- 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
Exploring endosomal signalling of G protein coupled receptors (GPCRs) using luciferase complementation and luminosensing optogenetics 31 May 2018
Recently G protein coupled receptors (GPCRs) have been shown to signal from intracellular compartments such as endosomes, as well as the cell surface. There is increasing evidence that some synthetic drugs can activate such intracellular located GPCRs directly, but the functional consequences of this activation have been difficult to isolate. This project will study endosomal signalling from engineered optogenetic receptors (OptoXRs), activated by light and coupled to the Gq pathway linked to calcium mobilisation. I will deliver the light stimulus to OptoXR using bioluminescence from a luciferase enzyme, in a local manner to receptors either at the cell surface or within endosomes. This will be achieved using a complementation system, in which luciferase fragments are tagged to the OptoXR and cell surface or early endosomal markers respectively. Active luciferase will therefore only be reconstituted to deliver the light when the OptoXR is present in the same location as the expressed marker. I will then be able to compare the effect of adding the substrate on OptoXR calcium responses in which case, to determine whether calcium signals are produced from receptors in different locations. This will be backed up by confocal and luminescence microscopy to identify reeptor and luciferase locations.
DNA-clustering in yeast 31 May 2018
We propose to describe mathematically the processes by which telomeres form clusters in the nuclei of yeast cells. It is thought that this is due to the presence of proteins which bind to the telomeres. We wish to investigate the processes which determine cluster size through: 1. forming a mathematical model of the attachment processes through which proteins bind to telomeres. This may involve a site exclusion process, whereby the protein binding rate slows due to the declining number of vacant protein binding sites as the telomere `fills up'; 2. constructing a mathematical model of the aggregation of telomere-protein complexes into clusters, and determine which rate parameters are key to controlling cluster size; 3. investigating the rate of equilibration of these processes. The models we construct will have the form of systems of coupled ordinary differential equations. Through a judicious choice of rate constants, explicit equilibrium solutions will be obtainable, allowing available data to be fitted. Numerical simulations of the process will also be performed, to verify that the equilibrium solutions are accessible within biologically realistic time periods.
Imaging Flow Cytometry 05 Jul 2018
We seek funding for an ImageStream X Mark II Imaging Flow Cytometer to be hosted by the University of Nottingham (UoN) Flow Cytometry Facility. This exciting technology combines the speed, statistical robustness and multi-parametric data of conventional flow cytometry with the detailed localisation information of microscopy. The machine has been specified following a trial demonstration and in consultation with our extensive user group and co-applicants. It includes five lasers, high magnification, extended depth of field and automated sampler options that would make it one of the most capable machines in its class in the UK. An imaging cytometer would significantly enhance research capability by offering unique high number and high content experiments. Novel applications include the ability to detect, enumerate and characterise rare cells; the ability to dissect cell signalling events through precise measurements of nuclear translocation events or co-localisation; the study of extracellular vesicles in both eukaryotic and prokaryotic systems; precise quantification of drug delivery/internalisation; and the study of cell death mechanisms. We have established mechanisms for access, charging and an extensive client list that have indicated keen interest in this technology, and have secured a 30% financial contribution from the University demonstrating the strategic importance of this equipment.
Myotonic dystrophy type 1 (DM1) is the most common form of muscular dystrophy in adults. It is a highly debilitating condition affecting more than 100,000 patients in developed countries with an average life expectancy of 58 years. DM1 is primarily a neuromuscular disorder, which also affects a range of other systems including the heart, brain, endocrine and digestive systems. Patients may also show specific patterns of psychological dysfunction and personality traits, cognitive impairment/mental retardation and excessive daytime sleepiness. All features show an obvious deterioration with time and difficulty swallowing and sucking food into the lungs in the later stages of the disease contribute towards chest infections and represent a major cause of morbidity and mortality. There is no treatment for DM1. DM1 is caused by a repeat expansion mutation in the 3' untranslated region of the DMPK gene. Unaffected people have 5 to 30 copies of this sequence whereas patients may have hundreds or sometimes thousands of copies. When expressed the DMPK expansion transcripts remain in the nucleus where they form distinct spots or foci. Professors Chris Hayes and David Brook at the University of Nottingham developed an assay to screen for compounds that might provide a treatment for DM1. They identified small molecules that target a novel protein and destroy the spots in DM1 cells, thereby leading to a significant reduction in the faulty RNA and other molecular features of the disorder. Their drug discovery approach, in collaboration with Argenta, a Charles River company, is based on targeting this novel protein, by refining the chemical starting points to make them more selective and more suitable for oral administration to patients. The multisystem nature of DM1 provides particular challenges but Professors Hayes and Brook anticipate that a successful drug would target most/all features of the disease
Vacation Scholarship Institutional Award
Modelling Antimicrobial Resistance 27 Apr 2017
The Vacation-Scholarship project will develop and explore mechanistic mathematical models relevant to antimicrobial resistance within a bacterial population. Specifically, the regulation of efflux-pump expression, antibiotic uptake and the competition between resistant and susceptible strains within a population will each be modelled and the models integrated to investigate computationally the potential efficacy of combined treatments involving efflux inhibitors as well as antibiotics. The work will focus on multiscale ordinary-differential-equation formulations, but will also involve limited partial-differential-equation investigations of spatial effects. The goals of the work are thus to gain insight into the effectiveness of efflux-inhibitors/antibiotics combinations in mitigating resistance, to provide further evidence for the applicability of modern mathematical methods in such contexts and to introduce the student to systems-biology approaches as applied in the study of a crucial area of healthcare.
Stochastic modelling for detecting interactions and evidence of selection between phase variable genes of Campylobacter jejuni 27 Apr 2017
Recently, the laboratories of Dr. Mike Jones (School of Veterinary Medicine and Science, Nottingham) and Dr. Chris Bayliss (Genetics Department, Leicester) generated a large amount of in vivo data collected from two populations of birds at three time points during colonisation of chickens by Campylobacter jejuni. One of the populations was immunised prior to administering Campylobacter jejuni to the birds while the other population was not immunised and serves as a control group. The immunisation was with a whole cell lysate of the bacterium, which elicited C. jejuni-specific serum antibodies but did not prevent colonisation. The observation points include inoculum (which was the same for both groups), one time point during the life of a bird (anal swabs) and a final time point when swabs and post-mortem caecal samples were collected. This project aims to determine how immunisation affects specific patterns of switching in the phase-variable genes of Campylobacter jejuni occurring during host adaptation of this foodborne pathogen. To analyse the data and make qualitative and quantitative conclusions, a variety of statistical techniques will be used in combination with mutation only and mutation-selection models.
The use of equine hepatocytes for the prediction of the pharmacokinetics and potential drug-drug interactions of Meloxicam and the nutraceutical Quercetin 27 Apr 2017
There is limited information available about equine drug metabolism to ensure optimal dosing, decrease risks of drug-drug interactions and to predict the effects of treatments. New findings are particularly important in the horse racing industry and for equine welfare. As there are ethical issues associated with in vivo experimentation, using equine hepatocytes to develop a highly predictable horse in vitro model to estimate the in vivo clearance and potential drug-drug interactions may be a way forward. This would reduce costs and reduce and refine animal testing. More research is needed to understand its benefits and limitations. This project is designed to use fresh and cryopreserved equine hepatocytes to predict the in vivo pharmacokinetics of Meloxicam, commonly used for the treatment of inflammation. This drug will be used as its pharmacokinetic data in the horse is known. Furthermore, potential drug-drug interactions of Meloxicam with Quercetin, a flavanol, used in equine nutraceutical products as an anti-inflammatory will be assessed. The main experimental aims are to compare metabolism (enzyme kinetics) of Meloxicam in fresh and cryopreserved equine hepatocytes and scale results physiologically to estimate hepatic clearance. These results will then be compared with measured pharmacokinetic blood clearance obtained from in vivo pharmacokinetics.
Attention Deficit/Hyperactivity (ADHD) disorder is typified by impairments in control of attention and/or inhibitory motor control, both of which are related to gaze control: visual attention requires control of gaze direction; and gaze control is itself a motor control skill. Our goal is to develop a suite of computer-based games using eye-tracker technology, in which the trainee’s own eyes become the game controller: to advance in the games, the trainee must acquire increasing control of gaze direction. The training games will be designed to improve fixation in the face of irrelevant distractors, and while monitoring the periphery for relevant information. The games will also train the timing of saccadic responses, in line with evidence that motor timing may be a related impairment in ADHD. The money requested will fund the purchase of two high quality eyetracker systems, enabling games to be prototyped using MatLab in Nottingham, in consultation with young people and adults with ADHD, while in parallel game prototypes are developed and programmed using a standard gaming platform with visual and auditory effects. The deliverable will be a trial-ready suite of training games, which will be evaluated by students and teachersin local schools.
A new way to be deaf? Causes of congenital deafness in the world’s least diverse population 14 Dec 2015
Multidisciplinary teamwork between the PI (a population geneticist revisiting human genetics) and an anthropologist (the named PDRA) has produced exciting preliminary data from South Sinai, Egypt. Our research shows that South Sinai Bedouin tribes, who favour consanguineous marriage, are the world's least genetically diverse populations, and that the Mzeina tribe experiences extraordinary levels of profound non-syndromic deafness (~8%), not attributable to the common causal mutations found in Europe. We aim to build upon these findings with two aims: 1: to establish the underlying genetic basis of deafness in the Mzeina; 2: to determine any socio-environmental factors that may predispose them to deafness. The PI will analyze samples from deaf and hearing individuals to establish the likely genetic basis of the condition, potentially describing a new way of being deaf. The PDRA will interview Mzeina families, aided by Bedu and signers, to establish inheritance patterns for deafness and demographic, social and environmental factors. She will take the tissue samples. Through understanding both genetic and socio-environmental components, we expect to identify both a new form of deafness which may also occur elsewhere, and possible preventative measures. The study has potential to achieve widespread impacts, both social and therapeutic, and generate further research.
Structure-aided discovery of kinase inhibitors as targeted therapeutic agents for breast cancer 30 Jul 2016
Kinases are important targets for blocking cancer progression. However, many remain to be exploited. For example, no drugs are yet available to specifically inhibit any kinase which is switched on by a regulatory protein called calmodulin. Nonetheless, faulty expression of these “CaMK” enzymes is now thought to play a key role in breast cancer progression. The Wellcome Trust has funded the CAMSEED consortium to discover small molecule inhibitors for a CaMK protein involved in basal-like breast cancer. The three dimensional structure of this target has been solved by the Structural Genomics Consortium and Professor Stefan Knapp at the University of Oxford. Interactions with small molecules are being screened by Professor Michael Overduin’s lab at the University of Birmingham using superconducting magnets and high throughput robots at the national HWB-NMR facility. The design of improved inhibitors that can enter cells and selectively block the oncogenic state is being led by Professor Peter Fischer at the University of Nottingham, with Colin Kenyon at CSIR, Pretoria, designing deuterated analogs for enhanced activity. The result of the two year project is expected to be a set of lead molecules for development as potential therapeutic agents for breast cancer, and may yield a new approach for using nature’s own inhibitory mechanisms to block cancer-causing kinases.
This application seeks to fund service user participation at a major conference on mental disability law, to be held in Nottingham on 30 June and 1 July 2016.
Moving functional brain imaging into the real world: A wearable, cryogen-free, magnetoencephalography (MEG) system 05 Jul 2016
Functional neuroimaging systems currently comprise cumbersome equipment built around a small bore into which a participant’s head is gently clamped. This artificial environment restricts both the subject groups that can be scanned, and the experimental questions that can be addressed. Here, we aim to develop a new type of magnetoencephalography (MEG) system which will be worn on the subject’s head, allowing them to move freely whilst being scanned. This is possible due to the introduction of new quantum sensors which do not rely on superconducting technology. The new system will offer higher spatial resolution and 5-10 times the sensitivity of current state-of-the-art MEG instrumentation because the sensors can be placed directly on the scalp. This will allow non-invasive electrophysiological measurements to be made with unparalleled spatiotemporal accuracy while participants operate in an unconstrained environment, e.g. navigating in space, or interacting with other people. Importantly, the device opens up the possibility of scanning almost any subject or patient group ranging from children requiring pre-surgical epilepsy evaluation, to patients with movement disorders. The low cost, wear-ability and enhanced sensitivity of the new system will be transformative, offering new opportunities to study ‘real world’ human brain function in health and disease.
In vitro cross-species membrane insertion of Neisserial Nalp by Campylobacter jejuni-BamA 01 Apr 2016
Outer membrane proteins (OMPs) play a fundamental role in physiological functions, multidrug resistance and virulence in Gram-negative bacteria making them essential for cell viability in a number of pathogens. Biogenesis of such beta-barrel proteins is facilitated by BamA, the main hub of the Barrel Assembly Machinery (BAM) complex. While structural characterisation of the MAS complex is yielding first results on the E. coli system (Backelar et al 2016), mechanistic details remain notoriously difficult to obtain. We propose to investigate the role of BamA from Campylobacter jejuni, an important pathogen and food spoilage organism, as a facilitator of membrane insertion of an autotransporter translocator domain from Neisseria meningitidis, NalP. NalP is a beta-barrel, which can insert at low levels spontaneously into lipid membranes in a membrane composition-dependent way. We propose to use our homology model of C. jejuni, annealed by molecular dynamics simulations (MD), to perform in silico CHARMM docking of NalP. We will investigate experimentally the degree of NalP insertion into model lipid membranes without (liposomes) or with BamA (proteoliposomes). We will use gradient separation and SDS-PAGE to quantify insertion, as well as solid state and DNP NMR to confirm insertion and to investigate specific interactions.
Congenital heart disease (CHD) affects around 1 in 133 newborns. The majority of cases are sporadic but there is increasing evidence of a significant genetic contribution. To identify genes involved in CHD the Brook lab has conducted exome sequencing in collaboration with Matt Hurles at the Wellcome Trust Sanger Institute. This has led to the identification of several novel genes involved in syndromic CHD, including the protein kinase D1 (PRKD1). Genetic network analysis suggests PRKD1 is linked to other genes implicated in CHD. In this study PRKD1 will be cloned and CHD-causing mutant versions of the gene generated using site-directed mutagensis. HDAC4 and HDAC5 are known to be targets for PRKD1 phosphorylation. Kinase assays will be established to compare the mutant proteins' capacity to interact with and phosphorylate HDAC4 and HDAC5, compared to the wild-type version of PRKD1. This will provide valuable insights to the role of PRKD1 in heart development.
Our vision is to develop and evaluate an innovative model of public engagement inhealth-related research called Café Connect. Uniquely the café will engage the public in ALL stages of the research life cycle, not just dissemination. The public will be invited to contemplate, discuss and react to issues in health-related research in Nottingham Contemporary café (a relaxed, non-intimidating environment) in the heart of the community. We will run one large flagship café event aimed at the general public. Three smaller events will follow-on (quarterly) with activities focusing on specific topics - self-harm, older adult drinking and emotions in food choice behaviour. During the flagship event, focus groups will identify key issues relevant to these topics shaping project design. Focus group participants will become ‘participant-engagers’ and gather data in the next two café events supported by a project manager and ‘engagement interns’. The final event will discuss project outcomes and disseminate findings. A spoken-word event will reflect on findings from the self-harm project. Engagement experiences will be captured in responses elicited via a video booth, surveys and via observation, all synthesised by an external evaluator. The applicants are experienced in public engagement and successfully piloted the Café Connect idea in 2015.