Diagnostic Significance of Exosomal miRNAs in the Plasma of Breast Cancer Patients

Poster Session AbstractsBackground and Aims: Emerging evidence that microRNAs (miRNAs) play an important role in cancer development has opened up new opportunities for cancer diagnosis. Recent studies demonstrated that released exosomes which contain a subset of both cellular mRNA and miRNA could be a useful source of biomarkers for cancer detection. Here, we aim to develop a novel biomarker for breast cancer diagnosis using exosomal miRNAs in plasma. Methods: We have developed a rapid and novel isolation protocol to enrich tumor-associated exosomes from plasma samples by capturing tumor specific surface markers containing exosomes. After enrichment, we performed miRNA profiling on four sample sets; (1) Ep-CAM marker enriched plasma exosomes of breast cancer patients; (2) breast tumors of the same patients; (3) adjacent non-cancerous tissues of the same patients; (4) Ep-CAM marker enriched plasma exosomes of normal control subjects. Profiling is performed using PCR-based array with human microRNA panels that contain more than 700 miRNAs. Results: Our profiling data showed that 15 miRNAs are concordantly up-regulated and 13 miRNAs are concordantly down-regulated in both plasma exosomes and corresponding tumors. These account for 25% (up-regulation) and 15% (down-regulation) of all miRNAs detectable in plasma exosomes. Our findings demonstrate that miRNA profile in EpCAM-enriched plasma exosomes from breast cancer patients exhibit certain similar pattern to that in the corresponding tumors. Based on our profiling results, plasma signatures that differentiated breast cancer from control are generated and some of the well-known breast cancer related miRNAs such as miR-10b, miR-21, miR-155 and miR-145 are included in our panel list. The putative miRNA biomarkers are validated on plasma samples from an independent cohort from more than 100 cancer patients. Further validation of the selected markers is likely to offer an accurate, noninvasive and specific diagnostic assay for breast cancer. Conclusions: These results suggest that exosomal miRNAs in plasma may be a novel biomarker for breast cancer diagnosis.link_to_OA_fulltex

Key performance indicators for sustainable manufacturing evaluation in automotive companies

The automotive industry is regarded as one of the most important and strategic industry in manufacturing sector. It is the largest manufacturing enterprise in the world and one of the most resource intensive industries of all major industrial system. However, its products and processes are a significant source of environmental impact. Thus, there is a need to evaluate sustainable manufacturing performance in this industry. This paper proposes a set of initial key performance indicators (KPIs) for sustainable manufacturing evaluation believed to be appropriate to automotive companies, consisting of three factors divided into nine dimensions and a total of 41 sub-dimensions. A survey will be conducted to confirm the adaptability of the initial KPIs with the industry practices. Future research will focus on developing an evaluation tool to assess sustainable manufacturing performance in automotive companies

A Systems Genetics Analysis of Metastatic Mammary Cancer Development in Mice Fed Varying Levels of Dietary Fat

High dietary fat intake and/or obesity may increase the risk of susceptibility to certain forms of cancer. To study the interactions of dietary fat, obesity, and metastatic mammary cancer, a population of F2 mice cosegregating obesity quantitative trait loci (QTL) and the MMTV-PyMT transgene was created. The mice were fed either a very high-fat or a matched-control-fat diet, and evaluated for growth, body composition, age at mammary tumor onset, tumor progression, and pulmonary metastases development. Single nucleotide polymorphism (SNP) genotyping across the genome facilitated analyses of QTL and QTL by diet interaction effects. To further investigate the complex genetic architecture that modifies mammary cancer and metastasis, expression profiles of axillary tumors were characterized with the Illumina Mouse-6 whole genome sentrix arrays. Using a systems-based analysis pipeline developed in R, we conducted a genome-wide expression QTL (eQTL) analysis was conducted. In addition, network and pathway QTL analyses for mammary tumors that have developed in the presence of varying degrees of obesity, and during exposure to high or normal fat diets. Results demonstrated that mice fed a high-fat diet are not only more likely to experience decreased mammary cancer latency but they also have increased tumor growth and occurrence of pulmonary metastases over an equivalent time. We identified 25 modifier loci for mammary cancer and pulmonary metastasis, likely representing 13 unique loci after accounting for pleiotropy, as well as novel QTL x diet interactions at a majority of these loci. Transciptome mapping revealed several candidate genes potentially underlying both tumor and metastasis QTL. These candidates were subsequently prioritized using multiple analytic approaches, including but not limited too causality testing, copy number variation analysis and database evaluations

Biocompatible tumour implant systems: towards an integrated biophotonic system

There is a need to perform comprehensive cell biology studies transferable across culture platforms using innovative cellular models. The higher purpose is to bridge the gap between in vitro cell culture and in vivo models. In this thesis a significant advance is presented in the embedding of an innovative optical biophotonic capability for the dynamic interrogation and single cell tracking of human osteosarcoma cells encapsulated in the hollow fiber (HF) platform. Two approaches have been implemented: quantum dot (QD) nanoparticles providing proliferative and cell cycle readouts and an in-fiber light illumination providing global features of particle and cell density. An in vitro HF encapsulation model was developed and characterised against standard two-dimensional tissue culture (TC) using the human osteosarcoma U-2 OS cell line expressing a cell cycle fluorescent reporter (cyclin Bl-GFP). Analysis of the packing and orientation of cells in the HF revealed that they grow like an anchorage dependent adherent layer. Overall cells in the fiber displayed a slower cell cycle traverse and a differential sensitivity to clinically relevant doses of the anticancer mitosis-inhibiting agent Taxol compared to cells under normal TC conditions. Comprehensive gene profiling, with bioinformatics and ontology network analysis, showed that the HF cells presented high steroid related but low differentiation gene expression. Specific biomarkers were indentified, and it is suggested that the HF model displays features that are closer to an in vivo tumour. A flow cytometry cell-tracking approach using QD labelling was validated and applied to the HF model for the first time. This represents an "embedded" biophotonic system where the QD sensors are integrated directly into the seeded cell population and then redistributed through the daughter cells, thus reflecting patterns of lineage expansion. This provides sub-population parameterized information on cell-cell heterogeneity and cell division. A biophotonic HF prototype comprising the integration of direct coupled-light excitation in the HF was conceived, this revealed the potential and limitations to detect die presence of cells inside the HF lumen by analysing light attenuation changes. Finally a "systems cytometry" acquisition concept has been proposed, comprising the use of embedded engineered nanoparticles as single cell "nano-memory" biophotonic intracellular probes

Primary sclerosing cholangitis: from genetic risk to disease biology

Primary sclerosing cholangitis: from genetic risk to disease biology Elizabeth Claire Goode One in 10,000 people in the Western world lives with Primary Sclerosing Cholangitis (PSC), an immune-mediated, inflammatory disease of the bile ducts that is highly co-morbid with inflammatory bowel disease (IBD). PSC confers risk of serious disease sequelae including hepatobiliary malignancy and progression to end-stage liver failure, for which the only treatment option is liver transplantation. The absence of effective medical therapies for PSC reflects our current limited understanding of the disease’s aetiology and pathogenesis. Our DNA, laid down at conception, gives us an unrivalled opportunity to understand the underlying causal biology of disease. This is because the genetic variants associated with disease susceptibility perturb genes and biological pathways that contribute to disease causality. Twenty-two regions of the genome, outside of the HLA, have been associated with PSC risk. These loci offer the potential for huge insight into the causal biology of PSC, if only we can robustly identify the true causal variants driving these loci and the genes they perturb. However, this is complicated by several scientific challenges. Firstly, the majority of disease-associated risk loci occur within non-coding regions of the genome. Secondly, patterns of correlation between variants within a risk locus means that the true causal variant driving the signal could be any of those highly correlated with the variant with the smallest p-value. In this thesis, I present analyses aimed at identifying the true genes and causal variants underlying each of the twenty-two PSC risk loci. Many non-coding risk variants associated with complex disease exert a quantitative affect upon gene expression i.e. are expression quantitative trait loci (eQTLs). Colocalisation assesses the evidence that a single shared causal variant is responsible for driving PSC risk and gene expression via an eQTL. In order to assign dysregulated genes to PSC risk loci, I perform colocalisation with eQTLs mapped in multiple cell-types and tissues mechanistically relevant to PSC. Because PSC is rare, eQTLs have not previously been mapped in all cell-types most relevant to this disease. In addition, I therefore map eQTLs in six peripheral blood T-cell subsets (including the rare CCR9+ gut-homing T-cells) from 80 patients with PSC and IBD. With colocalisation, I assign causal genes to five PSC risk loci, and assign other epigenetic regulatory features including methylation or histone modification, to six risk loci. Statistical fine-mapping of each risk locus in both the GWAS and eQTL data enables me to resolve three PSC risk loci to a single causal variant and nine loci to 95% credible sets containing ten or fewer variants. The results presented in this thesis identify three genes (PRKD2, ETS2 and UBASH3A), causal in the pathogenesis of PSC, which are currently the target of existing or experimental therapeutic agents. Firstly, reduced expression of PRKD2 causes excessive cell-autonomous T-follicular helper cell development and B-cell activation, and is associated with increased risk of PSC. Several studies are investigating the therapeutic effects of increasing the kinase activity of PRKD2. ETS2 is involved in the induction of pro-inflammatory cytokine release from macrophages and IL-2 regulation in Th to Th0 transition. ETS2 inhibitors are currently the subject of early therapeutic trials. Finally, UBASH3A attenuates the NF-kB/I-KKb pathway, an inflammatory pathway that is already targeted by proteasome inhibitors and acetylsalicylic acid, both of which could be potentially therapeutic in PSC. PSC is a debilitating disease with serious disease sequelae, for which new therapeutic options are urgently needed. In this thesis, I elucidate multiple genes with a causal role in PSC pathogenesis, several of which are potential candidates for future therapeutic target