- Total grants
- Total funders
- Total recipients
- Earliest award date
- 10 Apr 2001
- Latest award date
- 30 Sep 2018
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
Human Fcgamma receptors (FcgammaRs) are proteins found on the surface of immune cells. They bind to antibodies, which are produced by the body, in response to infection. Some antibodies produced recognise their own tissues and are found in many diseases, including rheumatoid arthritis and lupus. It has been shown that genetic changes in the FcgammaRs are found more frequently in rheumatoid arthritis sufferers compared to healthy individuals. This project will focus on FcgammaRIIa, which is present on cells which are responsible for the destruction of many antibody-bound objects. Through a combination of cutting edge techniques, spanning physics, biology, immunology and medicine, we will uncover fundamental information within this field. This information would aim to inform the production of effective therapies to treat diseases such as arthritis, which put a huge strain on the NHS every year.
A structural investigation into the action of and resistance to ribosome-targeting antibiotics 30 Sep 2018
Antibiotics are crucial to modern medicine, allowing treatment of life-threatening bacterial infections and making many surgeries like transplantations possible. However, pathogenic bacteria are rapidly evolving to resist their effects. Protein synthesis is one of the main antibiotic targets in bacterial cells. I will use structural biology techniques, principally cryoEM and single particle image processing, to understand how both novel natural products and clinical antibiotics bind to the ribosome to bring about their inhibitory effects on protein synthesis. Furthermore, I will investigate the cause of toxicity of certain ribosome-binding antibiotics by examining how they bind to the mammalian mitochondrial ribosome. Finally, I will use a combination of cryoEM and protein X-ray crystallography to elucidate how certain ribosomal-protecting proteins form complexes with the ribosome in order to bring about antibiotic resistance. On an individual level, these studies will allow an assessment of the viability of novel natural products as suitable clinical antibiotics. More generally, they will contribute to our knowledge of how different classes of antibiotics target the ribosomes of pathogenic bacteria, and how these bacteria evolve resistance. This knowledge will help the development of methods to rationally design new ribosome-targeting antibiotics that are able to overcome or circumvent resistance.
The proposed research uses standard molecular biology, protein purification and biophysical structural analysis methods in a focused series of experiments that comprise a complete 6-week project. This builds on existing molecular genetics studies that have identified novel missense mutations in KMT2D (also known as MLL2) as the cause of a unique phenotype (renal tubular dysgenesis, choanal atresia and athelia). Previous studies have identified KMT2D mutations as a major cause of Kabuki syndrome, a comparatively common autosomal dominant congenital mental retardation syndrome. The missense mutations occur in a central region of the KMT2D protein (2841-3876) that does not have variants associated with Kabuki syndrome. This central region contains a series of coiled-coil domains that are likely to mediate protein-protein interactions. However, the effect of the missense mutations on KMT2D structure and interactions is completely unknown. This project will determine the structure-function relationships between KMT2D and a unique phenotype that are likely to be caused by altered protein-protein interactions, as well as describing the broader genotype-phenotype correlations in this important gene. The approach described in the proposal is the only tractable way to understand possible structure-function relationships, given the large size of the gene and encoded protein.
The diffusion of chemokines in the extracellular matrix is a requirement for the formation of chemokine gradients that guide immune cell migration to sites of inflammation, and controlled by matrix glycans of the glycosaminoglycan family. The focus of this research is to use well-defined models of the extracellular matrix to probe the interaction between the chemokine CXCL12 and the glycosaminoglycan heparan sulphate, and how this defines the mobility of CXCL12. The first key goal of the project is to design and produce a fluorescently-tagged CXCL12 mutant with modulated glycosaminoglycan binding which can be compared against the wild-type chemokine and other mutants already available. The second key goal is to use the biophysical method of fluorescence recovery after photobleaching to characterise the differential diffusion of mutant and wild-type CXCL12 in glycosaminoglycan-rich matrices. This project thus combines biochemistry and biophysics to gain a better understanding of the molecular mechanisms underpinning the formation of chemokine gradients in extracellular matrix.
Historically, ribosomes have been viewed as unchanged homogeneous units with no intrinsic regulatory capacity for mRNA translation. Recent research is shifting this paradigm of ribosome function to one where ribosomes may exert a regulatory function or specificity in translational control. Emerging evidence has identified heterogeneity of ribosome composition in specific cell populations, leading to the concept of specialised ribosomes. Specialised ribosomes may therefore exhibit control and regulation over the translation of specific mRNAs, resulting in a substantial impact on how the genomic template is translated into functional proteins. Due to the emerging concept that cells can control the composition of ribosomes to regulate protein expression, it would seem highly likely that viruses could also manipulate host cell ribosome compositions to enhance the production of viral proteins. We have quantitative proteomic and ribosomal profiling data suggesting Kaposi's sarcoma-associated herpesvirus (KSHV) manipulates ribosomal biogenesis. Firstly, we will investigate changes in composition and stoichiometry of proteins within the ribosome, driven by KSHV. We will isolate ribosomal complexes by tandem affinity purification, during KSHV infection and analyse changes by LC-MS/MS and cryo-EM. We will elucidate how these changes exert ribosome-mediated specificity to promote KSHV lytic infection using a number of cellular and molecular techniques.
We will be investigating the viability of using cyanobacteria as a model for our own by exploring the evolutionary links as well as the similarities between human cells and cyanobacteria cells in terms of the communication and cell differentiation. This will allow us to use the cyanobacteria as a model for human stem cells. There are 3 cases which will be investigated: metabolism of retinoic acid, nitrogen-fixing cells and prostaglandin cell signalling. In each case, we will be blocking the signal, modifying the bacteria and studying how this affects the bacteria. The production of proteins and the chemical signalling are amongst the several responses we will be monitoring. Using information gained from this we will be able to see if there is a viable link that can be used to monitor cyanobacteria that have human orthologues spliced into it.
Cancers develop as a result of many interacting factors. Two such factors are cell stress and microRNA (miRNA) expression. Cell stress causes fluctuations in protein levels, which can perturb the proper functioning of the cell. miRNAs silence specific genes, and therefore can induce changes within the cell which cause them to become cancerous. However, little is known about how miRNA expression is altered. I aim to investigate a novel mechanism of miRNA regulation, which may be perturbed by cell stress. I will determine how the levels and activity of key components in miRNA biogenesis are altered in cells expressing different proteins and which have been subject to different stress conditions, using a range of in vitro, cell-based and biophysical approaches. I will also perform several screens to identify key microRNAs regulated by this mechanism, and how their expression changes with cell stress. This work will reveal new avenues for cancer therapy and help us to target cancer with a fresh perspective.
Obesity causes brain insulin resistance and prevents the brain from regulating metabolic responses, maintaining energy balance and controlling the nutritional status of an individual. Restoring the brain’s ability to modulate metabolic functions could be an important intervention to prevent the negative outcomes of obesity and diabetes. The Dorsal Vagal Complex (DVC) in the brainstem senses insulin to regulate glucose metabolism, food intake and body weight in rodents. Three days of high-fat diet feeding is sufficient to completely disrupt the insulin response in the DVC, thus causing an increase in blood glucose levels and excessive eating. Recently, I discovered that increased mitochondria fission and ER stress in the DVC can cause insulin resistance and affect the ability of the DVC to regulate blood glucose levels. I aim to understand whether increased mitochondria fission in the DVC can affect food intake and body weight in rats. Using in vivo and in vitro experiments, I aim to uncover the mechanism by which changes in mitochondria shape and size affect DVC insulin sensing and eating habits in rodents. This project could lead the way for the development of novel approaches that target the brain to regulate food intake and body weight in obese subjects.
Peripheral gate in somatosensory system 17 Jul 2018
Peripheral nerves are responsible for haptic, somatic and visceral sensations including that of pain. Healthy nerves conduct action potentials from their peripheral endings to the dorsal spinal cord, where synaptic transmission first takes place. It is assumed that the peripheral somatosensory signals are first integrated in the spinal cord and subsequently analysed in the brain. Our recent findings has challenged this view and suggested that peripheral somatosensory ganglia (such as dorsal root ganglia, DRG) are capable to regulate pain transmission utilising GABAergic somatic cross-talk mechanisms. I hypothesize that somatosensory ganglia represent a new type of a ‘gate’ within the somatosensory system. My overarching goal is to develop a comprehensive mechanistic understanding of the peripheral somatosensory gating. I will use in vivo electrophysiology, mouse transgenics, chemo- and optogenetics, behavioural models and other cutting-edge approaches to address the following specific aims. (1) Secure direct in vivo evidence for peripheral somatosensory integration at the DRG. (2) Moving beyond GABA: identify other major ganglionic communication mechanisms. (3) Elucidate physiological context of signal integration in the DRG. (4) Identify subcellular structures involved in somatic integration. These studies will change current understanding of somatosensory processing and will provide new ideas for pain treatment at the periphery.
Non-genetic oncogenesis in adenocarcinoma of the oesophagus and gastro-oesophageal junction: characterising the stress-inducible FGFR2-GRB2-miRNA axis. 30 Sep 2018
Oesophageal adenocarinoma (OAC) is a type of cancer affecting the lower part of the oesophagus (the gullet). The number of patients diagnosed with OAC has increased substantially over the past three decades. As a result, OAC is now the most common type of cancer affecting the oesophagus. Unfortunately, there are currently few effective treatments for OAC. Many patients with OAC will however first be diagnosed with Barrett's oesophagus. This is a condition in which the normal lining of the lower oesophagus becomes glandular and starts to grow abnormally. It is not clear why this happens but it may be related to reflux of stomach contents (such acid and bile) into the oesophagus (as happens in patients who frequently experience heartburn). Our research suggests that bile and acid may be able to change the concentrations of proteins within the glandular cells. The altered levels of these proteins may then contribute to the cells becoming cancerous. This work will look to see if this is the case and ma help us to find new drug targets to prevent and treat Barrett's and OAC.
The majority of small molecule drugs on the market only target a very small range of potential targets. They function by their binding event causing a direct modulation of their target protein’s function, however it is not clear that all proteins involved in disease can be targeted in such a manner. In this project, I aim to develop drug molecules with a different mechanism of action. One half of the molecule will be able to bind to a protein involved in a disease pathway and the second half of the drug would be capable of dragging the molecule to an enclosed cell compartment, known as the peroxisome. Such re-localisation will trap the disease pathway associate protein making it unable to carry out its function providing a new strategy for therapeutic intervention.
Lead identification of a novel anticoagulant agent targeted against activated factor XII for the prevention of thrombosis without the risk of bleeding associated with current antithrombotics 01 Oct 2017
At the University of Leeds, Dr Helen Philippou (PI) and Prof Robert Ariens (Faculty of Medicine and Health), with colleagues (Faculty of Mathematics and Physical Sciences) Dr Richard Foster and Prof Colin Fishwick with Prof Gregory Lip (University of Birmingham) have secured a £3,021,002 Wellcome Trust Seeding Drug Discovery award to develop new safer anticoagulants with minimal risk of bleeding. The medicines available to treat patients who suffer from an increased risk of blood clotting are effective but carry a significant risk of causing bleeding. Current treatments therefore involve a balance between reducing blood clots with inducing bleeding. The objective of this proposal is to discover highly specific compounds which block a protein, Factor XIIa, which is involved in clot formation, and for which there is good evidence to suggest inhibition will not cause bleeding. This will allow patients to be treated more safely with minimal risk of bleeding without needing therapeutic monitoring. The aim is to produce an agent which will be given by mouth daily.
A temporary exhibition on cancer: Exploring understandings of and relationships with cancer 18 Jun 2018
The temporary exhibition on cancer aims to showcase how understandings and experiences of cancer have changed over time including addressing the medical, social and ethical complexities associated with current research developments. This includes an examination of the impact of developments in scientific research such as genomic medicine on perceptions of what cancer is, how it is managed and what it might look like in the future. The researcher embedded in the project team will conduct the research and development that would shape the overarching narrative, content and interpretation strategy and learning outcomes of the exhibition; with the option of focusing on a key area identified during the development process, for example, exploring challenges and opportunities associated with generating and interpreting genomic data. This would include interviews with those affected by cancer as well as experts in the field of cancer research including scientists, data specialists and healthcare practitioners. The researcher will also develop a public participation project which would take the form of a public consultation exercise centred on the contemporary patient experience, with the objective of generating public dialogue and policy recommendations for relevant organisations.
This project examines ‘Chronic Traumatic Encephalopathy’ (CTE), a form of dementia which arises from concussive and sub-concussive blows to the head. The majority of cases of CTE result from head impacts suffered during sporting activity. Given the large number of sports associated with a risk of CTE, there are increasing concerns about a ‘silent epidemic’ of dementias which may affect both amateur and professional athletes. These concerns have led to diverse calls for technological innovation, rule change, and legislation to ward against the disease. Drawing upon elite interviews and ethnography conducted with three sporting communities (American football; professional wrestling; age group rugby) this project will ask: How is CTE rendered intelligible within diverse sporting contexts? How do practitioners understand themselves, their brains, and their conduct in the context of the risk of CTE? Finally, how is knowledge of the brain and dementia entangled with classed, raced, and gendered sporting activities? This ambitious study will be amongst the first in Medical Sociology and Science and Technology Studies to consider CTE and will yield novel empirical and theoretical findings relating to the social shaping of this crucial, emerging diagnosis and its place within contemporary society.
How and why proteins aggregate is an important fundamental question. It also has far-reaching biomedical importance given the increasing prevalence of amyloid-diseases in today’s ageing population. Whilst some amyloid precursors are intrinsically disordered, others are natively folded. Each must undergo major conformational changes to form amyloid fibrils. Defining these changes is key to developing therapeutic strategies. This proposal aims to achieve this by focusing on two overarching questions: What is the nature of the protein-protein interactions that initiate amyloid formation; Can we use this knowledge to develop molecules able to control aggregation in vitro and in vivo? The challenges in answering these questions lie in the heterogeneity of aggregating species, their transient/weak interactions, and the fact that amyloid precursors adopt non-native, dynamic structures. We will meet these challenges: exploiting cutting edge structural methods to map the protein-protein interactions that commit proteins to aggregate and will use the knowledge gained to inform the design of small molecules/artificial proteins able to control aggregation by targeting these surfaces. Focusing on islet-amyloid-polypeptide (involved in type II diabetes) and beta2-microglobulin (involved in systemic amyloidosis) the goal is to enhance our fundamental understanding of protein aggregation and to inspire new strategies for intervention in disease.
We have recently shown that fibrin, but not its precursor fibrinogen, activates platelets through a receptor complex, GPVI-FcRg-chain, which was identified by one of us (SPW) in the 1990s, and is recognised as the primary signalling receptor for collagen. The paradigm-changing observation that GPVI is a receptor for fibrin establishes a role for GPVI not only in initiation (via collagen) but also in propagation (via fibrin) of thrombus growth and may explain the increase in embolisation in thrombosis models in GPVI-deficient mice. We propose that the interaction of fibrin with GPVI represents a target for a new class of antiplatelet agent that may have benefits over current antiplatelet drugs. To investigate this we will map the site of interaction of fibrin with GPVI and develop agents (inhibitors and mouse models) that block this interaction but preserve the activation of GPVI by collagen. We will use these to determine the importance of GPVI-fibrin in haemostasis and thrombosis, and in other vascular pathways where GPVI is known to play a role including maintenance of vascular integrity.
We request funds to purchase equipment to perform correlative light and electron microscopy (CLEM) and electron microscopy of cells & tissues at the University of Leeds. This will include: a high-pressure freezer to allow rapid freezing of large and/or labile specimens whilst minimizing structural deformation; a freeze-substitution unit for low-temperature substitution of water by dehydrating agents and fixatives, avoiding the damaging effects of room-temperature dehydration; a modern cryo-ultramicrotome, for room-temperature and cryo-sectioning of samples for (cryo-)EM imaging; and a cryo-CLEM system, to image cryo-fixed samples by fluorescent microscopy, identifying interesting biological events for further analysis at high-resolution by cryo-EM. This equipment will build upon the outstanding infrastructure for super-resolution light microscopy and cryo-EM in Leeds. The combination of new and existing equipment will give us an unprecedented ability to integrate high-resolution structural information into its cellular/tissue context. This will help us to tackle the most challenging projects in biomedical discovery, and understand the molecular basis of diseases such as infections, cardiovascular disease, neurodegeneration and cancer. We request £429k of Wellcome Trust funds, complementing the University’s contribution of ~£192k in capital and support salary costs, towards a total project cost of ~£621k.
Mass spectrometry infrastructure underpinning research into the molecular basis and biological mechanisms in health and disease 06 Jul 2017
Funding of £715k is requested as a 50% contribution to the purchase of three mass spectrometers in Leeds: 1) Orbitrap Q-Exactive Plus-EMR (collaboration with Thermo Scientific; high-mass modified for membrane protein/lipid studies and top-down protein characterization); 2) Tofwerk high-resolution ion-mobility platform, allowing acquisition of unprecedented conformational detail of dynamic protein structure and ligand interactions; 3) Orbitrap Q-Exactive Plus for providing a major upgrade to the proteomics capabilities. The instruments will include associated nano-LC systems, software, processors and extended maintenance contracts. This equipment will revolutionise current capabilities in biomolecular characterization and quantification using mass spectrometry (MS)-based technologies, impacting on translational medicine through mechanistic studies, biomarker discovery and drug development. The new state-of-the-art platforms will extend analysis capabilities in membrane protein structure, "top-down" analysis of post-translational modification and ligand binding, and conformational and (mis-)folding studies. The replacement of an aged unreliable Orbitrap MS will allow considerably faster proteomic data acquisition with higher resolution and sensitivity and lower sample requirements, and exploratory metabolomics. Forming part of a virtual, Leeds-wide facility for MS with complementary centres of expertise at the Astbury Centre on the main campus and at the St James’s University hospital (SJUH) campus, beneficiaries span the Life, Medical and Physical Sciences.