- Total grants
- Total funders
- Total recipients
- Earliest award date
- 06 Jan 2017
- Latest award date
- 31 Dec 2017
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
Investigating the role of folate metabolism in driving mitochondrial DNA mutations and ageing. 30 Sep 2017
The central research question is whether folate metabolism has a direct impact on the integrity of the mitochondrial genome during ageing. The mitochondrial genome has many copies within the cell and somatic mutations are known to occur in mitochondrial DNA (mtDNA) at a higher rate than the nuclear genome. The term heteroplasmy is used to describe the ratio of mitochondrial DNA (mtDNA) type i.e., ratio of normal to mutant mtDNA. The percentage of heteroplasmy increases during one’s lifetime and may be a contributor to disease onset and ageing. We propose that prolonged suboptimal folate status leads to increased mtDNA heteroplasmy. Folate should be considered a key nutrient, not just for the prevention of birth defects but also for healthy ageing. The overarching aim of my project is to assess the relevance of folate metabolism and ageing on mtDNA heteroplasmy using (i) Human samples of varying ages with measurements of folate status and (ii) Mouse and zebrafish models of compromised folate one carbon metabolism. Objective 1: Assessment of the impact of age and folate status on mtDNA heteroplasmy in human subjects. Objectives 2 & 3: An analysis of mtDNA heteroplasmy and copy number in mouse and zebrafish models.
This research investigates the relationship between 5-HT neurons and two key brain functions: sleep-wake changes and fear learning. Of particular interest is the role in these processes of glutamate co-released from 5-HT neurons. Two approaches will be used. One approach is based on recent evidence that glutamate is preferentially released from 5-HT neurons firing at low frequencies, whereas 5-HT is preferentially released at higher frequencies. The second approach is based on a mouse with glutamate deficient 5-HT neurons.In Oxford the research will have 3 main goals:i) Measurement of firing of identified 5-HT neurons during the sleep-wake cycle. This will be achieved using optotagging to record for the first time, the firing of identified 5-HT neurons during changes in sleep-wake activity.ii) Optogenetic manipulation of 5-HT neurons on sleep-wake activity. These experiments will optogenetically manipulate 5-HT neurons (using different stimulation frequencies) to establish the causal relationship between changes in 5-HT firing and sleep-wake activity.iii) Optogenetic manipulation of 5-HT neurons in mice with glutamate-deficient 5-HT neurons. These experiments will use a mouse with glutamate-deficient 5-HT neurons to further test the role of glutamate co-release.I will follow a similar research strategy to investigate the role of glutamate co-released from 5-HT neurons in emotional learning at NIH.
Analysis of neurodegenerative diseases progression, at a single cell level, with Deep Neural Networks 30 Sep 2017
The aim of the project is to investigate neurodegenerative diseases, by determining novel neuronal cell phenotypes, classify them and to develop a step wise neurodegenerative diseaseprogression map. Stem cells differentiation technology allows scientists to access neurons involved in pathology of various neurodegenerative disease, however, there is big gap in good methods to investigate diseases at the single cell level. Thus, I will imagine organelles of iPSC derived neurons at a super-resolution level with 3D-SIM. Cellular heterogeneity will be investigated with the deep neural networks (DNN), which can detect numerous phenotypes of neurons and classify them into various heath states, this will allow us to produce a step wise disease progression map for PD and FTD in vitro models. Key goals:Goal 1: Develop and train DNN to detect cellular phenotypes (University of Oxford);Goal 2: Develop neurodegenerative disease progression map (University of Oxford)Goal 3: Understand why DNN grouped neurons into the particular classes (NIH);Goal 4: Undertake CRISPRi screens and DNN will be used as a readout (NIH).This project could set a scene for developing novel analysis methods for neurodegenerative diseases, it can produce a step wise disease progression map and reveal novel interventiontargets.
UK Biobank 30 Sep 2017
As summarised in the extracted text below from our full application (page 5 "Proposed work") in Section 10 of this application form (and described in more detail on pages 5-16 of Section 10). During the next 5 years, it is intended to: Extend cohort-wide linkage to primary care health records, while maintaining and updating health outcome data from existing linkages to death, cancer and hospital records (and explore the added value of linkages to additional healthcare datasets); Further develop and implement large-scale identification and characterization of many different types of health outcome; Streamline the research access systems, and improve methods for processing, presenting and providing linked healthcare data to researchers; Maintain the sample resource, and make increasing amounts of different types of genotype and biomarker data available; Conduct imaging assessments in 100,000 participants, and develop and implement further enhancements (such as cardiac monitoring and further sample collection and assays). Efforts will continue to be made to ensure that researchers from around the world are suitably well informed about the resource so that the effective use of it increases substantially, leading to novel findings that have a major impact on human health.
Currently there is no cure for MS and therapies focus on symptom management. Current treatments involve Sativex, a combination of plant-derived cannabinoids, tetrahydrocannabinol (THC) and cannabidiol (CBD), to alleviate MS-associated spasticity/pain. However, some patients do not respond to therapy and side effects have been identified. Hence, new data is needed to refine cannabinoid therapies, and by determining the precise cellular and molecular mechanisms of action of cannabinoids in blood cells isolated from patients, this proposal will assist the development of new targeted cannabinoid-based treatments that lack off-target effects. The proposal is designed to determine the cellular action of cannabinoids in patient cells by targeting innate immune receptors, including viral toll-like receptor (TLR)3 and bacterial TLR4 signalling mechanisms. The hypothesis is that targeting innate immune receptor signalling by plant-derived cannabinoids is a therapeutic avenue that may be exploited in the treatment of MS. Key goals: This work will contribute to the understanding of the inflammatory signalling mechanisms driven by TLR3 and TLR4 activation in human blood cells. This study will also determine if innate immune signalling responses are dysregulated in MS patient cells. Determine if plant-derived cannabinoids target inflammatory signalling mechanisms with relevance to MS at a cellular level.
Understanding the multidirectional axes of communication between the gut microbiome and the brain in acute stress 27 Apr 2017
Different rodent strains exhibit different susceptibility to stress and release distinct levels of stress hormones into the gut during psychological stress1, this research will interrogate whether animal strain-dependent stress-response is reliant on gut microbiota-host interaction. We aim to identify a mechanism by which host genetic background interacts with the gut microbiota to generate a distinct animal stress-response phenotype. Host-derived neuroactive molecules are important signalling molecules in host-microbiota interaction along the microbiota-gut-brain axis2. Noradrenaline, a key catecholamine released into the intestinal lumen during stress-response, stimulates bacterial growth whereas microbe-derived products stimulate host enteric afferent nerves to influence host behaviour under stressful conditions3,4.Although the exaggerated stress-response of germ-free rodents (i.e. sterile animals without a microbiome) is normalized following microbial colonization6, it is unknown how host genetics influence host-microbe interaction to confer stress resilience. We will test the hypothesis that host genetic background is critical in determining whether the gut microbiota effect changes in host stress-response along the microbiota-gut-brain axis. We have established the following Key Objective to interrogate our hypothesis within an 8-week time-period: Define host-microbe interaction during psychological stress by characterizing microbial- and host-derived molecules and signalling along the microbiota-gut-brain axis in rodents.
Design, synthesis and evaluation of novel analogues of [Ser14]mitoparan, a new class of broad-spectrum antibiotic 27 Apr 2017
Background: The recent emergence of resistant strains of Gram-negative bacteria, including Acinetobacetr baumannii, is the major challenge for the development of new antibiotics. Derivatives of the cationic peptide mastoparan (INLKALAALAKKIL), a mast cell (MC) degranulating peptide derived from wasp venom, display broad spectrum antibiotic activities but are clinically challenged by eukaryotic cytotoxicity and the potential to promote MC secretion. Comprehensive studies of MP analogues recently identified [Ser14]Mitoparan (INLKKLAKL(Aib)KKIS; Aib = aminoisobutyric acid) a novel broad spectrum antibiotic, active against A. baumannii and other human pathogens but lacking both eukaryotic cytotoxicity and MC degranulating activity. Objectives: Structure-activity relationships confirm that [Ser14]MitP is a versatile template for the generation of antibiotic peptides. The major objective is to design and evaluate protease-resistant structural analogues of [Ser14]MitP with enhanced toxicity against human pathogens. A key goal is to identify [Ser14]MitP analogues suitable for early-stage clinical trials. Methodologies: Rationally-designed analogues of [Ser14]MitP will be prepared using microwave-enhanced solid phase peptide synthesis. Structural modifications to improve biological properties will include the substitution of unnatural amino acids and changes to N- and C-termini. Cytotoxicity assays will utilise a range of pathogenic bacteria and fungi both in RIHS and in collaboration with the University of Melbourne.
In this project I will measure the expression of various proteins involved in the DNA damage response (DDR) in extracts of a novel cell line that spontaneously immortalised from a primary culture of ascites cells from a patient with clear cell ovarian cancer. I will also determine the sensitivity of this cell line to DNA damaging anticancer drugs and ionising radiation, potentially also in combination with inhibitors of the DDR, e.g., ATR, PARP, using both growth inhibition and cytotoxicity assays. If time allows, I will also measure cell cycle perturbations after DNA damage. At the end of the project I will compare the cytotoxicity data with the protein expression data (and potentially the cell cycle analysis) to determine if expression levels of DDR proteins can be related to sensitivity.
Mapping the developmental time course of social behaviours in a rat model of Fragile X syndrome. 27 Apr 2017
Fragile X syndrome (FXS) is the most common form of inherited intellectual disability. It is caused by a single gene (FMR1) mutation resulting in the absence of the FMR1 protein. Children with FXS display learning impairments, attention deficits, anxiety, and difficulty with social interactions. The pathophysiology of FXS has been investigated using rodents lacking the FMR1 gene. Recent research has demonstrated that FRM1 KO rats have selective cognitive deficits that normally develop in the late postnatal period. Treatment with potential therapeutics between 5 and 9 weeks old prevents these deficits emerging and the beneficial effects last for at least 14 weeks following the end of treatment. This project will investigate whether FMR1 KO rats show abnormal or delayed development of social behaviours compared to wild type control rats. The objectives are: 1) to characterise the normal development of social behaviours from shortly after weaning (when the young pups become independent of their mother) until adulthood, and 2) to determine the developmental time course of any differences in social behaviour in FMR1 knockout rats. Defining the age at which social deficits occur will provide the basis for future preclinical studies testing whether potential therapeutics can prevent or reverse social impairments.
Application for Summer 2017 Vacation Scholarship 27 Apr 2017
Anaemia is one of the major health complications in patients suffering from end-stage chronic kidney disease and its prevalence is further accentuated by administration of dialysis as treatment. Anaemia contributes significantly to both the morbidity and debilitating condition of these patients. Iron supplementation to treat anaemia is beset by the consequences of toxicity of excess free iron. Histidine had previously been identified as an inert adjunct that could be employed with iron in the treatment of anaemia of kidney disease (KD). However, none of these studies adequately investigated the molecular mechanisms by which L-histidine could be exerting iron detoxifying properties in the management of anaemia of KD. The project entitled "L-histidine ameliorates iron-induced oxidative stress damage in the kidney" aims to investigate the mechanism by which L-histidine and its derivatives could be used as co-supplements with iron to ameliorate iron toxicity in the treatment of anaemia of kidney disease. The overall hypothesis of the proposed study is that histidine, as an iron chelating buffer, exerts cytoprotective amelioration on iron-induced oxidative stress in vivo in mouse kidney and in vitro in HK-2 cells.
This research project is designed to interrogate an alternative therapeutic for age-related macular degeneration. FKBPL is an anti-angiogenic protein that has a mechanism of action independent of the VEGF pathway. As such this project will deliver DNA than encodes the FKBPL protein in vitro and the molecular markers of angiogenesis will be measured. The novel peptide delivery sequence, termed RALA (WO2014087023) will be used to deliver the FKBPL DNA to the nucleus of the cells where the therapeutic protein can be made. Key Goals include: Characterisation of the RALA/FKBPL nanoparticles In vitro culture of human retinal pigment epithelial cells (ARPE-19), ACBR 183 primary human retinal pericyte cells, ACBR181 primary human retinal microvascular endothelial cells and choroidal endothelial cells (RF6A) Learning a range of molecular techniques including FACS analysis, rtPCR, IHC, western blot and migration/tubule formation assays Development of transferable skills such as health and safety knowledge in a laboratory, presentation skills, data analysis, experimental design, critical thinking and scientific report writing skills. It is anticpated that at the end of this placement that the student will have acquired an in depth knowledge of what a career in research entails and have learnt several techniques that can be employed post degree.
The Effect of Long Acting Muscarinic Antagonists on Corticosteroid Action in Airway Smooth Muscle Cells in Asthma 27 Apr 2017
Severe asthmatics are less responsive to the positive therapeutic effects of corticosteroid therapy. Identifying therapies to overcome this steroid resistance is a key goal of research. Current research has shown that molecular interactions between glucocorticoid receptors (GR) and G protein-coupled beta receptors exist. However, less is known regarding the interaction between GR and muscarinic receptors (M2, M3) and on the effect of long-acting muscarinic antagonists (LAMAs) on corticosteroid insensitivity in severe asthmatics. This proposal aims to ascertain whether corticosteroids used in conjunction with muscarinic antagonists will enhance the suppressive effect of steroids on inflammatory mediator release from airway smooth muscle cells (ASMC) in severe asthmatics. By measuring inflammatory mediators using ELISA, and investigating gene expression via real-time qPCR of isolated RNA from ASMC of severe asthmatics; the effects of i) muscarinic antagonists alone, ii) corticosteroids alone and iii) muscarinic antagonists given in conjunction with corticosteroids, can be compared. The results will then be statistically analysed to compare and identify any significant differences in inflammatory mediator release when using muscarinic antagonists and corticosteroids together, and when using them separately. The objective of this project is to advance current knowledge of the therapeutic benefits of these treatments for asthma.
IgA nephropathy (IgAN) is, globally, the most common primary glomerulonephritis, and is characterised by the deposition of IgA in the renal mesangium resulting in mesangial cell proliferation and extracellular matrix synthesis. The condition has a variable prognosis; the severity of which does not correlate with the extent of IgA deposition. An IgAN genome wide association study identified a risk haplotype in the coding region of the non-redundant innate immune system protein, CARD 9. CARD 9 is known to play a pivotal role in the pathways linking extra cellular and intracellular stimulus to the production of pro-inflammatory cytokines. A number of groups have shown human mesangial cells (hmc) produce pro-inflammatory cytokines, when incubated with IgA in vitro. The Leicester group has also shown increased expression of CARD 9 in these cells and renal biopsies from patients with the more severe from of IgAN. This project will: 1. Investigate changes in CARD9 expression by hmc afer incubation with serum IgA1 IgAN patients and healthy controls. 2. Investigate changes in cytokine synthesis by hmcs incubated with IgA1 with and without CARD 9 siRNAs; 3 .Determine if spleen tyrosine kinase has a role to play in CARD 9 mediated IgA1 stimulation of hmcs.
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.
Understanding the role of autophagy in the pathogenesis of AMD using a patient specific iPSC model 27 Apr 2017
AMD remains the most important cause of blindness in the elderly. The number of AMD affected people in UK is expected to rise to 1.3 M by 2050 with healthcare costs rising to £16.4 billion during 2010-2020. Currently there is no effective treatment hence a huge unmet need for investigations into therapies. This project aims to contribute to the identification of the underlying causes of AMD by interrogating a new cell model created by the host lab. Preliminary data suggest that autophagy, a process by which the cells remove waste products, is impaired in retinal pigmented epithelial cells derived from patients with AMD, leading to deposition of waste products and increased stress. Using this model, I will investigate which step of the autophagic process is not functioning properly in retinal cells, thus leading to a better understanding of AMD etiology. To date, small molecules that regulate autophagy are used in clinical trials of other neurodegenerative diseases. By the end of this project we will find out if the existing small molecules restore the autophagy in retinal cells of AMD patients. This will contribute to the design of preclinical studies for preventing further progression of AMD and restoring sight loss.
Investigating the role of cyclin-dependent kinases in the regulation of DNA repair by non-homologous end-joining 27 Apr 2017
DNA double-strand breaks (DSBs) are highly toxic lesions that can drive genome instability and cancer. In human cells, most DSBs are repaired by the non-homologous end joining (NHEJ) pathway, except when they occur during DNA replication or mitosis when NHEJ is toxic and can generate chromosomal translocations. NHEJ must therefore be tightly regulated to maintain genome stability. However, little is understood about how the core NHEJ repair machinery is regulated. PAXX is the last core NHEJ factor to be identified, and may be a substrate for cyclin-dependent kinases (CDKs). A highly conserved residue in PAXX, S148, matches the consensus for phosphorylation by CDK1/2, and preliminary data indicate that it is indeed phosphorylated by these kinases. The aim of my project is to generate human cell lines expressing PAXX-S148 mutants that either prevent or mimic phosphorylation (Ser>Ala or Ser>Glu) on this site and investigate how expression of these PAXX variants impacts on DSB repair. Key goal 1: produce human cell lines expressing wild-type or mutant fluorescent PAXX variants and examine their DSB repair capacity and sensitivity to ionising radiation Key goal 2: investigate when CDK-dependent phosphorylation of PAXX occurs during the cell cycle
Pigment epithelium-derived factor (PEDF) in the therapeutic response to Electroconvulsive Therapy 27 Apr 2017
Depression is one of the leading contributors to the global burden of disease. One third of depressed people have treatment-resistance and don’t respond to standard antidepressant medications and/or psychological therapies. Such patients may benefit from electroconvulsive therapy (ECT), the most effective treatment for severe depression. Changes in brain plasticity are implicated in depression and the mechanism of action of antidepressant treatments. We recently discovered that a neurotrophic factor called PEDF (pigment epithelium derived factor) is increased in the blood of severely depressed patients following ECT. Interestingly, we also found similar changes in rat brain following chronic treatment with electroconvulsive stimulation. To extend these novel findings, we now aim to explore how blood levels of PEDF might be associated with remaining well following successful ECT and if this might be useful for predicting depression relapse. We already have clinical details and blood samples from over 100 patients with severe depression who participated in a now-completed randomised ECT trial (The EFFECT-Dep Trial). At present, we have no clinically useful "blood test" to help us in managing depression. So, any practical development in such a test would be an important clinical advance for patients with depression.
Parkinson’s Disease (PD) is the most common motor-related neurogenerative disease, with numbers expected to increase annually due to an ageing population. PD patients suffer with motor dysfunction due to a reduction of brain dopamine levels. Dopaminergic neurone degeneration occurs in tandem with alpha-synuclein protein aggregate deposition. These alpha-synuclein protein aggregates are thought to contribute to neuroinflammation, and a neurodegenerative phenotype in PD. The aim of this project is to compare the prefrontal cortex brain tissue of PD patients with that of age and sex matched control subjects to investigate inflammation levels. This will be achieved by quantifying the levels of proinflammatory factors IL-1, TNFalpha, IL-6, IL-8 and NLRP3 inflammasome components present in control vs. PD tissue. Quantification of the inflammatory factors will be achieved using ELISAs and Western Blotting techniques. The discovery of evidence demonstrating increased inflammation in PD brain tissue would further reinforce the justification for current, and future, use of anti-inflammatory agents to reduce damaging neuroinflammation.
The project aims to utilise mutations identified in congenital myasthenic syndromes to study the interactions of the muscle acetylcholine receptor (AChR) and its anchoring protein, Rapsyn.Widely quoted publications suggest that Rapsyn interacts with the M3-M4 intracellular loop of the AChR alpha, beta and epsilon subunits, however we have identified several kinships in which mutations in the M3-M4 intracellular loop of the AChR delta subunit underlie a phenotype that mimics myasthenic syndromes caused by mutations in RAPSN. The project will use in vitro mutagenesis and cell culture experiments to investigate how mutations in CHRND impair agrin induced-AChR clustering in C2C12 myotubes. Variants identified in CMS patients, ie. p.(Glu381Lys) or p.(Arg376His) will be incorporated into expression constructs and transfected in chrnd-/- C2C12 cell that have been created using CRIPR/Cas9 techniques. Similar experiments will be performed following in vitro mutatagenesis that is designed to disrupt potential PKA, PKC and tyrosine kinase phosphorylation sites within the M3-M4 intracellular loop of the delta subunit. The effects of the mutations on agrin-induced AChR cluster formation will be assessed by using fluorecent-labelled alpha bungarotoxin and microscopy. This short project will provide novel data on how mutations within the AChR itself can impair the cluster formation.
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.