- Total grants
- Total funders
- Total recipients
- Earliest award date
- 21 Jan 2017
- Latest award date
- 11 Dec 2017
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
TRAF6 is essential for many biological processes, including those activated by interleukin-1 (IL-1) family members, Toll-Like-Receptor (TLR) ligands, and TNF superfamily (TNFSF) members. We have unexpectedly discovered that many of the essential roles of TRAF6 are independent of its E3 ubiquitin ligase activity and that, surprisingly, a key role of the TRAF6 E3 ligase is to restrict T cell activation and prevents autoimmunity. Building on these and other exciting findings we have made, the aim of the proposed reseach is to solve three major questions:- 1.What are the molecular mechanisms by which TRAF6 mediates signaling by the TNF superfamily members RANKL, CD40 and BAFF? 2. What are the molecular mechanisms by which the TRAF6 E3 ligase restricts T cell activation, and will they open up new opportunities to unleash the power of cytotoxic T cells to treat cancers and other diseases? 3. How does the TRAF6 E3 ligase activate a novel IL-1-dependent signaling pathway we have identified, which triggers activation of the "master" kinase TAK1 in the absence of its ubiquitin-binding subunits TAB2 and TAB3, and does this pathway have important roles in other TRAF6-dependent processes?
The control of cell division orientation impacts on the position and function of cells. Here we focus on the genetic and mechanical control that establishes the orientation of polarity and division in stem cells. Using neural stem cells, called neuroblasts, of the developing Drosophila brain, we aim to understand how stem cells remain within niche signalling range when actively proliferating. We hypothesised that the outcome of neuroblast division provides a polarity landmark for division orientation in the next cell cycle. Consistently, we found that a neuroblast daughter cell derived signal influences Par polarity module localisation when neuroblasts divide again. We now want to identify the genes behind this symmetry-breaking event. Our results suggest that midbody-organised structures link symmetry-breaking to cortical tension anisotropies and want to test if deforming the cortex using optical tweezer microscopy can bias the orientation of Par polarity. Downstream of Par complex localisation, aPKC coordinates polarity. To dissect the pleiotropic effects of aPKC, we have generated ATP analog sensitised alleles, which are homozygous viable and can be specifically inhibited. This enables us for the first time to dissect the functions of aPKC in a temporally resolved manner that establish asymmetric cell fate localisation and cell size asymmetry.
The overarching aim of our research is to understand tissue development and disease mechanisms. To develop new tissue imaging capabilities, we request a Zeiss LSM 880 confocal system with Airyscan module. The Airyscan detector replaces the pinhole with a sensitive 32-element detector - each element acting as a minimum sized pinhole. By combining the signals from the 32 elements the signal to noise is sufficiently improved to produce an increase in resolution in all spatial directions of 1.7 (140nm laterally and 400nm axially). This will permit super-resolution microscopy of thick tissue sections that cannot be achieved by conventional confocal microscopy, multi-photon microscopy or existing OMX Structured Illumination Microscopy (SIM). Overall the Airyscan will aid the specific scientific objectives of the applicants that span fundamental knowledge on cell division and neurobiology including deciphering kinase mechanisms of Parkinson's disease in the brain (Miratul Muqit); understanding the regulation of mitophagy in vivo (Ian Ganley); probing the regulation of neurogenesis in chick/mouse including altered subcellular structures and live signalling (Kate Storey); understanding kinetochore-microtubule interactions in human cells (Tomo Tanaka); elaborating new mechanisms that drive mitotic chromosome segregation (Jason Swedlow) and discovering new determinants of cell polarisation (Jens Jansuchke).
Genomic and experimental investigation of inter-bacterial competitive mechanisms and protein secretion systems in clinical isolates of Serratia marcescens 31 Jan 2017
Research into bacterial pathogenesis has long focused on ‘classical’ anti-host virulence factors. However given the current appreciation of the polymicrobial nature of infections, inter-bacterial competition mechanisms should also contribute significantly to a pathogen’s ability to colonise a niche within the host and cause infection. We have observed that the opportunistic bacterium Serratia marcescens, an important nosocomial pathogen, is highly adept at inhibiting rivals. Additionally, comparative genomics of a small number of clinical isolates has indicated the possession of multiple strain-specific inter-bacterial competition mechanisms in S. marcescens. We will investigate the spread of interbacterial competition mechanisms, including protein secretion systems, within a collection of clinical isolates of S. marcescens, using whole genome sequencing (WGS) and comparative genomics. Novel or previously unidentified competition mechanisms in S. marcescens, identified from comparative genomics will be tested for their ability to confer bacterial killing or inhibition in vitro and subsequently in vivo in a mouse model of pneumonia. Through this work, we aim to elucidate the extent and contribution of inter-bacterial competition to S. marcescens pathogenic success, and by extension, to that of opportunistic Gram-negative bacteria more generally. This may pave the way for future work aiming to combat hospital-acquired infections.
Pluripotency maintenance and transition to differentiation require tight regulation of gene expression at the transcriptional and post-transcriptional level. The mRNA cap consists of a series of co-transcriptional modifications of the nascent transcript that are important for its processing and translation initiation. The mature cap structure requires methylation by the complex RNMT-RAM. A recent study showed that RAM, the mRNA cap methyltransferase activating subunit, is highly expressed in embryonic stem cells (ESCs) and required for the expression of pluripotency-associated genes such as Oct4 and Sox2. During the process of neurodifferentiation, degradation of RAM is required for the loss of pluripotency and expression of neural-associated genes. Given the importance of mRNA cap methylation in pluripotency maintenance, in this project we will investigate the role of mRNA capping in the process of ESCs differentiation and early embryo development. We will analyse the expression patterns of the capping enzymes in both embryoid bodies (EBs) and blastocysts along with pluripotency and lineage-specific markers. We will perform this analysis in wild-type and capping enzyme knockout-derived EBs and blastocysts. Depending on the observed expression patterns, we will pursue any interesting emerging phenotypes that may suggest the involvement of regulated mRNA capping in the differentiation process.
Intraepithelial lymphocytes (IEL) are the first immune cells that pathogens encounter in the gut. These lymphocytes lie within the epithelial layer, and are central to controlling infection, stress or transformation of the gut epithelium. Despite their importance, we have a poor understanding of how IEL sense and respond to stress in epithelia, partly due to the lack of genetic tools that specifically target IEL. To get unbiased insights into the complex biological processes underlying IEL function, I determined the quantitative proteomes of three main intestinal IEL subsets. This unique dataset reveals that IEL are phenotypically and functionally distinct from conventional T cells. The IEL proteomes provide unprecedented insights into the effector functions of intestinal IEL and suggest new hypotheses of how their function might be regulated. Based on these data, I will (1) explore the apoptotic and non-apoptotic functions of abundantly expressed granzymes in IEL, (2) develop novel genetic tools to dissect the role of IEL in the early immune response to pathogens, and (3) address the contributions of inhibitory receptors and their signalling pathways to IEL homeostasis and sensing of epithelial dysbiosis. In summary, the proposed research will reveal how IEL communicate and respond to bacterial and parasitic infections.
Development of Cardiovascular Disease (CVD) is preceded by endothelial dysfunction, inflammation and infiltration of vessel walls by pro-inflammatory leukocytes, resulting in formation of atherosclerotic plaques. Recent in vivo studies have suggested a role for the Salt Inducible Kinase-2 (SIK2) enzyme in regulation of inflammatory cytokine production. SIK2 phosphorylates CRTC3, promoting cytosolic retention. Inhibition of SIK2 prevents CRTC3 sequestration and promotes nuclear accumulation where it associates with CREB and promotes transcription of IL-10, IL-1ra and the anti-atherosclerosis associated gene, Nur77. Furthermore, macrophages from SIK2 kinase-null mice produce significantly less pro-inflammatory IL-6, IL-12 and TNF-a. The precise role of SIK2 in CVD is unknown. Using femoral artery cross-sections and subcutaneous blood vessels collected from diabetic humans following elected above knee amputations, we aim to characterise the expression and function of SIK2 in the development of atherosclerosis. Furthermore, we aim to compare these findings with blood biomarkers of inflammation and kidney function. Expression of SIK2 shall be investigated using RT-PCR gene expression analysis and related to enzyme function using western blot analysis of protein phosphorylation, focussing on the TLR4-MAPK-MSK-CREB-CRTC3 signalling axis. Identification of SIK2 as a regulator of atherosclerotic development could provide a novel drug target for the prevention and management of CVD.
Protein O-GlcNAcylation is an essential posttranslational modification regulated by two opposing highly conserved enzymes, O-GlcNAcase (OGA) and O-GlcNAc transferase (OGT), influencing a wide range of biological processes. The dysregulation of the O-GlcNAc system is linked to neurodegeneration, cancer and diabetes. OGT consists of a catalytic domain, and an N-terminal domain comprising 13.5 tetratricopeptide repeats (TPRs). This TPR domain is believed to mediate protein-protein interactions. The OGT gene is located on the Xq13.1 band. Four XLID causing mutations were identified in the OGT gene so far. Residing in the TPR region, they lead to a decrease of steady-state global OGT protein levels and effects on the O-GlcNAc proteome. In my summer research project, I would like to focus on the A319T (c.1193G>A) missense mutation. Having been found in a French family with three XLID affected sons, this mutation is regarded as the possible pathogenic factor in this particular case. My key goals include the production of the A319T mutated OGT in E. coli, to be further on purified and characterised in vitro. The structural integrity will be checked and assays screening the enzymatic activity will be carried out. I hope to detect differences in enzyme kinetics caused by the structural changes.
The protease, BACE1 (beta-site amyloid precursor protein (APP)-cleaving enzyme 1) is rate-limiting in amyloid beta (Ab) production. Ab peptides are prone to aggregation causing amyloid plaques, a process strongly linked to Alzheimer’s disease (AD). People with AD have an increased risk of T2D (Janson et al., 2004) and people with peripheral insulin resistance correlate with AD risk (Li & Hölscher 2007). Central obesity also increases dementia risk, independently of diabetes (Luchsinger 2010). Thus, the pathological processes underlying AD are linked with the metabolic disturbances associated with T2D and obesity. We hypothesise that raised BACE1 activity driven by high fat feeding results in the induction of central insulin and leptin resistance and disordered glucose and lipid metabolism, increasing susceptibility to diabetes and cardiovascular disorders. Key Goals By proteomics determine which brain proteins in hippocampus and hypothalamus are up- or down-regulated in a BACE1-dependent manner following chronic high fat feeding. Validate the top proteomic hits by immunoblotting, mRNA and/or ELISA analysis in the appropriate brain areas. Attempt to correlate changes in key signalling pathways with the phenotypic and metabolic characteristics of BACE1 null (resistant to obesity and diabetes) and BACE1-knock-in (increased susceptibility to diabetes) mice.
Patients with chronic respiratory diseases such as bronchiectasis and COPD suffer from frequent exacerbations (chest infections) that are typically treated with antibiotics +/- corticosteroids. The goal of treatment with preventative measures is to reduce the frequency of chest infections and this is the major end-point in most randomized trials of new drugs in these diseases. Previous dogma held that the airways were sterile and that exacerbations were caused by exogenous acquisition of bacteria from the environment. More recent work suggests that the bacteria isolated during exacerbations are often found in the lungs when patients are well and that they may represent an overgrowth or alteration in the natural microbial flora of the lungs (the microbiome). We hypothesise that if this is true, the presence of pathogenic bacteria at low levels in the lung when patients are well will predict future bacterial chest infections. 1. To test the hypothesis that the presence of bacteria known to be associated with exacerbations(particularly Haemophilus influenzae and Pseudomonas aeruginosa) in the sputum microbiome is associated with the future risk of exacerbations of bronchiectasis over 1 year follow-up 2. To test whether the bacterial load of these pathogens is associated with an increased risk of exacerbations.
The ultimate goal of the research is to develop a microfluidic system, usable in labratory and field situations, for the detection and quantification of the viral diversity in a sample. The initial stage will focus on analysing viral properties in order to differentiate the viruses into their different families. This data, which focuses on the size, shape and deformability of the viruses, will be used to inform mathematical modelling which will lead to the selection of nano scale filters to be used for testing. Analysis will then be performed on microfluidic experiments to validate models and test the effectiveness and efficiency of the filters when used for viral separation. For this research we will use easy to handle and non pathogenic viruses, as well as control substances with known proportions of these different viruses to allow us to effectively evaluate our initial systems. This will also be complemented by modelling of the particles as they interact with the filters to further investigate the dynamics and properties of separation. A key aim of this research is to generate a wealth of data which will have applications in the viral detection at low concentrations in bodily fluids and unbiased characterisation of viruses.
To support the development of the WASSUP project.
Beyond Transitions 21 Jan 2017
Volunteering Matters will use the grant to support younger people with disabilities in Blaenau Gwent by providing them with skills and empowering them during the transition from education to further education, employment or training. This grant for Â£5,000 will fund volunteer expenses, mentor training, stationery and telephone costs, promotional materials, evaluation, and administration support.
The organisation provides support for the Armed Forces community and their families. The project is using £10,000 to refurbish the toilet facilities in their hall so more people can take part in activities.
Any grant will support veterans who need help with improving their lives at a difficult time or in challenging circumstances.