Patient-level evidence obtained from clinical trials is essential in assessing the cost-effectiveness of health care technologies. Given the increasing demand for primary evidence and limited public resources for health care research, research funding organisations are routinely called to make decisions on which clinical trials to fund. Such decisions need to be informed by evidence on the likely costs and benefits of competing research programmes. Two main analytic approaches have been proposed to provide such evidence, ‘payback of research’ and ‘value of information’. This work applied the ‘payback’ and ‘value of information’ methodologies to case studies representing proposals for clinical trials in cancer. This application gave estimates of the value of undertaking the trials and offered an insight into the strengths, limitations and usefulness of the methods. ‘Payback of research’ and ‘value of information’ can help with different funding decisions in the context of different funding streams, they are practical to undertake and can be readily incorporated into the existing research funding processes. It is suggested that the methods should be used as part of existing deliberative processes, to provide additional assurance that limited public resources are allocated to clinical trials which are likely to result in benefits to the population.

We examined both the chromosomal copy number changes and differential RNA expression profiles in Nasopharyngeal carcinoma (NPC). Gene expression profiles identified a large number of differentially regulated genes involved in diverse functional processes, while genetic analysis detected extensive genomic abnormalities including large and small, discrete regions of copy number change and loci that exhibit uniparental disomy (UPD). The relationship between chromosomal copy number and level of gene expression were analysed. This revealed that the direct copy number/expression link applies in about 60% of the instances of copy number loss/down-expression and less than 35% of instances of copy number gain/up-expression that were examined. Signalling pathway analysis revealed that numerous components involved in the TGF-β, Wnt/β-catenin and Hedgehog pathways were universally upregulated in NPC tumours, and gene expression pattern of the C666-1 cell line approximated to other NPC tumours, indicating that it is a good tumour model. A preliminary in vitro investigation of signalling pathways revealed that the C666-1 cell line is intact in the activin and Hh signalling pathways but not in the TGF-β pathway. However, the C666-1 cells appear to resist activin-mediated cell growth inhibition

This Birmingham Prostatic Neoplasms Association Study (BiPAS) was initiated to investigate determinants of prostate cancer. The study recruited 314 prostate cancer patients, 381 active surveillance patients, 201 hospital controls and 175 population controls. By comparing groups of varying risk, the aetiology of the disease was investigated. Within the BiPAS dataset, sun exposure, physical activity and obesity were analysed. The association with occupation was assessed by performing a meta analysis of 7, 762 cases and 20, 634 controls. Finally, a replication study on genetic polymorphisms on 8q24 using 277 cases and 282 controls from the Netherlands Cohort Study (NLCS) is presented. A protective effect was observed for high sun exposure in early adulthood and high intensity exercise. An increased risk was observed for low intensity exercise and men classed as obese at age 20. The meta analysis suggested moderately increased and decreased risks associated with a number of job titles, however none were statistically significant. The results for allele A on the single nucleotide polymorphism rs1447295 were replicated; however a decreased risk was detected for allele -8 on the microsatellite DG8S737. No significant difference was detected for analysis comparing prostate cancer or high PSA cases.

TRAIL is a member of TNF superfamily and binding of TRAIL to its receptors induces cell death with apparent specificity for carcinoma cells compared with normal epithelium. On this basis, a number of TRAIL-targeted therapies are currently under investigation as anti-cancer agents. Recent studies in our lab have shown that membrane bound forms of both FasL and CD40L are more potent in cell death induction, suggesting that a membrane bound TRAIL could be more effective in cell death induction compared to the wild-type or the soluble counterparts. TRAIL gene therapies have demonstrated that ex vivo infection of cancer cells with a recombinant adenovirus expressing TRAIL (RAdTRAIL) enhances apoptosis and promotes tumour regression. In the present study, we have generated two adenoviral vectors. The first one expressing wild-type TRAIL (RAd wtTRAIL) that subject to cleavage from cell membrane and the second adenoviral vector expressing fusion CD40LTRAIL protein that is resistant to metalloproteinase cleavage (RAd CD40LTRAIL). The direct effects of these viruses were examined on TRAIL receptor positive carcinomas either alone or in combination with different chemotherapeutic drug. The RAd CD40LTRAIL that expressed the membrane bound CD40LTRAIL was found to exhibit more cell death than the RAd wtTRAIL. This cell death was through the activation of caspase 3/7.

Thyroid cancer, the most common endocrine malignancy has a rising incidence worldwide. Radiation damage is a known aetiological factor in thyroid tumourigenesis, particularly in children. Pituitary tumor transforming gene (PTTG) and its binding partner (PTTG binding factor; PBF) are overexpressed in thyroid cancers. Critically, PTTG and PBF have been shown to be independent markers of poor prognosis in thyroid cancer. PTTG, a human securin, has multifunctional roles in mitotic control, DNA repair, apoptosis, cell transformation and genomic instability. PBF, which has independent tumourigenic and transforming actions, binds to PTTG, transports it to the nucleus and facilitates its actions within the nucleus. Taken together, the above implicate the functional role of PTTG and PBF in genetic instability. This thesis describes the generation of a transgenic murine model of thyroid cancer which overexpressed both human PTTG and human PBF in the thyroid gland (BI-Trans). The BI-Trans murine model developed goitres from a young age in both genders. Additionally, they develop thyroid adenomas at a later age which had a female preponderance. Unexpectedly, this BI-Trans murine model did not develop cancers. We next measured the index of genetic instability (GI) in the thyroids of our transgenic murine models with fluorescent inter-simple sequence repeat-PCR (FISSR-PCR). This technique was refined to measure GI with small quantities of DNA obtained from murine thyroids. We performed microarray analysis on our murine thyroids with and without ionising radiation to elucidate the mechanism by which PBF and PTTG induced genetic instability.

Casein Kinase 2 (CK2) is a ubiquitous serine/threonine kinase. Due to its pleiotropic nature CK2 is involved in a multitude of cellular pathways including cell survival, proliferation and apoptosis. Therefore it has come as no surprise that a requirement for CK2 activity has been identified during the repair of DNA damage. Here we have described a novel role for CK2 in the response to DNA double-strand breaks. We have shown the mediator of DNA damage checkpoint 1 (MDC1) is constitutively phosphorylated by CK2, which is required for an interaction with the MRE11/RAD50/NBS1 (MRN) complex, via the FHA domain of NBS1. Moreover, disruption of this interaction resulted in loss of MRN foci following ionizing radiation and a partial G2/M checkpoint defect. Furthermore, the identification of three siblings presenting NBS/Seckel-like phenotypes with a unique mutation in the BRCT domain of NBS1, that phenocopied some of our observations, provided additional evidence for the importance of phospho-dependent interactions within the cell. Lastly, the identification of putative CK2 target residues in MRE11 and our preliminary data suggest that the kinase may play further roles in regulating the activity of the MRN complex that lie outside its activity in DNA double-strand break repair.

Epstein-Barr virus (EBV) has been etiologically associated with Burkitt lymphoma (BL) since its discovery 50 years ago, but despite this long-standing association the precise role of the virus in the pathogenesis of BL remains enigmatic. EBV can be lost spontaneously from EBV-positive BL cell lines, and these EBV-loss clones have been reported to exhibit increased sensitivity to apoptosis. We have confirmed and extended those observations and report that sporadic loss of EBV from BL cells is consistently associated with enhanced sensitivity to apoptosis-inducing agents and conversely, reduced tumorigenicity \(in\) \(vivo\). Importantly, reinfection of EBV-loss clones with EBV can restore apoptosis protection, although surprisingly, individual Latency I genes cannot. We also used inducible pro-apoptotic BH3 ligands to investigate Bcl-2-family dependence in BL clones as well as profiling gene expression changes in response to apoptosis induction in EBV-positive versus EBV-loss clones. We found that EBV-loss was consistently associated with enhanced sensitivity to BH3-ligand-induced death and increased activation of apoptosis signalling pathways, although no individual apoptosis-related gene was responsible. Instead we find that Latency I EBV genes co-operate to co-ordinately repress the BH3-only proteins Bim, Puma and Noxa to inhibit apoptosis in BL.

Despite improvements in early diagnosis and prevention, late stage breast cancer is often incurable due to metastasis, tumour relapse, resistance and incomplete response to treatments. Metabolic reprogramming has been recognised as a critical element for cancer cells to grow under hostile conditions and this is likely to contribute towards resistance against chemotherapeutics. This thesis therefore aimed at deciphering the metabolic phenotype of residual breast cancer which survived docetaxel treatment, in vitro and in vivo, quantifying polar metabolite levels and conducting pathway tracing and metabolic flux analysis using stable isotope (¹³C) labelled tracers. \(In\) \(vitro\) residual cells presented a hypermetabolic phenotype characterised by significant accumulation of essential and non-essential amino acids, together with an elicited Warburg effect and an increased antioxidant response based on glutathione production, while in growth arrest. A method to carry out in vivo tracer-based metabolic studies was successfully developed using a breast cancer mouse model. Although the metabolite accumulation outlined in vitro was not observed in vivo, a protective phenotype against oxidative stress was supported by increased flux through the oxidative branch of the pentose phosphate pathway. In conclusion, this thesis demonstrated that metabolic phenotyping is a valid approach to uncover key metabolic alterations in residual tumours both in vitro and in vivo, and could be further exploited to design personalised treatments aimed at restoring sensitivity to therapies.

Proteins which regulate the expression and activity of the small oncogenic lipids, LPA and S1P are important in the development and progression of cancers. This study set out to explore the expression of lipid signalling molecules in two types of lymphoid malignancies and a few epithelial malignancies and correlate their association with clinicopathological parameters. The expression of SPHK1, S1PR1, ABCC1 and ATX in epithelial malignancies were studied which included breast, bladder and lung cancer. It is shown that S1PR1 is a poor prognostic marker in breast cancer and ATX is a poor prognostic marker in both breast and lung cancers. The expression of lipid signalling proteins and mRNA showed that S1PR1 was commonly expressed in HL, while S1PR3 was expressed in all HL cases suggesting its role in lymphomagenesis. L591 Hodgkin lymphoma cells treated with anti-S1P, Sphingomab showed restoration of four B cell receptor genes namely BCL10, RAPGEF1, NFATC3 and MALT by gene expression microarray. Sphingomab also showed downregulation of S1PR3 and S1PR5 genes which may be a possible therapeutic target in cancer. The incidence of EBV+ DLBCL in this series is 14/300 (4.7%). S1PR1 was commonly expressed in DLBCL and patients with S1PR1 expressing tumours were more likely to have more than one extranodal site involvement. EBV positivity, presence of B symptoms, bulky disease, BCL6 positivity and high IPI were found to be poor prognostic factors in DLBCL.

Eph receptor tyrosine kinases and their ligands ephrins are involved in many physiological processes, including tissue homeostasis, vascularisation and development. They are also involved in promoting angiogenesis and metastasis in many cancers, including breast, colon and lung. Upon interaction with EphB receptors, ephrinB ligands signal through SH2- and PDZ-interacting cytoplasmic adaptors. To date, little is known on PDZ-mediated ephrinB signalling. The aim of our study was to determine the role of PDZ domain-containing proteins in modulating ephrinB2 signalling and trafficking pathways in the context of epithelial breast cancer cells, with a specific focus on the scaffolding protein syntenin-1. We also endeavoured to determine whether the ephrinB2 – syntenin-1 axis affects breast cancer tumourigenesis. Our findings demonstrate that syntenin-1 modulates ephrinB2 internalization upon receptor-induced stimulation and that this affects ephrinB signalling. Furthermore, we found that phospho-Tyr330 on ephrinB2 increases binding to the PDZ domain-containing proteins syntenin-1 and PAR3. Finally, we report that ephrinB2 drives MCF7 colony growth in 3D cultures and that syntenin-1 is involved in boundary formation between ephrinB2 and EphB4 expressing cells. These findings describe, for the first time, the role of syntenin-1 in ephrinB2 signalling and the functional relevance of the ephrinB2 – syntenin-1 axis in epithelial breast cancer pathways.