NIHRDataInsights/HRCSData huggingface dataset:
Title: Characterisation of deregulated miRNAs during colorectal cancer progression - Interplay between tumour stroma and malignant epithelium
Funder: Medical Research Council

Abstract: Introduction: Metastases are the principle cause of death in colorectal cancer (CRC). An emerging theme in metastasis development is the role of non-coding microRNAs (miRNAs). An additional area of interest is the role of stromal cells and their interplay with cancer epithelium in dynamically influencing this process. The aim of this project is to establish whether stromal and epithelial miRNA expression patterns have important consequences for the development of metastases in CRC. Methods: MiRNAs highly differentially expressed in advanced stages compared with early stage disease have been selected as candidates for study. The molecular targets of candidate miRNAs will be identified using Western Blotting and 3'UTR reporter gene assays. Biological significance will be assessed in cultured cell lines in which miRNA expression has been selectively induced or knocked-down. Standard in-vitro functional assays will be used in addition to bespoke organotypic models in which malignant epithelial cells and ex-vivo colonic fibroblasts are juxtaposed in a configuration that closely resembles in-vivo conditions. Preliminary work: CRC specimens of different, clinically relevant pathological stage have been subjected to Laser Microdissection (LMD) in order to separately profile miRNA expression in stromal and epithelial layers. Stromal expression successfully separated stage-I/II tumours from stage-IV tumours based on a panel of miRNAs, illustrating the potential relevance of stromal candidates. Candidates are currently being tested for functional significance and 3D organotypic models reveal that miR-153 in the epithelium and miR-21 in the stroma are important regulators of CRC invasion. Applications: Identification of clinically relevant miRNAs in the CRC stroma is novel and may improve staging, guide therapy and reveal new therapeutic targets. Added value is provided as the distinctive biology of CRC stroma has not been reflected in miRNA profiling studies to date.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Exploring the molecular interface between the HIF/hypoxia signalling pathway and mitochondria in cancer
Funder: Medical Research Council

Abstract: Hypoxia-inducible factors (HIF) are crucial, evolutionarily conserved components of the cellular oxygen sensing machinery that are activated when oxygen levels are low (hypoxia). Recent studies have highlighted an important role for mitochondria in relaying hypoxic signals to regulate HIF-1 in mammalian cells. For example, reactive oxygen species (ROS) produced in mitochondria, have been proposed to play a role in HIF-alpha stabilization during hypoxia. Pharmacological inhibitors of the respiratory chain, or genetic knockdown of cytochrome C or mitochondrial complex components have been shown to suppress hypoxia-induced ROS production and HIF-1alpha stabilization. Furthermore, HIF-1alpha can regulate mitochondrial function to optimize the efficiency of respiration in hypoxic cells by modulating the expression of cytochrome oxidase subunits. Thus, several studies connect mitochondria and components of the respiratory chain with HIF-1alpha induction and the hypoxic response, and vice versa. However, the molecular mechanisms underlying precisely how the HIF/oxygen sensing machinery interface with mitochondria remain unclear and this has been a long-standing question in the field. Using a bioinformatics approach, we have identified a new family of mitochondrial proteins (CHCHD4.1 and CHCHD4.2), that interact with HIF-1alpha under physiological conditions and are important regulators of HIF-1alpha-hypoxia signalling in vivo. CHCHD4.1 is identical to MIA40, which in yeast, functions to import proteins into the mitochondrial intermembrane space. The proposed work will investigate the role of the HIF-1alpha/hypoxia signalling pathway in directly regulating mitochondrial function and metabolism in tumour cells (in vitro and in vivo), with particular focus on exploring the relationship between HIF-1alpha and CHCHD4 proteins.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Modulating Beta-1 Integrin Activation as a Novel Therapeutic Strategy in Malignant Mesothelioma
Funder: Medical Research Council

Abstract: Hypothesis: This fellowship will test the hypothesis that in MM cells galectin-3 and CD98 promotes proliferation and resistance to chemotherapy in MM cells and the permissive inflammatory tumour environmental in a beta1 integrin dependent manner Aim: To determine the impact of galectin-3 and CD98 on beta1 integrin-mediated functions in MM cells in vitro and in vivo. Key beta1-integrin functions will be examined: cell proliferation, chemoresistance, ECM secretion and fibronectin matrix assembly. Manipulation of galectin-3/CD98/beta1 integrins in MM cells prior to co-culture with macrophages on macrophage phenotype will be examined. These results will inform the experimental crux of this fellowship - to determine the effect of intrapleural administration of an inhibitory agent (defined by the in vitro experiments) on mesothelioma growth and associated inflammation and fibrosis and mesothelioma sensitivity to chemotherapy . Efficacy of this agent will be assessed by PET/MRI in a murine model. We will address molecular mechanisms by examining in vitro cell signalling cascades and in vivo by novel imaging techniques. Medical Potential: to identify key molecules driving the inflammatory microenvironment and tumour propagation and to pharmacologically/ genetically inhibit these pathways. To drive this research forward to the creation of a novel treatment for malignant mesothelioma and to potentially improve the efficacy of existing treatments.


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Mechanism of transcriptional gene silencing induced by convergent transcription in human cells and its application in breast cancer.
Funder: Medical Research Council

Abstract: All organisms consist of cells that multiply through cell division. The genetic code, which defines each cell type, is stored within DNA molecules. DNA contains coding regions called genes. Genes are transcribed into messenger RNA (mRNA) molecules, which are in turn translated into proteins, a process called gene expression. It is important that each cell makes the right levels of proteins, which is achieved by controlled gene expression. Uncontrolled gene expression can lead to cancer or cell death. One of the most important mechanisms for the regulation of gene expression is gene silencing. Gene silencing in human cells requires the presence of small RNA molecules (siRNA), which activate the RNAi pathway. This leads to recognition and distruction of target mRNA in the cytoplasm. Another type of gene silencing is the establishment of specific modifications on DNA, which leads to inhibition of gene transcription, also called transcriptional gene silencing (TGS). Little is understood about the mechanism of TGS in humans. The work proposed in this fellowship application will use available knowledge about TGS from other organisms to investigate and explore the details of how TGS can be induced in humans and by what mechanism. This work has important implications given that it can lead to development of new research tools with a great potential for therapeutic application.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: MICA:Discovery of ERK5 inhibitors for biomarker-driven clinical evaluation in cancer
Funder: Medical Research Council

Abstract: Extracellular regulated kinase 5 (ERK5, BMK1, MAPK7) is the terminal kinase in one of the 4 major mitogen activated protein (MAP) kinase pathways. MAP kinase pathways are central to cancer, as well as numerous other pathologies. Recent success with B-RAF inhibitors in melanoma, and ongoing clinical trials with MEK1/2 inhibitors, has demonstrated that these pathways are drug-able. Correlative clinical data have identified ERK5 as a target in a number of tumour types, including prostate, breast and liver cancer, and mechanistic studies have demonstrated a tumour-type dependent role for ERK5 in cell proliferation, survival, invasion, and/or angiogenesis. Importantly, recent encouraging pre-clinical data using a small molecule ERK5 inhibitor (XMD8-92) demonstrated pronounced inhibition of in vivo tumour growth. In collaboration with the CR UK Beatson Institute (Prof. Hing Leung), the Babraham Institute (Dr Simon Cook) and Cancer Research Technology, Newcastle has identified two novel series of potent and highly selective ERK5 inhibitors. The lead series has low MW and high ligand efficiency, sub-microM activity against ERK5, no activity against 131 other kinases in a screen which included 10 ERK/p38/JNK isoforms, low microM inhibitory activity in a cell-based ERK5 reporter gene assay, together with excellent solubility, in vitro metabolic stability and plasma protein binding properties. The lead compound series was identified by a high-throughput screen (HTS) of ~160,000 compounds, which has also identified a back-up series. The current application is for support to undertake lead optimisation in the primary lead series, and hit validation and hit-to-lead studies on the back-up series, supported by structure-based drug design. In addition to identifying a clinical trial candidate, a comprehensive portfolio of invasive and non-invasive biomarkers will be developed to facilitate hypothesis-testing early phase clinical trials.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Translational Termination and its Regulation by Hydroxylation
Funder: Medical Research Council

Abstract: Deregulated protein synthesis and translational control are crucial aspects of cancer initiation and progression. As such protein translation is attracting increasing interest as a source of novel cancer targets. I have recently discovered that the molecular machinery controlling termination of protein synthesis is targeted by JMJD4, a completely uncharacterised 2-oxoglutarate-dependent oxygenase related to the HIF hydroxylases and JmjC-histone demethylases. I have shown that JMJD4 is overexpressed in tumours and required for the viability of cancer cells. JMJD4 hydroxylates K63 of the translational termination factor eRF1 within a highly conserved functional motif involved in stop-codon recognition: JMJD4 inhibition causes ribosomal readthrough of stop codons. I hypothesise that inhibition of eRF1 hydroxylation causes aberrant protein synthesis, re-expression of nonsense mutated tumour suppressor genes and reduced tumuourigenesis. This proposal aims to test these hypotheses using a multidisciplinary approach with the ultimate aim of validating JMJD4 and the translational termination pathway as a novel and druggable anti-cancer target.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Evaluation of non-invasive metabolic imaging biomarkers for novel RAF/MEK1/2-targeted anti-cancer agents
Funder: Medical Research Council

Abstract: RAF-MEK1/2-ERK1/2 signalling is a focus for targeted cancer drug development with many RAF-MEK1/2 inhibitors now in clinical trials. Detecting biomarkers of pathway blockade, particularly those that are non-invasive, is key for assessing target modulation and drug efficacy and thereby aiding identification of promising pharmaceuticals in early stage trials to prioritise for pivotal trials. Using magnetic resonance spectroscopy (MRS), we have shown a reduction in lactate levels in BRAF mutant human melanoma cells and xenografts following treatment with MEK1/2 selective inhibitors. Here we will a) identify and assess the metabolic biomarkers of response to novel classes of RAF/MEK1/2 inhibitors (BRAF-selective, MEK1/2 selective and RAF+MEK1/2 selective) in human cancer cells with varying BRAF and RAS mutation status, b) investigate the metabolic mechanisms involved in response and resistance to treatment and c) assess the translatability of any metabolic biomarkers in in vivo tumour models and patient tumour tissues, to identify the value of incorporating in vivo measurements into clinical trials. We will assess the effect of RAF/MEK1/2 signalling inhibition in human cancer cells with varying BRAF and RAS mutation status in vitro using 1H, 31P and 13C MRS combined with molecular and biochemical assays to investigate changes in metabolism and metabolic pathway fluxes and enzyme activities associated with response and resistance to treatment. These investigations will then be extended to a) tumour models in vivo where we aim to delineate the impact of any therapy-driven physiological changes (using MR imaging) on metabolism and b) RAF-MEK1/2 inhibitor-treated patient samples to assess the clinical relevance of our findings. This work will define the potential of metabolic changes as non-invasive imaging biomarkers for the action of RAF/MEK1/2-targeted agents and will be a major step in translating findings from the laboratory to the clinic.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Estimation of the genetic correlation among human cancers and identification of pleiotropic cancer loci
Funder: Medical Research Council

Abstract: This project proposes to develop methodology for the estimation of the genetic correlation between two traits. These traits could be two diseases, a clinically relevant intermediate trait and a disease or two clinically relevant continuous traits. The methods are targeted to exploit the use of unrelated samples of the population. We will mainly focus on the most challenging scenario where both traits are not observed/measured on the same individuals. The methods will allow obtaining estimates of the degree by which diseases share susceptibility loci and gene pathways represented in current genotyping arrays. The project will set up a general analytical framework for complex diseases and an approach to combine different sources of information where missing data is a problem. For instance, when the clinical intermediate phenotype has been measured only in a subset of samples or when repeated measures are difficult to obtain (eg, measure cognitive ability in childhood and old age). We will use cancer as a paradigm of correlated disease. The incontrovertible evidence for genetically determined susceptibility to cancer has spurred a large number of genome-wide association studies (GWAS) to identify susceptibility loci. Despite their arguable success, the current approach is unsatisfactory in that uses a 'one SNP at a time' approach which might miss combinations of common and/or rare variants at particular loci and that it models 'one trait at a time' thereby missing vital information about the correlation structure of diseases and clinical traits. Our methodology has the potential to overcome the limitations of the current approach and identify some of the large proportion of genetic variation that remains unexplained. We will use restricted maximum likelihood (REML) in a mixed-model framework to estimate the genetic correlation captured by SNP arrays at a global and local genomic level.


Research activity codes: 8.4 RESEARCH DESIGN AND METHODOLOGIES (HEALTH SERVICES)
Health categories: CANCER AND NEOPLASMS
Title: MICA: Determining the therapeutic potential of targeting mTORC-1/2 in chronic lymphocytic leukaemia - a pre-clinical study
Funder: Medical Research Council

Abstract: Chronic lymphocytic leukaemia (CLL) is refractory to current chemotherapeutic regimes, highlighting the need for novel therapies. Recent research has established that in addition to cellular accumulation due to increased expression of anti-apoptotic proteins, CLL cells also exhibit enhanced proliferation within patient lymph nodes (LN), promoting disease progression. Targeting survival and proliferation signals represent a promising way to deliver a curative strategy for CLL. The protein kinase mammalian target for rapamycin (mTor) forms two distinct complexes mTorc-1 and mTorc-2, which promote cell survival and proliferation. Our preliminary data demonstrate that mTorc-1 and -2 substrates are activated both in ex vivo and proliferating human primary CLL cells, and in cells derived from a CLL mouse model. While the mTorc-1 inhibitor rapamycin inhibits CLL proliferation, it is not cytotoxic to CLL cells, leading us to hypothesise that dual targeting of mTorc-1/2 will preferentially disrupt proliferation/survival pathways present in CLL patient LN microenvironment to induce apoptosis. To delineate the functional and molecular roles played by mTorc-1/2, we will utilise three complementary experimental systems: - Primary CLL LN tissue biopsies, to determine the activation status of mTorc-1/2 in a biologically-relevant setting in vivo; - CLL samples cultured in in vitro experimental systems that mimic in vivo survival- or proliferative signals, to assess the impact of manipulating mTor signalling with shRNA techniques or dual mTorc-1/2 inhibitors; - CLL mouse models, to assess the therapeutic response of mTor inhibition in vivo. These model systems will enable us to define the importance of mTor as a valid therapeutic target for CLL. Moreover, elucidating the molecular signatures that accompany mTor inhibition in CLL cells may lead to the identification of potentially novel biomarkers, which will assist in future clinical trial design.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: MICA: The role of Src in cancer-induced bone pain
Funder: Medical Research Council

Abstract: Bone pain is a significant clinical feature of several common cancers, including breast, prostate and lung cancers, which all have a tendency to spread to bone. Adequately control of bone pain is a significant unmet clinical need in almost half of cancer patients with bone metastases, and therefore understanding the mechanisms of cancer-induced bone pain would be helpful in developing rationale-based therapy. The current proposal is a collaborative project between basic and clinical scientists at the University of Sheffield and a commercial partner, AstraZeneca, to test the hypothesis that the non-receptor tyrosine kinase Src is a critical component of cancer-induced bone pain, and that its inhibition represents a novel analgesic strategy for controlling bone pain in cancer patients. A series of preclinical studies using an animal model of bone cancer pain will investigate the effects of inhibiting Src on pain-related behaviour, spinal cord neuron phosphorylation and signalling as well as bone resorption to identify potential analgesic mechanisms of Src inhibition. We will also translate these studies into patients by performing a randomized controlled trial of the Src inhibitor Saracatinib, to investigate whether it has a role as an analgesic in cancer patients with bone metastases. The results of this study will elucidate the role of Src in bone cancer pain, and they will confirm whether Src inhibition shows clinical promise as a mechanism of analgesia.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: MICA: GSK-3 as a multifunctional target for glioblastoma treatment; Hitting multiple tumour hallmarks with a single drug.
Funder: Medical Research Council

Abstract: New treatments are badly needed for the aggressive brain tumour glioblastoma multiforme. We have identified a potential strategy based on inhibition of GSK-3 that may be of benefit to patients through effects on multiple tumour hallmarks, including invasion and angiogenesis. There is also evidence that GSK-3 inhibition may enhance the effectiveness and increase the tumour selectivity of the current standard of care, temozolomide plus radiation. The potential of the novel GSK-3 inhibitor AZD1080 for glioblastoma will be tested using a range of in vitro assays, molecular readouts, and studies on animal models. In Aim 1, a panel of migration assays (transwell, 3D spheroid, and a novel 3D nanofiber-based assay) will be used in glioblastoma cell lines to determine the effects of AZD1080 on tumour cell migration. We will also examine cell viability parameters in combination with radiation and temozolomide, as well as assays on endothelial cell migration/tube formation and glioma initiating cell differentiation. In Aim 2, analysis of signaling pathways will be performed to determine the downstream effectors of AZD1080 treatment in glioblastoma cells. This will be done by combining global gene expression profiling data and phosphoproteomic data with readouts from luciferase reporter constructs measuring the activities of specific transcription factors regulated by GSK-3 in cancer cells. Finally in Aim 3, we will perform studies on intracranial mouse tumour models of glioblastoma. Dosing will be established followed by survival studies, using AZD1080 alone, and also in combination with radiation and temozolomide. Detailed analysis of the phenotypes of treated tumours will be carried out using immunostaining for tumour proliferation, apoptosis and tumour associated blood vessels. These experiments will determine whether AZD1080 is a suitable drug to bring forward for the treatment of glioblastoma in human patients.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS