- Total grants
- Total funders
- Total recipients
- Earliest award date
- 20 Nov 1998
- Latest award date
- 17 Apr 2020
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
Wellcome Trust Clinical PhD Programme at the University of Dundee: "Dissecting folate metabolism in Trypanosoma brucei." 14 Apr 2014
The aim of the project is to further dissect folate metabolism in T.brucei, the causative agent of human African trypanosomiasis (HAT). An understanding of folate transport, the distribution of intra-cellular folates and the essentiality of enzymes in folate metabolism will not only provide a better understanding of folate metabolism but also inform our understanding of anti-folate drug mode of action and mechanisms of resistance. Specifically, we aim to: - Assess the role of thymidine kinase (TK) in modulating sensitivity to anti-folates through gene deletion - Determine how folate and its derivatives are taken up and further metabolised, and identify the genes involved using an RNAi-knockdown approach - Determine the effect of anti-folates on intra-cellular metabolite distribution - Identify mechanisms of resistance to anti-folates using a genome-scale tetracycline inducible RNAi library screen and compare these to experimentally derived resistant lines Throughout the project we will target key enzymes in the folate metabolic pathway (e.g. methionine synthase, folylpolyglutamyl synthase) to determine their essentiality and their potential as therapeutic drug targets, by creating conditional knockouts and over-expressers.
Wellcome Trust Clinical PhD Programme at the University of Dundee: "Genomic investigation into evolution of Staphylococcus aureus and the micro-epidemiology of Staphylococcus aureus in atopic eczema." 14 Apr 2014
Across all continents Staphylococcus aureus is the commonest cause of skin and soft tissue infection. Surface displayed and secreted proteins are fundamental for the ability of this bacterium to colonise and infect human hosts. Such secreted factors confer this organism’s ability to adhere to host cells and evade the immune system. The Type VII protein secretion system (T7SS) in S. aureus is emerging as an important system that contributes to disease-causing mechanism for this human pathogen. It has previously been shown that inactivation of the Type VII system in S. aureus (or equivalent in Mycobacterium) reduces pathogenicity. As of yet the precise function of this secretory system is still not fully understood. The aim of this work is directed towards determining the role the T7SS plays in human skin colonisation and the potential role it has in infection. This will be achieved by studying both T7SS wild type and mutant strains in models of skin infection (murine) and colonisation (ex-vivo human tissue culture). Once the involvement of the T7SS in these models in confirmed we will examine the contribution of the secreted substrates with human cells.
Musculoskeletal Study of Limb Development in Vertebrates by Magnetic Resonance Imaging Microscopy. 13 Dec 2006
Magnetic resonance imaging (MRI) microscopy will be used to study embryonic development in vertebrates, in particular chicks, as a model for the investigation of human limb deformities. The chick embryo will be studied from the appearance of limb buds in Day 3 embryos through to wing formation. The anatomical changes during development will be characterised, with particular reference to organogenesis of the limbs. MRI parameters such as relaxation rates will be quantified to give insights into t he tissue s microscopic morphology. The rate of the limb outgrowth and rate of bone ossification will be measured. MRI will also be used to observe the formation, differentiation and striation of muscles and growth of tendons and blood vessels. Once the three-dimensional chick anatomy has been systematically characterized, the anatomy will be compared with experimentally manipulated chicks. Ultimately the MRI of the development of live chick embryo will be studied overtime as the embryo grows in side the egg. MRI contrast agents will be used to follow specific biological processes. Chick embryos are uniquely suited for testing new contrast agent experiments. This project will be to open up novel ways of looking deeper into the developing embryo.
The proposal requests funding for Flow Cytometry at the College of Life Sciences at the University of Dundee. The main users of the Flow cytometry facility are applicants studying Dendritic cell biology, Lymphocyte Signal transduction and Carbohydrate recognition in the immune system. Immunology Research in Dundee is embedded in the College of Life Sciences . The acquisition of flow cytometry equipment for immunology was a catalyst that promoted the use of flow cytometry by 19 non immunological research groups with diverse interests including parasitology, gene transcription and developmental biology. A common theme is to use flow cytometry for cell cycle analysis; apoptosis analysis; single cell analysis of protein phosphorylation; single cell analysis of the expression of integrins, cytokine and chemokine receptors and adhesion molecules inc ells of the adaptive and innate immune system. There is also a need to analyse expression patterns of cell surface and transfected proteins f ollowing diverse immunological stimuli, to cell sort rare cell subsets based on antigenic profiles. Flow cytometry is also used to isolate subpopulations of cells transfected with green fluorescent protein (GFP) tagged proteins prior to single cell analysis by confocal microscopy.
Identification of substrates and roles in virulence of the Staphylococcus aureus Ess protein secretion system. 02 Dec 2015
The interplay between the Notch and Wnt signalling pathways in somitogenesis This project aims to investigate the potential crosstalk between the Notch and Wnt signalling pathways in the important process of somitogenesis. It is well established now that the Notch pathway plays a crucial role in somitogenesis during vertebrate embryogenesis in a variety of species. More recently it was shown that the Wnt pathway is also required for this process in the mouse embryo. It was also reported that Wnt may play a role upstream of Notch in this process. The first aim of the project is to confirm and extend preliminary findings that Notch may be upstream of Wnt in chick somitogenesis. The second aim of the project is to examine the level of interplay between the two pathways in both the mouse and the chick embryo. Establishing how the Notch and Wnt pathways control somitogenesis will be an important step in understanding how this process is regulated.
The role of LKB1 in T-Lymphocytes. 20 Dec 2006
The serine/threonine kinase LKB1 was first identified as a tumour suppressor from genetic linkage analysis of mutations in Peutz-Jeghers Syndrome. LKB1 has now been identified as an essential multifunctional kinase, with LKB1 null mice dying midgestatin. We have previously demonstrated that LKB1 has an essential role in T-lymphocytes via the generation of mice with a conditional LKB1 gene deletion in T-cell progenitors in the thymus. Thesemice have effectively no peripheral T-lymphocytes, with the mice exhibiting a block in theymocytic development at the pre-T (double negative 4 - DN4) stage. Recently we have shown that these DN4 cells will proliferate, but not differentiate, when grown on OP9-DL1 stromal cells, allowing the generation of large numbers of cells for analysis. The binding partners and localisation of LKB1 in T-lymphocytes have yet to be defined, however affinity purification studies in other cell types suggest that the localisation of endogenous LKB1 is dependent on attached proteins. It has been observed that LKB1 localises to the nucleus unless complexed with the pseudokinase STE20-related adaptor (STRAD) and mouse protein 25 (MO25), in which case the complex localises to the cytoplasm. Additional binding partners for LKB1 have also been reported. Importantly, the presence of STRAD and MO25 increases the catalytic activity of LKB1 about 10 fold. The aim of this project is therefore to investigate the role of LKB1 in lymphocytes via the study of LKB1 localisation, characterisation of existing LKB1 null DN4 thymocytes and the generation of lymphocytes with tamoxifen-inducible LKB1 gene deletions. These cells will be utilised to investigate the role of LKB1 in the polarisation of lymphocytes during both migration and CTL degranulation.
Characterization of the enzymes of the Kennedy pathway for PE formation in Trypanosoma brucei. 20 Dec 2006
Trypanosoma brucei is the causative agent of African sleeping sickness, a vector-borne parasitic disease that threatens millions of people in sub-Saharan Africa. Ethanolamine (tN) is a major component of the trypanosome membrane phospholipids, in the form of phosphatidylethanolamine (PE). PE constitutes 25-30% of the total membrane phospholipid of T. brucei and for this reason it has a very important role in maintaining membrane homeostasis; because membrane phospholipids determine membrane fluidity and the surface charge of cell surfaces, ethanolamine utilization and PE biosynthesis are likely to control a variety of cellular processes. In addition, EtN is found as an integral component of the glycosylphosphatidylinositol (GPI) anchor that is required for membrane attachment of cell surface proteins, most notably the Variant Surface Glycoproteins (VSG), which in the bloodstream form of the parasite plays the essential role of protecting the parasite from the host's immune system. Consequently, a trypanosome which can no longer synthesize or replace its surface glycoprotein coat would be cleared by the immune system and accordingly, the biosynthesis of the conserved GPI-anchor has previously been genetically and biochemically validated as a drug target in bloodstream form T. brucei. The analysis of how ethanolamine is metabolized in T. brucei could unravel novel targets for the development of chemotherapeutic drugs, as well as novel insights on how eukaryotes assemble glycolipid components of membranes and membrane-bound proteins at the cell surface.
Previous studies have shown that a network of serine kinases has essential functions in T cells. Accordingly, we plan to establish the molecular basis for the actions of four key serine kinases in T lymphocytes. Genetics, biochemistry, immunology and cell biology will be used to characterise the role of Phosphoinositide dependent kinase 1 ( PDK1), LKB1 and the AMP dependent protein kinase in peripheral T cell activation and function and to analyse the role of LKB1 and AMPK in regulating T lymphocyte metabolism. Diacylglycerol binding kinases of the Protein Kinase D family are specific targets for antigen receptors in lymphocytes and temporally and spatially disseminate antigen receptor signals away from the plasma membrane into the cell interior. Functions for PKDs in lymphocytes include the control of the phosphorylation and localization of class II histone deacetylases, key regulatory enzymes that control chromatin and repress gene expression. . We have also identified actin regulatory proteins and adapter molecules as PKD substrates. The key issues we will address are the mechanisms that regulate the subcellular localisation of PKDs; the role of PKDs in regulating the lymphocyte cytoskeleton; the function of different PKD isoforms in lymphocytes.
This is an application to support the purchase of a Fluorescence Activated Cell Sorter (FACS) housed in a Biological Safety Containment Level 2 cabinet and to support the salary of a Flow Cytometry Operator. This equipment will support the research of approximately 30 academic research groups within the School of Life Sciences including 21 Wellcome Trust funded Principal Investigators. The research by these groups includes cell and molecular biology, bacteriology, immunology, mammalian developmental biology, parasitology and drug discovery. A dominant focus is on cell signaling, particularly in the context of the immune system, the regulation of cell metabolism and neuro-degeneration. There are also projects involving bacteria, yeast, trypanosomes and leishmania, as well as multiple non immune mammalian cells. The requirement for a new FACS stems from increased usage in research projects, a large increase in demand for use of single cell sorting cell sorting as part of efficient and CRISPR/Cas9 gene editing protocols. There is also the need to use Biological Safety Containment level 2 for cell sorting to allow translation of research ideas generated using model organisms to studies of freshly isolated human blood derived lymphocytes and other human cells.
IgA immunotherapy of tuberculosis. 24 Apr 2007
of 2E9/IgA1 to modulate infection with tubercle bacilli of monocyte-derived cells and macrophage cell lines. In addition, we will investigate the capacity of 2E9/IgA1 to mediate ADCC against Mtb infected macrophages. The protective effect of passive IgA therapy in vivo will be assessed in mice transgenic for human FcalphaRI/CD89. Protection will be optimized by co-inoculation of rIFNgamma and anti-IL-4 antibody. To develop a deeper understanding of the mechanisms involved, we will investigatethe influence of 2E9/IgA1 on Mtb-induced phagosomal membrane changes using confocal microscopy and real time imaging, and we will assess the 2E9/IgA1's ability toenhance antigen presentation.
Molecular mechanisms of O-GlcNAc signalling. 03 Dec 2013
Post-translational modification of eukaryotic protein serines/threonines with N-acetylglucosamine (O-GlcNAc) was discovered 20 years ago. Subsequent work has shown that O-GlcNAcylation is regulated by a tranferase (OGT) and a hydrolase (OGA), both single, essential genes, conserved from C. elegans to humans, and that a plethora of proteins in the nucleoplasm are O-GlcNAcylated. Excitingly, there are examples of O-GlcNAcylation having interplay with phosphorylation, including competition for the same serines/threonines, giving rise to the Yin-Yang hypothesis , that proposes O-GlcNAc as a global, glucose dependent, mechanism to control protein phosphorylation. I aim to use a multi-disciplinary approach to uncover the molecular mechanisms governing O-GlcNAcylation and its importance in a number of signal transduction pathways. Capitalising on a significant body or preliminary data, the aims are to: 1) Synthesize/discover OGA/OGT inhibitors and substrate analogues, including glycopeptide s. 2) Determine the structures of human OGA/OGT, including inhibitor/protein substrate complexes and probe mechanism/specificity with mutagenesis and in vitro assays. 3) Understand the effect of O-GlcNAc on phosporylation/activity on key signalling proteins in pathways involve in the insulin response, neuronal development and energy stasis. 4) Investigate whether orthologues of these enzymes in pathogenic bacteria are involved in prokaryotic O-GlcNAc and/or are virulence factors targetting ho st proteins.
The role of mitogen activated protein kinases in the regulation of micro RNA (miRNA) gene transcription and miRNA dependent gene expression. 23 Jan 2006
The role of Mitogen Activated Protein Kinases in the Regulation of micro RNA (miRNA) Gene Transcription and miRNA Dependent Gene Expression
Mechanisms for Remodelling Chromatin. 13 Apr 2011
All eukaryotes have developed an assortment of different strategies by which chromatin structure can be altered. We aim to gain insight into this fundamental process with special emphasis on the action of Snf2 related chromatin remodelling enzymes. Specific areas of interest include: 1) The structure and mechanism of action of Snf2 related enzymes. We aim to build a structural model for an intact remodelling enzyme and to establish how this interacts with nucleosomes. The experimental approa ches we use will then be directed at understanding how energy derived from ATP hydrolysis is converted into structural changes in chromatin. With the aim of providing insight into the diverse functions of Snf2 related proteins, we will study the structure and mechanism of a carefully selected subset of these enzymes. 2) The functions of ATP-dependent chromatin remodelling enzymes. In order to understand how the alterations to chromatin structure directed by these enzymes are used by cells it is important to gain insight into the biological pathways in which they are involved. One of the strategies we will adopt will be the application of genomics to catalogue the genes different enzymes interact with and the nature of the chromatin alterations they direct.