- 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
Uncovering the Molecular Mechanisms of Asymmetric Cell Divisions in Mammalian Adult Epithelia 04 Dec 2017
Loss of asymmetric cell divisions (ACDs) regulation in the normal self-renewing stem cells is entwined with the growth and progression of poorly differentiated cancers. Yet, the molecular mechanisms controlling the execution of symmetric versus asymmetric divisions in adult epithelia are still unfolding. We recently demonstrated that KIF5/kinesin-1 is essential for mammary epithelial cell divisions and cytoarchitecture by governing the trafficking of the spindle orientation and apical polarity components, respectively. Whether KIF5/kinesin-1 couples spindle orientation and polarity machineries during mitosis to promote ACDs; and whether other mechanisms are involved remain open questions. Here, we will address these questions in mammary 3D organoids. We will use CRISPR/Cas9 gene editing to generate cells expressing AID-tagged endogenous KIF5/kinesin-1 to allow inducible and rapid degradation of the microtubule motor specifically during mitosis, for a precise evaluation of the role it plays in ACDs. We generated cells expressing GFP-tagged LGN –a key player in the spindle orientation machinery–to purify and analyse the LGN-containing complex by LC-MS/MS mass spectrometry and identify novel factors that participate to ACDs. These studies will elucidate the molecular mechanisms of ACDs in the mammary epithelia and provide rational for subsequent detailed investigations in vivo of their precise roles in development and homeostasis.
Role of Inhibitor of Kappa B Kinase Epsilon in Sex-dependent Differences in Metaflammation 19 Apr 2017
With the failure in curbing the global obesity epidemic, there is an urgent need to develop therapies to treat obesity-associated metabolic diseases. This requires a better understanding of the molecular mechanisms that not only promote, but also prevent metabolic disease development. Although, chronic low-grade inflammation ("metaflammation") is an important causative factor in metabolic diseases, the anti-inflammatory effects of estrogens may provide important insights into disease prevention. Indeed, pre-menopausal women are protected from obesity-linked metabolic diseases. Previously, I have identified immunometabolic proteins, e.g. IKBKE, that limit metabolic stress-induced inflammation and these are influenced by sex. However, it remains unclear whether, and how, estrogens alter innate immune sensitivity to metabolic stress. This project will address this and test the hypothesis that both estrogen- and IKBKE-dependent pathways function in a common signalling network to regulate energy balance and metaflammation. I will investigate, invitro and invivo, the synergies that exist between estrogen- and IKBKE-depended actions in metaflammation. Ultimately, the knowledge gained may support the notion that estrogens and IKBKE are key in raising the threshold for metabolic stress-induced inflammation. This may identify novel therapeutic targets to treat obesity-linked metabolic diseases and shed light on the immunometabolic changes that accompany menopause in all ageing women.
This collaborative research programme will combine empirical bioethical research, conceptual, and theoretical analysis to examine the issue of ethical preparedness in the context of genomic medicine, and to inform and develop relevant policies for practice. We will conduct our research as genomic approaches to diagnosis and treatment, such as the 100,000 genome project, become embedded within health care. We will focus on the extent to which professionals are prepared for navigating the ethical issues within the new working environment of clinical genomics; one where research and clinical practice are more co-dependent than in the past, and where responsibilities of care, both to one person over time as well as to their current and future relatives are changing. The programme will utilise a range of research methods across a variety of settings to map the experience of practitioners, patients and participants in genomic medicine, the ethical issues they confront, and the impact on practice when these are challenging of their established practice, be that at the stage of recruitment/ introduction, diagnosis (or lack of), treatment, surveillance, longer term contact or need for contact of other .
This five-year collaborative programme will develop approaches for understanding laboratory animal research as a nexus, asking how reconceptualising connections and generating communication across different perspectives can contribute to improving the future of animal research. New research will draw attention to historical independencies between science, health and welfare; identify challenges emerging at the interfaces of animal research and create opportunities for informing policy and public engagement. We suggest collaborative approaches are essential for understanding how rapid transformations across science and society are changing the patterns of responsibility, trust and care which hold together, or constitute, this nexus. We will deliver new: integrated research across the social sciences and humanities, using historical research to inform understanding of present challenges and create new engagement opportunities for the future; interactive research projects, co-produced with researchers, animal suppliers, veterinarians, publics and patients, to investigate the contemporary dynamics of animal research; interfaces for generating cultures of communication with publics, policy-makers and practitioners across the animal research nexus. This programme brings together five leading researchers on the social and historical dimensions of animal research, uniting the strengths of five institutions, engaging creative practitioners, and advancing the work of five early career researchers and three PhD students.
Macrophages represent the most abundant antigen presenting cells in inflammatory infiltrates and tumors. An important immunoregulatory function of macrophages is to present antigen to cognate T cells. Antibody (IgG)-antigen complexes are internalized into macrophages through potential interactions with multiple different Fc receptors and are subsequently delivered to subcellular compartments where peptides derived from the proteolyzed antigen can be loaded onto MHC molecules. However, the multitude of possible Fc receptor interactions with the antibody-antigen complexes influences the spatial and temporal behavior of antigen and consequent peptide-MHC repertoire through a complex, and poorly understood, network of subcellular trafficking pathways. We have developed advanced microscopy tools to enable the analysis of three-dimensional, dynamic cellular processes at unprecedented levels of spatiotemporal resolution. We will combine these approaches with antibody engineering and analyses in mouse models of cancer and autoimmunity to elucidate how the combination of antigen delivery vehicle, microenvironmental factors and macrophage phenotype interplay to result in specific antigen presentation outcomes. The two long term goals of these studies are: one, to develop novel mechanistic insights into the fundamental cell biological processes determining the repertoire of presented antigens; two, to define how to deliver antigen to macrophages to orchestrate a predictable immune response.
Kinases and their antagonistic phosphatases both provide a key regulatory mechanism controlling many cellular events. Yet our molecular understanding of phosphatases lags behind. Recently we described an interaction between Protein Phosphatase 4 (PP4) and Drosophila centromeres, which when disrupted affects the mitotic centromere integrity and spindle assembly checkpoint activity. The details of this pathway remain to be elucidated. Nor is it known how this finding relates to PP4's de-regulation in many cancers. I will therefore turn to human cell line model to address these questions. Towards determining human PP4’s mitotic functions, CRISPR/Cas9 gene editing will be used to generate cell lines expressing endogenous PP4 fused with the AID tag allowing for inducible and rapid degradation of the phosphatase. This approach will facilitate PP4 removal specifically in mitotic cells, the effects of which will be characterised by cell imaging. Parallel studies, including proteomics, will deliver precise information on the human PP4 interaction network and behaviour during mitosis. Together these data will reveal the affected processes and provide a blueprint for subsequent detailed investigations that identify the involved pathways, PP4-specific substrates and their phospho-regulatory sites. The results of this study will be later placed within a larger investigatory framework dissecting chromosome segregation.
Vacation Scholarships 2017 - University of Southampton
Internal daily timekeeping systems known as circadian clocks are used by all types of organisms to regulate their metabolism. For example, the clock-controlled scheduling of food intake and use affects health and longevity in both mammals and insects. Time-restricted or nutrient-restricted feeding improves the health of both mice and fruit flies (Panda, Science 2016). The period gene, which plays a key role in the circadian clocks of animals, provides a link between daily timekeeping and metabolic health. Flies lacking the period gene store less glycogen and triglycerides in spite of ingesting more food and are, therefore, more sensitive to starvation. I propose to test where and how the period gene provides this metabolic function. In particular, I will ask whether its role in starvation resistance is due to its control of sleep/wake rhythms or its function in periperhal clocks of metabolic tissues such as the fat body. Moreover, Dr Wijnen's laboratory recently showed that the period gene is induced at colder temperatures (Goda et al., Proc Biosci 2014) and I will test whether period-mediated starvation resistance is found preferentially at colder temperatures.
Biofilm formation and dispersal are controlled by the secondary messenger bis-(3’-5’) cyclic dimeric guanosine monophosphate (c-di-GMP); high levels are associated with biofilm formation, while a reduction induces dispersal. The enzymes catalysing formation of c-di-GMP are diguanylate cyclases, while phosphodiesterases catalyse the breakdown of c-di-GMP. Structural studies of the EAL type phosphodiesterases were observed in the host laboratory in the presence of substrate c-di-GMP, suggesting that these structures would require further steps to attain full catalytic activity. The host laboratory have recently demonstrated that dimerization and active site formation as well as formation of three metal binding sites are distinct activation steps required to tune the enzymatic activity of phosphodiesterases (Scientific Reports 2017). The proposed project deals with the motility regulator protein, MorA, and the mucoid alginate regulator, MucR, from Pseudomonas aeruginosa. Both proteins have dual activity as diguanylate cyclase and phosphodiesterase. Interestingly, the two linked catalytic domains influence each other's activity; different levels of activity are observed for the phosphodiesterases when comparing isolated domains with the linked conformation. The objective of the project is to purify these proteins and determine their catalytic activities, comparing the isolated domains, mixtures of the individual domains, and the cyclase-phosphodiesterase domains in linked conformation.
Vascular endothelial growth factor (VEGF) has important roles in the blood vessels of adult tissues and in disease. Dr Clarkins laboratory have found that deletion of VEGF by bone forming osteoblast (osteocalcin; Ocn expressing) cells results in severe bone porosity similar to osteoporosis. To investigate how vascular endothelial cell- factors contribute to this porous phenotype, further experiments from Dr Clarkins lab collected conditioned media (CM) from osteoblasts (OBs) of mice with VEGF removed and added to vascular endothelial cells. In the absence of VEGF, endothelial cells were found to upregulate inhibitors of mineralisation including sclerostin which may contribute to osteoporotic bones of VEGF KO mice. The goal of my studentship project is to investigate how endothelial cells produce sclerostin, which is important as sclerostin has previously been reported to be secreted only by bone cells. I will investigate this by immunolabelling of sections of VEGF KO and WT long bones for sclerostin and a blood vessel marker CD31. I will also treat endothelial cells with CM from VEGF WT and KO cells and undertake Western blotting for sclerostin. Inhibiting blood vessel anti-mineralisation factors could provide a new a means to treat degenerative bone disease such as osteoporosis.
Brain tissue from adult mice has already been collected, with a control group treated with saline and another treated with an attenuated strain of Salmonella typhimurium, triggering a systemic immune response including production of pro-inflammatory mediators that communicate with the brain triggering neuroinflammation and neuronal dysfunction. Selected groups of mice have been treated with two different inhibitors that block the activity of the cytokine IL-1beta, in order to investigate if this pro-inflammatory cytokine plays a direct role in neuroinflammation during systemic infection. In this project we will compare a small molecule with a biologic, both targeting cytokines, but via different mechanisms. The brain tissue will be examined using antibodies to stain activation markers of microglial cells (CD11b, FcRI, MHCII) and activation markers of cerebral endothelial cells (VCAM, ICAM, MHCII). Synaptic activation markers, from both the pre- and post-synaptic membranes, will be investigated by isolated mRNA from hippocampal enriched tissue. This investigation into phenotypic changes in microglial, cerebral endothelial cells and BBB function will be used to determine if the inhibition of IL-1beta is beneficial in reducing neuroinflammation as a result of systemic infection. This could later be applied to AD models, taking into consideration behavioural changes.
The rise of antimicrobial resistance (AMR) threatens many of the major advances in modern healthcare as the treatments rely on the implicit ability of the antibiotic to allow the body to heal from a serious infection. Antibiotic efflux transport is central to the development of AMR. At the single cell level, it is becoming apparent that the use of efflux pumps is the first line of defence against an antibiotic. These pumps decrease the intracellular level of antibiotic while the cell activates the various other levels of protection. This frontline of defence involves a coordinated network of efflux transporters. In the future, inhibition of this efflux transporter network, as a target for novel antibiotic therapy, will require the isolation and then biochemical/biophysical characterisation of each pump against all known and new antibiotics. This depth of knowledge is required so that we can fully understand and tackle the mechanisms of developing antimicrobial resistance. The key goals of this project are to over-express, IMAC purify a hexahistidine-tagged versions of a number of the efflux transporter and then characterise ligand binding against a set of clinically relevant antibiotics. The method used to measure ligand binding is thermophoresis (Nonotemper, Monolith).
How does Serum Amyloid-P component contribute to the cognitive impairment observed in Alzheimer’s Disease? 27 Apr 2017
The neuropathology of Alzheimers Disease (AD) is associated with the formation of insoluble amyloid deposits containing fibres of amyloid-beta and tau protein. Despite this some patients exhibit high amyloid load yet little loss in cognitive performance. Recent studies have shown particular polymorphs of amyloid beta fibres are correlated with AD phenotypes, raising the possibility that factors that influence the dominant polymorph formed may determine the level of cognitive impairment experienced by the AD patient. Comparison of patients exhibiting similar amyloid load but significantly different cognitive outcomes have revealed that poor cognitive performance is associated with elevated levels of serum amyloid-P component (SAP) within the brain. Given SAPs role in the growth and persistence of amyloid deposits, this research project seeks to determine if SAP can influence the polymorph of amyloid-beta fibre formed, thereby influencing the level of cognitive impairment. To achieve this amyloid-beta fibres will be grown in with and without SAP, and the resulting fibres studies by solid-state magic-angle spinning to assess if the SAP has influenced the structure of the amyloid fibre formed. The data obtained will be compared with published data to assess if the structures formed can be related to particular AD phenotypes.
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.
Our aim is to provide financial and emotional support to local veterans and their dependants, being there for them in their time of need.