- Total grants
- Total funders
- Total recipients
- Earliest award date
- 17 Oct 2005
- Latest award date
- 28 Apr 2020
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
Biochemical examination of metabo-synaptic coupling in a model of early cognitive decline 01 Apr 2016
Recently, interest has grown regarding the role of amyloid beta (Abeta) in impairing synaptic function in the early stages of Alzheimer’s disease (AD). These specialised areas of cell-to-cell communication play a vital role in memory formation but also represent a significant metabolic burden to neuronal cells. Notably, dysfunction in two key neurotransmitter subtypes (acetylcholine and glutamate) occurs early on in AD progression and these are the target of two classes of drugs to treat AD (cholinesterase inhibitors and memantine respectively). Additionally, functional brain imaging has shown that impaired use of glucose (the predominant fuel source for neurotransmitter production in the brain) is an early and potentially causative change occurring in AD pathophysiology. However, our understanding of how this coupling of metabolism and synapse activity becomes dysfunctional remains limited. To address this, the protein expression and activity of key pathways involved in both glucose metabolism and neurotransmitter release/recycling will be assessed in AD-relevant brain regions (cortex and hippocampus) following acute Abeta overexpression using Western blotting and enzyme-linked immunosorbent assays (ELISAs). Assessment of these changes will highlight where in these pathways becomes dysfunctional first, providing novel targets for early, potentially disease-modifying treatments.
Challenging the dogma of latency in Epstein-Barr virus associated diseases: are there more ‘latent’ genes than we thought? 01 Apr 2016
This project aims to address a fundamental question in the biology of Epstein-Barr virus (EBV) infection; namely, how do we define which viral genes are associated with viral latency? It is well established that the EBV genome codes for around 90 genes, a subset of which (known as the latency genes) are critical for EBV-driven growth transformation of primary B cells in vitro. The remaining genes are involved in viral DNA replication, virus assembly and virus egress and are only thought to be expressed in productively infected cells. However, we and others have recently shown that the pattern of EBV transcription in apparently latently infected cells is more complex than was previously thought and extends beyond the subset of traditional latency-associated genes. Here we shall focus on three of these recently identified genes, LF1, LF2 and LF3, which are poorly characterised. Using a combination of in situ hybridisation and flow cytometry, we shall screen for LF1-3 transcripts in EBV-infected cells and virus-positive tumour lines at the single cell level and ask whether these transcripts really are expressed in the absence of other markers of lytic replication.
Key Goal : Investigation of Clinical Relevance of Exosome-encapsulated microRNAs (ExomiRs) in Circulation of Breast Cancer Patients MicroRNAs(miRNAs) are small non-coding RNA molecules known to regulate gene expression at a posttranscriptional level. In cancer, they play a pivotal role as either oncomirs or tumour suppressors, and are also detectable in the circulation, raising their potential as biomarkers of disease. miRNAs are now known to be secreted by cells, and recent studies have shown that this involves encapsulation into exosomes. The majority of miRNAs in whole blood are likely not reflective of the disease due to the confounding factors present. Recent studies support the fact that exosomes are reflective of their cells of origin, but this has not been investigated in any depth on a miRNA level. If the miRNA signature of exosomes (ExomiRs) were also reflective of the cell of origin, this could have important implications in their use as biomarkers, and potential indicators of tumour phenotype. The Discipline of Surgery Biobank currently contains >1200 serum samples, from which exosomes can be isolated. This is an invaluable resource and is linked to complete clinicopathological, and treatment details of patients. ExomiRs may represent robust biomarkers reflective of tumour characteristics.
Antibody mediated induction of type 1 interferon responses in the central nervous system 01 Apr 2016
Intrathecal synthesis of lipid-specific IgM antibodies in patients with multiple sclerosis (MS) is associated with a decreased risk of developing progressive multifocal leukoencephalopathy (PML), a potentially fatal opportunistic virus infection of the central nervous system (CNS). Exciting preliminary studies suggest this reflects induction of an interferon-alpha/beta receptor (IFNAR1)-dependent anti-viral response by an IgM response directed against sulphatide, a glycosphingolipid is highly enriched in CNS myelin. This project will build on these observations using a sulphatide-reactive IgM monoclonal antibody (mAb) O4 to explore how this suppresses replication of Semliki Forest Virus (SFV; a model neurotropic virus) in myelinating cultures derived from in wild type and IFNAR1-/- mice. Specifically we will: (1) Map the time course of infection to determine its cellular tropism, how this is effected by mAb O4 and dependency of these effects on IFNAR1 (2) Determine the time frame in which the mAb O4 mediates it protective effect, in particular whether we can exploit this therapeutically, as well as prophylactically Effects on viral replication will followed and quantified using a combination of immune fluorescence microscopy (to identify and quantify SFV infected cells) and qPCR to determine changes in copy numbers of viral transcripts.
Identification of interferon-stimulated antiviral genes that contribute to the HIV-1 transmission bottleneck 01 Apr 2016
Identification of immune mechanisms of HIV-1 control at mucosal transmission sites is of importance to facilitate the development of strategies to block infection prior to systemic virus spread. My host lab have shown that HIV-1 transmitted founder viruses are relatively resistant to type I interferon (IFN)-mediated antiviral activity (as compared to viruses replicating in chronic infection), suggesting that type I IFNs make an important contribution to the HIV-1 transmission bottleneck. This project will contribute to ongoing work aiming to identify the IFN-stimulated antiviral genes that drive the IFN-resistance of founder viruses. My first aim will be to confirm preliminary results indicating that high concentrations of type II and III IFNs also inhibit HIV replication and that founder viruses are relatively resistant to their activity; and to test the effects of lower IFN concentrations. Findings from these experiments together with results from the lab’s microarray and qPCR-based analysis of common/differential gene up-regulation by high/low concentrations of type I, II and III IFNs will then be employed to identify genes to which founder viruses may be preferentially resistant. My second aim will be to use siRNA knockdown to test the role of 1-2 selected genes in founder virus IFN-resistance.
Factors Influencing HSP20 Turnover 01 Apr 2016
HSP20, a chaperone protein which has been shown to have extensive cardio-protective properties, is upregulated in a variety of tissues in response to stressful stimuli. Research has demonstrated the anti-apoptotic, anti-ischaemic and anti-hypertrophic effects of HSP20. Little is known about the turnover of HSP20 in cells; recent research has highlighted the possible involvement of the Ubiquitin proteasome system in degradation of HSP20. This project will determine the involvement of this system by firstly, measuring HSP20 half-life by cycloheximide chase, secondly, inhibiting of the suspected proteasome component and studying the ubiquitination of HSP20 and finally, determining binding of the proteasome to HSP20 by immunopurification and peptide array. This project will uncover important details about HSP20 and its turnover and the research gathered will aid progress in development of therapeutic strategies to inhibit the turnover of HSP20 in order to utilise its protective properties.
Synthesis and characterisation of a cyclic peptide protein-protein interaction inhibitor. 01 Apr 2016
The student will synthesize analogues of a cyclic peptide protein-protein interaction inhibitor, that selectively disrupts dimerization of C-terminal binding protein (CtBP), a transcriptional co-repressor that plays a key role in tumour development. The synthesized molecule is expected to have improved potency based on a crystal structure of the peptide/protein complex.They will express the target protein and assess the binding affinity by ITC and MST. This will provide experience in a range of multidisicplinary techniques.
Previous research has shown that the metabolic profiles in the blood of rheumatoid arthritis patients differ significantly from controls and relate closely to the level inflammation. Changes observed are likely to result from the high number of active immune cells present in the arthritic joint. The switch between different metabolic states of the cells is governed by a number of signalling pathways that ultimately involve AKT/PI3 kinase regulation of mTOR. It was shown that overexpression of PTPN22, which encodes the lymphoid-specific phosphatase Lyp, led to changes in phosphorylation of AKT and PI3 kinase. It may be the case that PTPN 22 differentially regulates this pathway and thus alters the switch between metabolic states, with the result that chronic inflammation is perpetuated. I will test the hypothesis that PTPN22 regulates the switch between glycolysis and oxidative phosphorylation. I will test this by using shRNA to knock down of Lyp expression in Jurkat cells and by using a Lyp inhibitor. Lyp phosphatase activity will be assessed using an activity assay. Whole cell phosphotyrosine immunoblots before and after activation will reveal if substantial changes in phosphorylation occur and possibly identify specific hyperphosphorylated targets of Lyp in the cells.
During this project, I will prepare and characterise a new potential hydrogelating compound which is a phosphate monoester derivative of a newly-discovered nucleoside organogelator containing a photoswitch. The organogelator was effective in solvent mixtures containing 50% water at concentrations as low as 2 wt%. It is hoped that the new phosphorylated material will gel water (perhaps combined with low concentrations of a non-toxic organic solvent such as DMSO) and thereby enable long duration delivery of nucleoside antiretroviral agents under conditions where adherence to complicated drug regimens based upon oral delivery is limited. I will seek to establish that: the phosphorylated material will gel aqueous solutions; this hydrogelator forms Watson-Crick base-pairs with a model nucleoside antiretroviral agent (AZT) and maintains its gel-forming capability at 37 °C; upon exposure to sunlight the gel morphology is changed; the changes in 3. above give rise to liberation of AZT examine the photophysical properties of the gels at different intensities of illumination at >400 nm; examine the thermal stabilities of gels containing different concentrations of drug and photoswitchable excipient.
Innate lymphoid cells (ILCs) are a recently discovered family of cells implicated in key immune processes such as barrier repair and tissue homeostasis. Despite enormous advances in the past few years in the characterisation of this family along with identification of several key functions, much of the basic biology of these cells remains unclear. In particular, how certain transcription factors underpin the functions of these cells is an emerging area of focus in the field, addressing regulation of key mechanisms as well as potential plasticity. Such studies are fuelled by mouse models that enable targeting of key transcription factors specifically within ILCs. The Withers lab has recently generated mice enabling the inducible deletion of RAR-related orphan receptors in mice, key transcription factors in ILC. The fundamental aim of this studentship is to perform an initial characterisation of mice in which the RAR-related orphan receptor alpha (RORalpha) is inducibly-deleted in ILC. How loss of this transcription factor impacts on the number, phenotype, cytokine production and plasticity of group 3 ILCs will be assessed in both lymphoid and non-lymphoid mouse tissues. These cells will be compared with group 2 ILCs where a key role for RORalpha in development has been identified.
Semi-quantitative characterisation of the inflammatory response in trypanosomiasis using immunohistochemistry and image analysis 01 Apr 2016
The goal of the propsed research project is to investigate the pathogensis of Trypanosome infection in working equids. The project aims at examining the immunophenotype of the lymphocyte infiltrate of brain tissues from animals infected with trypanosomiasis. We will use immunohistochemistry markers for B and T lymphocytes and measure the distance these cells are moving from the Virchow-Robin space.
Investigating the effects of ethanol exposure on the structure and function of the primary cilium 01 Apr 2016
Primary cilia are unique sensory organelles that project from the cell surface. They function primarily as the cells antenna - sensing and responding to stimuli from the surrounding cellular microenvironment. The primary cilium acts as a major signalling hub for the cell, regulating key signalling networks involved in embryogenesis including the sonic hedgehog (Shh) pathway. Thus, congenital diseases which stem from defects in primary cilia often have underlying Shh perturbation. Work in Dr. Brennan’s laboratory focuses on investigating the molecular mechanisms underlying Fetal Alcohol Syndrome (FAS), a condition which arises from maternal ethanol abuse during pregnancy. The syndrome is characterised by craniofacial, neurodevelopmental and ocular defects. Using a chick embryo model of FAS, studies have shown the effect of ethanol on early embryo development involves significant disruption to the developing retina1 and also results in decreased Shh protein expression2. Further preliminary studies suggest that this ethanol-induced disruption of the Shh pathway may be mediated through structural changes to the primary cilium. Thus, this project aims to further explore the effects of ethanol on the primary cilium using a retinal cell culture model. Research in this area could have wider implications in understanding the aetiology of fetal ethanol-related ocular defects.
Investigating the association between genetic and epigenetic variability in the 5-HTT and BDNF genes and depression in young adults. 01 Apr 2016
A high prevalence of psychopathology has been reported in students in higher education leading to significant impact on their academic success and ability to complete their studies. To assess biological and environmental factors contributing to mental health issues in students, first year students were asked to take part in the Ulster University Student Wellbeing Study. As part of this study, students provided a saliva sample to determine biological risk factors and were asked to complete an online survey on their physical and mental health. Depression was one of the most commonly reported mental health disorders but the biological basis of depression remains poorly understood. There are no clinically validated biomarkers for depression but genetic and epigenetic changes in the serotonin transporter gene (5-HTT), which plays an important role in regulation of serotonin and brain derived neurotrophic factor, which plays a vital role in neuronal survival and differentiation, have been associated with depression. The goal of this study is to determine whether polymorphism of the 5-HTTLPR, prevalence of the Val66Met polymorphism of the BDNF gene and DNA methylation in both 5-HTT and BDNF promoter regions are associated with depression in the student population.
Osteoarthritis (OA) is a degenerative joint disease and a leading cause of disability in the UK. There is currently no pharmaceutical cure for OA and pain relief is rapidly inadequate. The role of both microRNAs (miRs) and the circadian rhythm in OA have recently come to the fore, and a clear involvement for both areas in cartilage homeostasis has been identified. However, there are no data on the control of miRs in cartilage by circadian rhythms. This proposal aims to: (i) measure expression of cartilage-relevant miRs in synchronised chondrocyte cell cultures to identify those expressed in a circadian fashion; (ii) validate CLOCK as a direct target of miR-455, predicted by bioinformatics. This will provide initial data on the role of the circadian rhythm in miR expression in cartilage and inform on novel therapeutic strategies in OA.
The potential of combining novel silk biomaterial and electric field application for spinal injury repair 01 Apr 2016
Spinal cord injury (SCI) leads to impairment, often permanent, of motor and sensory function below the level of injury. In the UK about 50,000 people live with the consequences of spinal injury, with an additional 1000 each year. There is still no cure currently available for SCI and there is an urgent need for new approaches to improve functional recovery. We have previously shown that a novel silk biomaterial called Spidrex® (sourced from Oxfordbiomaterials Ltd) significantly promoted regeneration of rat peripheral nervous system and that electric fields can provide excellent guidance cues to neurons. Our more recent results from cells in culture demonstrated that Spidrex® also supported growth of mammalian nerve cells from the central nervous system (CNS). The environment at the injury site following SCI is very complex, thus no single therapeutic approach has been successful, showing that a combinatorial approach is more likely to be effective. We therefore propose to test the regenerative potential of combining Spidrex® biomaterial with electrical stimulation. This testing will give a preliminary understanding of how CNS neurons respond to a combination treatment and whether this is likely to be effective in combination with growth factors and in an in vivo injury model.
Diabetic kidney disease affects ≈40% of those with diabetes and is the leading cause of end stage renal disease. Current therapies focusing on regulation of glycaemia, hypertension and RAAS blockade are inadequate, merely slowing progression to eventual organ failure. Given the increased prevalence and devastating consequences of DN there is an urgent need to develop novel therapies. The development of fibrosis, a key pathogenic factor in DN, reflects an imbalance in the activities of endogenous drivers and suppressors of fibrosis typified by the BMP agonist-antagonist system. Prof.Godson’s group previously reported that the BMP antagonist Gremlin-1 is a key driver of DN in experimental models and human disease. Increased Gremlin-1 expression drives fibrotic responses of renal cells antagonizing the activity of BMP agonists. Prof.Godson's group have used in silico tools to design Gremlin-1 molecules which are functional dominant negatives[FDN]. These molecules are expected to bind Gremlin-1 and not agonists, therefore allowing local bioavailability of endogenously generated agonist, thus limiting fibrosis. Prof.Godson's group have expressed these molecules in mammalian cells. We propose to evaluate these novel FDN Gremlin-1 using established in vitro models of DN and renal fibrosis routinely used in Dr.Crean’s laboratory. Such data will identify their utility as therapeutics.
This research hopes to ascertain the effect of the Human T-Lymphotropic Virus Type 1 (HTLV-1) Basic Leucine Zipper (HBZ) protein on Interferon Regulatory Factor 7 (IRF7). The first objective is to determine the intracellular localisation of HBZ and IRF7 in HeLa cells. To achieve this HeLa cells will be grown on glass slides and transfected with plasmids encoding GFP-HBZ and FLAG-IRF7. IRF7 expression will be detected using an antibody against IRF7 followed by a secondary antibody linked to ALEXA-595 (Red). The localisation of proteins in the cells will be visualised using a fluorescent microscope. The second objective is to map the domains in HBZ that are involved in binding IRF7. To achieve this 293T cells will be transfected with plasmids encoding FLAG-IRF7 together with plasmids encoding HBZ-HIS wildtype or HBZ deletions HBZDAD-HIS, HBZDCD-HIS and HBZDbZIP-HIS. Co-immunoprecipitation assays will then be carried out using FLAG resin and precipitates will be analysed by Western Blot.
The impact of comorbidities: Innate immune reprogramming of atherosclerosis in response to infection. 01 Apr 2016
In previous published studies, the supervising group modelled the impact of the innate immune response on the cholesterol biosynthesis pathway. This was achieved by taking microarray measurements of bone marrow derived macrophages (BMDM) at half hour intervals for 12 hours post infection by murine cytomegalovirus. By using these measurements of transcriptional activity as crude relative approximations to protein abundance, the group was able to construct a time course of enzyme concentrations and introduce this into an ordinary differential equation (ODE) model of the pathway, using numerical integration to simulate the impact of the changing enzyme concentrations on the flux through the pathway. In this project, the student will extract measurements of the transcriptional activity of the genes coding for the proteins associated with atherosclerosis pathways from the same microarray datasets. These measurements will be used as crude estimates of protein abundance and will be incorporated into ODE models of the atherosclerosis disease process that have already been produced by the supervisor’s group. The resulting model will be numerically integrated to simulate the impact of the changing proteins concentrations on atherosclerosis disease progression as an indicator of the impact of innate immune regulation.
An investigation into the association between biofilm formation and adherence/invasion of human epithelial cells by Campylobacter jejuni 01 Apr 2016
Campylobacter jejuni is the leading cause of bacterial gastroenteritis in the world. Although the organism is highly pathogenic in humans it appears to act as a commensal in chickens where it is found in very high numbers as part of the microbiota of the chicken gut. This results in chicken meat being a major reservoir of infection for C. jejuni and recent EU studies have indicated that 97% of chicken meat in the supermarket is contaminated with C. jejuni. Several publications have highlighted biofilm formation as being an important survival mechanism for this pathogen and the O Croinin laboratory have generated unpublished data showing that strains freshly isolated from chicken meat display an increased ability to generate biofilms to survive. Given that attachment is one of the first steps in the formation of a biofilm these results could indicate that these strains are more prone to adhesion and possibly invasion of human epithelial cells which is the key virulence determinant for this important human pathogen. Tha aims of this study are thus to investigate whether a link exists between the ability of freshly isolated strains to form biofilms and their ability to attach to and invade epithelial cells.
Autophagy is an intracellular catabolic pathway that is required to maintain cell homeostasis during times of stress and starvation, as well as selectively degrade damaged organelles, protein aggregates, and intracellular pathogens. In order to develop future therapies that target this pathway to treat diseases such as cancer and neurodegeneration, further research is required to understand the mechanism of cargo selection, its mode of regulation, and its role in facilitating phenotypic alterations. Autophagy requires the integration of multiple signals, proteins, and lipids in order to identify the appropriate substrate, form the autophagosome, and fuse with the lysosome to degrade its cargo. This proposal will aim to elucidate the role of one such protein adaptor implicated in the autophagy pathway, Toll-interacting partner (TOLLIP), which has a crucial role in regulating both endocytic and autophagic cargo and their trafficking to the appropriate subcellular compartment. This project will utilise CRISPR-Cas9 gene knockout approaches and reconstitution experiments in an established human cell line to evaluate TOLLIP’s mechanistic role during autophagy.