Institute of Cancer and Genetics

Cardiff, United Kingdom

Institute of Cancer and Genetics

Cardiff, United Kingdom
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Duffy M.R.,University of Glasgow | Parker A.L.,University of Glasgow | Parker A.L.,Institute of Cancer and Genetics | Kalkman E.R.,University of Glasgow | And 5 more authors.
Journal of Controlled Release | Year: 2013

Due to many favourable attributes adenoviruses (Ads) are the most extensively used vectors for clinical gene therapy applications. However, following intravascular administration, the safety and efficacy of Ad vectors are hampered by the strong hepatic tropism and induction of a potent immune response. Such effects are determined by a range of complex interactions including those with neutralising antibodies, blood cells and factors, as well as binding to native cellular receptors (coxsackie adenovirus receptor (CAR), integrins). Once in the bloodstream, coagulation factor X (FX) has a pivotal role in determining Ad liver transduction and viral immune recognition. Due to difficulties in generating a vector devoid of multiple receptor binding motifs, we hypothesised that a small molecule inhibitor would be of value. Here, a pharmacological approach was implemented to block adenovirus transduction pathways. We developed a high throughput screening (HTS) platform to identify small molecule inhibitors of FX-mediated Ad5 gene transfer. Using an in vitro fluorescence and cell-based HTS, we evaluated 10,240 small molecules. Following sequential rounds of screening, three compounds, T5424837, T5550585 and T5660138 were identified that ablated FX-mediated Ad5 transduction with low micromolar potency. The candidate molecules possessed common structural features and formed part of the one pharmacophore model. Focused, mini-libraries were generated with structurally related molecules and in vitro screening revealed novel hits with similar or improved efficacy. The compounds did not interfere with Ad5:FX engagement but acted at a subsequent step by blocking efficient intracellular transport of the virus. In vivo, T5660138 and its closely related analogue T5660136 significantly reduced Ad5 liver transgene expression at 48 h post-intravenous administration of a high viral dose (1 × 1011 vp/mouse). Therefore, this study identifies novel and potent small molecule inhibitors of the Ad5 transduction which may have applications in the Ad gene therapy setting. © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

Du P.,University of Cardiff | Du P.,Institute of Cancer and Genetics | Du P.,Capital Medical University | Ye L.,University of Cardiff | And 6 more authors.
Journal of Experimental Therapeutics and Oncology | Year: 2012

Background: Metastasis suppressor 1 (MTSS1), a cytoskeletal associated protein, has been indicated in certain types of human cancers, but its role in kidney cancer remains unknown. We have investigated the expression of MTSS1 in normal and malignant human kidney tissues and its molecular interaction within kidney cancer cells. Materials and Methods: The expression of MTSS1 in human kidney tissues and kidney cancer cell lines was assessed at both the mRNA and protein levels using RTPCR and immunohistochemistry, respectively. Full-length MTSS1 cDNA expression vector was used to generate MTSS1 over-expressing cells. Effect of MTSS1 overexpression on cellular functions, was examined in kidney cancer cells of MTSS1 being over-expressed using a variety of in vitro assays. Involvement of Sonic Hedgehog (SHH) pathway was tested by using Shh small inhibitors. Results: Epithelial cells at proximal tubules of kidney tissues were stained positively for MTSS1, while the staining was weak or absent from cells at corpuscles and cancer cells of tumour tissues. Similarly, in kidney cancer cell lines, CAKI-2 and UMRC-2, expressed very low level of MTSS1. Over-expression of MTSS1 reduced the growth, invasion, adhesion and migration of kidney cell lines in vitro. Shh inhibitors diminished the inhibitory effect of MTSS1 on cell migration. Conclusion: MTSS1 expression is reduced in human kidney cancer cells. MTSS1 levels are inversely correlated with the growth, invasion, adhesion and migration of kidney cancer cells in vitro. MTSS1 suppresses migration of kidney cancer cells via SHH pathway, and is a putative tumour suppressor of kidney cancer. © 2012 Old City Publishing, Inc.

PubMed | University of Oxford, St James Hospital, University of Cardiff, University College London and Institute of Cancer and Genetics
Type: Journal Article | Journal: Journal of clinical pathology | Year: 2015

Molecular characterisation of tumours is increasing personalisation of cancer therapy, tailored to an individual and their cancer. FOCUS4 is a molecularly stratified clinical trial for patients with advanced colorectal cancer. During an initial 16-week period of standard first-line chemotherapy, tumour tissue will undergo several molecular assays, with the results used for cohort allocation, then randomisation. Laboratories in Leeds and Cardiff will perform the molecular testing. The results of a rigorous pre-trial inter-laboratory analytical validation are presented and discussed.Wales Cancer Bank supplied FFPE tumour blocks from 97 mCRC patients with consent for use in further research. Both laboratories processed each sample according to an agreed definitive FOCUS4 laboratory protocol, reporting results directly to the MRC Trial Management Group for independent cross-referencing.Pyrosequencing analysis of mutation status at KRAS codons12/13/61/146, NRAS codons12/13/61, BRAF codon600 and PIK3CA codons542/545/546/1047, generated highly concordant results. Two samples gave discrepant results; in one a PIK3CA mutation was detected only in Leeds, and in the other, a PIK3CA mutation was only detected in Cardiff. pTEN and mismatch repair (MMR) protein expression was assessed by immunohistochemistry (IHC) resulting in 6/97 discordant results for pTEN and 5/388 for MMR, resolved upon joint review. Tumour heterogeneity was likely responsible for pyrosequencing discrepancies. The presence of signet-ring cells, necrosis, mucin, edge-effects and over-counterstaining influenced IHC discrepancies.Pre-trial assay analytical validation is essential to ensure appropriate selection of patients for targeted therapies. This is feasible for both mutation testing and immunohistochemical assays and must be built into the workup of such trials.ISRCTN90061564.

Ge Z.,Institute of Cancer and Genetics | Ge Z.,Capital Medical University | Sanders A.J.,Institute of Cancer and Genetics | Ye L.,Institute of Cancer and Genetics | And 2 more authors.
Oncology Reports | Year: 2013

Death receptor-3 (DR3) plays controversial roles in cancer. Currently, DR3 is known to be a functional receptor of vascular endothelial growth inhibitor (VEGI). The role of DR3 in breast cancer remains unclear. The present study investigated DR3 expression in a clinical cohort of breast cancer patients and its role in breast cancer cells in vitro. The expression of DR3 was examined in a breast cancer cohort using quantitative PCR (Q-PCR) and immunohistochemistry (IHC) in comparison to the patients' data. In vitro function of DR3 was examined through the targeting of this molecule in MCF7 and MDA-MB-231 breast cancer cells using ribozyme transgene technology. Decreased DR3 expression was noted in breast cancer tissues compared to normal tissues and decreased expression of DR3 was generally associated with a poorer prognosis as well as a significantly shorter long-term survival (p=0.038). Targeting of DR3 in vitro in breast cancer cell lines resulted in impaired migratory rates compared to respective control cells. Collectively, these data suggest a complex role for DR3 in breast cancer development and progression. © 2013 Spandidos Publications Ltd. All rights reserved.

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