimental plant which is due to the high reactivity of the hydroxyl groups in its structure. Study revealed that flavonol isolated from Indigofera tinctoria hydromethanolic extract can alter various endogenous antioxidant enzymes, and lung specific markers significantly and thus can be further used as a promising treatment of pulmonary cancer after drug development.

Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis.

Conformational changes in proteins can lead to disease. Thus, methods for identifying conformational changes in proteins can further improve our understanding and facilitate detection of disease states. Here we combine limited proteolysis (LiP) with Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) to characterize breast cancer-related conformational changes in proteins on the proteomic scale. Studied here are the conformational properties of proteins in two cell culture models of breast cancer, including the MCF-10A and MCF-7 cell lines. The SILAC-LiP approach described here identified ∼200 proteins with cell-line-dependent conformational changes, as determined by their differential susceptibility to proteolytic digestion using the nonspecific protease, proteinase K. The protease susceptibility profiles of the proteins in these cell lines were compared to thermodynamic stability and expression level profiles previously generated for proteins in these same breast cancer cell lines. The comparisons revealed that there was little overlap between the proteins with protease susceptibility changes and the proteins with thermodynamic stability and/or expression level changes. Thus, the large-scale conformational analysis described here provides unique insight into the molecular basis of the breast cancer phenotypes in this study.

Return to work among self-employed cancer survivors.

The aim of this study is to investigate whether salaried and self-employed workers differ regarding factors relevant for return to work after being diagnosed with cancer. The possible mediators of an effect of self-employment on work ability were also investigated. A total of 1115 cancer survivors (1027 salaried and 88 self-employed) of common invasive cancer types who were in work at the time of diagnosis completed a mailed questionnaire 15-39months after diagnosis. Twenty-four percent of self-employed cancer survivors reported that they had not returned to work at the time of the survey, and 18% of those who were salaried had not. While 9% of the self-employed had received disability or early retirement pension, only 5% had received such a pension among salaried employees. Compared with the salaried workers, the self-employed people reported significantly more often reduced work hours (P<0.001), negative cancer-related financial (P<0.001), and occupational changes (P=0.005) and low overall health (P=0.02), quality of life (P=0.04), and total work ability (P=0.02). The negative effect of self-employment on total work ability seems to be mediated by reduced work hours and a negative cancer-related financial change. Compared with salaried, self-employed workers in Norway, they seem to struggle with work after cancer. This may be because the two groups have different work tasks and because self-employed people have lower social support at work and less legal support from the Working Environment Act and public health insurance. Self-employed people with cancer should be informed about the work-related challenges they may encounter and be advised to seek practical help from social workers who know about the legal rights of self-employed people.

Dissecting the biological relationship between TCGA miRNA and mRNA sequencing data using MMiRNA-Viewer.

BackgroundMicroRNAs (miRNA) are short nucleotides that interact with their target genes through 3′ untranslated regions (UTRs). The Cancer Genome Atlas (TCGA) harbors an increasing amount of cancer genome data for both tumor and normal samples. However, there are few visualization tools focusing on concurrently displaying important relationships and attributes between miRNAs and mRNAs of both cancer tumor and normal samples. Moreover, a deep investigation of miRNA-mRNA target and biological relationships across multiple cancer types by integrating web-based analysis has not been thoroughly conducted.ResultsWe developed an interactive visualization tool called MMiRNA-Viewer that can concurrently present the co-relationships of expression between miRNA-mRNA pairs of both tumor and normal samples into a single graph. The input file of MMiRNA-Viewer contains the expression information including fold changes between normal and tumor samples for mRNAs and miRNAs, the correlation between mRNA and miRNA, and the predicted target relationship by a number of databases. Users can also load their own input data into MMiRNA-Viewer and visualize and compare detailed information about cancer-related gene expression changes, and also changes in the expression of transcription-regulating miRNAs.To validate the MMiRNA-Viewer, eight types of TCGA cancer datasets with both normal and control samples were selected in this study and three filter steps were applied subsequently. We performed Gene Ontology (GO) analysis for genes available in final selected 238 pairs and also for genes in the top 5 % (95 percentile) for each of eight cancer types to report a significant number of genes involved in various biological functions and pathways. We also calculated various centrality measurement matrices for the largest connected component(s) in each of eight cancers and reported top genes and miRNAs with high centrality measurements.ConclusionsWith its user-friendly interface, dynamic visualization and advanced queries, we also believe MMiRNA-Viewer offers an intuitive approach for visualizing and elucidating co-relationships between miRNAs and mRNAs of both tumor and normal samples. We suggest that miRNA and mRNA pairs with opposite fold changes of their expression and with inverted correlation values between tumor and normal samples might be most relevant for explaining the decoupling of mRNAs and their targeting miRNAs in tumor samples for certain cancer types.Electronic supplementary materialThe online version of this article (doi:10.1186/s12859-016-1219-y) contains supplementary material, which is available to authorized users.

Abstract # 1769 Exposure during fire-overhaul period demonstrates an increase in disease risk and immune dysregulation for first responders with impact being reduced using a transitional technique

Exposure during firefighting activities to combustion products (including carcinogens and asphyxiants) has been shown to increase immediate and long-term health risks of first responders. Therefore, we investigated the role of smoke inhalation on increasing disease risk and immune dysregulation by placing mice in the overhaul period of a live-fire scenario for 15 min. During the attack, we used two separate firefighting techniques, interior vs transitional. Lung tissue was harvested 2 hours post-exposure and RNA extracted for RNA-seq analysis. There was a 10.3% reduction in the temperature between interior and transitional techniques amongst live-fire scenarios. Correspondingly, there was greater differential gene expression with the conventional interior technique when compared to the transitional attack (2666 vs 60 genes, respectively). Within the interior technique, gene pathways that were over-represented included rap1 signaling, asthma, B-cell receptor signaling, proteoglycans in cancer, and non-small cell lung cancer. Interior technique showed upregulation of carcinogenic proteins such as Rorc and capn11 with downregulation of tumor inhibitors such as S100a8 and S100a9 whereas transitional technique did not show similar gene expression changes. Consequently, we conclude that first responders face an increased risk of disease during overhaul as demonstrated by over-representation of cancer-related pathways and changes in immune regulation. Importantly, this data shows strong evidence for first responders using techniques that rapidly apply water to mitigate their risk of cancer and immune related diseases.

Abstract 3604: Validated structural variant detection with prioritisation of known cancer related changes

Abstract Reliable detection of structural variation (SV) is playing an increasingly important role in cancer diagnostics and treatments. We introduce a best practice integrative workflow for detecting large structural variations (SVs) such as deletions, duplications, inversions, fusions and translocations from standard DNA re-sequencing techniques, annotating and prioritising for clinically actionable events. While established RNA-Seq fusion workflows exist to detect and filter fusion events in tumors detecting large structural changes from exome- and WGS data remains challenging. Current DNA-based SV callers predict a large percentage of false positive events, and the resulting event lists are not prioritised for validation. To streamline follow-up studies we automate running multiple structural variant callers (Manta, WHAM, Lumpy, MetaSV), identifying potentially disruptive large scale events. We evaluate the integrated workflow sensitivity and specificity against the ICGC-TCGA DREAM Mutation Calling Challenge data set, optimise filtering criteria based on quality and supporting information, and annotate the filtered calls using known cancer related alterations from CIViC and publicly available gene lists from AstraZeneca. This enables us to prioritise known and potentially interesting events over intergenic events or non-whole exon deletions. We show validation results from test data and clinically actionable inversions, duplications and fusion events from cancer cell lines. Our approach is based on a two tier approach of first focusing on known events that yield fusion transcripts such as the TACC3-FGFR3 tandem duplication and ALK-EML4 inversion, and secondarily on events in oncogenes and tumor suppressors of interest. While whole genome sequencing is the preferred approach to detecting these events, we also show how by design or by the nature of the breakpoint being close enough to exons, hybrid capture data can also be surprisingly useful for SV inference. Citation Format: Miika Ahdesmäki, Brad Chapman, Sally Luke, Hedley Carr, Daniel Stetson, Oliver Hofmann, Justin Johnson. Validated structural variant detection with prioritisation of known cancer related changes. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3604.

Objective-subjective disparity in cancer-related cognitive impairment: does the use of change measures help reconcile the difference?

Studies to date have found little correlation between subjective and objective measures of cognitive function in cancer patients, making it difficult to interpret the significance of their cognitive complaints. The purpose of this study was to determine if a stronger correlation would be obtained using measures of cognitive change rather than static scores. Sixty women with early stage breast cancer underwent repeated cognitive assessment over the course of chemotherapy with a neuropsychological test battery (objective measure) and with the FACT-Cog (subjective measure). Their results were compared to 60 healthy women matched on age and education and assessed at similar intervals. We used multilevel modeling, with FACT-Cog as the dependent measure and ordinary least squares slopes of a neuropsychological summary score as the independent variable, to evaluate the co-variation between the subjective and objective measures over time RESULTS: Measures of both objective and subjective cognitive function declined over the course of chemotherapy in the breast cancer patients but there was no significant relationship between them, even when using change measures. Change in objective cognitive function was not related to change in anxiety or fatigue scores but the decline in perceived cognitive function was associated with greater anxiety and fatigue. The discrepancy in objective and subjective measures of cognition in breast cancer patients cannot be accounted for in terms of a failure to use change measures. Although the results are negative, we contend that this is the more appropriate methodology for analyzing cancer-related changes in cognition.

The perceptions of Australian oncologists about cognitive changes in cancer survivors.

Cancer-related cognitive changes (CRCC) can have a profound impact on a cancer survivor's quality of life. However, cancer survivors frequently report receiving limited information about their experience of CRCC from their oncology specialists. This qualitative study aimed to explore the perceptions of oncology specialists regarding CRCC and the potential for their views to influence their decisions about patient care. Thirteen medical oncologists and five radiation oncologists currently practising in Australia participated in this study. Data collection involved individual semi-structured interviews via telephone. Data were audio-recorded, transcribed verbatim and analysed using a thematic approach. Four key themes emerged: (1) beliefs about the impact of priming on cancer survivors' perceived cognitive function, (2) perceptions of who is more likely to raise concerns of cognitive change, (3) uncertainty of how to best manage CRCC, and (4) the perceived role of oncologists in the management of CRCC. CRCC and its impact on the cancer survivor's journey have been under-addressed by oncology specialists, and they are uncertain of potential management strategies. With cancer survival rates increasing, there is a need for specific interventions and management guidelines addressing CRCC and their effects on cancer survivors. Future exploration should focus on the survivor as central to their care and holistic approaches to CRCC management involving all members of the multidisciplinary team.

Complementary and Alternative Medicine Use at a Comprehensive Cancer Center.

Background. Complementary and alternative medicine (CAM) use is common among cancer patients, but the majority of CAM studies do not specify the time periods in relation to cancer diagnoses. We sought to define CAM use by cancer patients and investigate factors that might influence changes in CAM use in relation to cancer diagnoses. Methods. We conducted a cross-sectional survey of adults diagnosed with breast, prostate, lung, or colorectal cancer between 2010 and 2012 at the Lineberger Comprehensive Cancer Center. Questionnaires were sent to 1794 patients. Phone calls were made to nonrespondents. Log binomial/Poisson regressions were used to investigate the association between cancer-related changes in CAM use and conversations about CAM use with oncology providers. Results. We received 603 (33.6 %) completed questionnaires. The mean age (SD) was 64 (11) years; 62% were female; 79% were white; and 98% were non-Hispanic. Respondents reported the following cancer types: breast (47%), prostate (27%), colorectal (14%), lung (11%). Eighty-nine percent reported lifetime CAM use. Eighty-five percent reported CAM use during or after initial cancer treatment, with category-specific use as follows: mind-body medicine 39%, dietary supplements 73%, body-based therapies 30%, and energy medicine 49%. During treatment CAM use decreased for all categories except energy medicine. After treatment CAM use returned to pretreatment levels for most CAMs except chiropractic. Initiation of CAM use after cancer diagnosis was positively associated with a patient having a conversation about CAM use with their oncology provider, mainly driven by patient-initiated conversations. Conclusions. Consistent with previous studies, CAM use was common among our study population. Conversations about CAM use with oncology providers appeared to influence cessation of mind-body medicine use after cancer diagnosis.

Emerging from the haze: A psycho-educational series to combat cognitive complaints after cancer treatment.

230 Background: Cognitive dysfunction after cancer treatment can be distressing and may profoundly impact quality of life, feasibility of returning to work, and social reintegration. Meaningful, evidence-based options for cancer-related cognitive changes are limited and urgently needed. A standardized rehabilitation program targeting common cognitive problems faced by cancer survivors may represent a safe, low-cost, effective treatment that can be easily disseminated. Emerging from the Haze (Haze) is a 6 week multi-disciplinary clinical program that provides a psycho-educational series to patients who are experiencing changes in cognition or having difficult adjustment after cancer diagnosis or treatment. The Haze course covers topics including: mood and stress management, mindfulness meditation, countering negative automatic thoughts, compensatory strategies for attention, memory, and executive functioning/problem solving, pacing and balance in addition to wellness issues related to cognition such as sleep hygiene, nutrition, social isolation, and exercise. The purpose of this research was to assess changes in the Functional Assessment of Cancer Therapy Cognitive Scale (FACT-Cog) scores immediately after the intervention and at regular intervals until the 1 year mark. Methods: Patients were given questionnaires including the FACT-Cog on the first day of the Haze series. The FACT-Cog includes the following domains: Perceived Cognitive Impairment (PCI), Perceived Cognitive Abilities (PCA), and Impact on Quality of Life (QOL). The survey battery was repeated on the last day of the series, 3 months, 6 months and 1 year after the series. Results: Based on the first 110 patients with various cancer types that have completed Haze, highly statistically significant improvements have been found in PCI (p &lt; 0.001; Mean Δ = 8.71), QOL (p &lt; 0.001; Mean Δ = 2.09), and PCA ( p &lt; 0.001; Mean Δ= 2.83) based on a before and after comparison of the FACT-Cog. Conclusions: It appears that a multi-disciplinary psycho-educational clinical program to address cognitive changes and adjustment difficulties after treatment can have a positive effect on patients’ perceived cognitive functioning as measured by FACT-Cog.

Mutational spectrum of Barrett's stem cells suggests paths to initiation of a precancerous lesion.

The precancerous lesion known as Barrett's oesophagus can evolve to oesophageal adenocarcinoma in decades-long processes of regenerative growth. Here we report the isolation and propagation of distinct, patient-matched stem cells of Barrett's, gastric and oesophageal epithelia that yield divergent tumour types following in vitro transformation and xenografting. Genomic analyses reveal a broad mutational spectrum unique to Barrett's stem cells that likely reflects their risk for oncogenesis. Remarkably, 25% of cases show no cancer-related genomic changes, suggesting that Barrett's initiates without driver mutations. Most cases, however, sustain patterns of deletions almost identical to adenocarcinoma though tumour-associated gene amplifications were absent. Notably, those suspected of low-grade dysplasia have p53 mutations or undergo amplifications of proto-oncogenes and receptor tyrosine kinases, implicating these events in lethal transitions. Our findings suggest paths for the initiation and progression of Barrett's and define a discrete stem cell underlying its regenerative growth whose eradication could prevent oesophageal adenocarcinoma.

Comparing surface properties of melanoma cells using time of flight secondary ions mass spectrometry.

Various techniques have been already reported to differentiate between normal (non-malignant) and cancerous cells based on their physico-chemical properties. This is relatively simple when studied cancerous cells originate from distant stages of cancer progression. Here, studies on chemical properties of two closely related human melanoma cell lines are presented: WM115 melanoma cells were taken from the vertical growth phase while WM266-4 from the skin metastatic site of the same patient. Their chemical properties were studied by two techniques, namely time-of-flight secondary ion mass spectra (ToF SIMS) and photothermal microspectroscopy (PTMS), used to record mass and photothermal spectra of cells, respectively. In our approach, independently of the spectra type, its full range, i.e. masses and wavenumbers within the range 0-500 kDa and 500-4000 cm-1, underwent a similar methodology for principal component analysis (PCA). PCA outcome shows results groupped depending on the sample type (either WM115 or WM266-4 cells). The results are independent of the method applied to study chemical properties of melanoma cells, indicating that cancer-related changes are large enough to be identified with these techniques and to differentiate between cells originating from vertical growth phase and skin metastatis.

P124: Targeted bisulfite sequencing of potential prostate cancer cell-free DNA markers: novel approach to reveal diagnostically significant molecules

Cancer cells display altered methylation signatures distinguishing them from normal cells. Originating from all tissues and cells of the body cell-free DNA (cfDNA) including aberrantly methylated DNA reflect epigenetic aberrations occurs not only in tumor cells but also in tumor microenvironment. Actually, aberrantly methylated cfDNA has proved to be a promising biomarker for noninvasive detection of cancer with several clinically-certified tests (Epi-pro Colon®, Epi-pro Lung®, Cologuard®). However only one test (Epi-pro Colon®) uses blood plasma – the most convenient source of cfDNA. Input of tissue and age specific methylation along with unidentified reasons lead to the presence of molecules with every conceivable cytosine-methylation patterns which decrease probability of tumor DNA identification. Among numerous variants of cfDNA methylation patterns only few reflect cancer related changes. To identify those tumor-specific profiles single nucleotide resolution of methylated cytosine locations in the individual circulating DNA molecules is obviously required. We performed target bisulfite sequencing of potential prostate cancer (PC) cfDNA markers (GSTP1; RNF219) isolated from blood plasma of 18 healthy donors (HD), 17 benign hyperplasia (BPH) and 20 PC patients using MiSeq platform (Illumina). RNF219 gene was shown to have high diagnostic potential in our previous comparative study of cfDNA from HD, PC and BHP patients using HCGI12k microarrays (Cortese, et al., 2012). GSTP1 is a common pathological DNA methylation event in PC and is most widely studied and promising methylation marker in the cfDNA of PC patients (Wu, et al., 2011). Selected loci were amplified after bisulfite conversion (Zymo Research) of cfDNA with methyl-independent barcoded primers and sequenced with coverage ranging from 23509 to 143953. Identification of CpG methylation status in DNA fragments was performed with the BiQ Analyzer HT Software. All statistical analysis was performed using R Statistical Software (version 3.1.1) To reveal diagnostically significant differences, several approaches to data analysis were used. Conventional approach is a prediction of patient’s diagnosis based on differences in methylation level of CpG-sites. Another approach used in this study relies on discrimination of cancer-related correlation between methylation statuses of CpG-sites within individual molecules of cfDNA – Intramolecular Correlation of Methylation Statuses (ICoMS). Study population was randomly subsampled into training and test cohorts. The logit regression model based on methylation level of CpG-sites achieved an area under the ROC curve (AUC) exceeding 0.94 in both cohorts for GSTP1 gene and 0.81 – for RNF219 gene. A novel approach to identify diagnostic significance of cfDNA methylation was based on comparison of correlation matrices (pairwise phi coefficient between methylation statuses of CpG sites) for HD, BHP and PC groups. Binomial regression models with LASSO penalization were used to predict patient’s diagnosis. ROC curves for fitted models show AUC, specificity and sensitivity as 0.99, 100% and 92% for GSTP1 gene and 0.99, 100% and 90% for RNF219 (test cohort). The ICoMS approach estimates diagnostic value of target genes and reveals cancer-related cytosine methylation. That could potentially input in gene expression, identify the features of tumor physiology and on a par with conventional approaches provide helpful information for rational design of noninvasive, methylation-specific cfDNA-based diagnostics for PC. Moreover, the planned further evaluation of relationship between the revealed correlations associated with cancer and tumor biology could increase the basic knowledge of carcinogenesis.

Abstract 815: DTX1 is an epigenetically regulated tumor suppressor gene discovered by integrative analysis of epigenetic and transcriptional alterations in HNSCC

Abstract 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

Tissue-specificity of heparan sulfate biosynthetic machinery in cancer

Heparan sulfate (HS) proteoglycans are key components of cell microenvironment and fine structure of their polysaccharide HS chains plays an important role in cell-cell interactions, adhesion, migration and signaling. It is formed on non-template basis, so, structure and functional activity of HS biosynthetic machinery is crucial for correct HS biosynthesis and post-synthetic modification. To reveal cancer-related changes in transcriptional pattern of HS biosynthetic system, the expression of HS metabolism-involved genes (EXT1/2, NDST1/2, GLCE, 3OST1/HS3ST1, SULF1/2, HPSE) in human normal (fibroblasts, PNT2) and cancer (MCF7, LNCaP, PC3, DU145, H157, H647, A549, U2020, U87, HT116, KRC/Y) cell lines and breast, prostate, colon tumors was studied. Real-time RT-PCR and Western-blot analyses revealed specific transcriptional patterns and expression levels of HS biosynthetic system both in different cell lines in vitro and cancers in vivo. Balance between transcriptional activities of elongation- and post-synthetic modification- involved genes was suggested as most informative parameter for HS biosynthetic machinery characterization. Normal human fibroblasts showed elongation-oriented HS biosynthesis, while PNT2 prostate epithelial cells had modification-oriented one. However, cancer epithelial cells demonstrated common tendency to acquire fibroblast-like elongation-oriented mode of HS biosynthetic system. Surprisingly, aggressive metastatic cancer cells (U2020, DU145, KRC/Y) retained modification-oriented HS biosynthesis similar to normal PNT2 cells, possibly enabling the cells to keep like-to-normal cell surface glycosylation pattern to escape antimetastatic control. The obtained results show the cell type-specific changes of HS-biosynthetic machinery in cancer cells in vitro and tissue-specific changes in different cancers in vivo, supporting a close involvement of HS biosynthetic system in carcinogenesis.

Impaired Glucose Handling Is Revealed in Prostate Cancer Patients

Prostate cancer survivors have increased risk for diabetes and cardiovascular disease. Here, we evaluate glucose handling and lipid profiles in men with prostate cancer and compared them with 2 cancer-free reference groups to elucidate potential cancer-related changes in metabolism. Eight high-risk, newly diagnosed prostate cancer patients (>T3 or Gleason >8 or PSA >20ng/mL), 7 age-matched, cancer-free (HM) males and 10 young healthy (HY) males were included. Fasting blood samples were obtained for glucose- and lipid-related parameters followed by an oral glucose tolerance test (OGTT) to assess glucose handling, insulin and c-peptide response. There were no significant differences in fasting glucose between groups (patients: 4.87±1.18mM; HM: 3.65±0.21mM; HY: 3.70±0.46mM). However, patients demonstrated impaired 2-hr glucose concentrations (8.68±2.87mM where normal is defined as <7.8mM), which were significantly higher compared with HM and HY groups (HM: 5.21±1.92mM, p=0.002; HY: 3.24±1.05mM, p<0.001). Similarly, insulin and c-peptide responses demonstrated significantly greater 2-hr insulin and c-peptide concentration vs HM and HY groups. There were no significant differences in total cholesterol, HDL, LDL, triglycerides between the 3 groups. Thus, glucose handling, not the lipid profile, may be affected by prostate cancer, which may influence the risk of diabetes in survivorship. Funded by the Motorcycle Ride for Dad Foundation and the Canadian Institute of Health Research.

Independent overexpression of the subunits of translation elongation factor complex eEF1H in human lung cancer.

BackgroundThe constituents of stable multiprotein complexes are known to dissociate from the complex to play independent regulatory roles. The components of translation elongation complex eEF1H (eEF1A, eEF1Bα, eEF1Bβ, eEF1Bγ) were found overexpressed in different cancers. To gain the knowledge about novel cancer-related translational mechanisms we intended to reveal whether eEF1H exists as a single unit or independent subunits in different human cancers.MethodsThe changes in the expression level of every subunit of eEF1H in the human non-small-cell lung cancer tissues were examined. The localization of eEF1H subunits was assessed by immunohistochemistry methods, subcellular fractionation and confocal microscopy. The possibility of the interaction between the subunits was estimated by co-immunoprecipitation.ResultsThe level of eEF1Bβ expression was increased more than two-fold in 36%, eEF1Bγ in 28%, eEF1A in 20% and eEF1Bα in 8% of tumor specimens. The cancer-induced alterations in the subunits level were found to be uncoordinated, therefore the increase in the level of at least one subunit of eEF1H was observed in 52% of samples. Nuclear localization of eEF1Bβ in the cancer rather than distal normal looking tissues was found. In cancer tissue, eEF1A and eEF1Bα were not found in nuclei while all four subunits of eEF1H demonstrated both cytoplasmic and nuclear appearance in the lung carcinoma cell line A549. Unexpectedly, in the A549 nuclear fraction eEF1A lost the ability to interact with the eEF1B complex.ConclusionsThe results suggest independent functioning of some fraction of the eEF1H subunits in human tumors. The absence of eEF1A and eEF1B interplay in nuclei of A549 cells is a first evidence for non-translational role of nuclear-localized subunits of eEF1B. We conclude the appearance of the individual eEF1B subunits in tumors is a more general phenomenon than appreciated before and thus is a novel signal of cancer-related changes in translation apparatus.

PR86 Opening Pandora's box – are needs of information about cancer-related changes in sexuality met?

PR86 Opening Pandora's box – are needs of information about cancer-related changes in sexuality met?

Smell, Lung Cancer, Electronic Nose and Trained Dogs

Lung cancer (LC) represents a problem of high magnitude for the medical systems due to its morbidity and mortality, also because of the huge human and economical efforts and costs that it catalyzes. Only in Europe, the average life-time cost of lung cancer patients ranges between €46,000 and €61,000 per patient. Secondary prevention, which consists in mass-screening of the high-risk population, could result of high benefit. For this aim to become a reality in the future, a potentially inexpensive and non-invasive approach for LC (pre)diagnosing is emerging. Such possibility relies on the detection of volatile biomarkers emitted from cell membranes. Tumor growth is accompanied by gene changes that may lead to oxidative stress, and the peroxidation of the cell membrane species causes volatile biomarkers to be emitted. Some of these biomarkers appear in distinctively different mixture compositions, depending on whether a cell is healthy or cancerous. These volatile biomarkers can be detected, among others, through the analysis of the exhaled breath, because the cancer-related changes in the blood chemistry are reflected in measurable changes to the breath through exchange via the lung. Importantly, these volatiles or their metabolic products are transmitted to the alveolar exhaled breath through exchange via the lung even at the very onset of the disease. There are several methods that can be applied to analyses the exhaled breath for the identification of a specific pattern of volatile biomarkers related with the target medical condition. The feasibility of three strategies, and the possible synergic effect of their concomitant application as a powerful pre-estimation tool for mass-screening of LC high-risk population: