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Results

Regulation of CD25 expression and IBD risk 31 May 2018

The inflammatory bowel diseases (IBD) Crohn’s disease (CD) and ulcerative colitis (UC) are chronic disorders of the gastrointestinal tract affecting 1 in 400 people in the UK. Both have a complex aetiology involving genetic predisposition and environmental triggers which is not yet fully understood, but may result in persistent bacterial infection, defective mucosal barrier, or a dysregulated immune response. Genome-wide association scans (GWAS) have identified over 200 IBD-associated loci (Lui et al 2015). One locus encompasses the gene encoding the IL-2 receptor alpha chain (CD25). CD25 is expressed by activated T cells and important in signalling pathways that regulate the immune response. Our fine-mapping data (Huang et al 2017) show that the GWAS signal at IL2RA/CD25 is due to a single SNP associated with CD, in intron 1 of the gene and co-localising with a super enhancer that regulates expression (Fahr et al 2015). This SNP is also associated with Type 1 diabetes and multiple other autoimmune disorders. In this study we will identify additional recently recruited IBD patients who are homozygous for the CD risk allele at this SNP by genotyping and investigate alteration in the expression of IL2RA by qPCR in flow-sorted regulatory and effector T cells.

Amount: £0
Funder: The Wellcome Trust
Recipient: King's College London

Role of Protease Activated Receptor 4 Signalling in Regulating Differentiation and Proliferation of CD4+ T cells 31 May 2018

CD4+ T-cells are components of the adaptive immune system and are divided in Th1, Th2, Th17 and Tregs each with a defined function. However, their function is regulated by different innate receptors. While the way that T-cells are controlled by toll-like receptors and complement receptors has been demonstrated, very little is known about how another family of innate receptor, protease activated receptors (PARs) are regulating T-cells. PAR receptors are members of the innate G-protein-coupled receptors and while PAR1 has been already shown to modulate T-cell function, nothing is known about the role of PAR4. The host laboratory has already data showing that Treg function is increased in the absence of PAR4 expression. The hypothesis that will be tested is that PAR4 regulates T-cell differentiation. To address this, T-cells will be obtained from PAR4+ and PAR4- mice. T-cells from the spleen of these mice will be cultured in cocktail of cytokines known to differentiate CD4+ T cells in Th1, Th2, and Th17 and the phenotype, proliferation and cytokine production will be compared between PAR4+ and PAR4- T-cells. The results of this study will contribute to our understanding of the function of PAR expression on T-cells.

Amount: £0
Funder: The Wellcome Trust
Recipient: King's College London

Commercial healthcare brokerage: pilot research in Delhi and London 30 Sep 2018

This research examines commercial brokerage of access to healthcare. Healthcare brokerage is performed by actors spanning a range of formality and organisation. There are individuals who facilitate care for others in their communities in return for financial or social rewards, and companies that broker personalised healthcare packages for wealthy domestic and international healthcare users. Brokers are influential mediators in the process of accessing healthcare, with important implications for the forms of care that users seek and receive, yet their activities and evolution are poorly theorised. The research focuses on the companies providing healthcare brokerage services in two settings: Delhi and London. These settings offer a range of specialised healthcare services and are established destinations for domestic and international healthcare users seeking care otherwise unavailable in their home locality. The requested funds will be used to conduct preliminary research, aiming to examine the trajectories and practices of companies in these settings using interviews with representatives in the industry. The work will inform design of a Wellcome Trust fellowship application and will enable further elaboration of a conceptual framework to be written up and submitted to a peer-reviewed journal.

Amount: £31,715
Funder: The Wellcome Trust
Recipient: King's College London

Single Molecule insights into Nuclear Mechanotransduction 17 Jul 2018

Mechanical stimuli regulate a large number of cellular functions. However, how mechanical forces are channeled through the cytoplasm to eventually reach the nucleus and alter gene activity remains poorly understood. Here we propose to employ a combination of state-of-the-art nanomechanical techniques at different length-scales to uncover the molecular details underlying the main force propagation mechanisms of nuclear mechanotransduction. We will first study the dynamics under force of each individual protein of the LINC complex, which forms a long molecular tether between the cytoskeleton and the nuclear envelope. Secondly, we will investigate the emerging role of the nucleus as a mechanonsensor. In particular, we will test (i) the lipidome changes in the NE of cells exposed to mechanical stress; (ii) the mechanical effect of key post-translational modifications of cryptic sites in specific nuclear proteins after mechanical unfolding. Finally, we will use a combined mechanical-fluorescence approach to track the dynamics of nuclear shuttling of cytoplasmic transcription factors in order to directly test the hypothesis that mechanical unfolding of proteins accelerates their transport through the nuclear pore complex. Altogether, this multidisciplinary project will provide an integrated, mechanistic and quantitative view on how mechanical forces propagate to the cell nucleus, from a molecular perspective.

Amount: £1,337,758
Funder: The Wellcome Trust
Recipient: King's College London

Thinking forward through the past: Linking science, social science and the humanities to inform the sustainable reduction of endemic disease in British livestock farming 30 Jan 2018

Livestock disease today is a complex and pressing problem that threatens the development of more sustainable, ethical and efficient farming methods. This project will devise a fundamentally new approach to its investigation that advances understandings and informs responses. Cutting across the traditionally separate realms of nature and culture, science and society, human and animal, and past, present and future, experts in veterinary history, environmental economics, epidemiological modelling, human and animal geography, rural sociology and cultural history will work collaboratively across six institutions, and in close conjunction with stakeholders to investigate two major endemic health problems: Bovine Viral Diarrhoea in cattle, and lameness in cattle and sheep. Experiences of these problems in Britain since 1947 will be examined within four contrasting farming systems - upland and lowland ‘beef and sheep’, and indoor and pasture-based ‘dairy’ – to reveal how perceptions of, and responses to disease co-evolved with farming systems and communities, human-animal relationships, expert knowledge-practices, consumer attitudes, and wider political, economic and cultural contexts. Findings will inform - and be informed by - the development of epi-economic models that better predict the future incidence of, and farmer responses to disease, generating suggestions for how to mitigate its effects.

Amount: £322,279
Funder: The Wellcome Trust
Recipient: King's College London

Medical Humanities MSc 30 Jun 2018

In undertaking this MSc, I am keen to pursue my interest in the entanglements between subject/object, human/nonhuman, persons/things and how this impacts on the boundaries of what we conceive to be ‘human’. I am particularly interested in representations of the body and its role in breaking down these boundaries. With a focus on organ transplant, I want to explore how the body becomes a site whereby these binaries interact and their boundaries begin to dissolve. In my dissertation, I plan to look at how the ‘gift’ of the organ troubles notions of identity, the complex relationship between donor and recipient, and the location of the subjective ‘I’ when then body becomes strange and alien. I then want to explore how the transplant process can turn bodies into objects of global commodification. I intend to use Titmuss’s study of ‘The Gift Relationship’ to consider how for-profit donation plays in to debates about which bodies matter. I aim to consider how future possible biotechnologies will impact upon these debates, such as animal transplant and 3D Bioprinting. I want to look at both the potential of new technology to overcome these issues, but also the different kinds of questions they invite.

Amount: £26,900
Funder: The Wellcome Trust
Recipient: King's College London

Expanding the capabilities and use of the South West Regional Facility for High-Resolution Electron Cryo-microscopy 07 Dec 2016

State-of-the-art direct electron detectors (DEDs) and new image processing strategies enable electron cryo-microscopy (cryoEM) routinely to achieve near-atomic resolution of biological samples. CryoEM has thus become a primary imaging technique, increasing the need for research institutions to provide cutting-edge cryoEM equipment. The Living Systems Institute (LSI) at the University of Exeter is a brand new interdisciplinary research centre, which will develop strategies to study diseases and their prevention. As part of the GW4 group (also including Bristol, Bath and Cardiff), we seek to develop regional research infrastructure on a scale beyond the capabilities of the single institutions. Within this remit, the Wellcome Trust-funded South West Regional Facility for High-Resolution Electron Cryo-microscopy will be established in Bristol, with a 200kV cutting-edge cryo electron microscope at its core. To support this venture and significantly increase the capabilities of the facility for all users within GW4, we plan to contribute a state-of-the art K3 DED with energy-filter. We also plan to establish an entry-level multiuser cryoEM facility at the LSI, supporting the research needs of local users in order to provide samples for further high-resolution analysis in Bristol and at the Wellcome Trust-funded electron Bio-Imaging Centre (eBIC) at Diamond.

Amount: £1,000,000
Funder: The Wellcome Trust
Recipient: University of Exeter

Senior Fellowship (WHO Placement & Global Health Justice Research) 30 Sep 2017

This proposal seeks support for an 18-month senior fellowship. The fellowship enables (1) placement at WHO GHE unit to help develop an ethical framework for ageing policies (‘just global ageing policies’) through a global, collaborative and comprehensive multi-stakeholder and multi-sectoral process, and incorporating WHO policy related documents. The framework would be informed by the fellow's (Venkatapuram) expertise in health justice philosophy and the capabilities approach previously cited by the WHO in the 2015 World Report on Ageing and Health. An ethical framework for addressing the needs of a specific social group (elderly) would be a distinctly new type of output for bioethics and the WHO compared to ethical guidelines for specific disease interventions or events (epidemics). The fellowship also enables (2) philosophical research on global health justice. Venkatapuram seeks to extend his argument for a human right to the capability to be healthy (CH)--presented in Health Justice (Polity, 2011) and in the WT funded project Population Level Bioethics and CA (WT 094245))--to global health governance and institutions. The research outputs will include two journal articles, short commentaries, lectures, and a monograph. The monograph proposal is being developed with Harvard University Press. 40% of time will be spent on this research.

Amount: £151,279
Funder: The Wellcome Trust
Recipient: King's College London

Unravelling the physics of the pressure drop in blood flow constrictions 28 Nov 2017

A blood flow obstruction introduces an additional burden to the heart, currently characterised by the pressure drop through the obstruction. The pressure drop is only available through catheterised sensors, with associated costs and risks, or through Doppler echocardiography with limited accuracy and robustness. This programme investigates the causes of the additional cardiac burden in those conditions that experience an obstruction of the blood flow. The idea is to examine the physics of the blood flow and unravel the three components of the pressure drop (i.e. unsteady, advective and viscous). The enabling technology, to be further developed within the programme, combines advanced medical imaging and computational technologies. The goals are (1) to identify the pressure biomarkers that best predicts adverse events, and thus guide the optimal choice of therapy options; (2) to enable an easy, robust and non-invasive access to the key pressure biomarkers in clinical practice.

Amount: £1,097,073
Funder: The Wellcome Trust
Recipient: King's College London

Elucidating allosteric processes in immunoglobulin E 31 May 2018

Immunoglobulin E (IgE) is thought to be the first line of defence against parasitic pathogens, mediating immune reactions by binding to either of its two receptors, either the high-affinity FcepsilonRI receptor or the low-affinity CD23 receptors. While the IgE molecule was previously thought to exist in a primarily acutely bent conformation in solution, Drinkwater et al. (2014) found that IgE was able to exist in a fully extended conformation while Davies et al (2017) showed that omalizumab (XolairTM by Novartis) trapped IgE in a partially bent state to block its action on its FcepsilonRI receptors. The McDonnell Laboratory has derived a series of anti-IgE antibody Fab fragments, selected for their ability to affect IgE’s overall structure and dynamics and consequently to allosterically affect the binding to IgE’s receptors. In this proposed study, we will investigate how observed ligand-mediated changes in conformational dynamics manifest themselves as entropically-driven allosteric modulation. As a complement to NMR studies of ligand-mediated changes in protein dynamics, currently ongoing in the McDonnell Laboratory, direct measurements of the thermodynamic parameters of ligand binding will be performed using isothermal titration calorimetry.

Amount: £0
Funder: The Wellcome Trust
Recipient: King's College London

The development of insulin resistance and anabolic resistance during muscle disuse: what is the role of fuel integration? 08 Nov 2017

Skeletal muscle atrophy, which occurs during short-term disuse, is thought to be due to the development of anabolic resistance of protein metabolism and insulin resistance of glucose metabolism, although their cause is currently unknown. The primary research aim of this fellowship is to establish the role of muscle fuel availability and integration in disuse-induced insulin and anabolic resistance. In collaboration with the Medical School, I will perform two randomized, placebo-controlled studies in which young, healthy participants undergo 2 days of forearm immobilisation with placebo, Acipimox (to decrease plasma lipid availability), Formoterol (to stimulate glycolytic flux), or dietary branched-chain amino acid (BCAA) manipulation, to alter substrate availability. I will combine the arteriovenous-venous forearm balance technique, that I have recently established in Exeter, with stable isotope amino acid infusion and repeated forearm muscle biopsies to quantify muscle glucose, fatty acid, and BCAA balance, oxidation, and intermediary metabolism (including muscle protein synthesis), both fasted and during a hyperinsulinaemic-euglycaemic-hyperaminoacidaemic clamp. Two periods of research at the University of Texas Medical Branch will enable me to develop skills in mass spectrometry tracer analyses and develop a network of collaborators in the USA, both crucial for my future career investigating disuse-induced muscle atrophy.

Amount: £250,000
Funder: The Wellcome Trust
Recipient: University of Exeter

Neurobiological mechanisms of emotional relief in adolescents with a history of sexual abuse 06 Dec 2017

Adolescents who experienced childhood sexual abuse (CSA) engage in non-suicidal self-injury (NSSI) more frequently than peers exposed to other forms of abuse or no abuse. NSSI serves an important function of relief from acute negative affect. Despite providing temporary relief from distress, NSSI is also linked to higher rates of suicide and hospitalisations and the effectiveness of current clinical interventions is limited. This may be attributed to a lack of understanding the neurobiological and behavioural mechanisms that underlie NSSI as a relief function in particular in youth who experienced CSA. To address this gap, the study aims (1) to model brain activity during distress and emotional relief (i.e., NSSI) in adolescents with and without a history of CSA using functional magnetic resonance imaging and (2) to examine if adolescents with CSA select actions to 'escape' an aversive context more quickly and often compared to non-abused peers. The ultimate goal of this translational research is to understand the neurobiological and behavioural mechanisms that confer vulnerability to NSSI following CSA (Stage 1) in order to develop effective intervention and prevention strategies to keep vulnerable teenagers safe (Stage 2) . Keywords: sexual abuse, non-suicidal self-harm, relief, functional magnetic resonance imaging, translational research

Amount: £230,048
Funder: The Wellcome Trust
Recipient: University of Exeter

Open Access Awards 2017/18 30 Sep 2018

Not available

Amount: £211,709
Funder: The Wellcome Trust
Recipient: King's College London

Cryo focussed ion beam scanning electron microscope for correlative functional studies of biological systems in situ by tomography 05 Jul 2018

Dramatic advances in cryo-electron microscopy (EM) have greatly enhanced macromolecular structure determination. The current frontier is in situ cellular structure determination to understand the machinery of life operating in the native biological environment. Visualising the native structure of cells and tissues by cryo-electron tomography requires the preparation of cryo-samples ~100-600 nanometers in thickness. The vitrified tissue sections must be thin enough to allow penetration by the electron beam, yet thick enough to contain complete structures of biological relevance. Although cells and tissues can be sectioned in the vitreous state by cryo-ultramicrotomy, both section thickness and quality are severely limited by mechanical damage including deformation, non-uniform compression and crevassing. The only available method able to precisely modify the thickness of frozen-hydrated samples and balance these conflicting requirements is focussed ion beam (FIB) milling in a scanning EM. FIB milling provides access to the cell interior, within otherwise unperturbed cellular environments, yielding vitrified lamellae suitable for electron tomography and unaffected by compression forces. Here we request a cryo FIB field emission gun scanning EM (FEG SEM) to produce lamellae targeted to fluorescent features of interest for transfer into a cryo-transmission EM (TEM) for three-dimensional (3D) structure determination by electron tomography.

Amount: £820,000
Funder: The Wellcome Trust
Recipient: King's College London

Impact of missense mutations in recessive Mendelian disease: insight from ciliopathies 10 Apr 2018

Background: Over 1,800 autosomal recessive (AR) Mendelian-disease genes have been identified. Missense mutations account for 59% of protein coding region mutations, yet their precise functional effects remain largely uncharacterized. Two important questions in human genetics aim to explain interindividual variation in phenotypic severity and assign pathogenic mechanisms to different disease phenotypes (including independent phenotypes within the same syndrome). For AR disorders, it is thought that many missense mutations cause protein instability. For these hypomorphic mutations, disease phenotypes are defined according to quantitative genetic threshold effects within a protein interaction network. Project: We will focus on a subset of AR ciliopathies which are strongly enriched for missense mutations, where complete gene knockout is thought to be lethal (see Rationale-below). We will use gene-editing and quantitative protein-protein interaction analyses to systematically compare the phenotypic effects of frameshift (likely knockout/null) and missense mutations. We will test our hypothesis that these missense mutations disrupt only a subset of gene functions/protein interactions, using phenomics algorithms we have developed and unbiased phenotyping in cell lines and mouse models, allowing us to assign pathogenic mechanisms to disease phenotypes. In future, this approach could be extended to the estimated 10-20% of > 3,000 Mendelian-disorders enriched for missense mutations.

Amount: £206,112
Funder: The Wellcome Trust
Recipient: King's College London

Santorio Santorio and the Emergence of Quantifying Procedures in Medicine at the End of the Renaissance: Problems, Context, Ideas. 12 Jan 2015

While mechanics and astronomy have always been placed at the heart of the major narrative of the scientific revolution, historians are currently reconsidering disciplines or ways of thinking once regarded as peripheral, in particular medical disciplines such as anatomy and physiology. In this context, a study on the Italian physician Santorio Santorio (1561-1636) could be particularly noteworthy. Santorio is reputed to be the first to conceive and apply the quantification of the so called 'persp iratio insensibilis' in his major work 'Ars de statica medicina' (Venice 1614). Although developed in the context of traditional medicine, concepts such as 'weight', 'measurement' and 'certainty' became essential pillars of his thought and for this reason, Santorio invented many scientific devices, among which were the first graded thermometers. Despite his relevance, however, there are no recent studies on Santorio. The goal of my research would be to illustrate the medical impetus towards cont rolled experimentation and quantitative measurement in the cultural context of the end of the Renaissance. Considering Santorio's wide legacy across the Europe, the research would also show how the development of instruments has driven medicine in the late seventeenth and early eighteenth centuries.

Amount: £127,884
Funder: The Wellcome Trust
Recipient: University of Exeter

Synaptic and Neuronal Autoantibodies in Psychosis (SNAP): clinical significance, neuroimaging correlates, infective/immune risk factors. 13 Nov 2014

Anti-neuronal surface autoantibodies (ANSAs) such as antibodies to the NMDA receptor can cause encephalopathies that feature psychosis alongside neurological symptoms. ANSAs may also cause a purely psychiatric phenotype that is a phenocopy of first episode psychosis (FEP). Moreover, ANSA prevalence in FEP cohorts is between 5-10%, suggesting they might underlie a significant proportion of FEP. These findings have however often been derived from relatively small or heterogenous samples, and the c linical significance of ANSA-positivity in psychosis remains uncertain. I will address this by assessing the prevalence of multiple ANSAs in large samples of FEP patients and individuals at ultra-high risk (UHR) for psychosis. I will examine how ANSA status relates to i) presenting symptomatology; ii) the main clinical outcomes in these groups: onset of psychosis in UHR subjects and response to treatment in FEP; iii) neuroimaging measures of brain structure and chemistry. In order to investiga te a neurodevelopmental model of autoimmunity in psychosis, markers of previous infection with neurotropic viral/other organisms will be measured, along with measures of blood-brain barrier dysfunction. Finally, I will assess the utility of using these autoimmune markers as a means of stratifying patients in the early phase of psychosis according to future clinical outcomes.

Amount: £233,755
Funder: The Wellcome Trust
Recipient: King's College London

Detecting inversion polymorphisms from SNP data 18 Sep 2015

The overarching aim of this project is to determine the most powerful method for detecting and genotyping single-mutation inversion polymorphisms from SNP data. The PI has conceived two such methods that will be developed to completion in the project because: (1) the PI considers them likely to out-perform the alternatives, and (2) the PI plans to extend them further to detect more complex inversions in the larger study. It is presently unclear which SNP-based inversion detection method has grea test power, and so before embarking on costly validation via sequencing in a larger study, we perform a comprehensive comparison study here to determine the most powerful method. While the PI is ideally placed to perform such a study given his development of the invertFREGENE inversion simulation software, such a rigorous comparison is extremely computationally and time-demanding and thus represents the primary focus for this project, requiring the postdoctoral researcher. The methodology develo pment, comparison study and associated software tool produced by this project, together provide a perfect platform for a larger study and application planned subsequently to investigate the impact of inversions on disease, the genome and the evolution of our species.

Amount: £97,799
Funder: The Wellcome Trust
Recipient: King's College London

Optogenetic dissection of NF-kB function in controlling macrophage behaviour during tissue regeneration. 27 Apr 2015

Transcription factor Nuclear Factor (NF)-kB is a critical regulator of macrophages and is implicated in several chronic diseases affecting muscle. Macrophages migrate into injured muscle during regeneration; this coincides with NF-kB activation in many cell types, but how NF-kB controls macrophage behaviour in this context is not understood. Based on our characterisation of muscle regeneration in zebrafish, we propose that NF-kB activity directs macrophage migration and that this is important fo r effective regeneration. This proposal therefore aims to determine how NF-kB signalling regulates macrophage responses to muscle injury in vivo. A combination of advanced genetic and optical techniques will be employed in the zebrafish to manipulate NF-kB activity and show how this affects macrophage cell behaviour in the context of tissue regeneration. To achieve this we will build a novel optogenetic system for rapidly activating NF-kB in a highly localised manner. This will permit direct v isualisation of macrophage cell responses to activation of NF-kB. Specific goals for this project are: 1. show if macrophage migration to wound sites requires NF-kB signalling. 2. develop a light-controlled optogenetic system for control of NF-kB activity in individual cells. 3. show if macrophage behaviour is affected by optogenetic activation of NF-kB signalling.

Amount: £99,511
Funder: The Wellcome Trust
Recipient: King's College London

Discovery and development of novel small molecule inhibitors of the human Hyperpolarization activated Cyclic Nucleotide-gated 2 (HCN2) ion channel for the treatment of inflammatory and neuropathic pain 25 Nov 2014

Treatments for inflammatory pain (IP) and neuropathic pain (NP) are frequently ineffective and have many side effects. Scientists in Professor Peter McNaughton's laboratory at the University of Cambridge have discovered that both IP and NP are abolished in mice when an ion channel is genetically deleted. This suggests that drugs blocking this ion channel will have value as novel analgesics. IP is associated with injury, infection or chronic conditions such as arthritis; and NP is caused by nerve damage in conditions such as post-herpetic neuralgia and diabetic neuropathy. Both IP and NP can impose major limitations on lifestyle and working patterns and currently available treatments have major drawbacks. For example, non-steroidal anti-inflammatories cause gastric and renal damage; and opioids cause constipation and problems with tolerance and addiction. The team aims to develop selective ion channel blockers, which avoid those that play essential roles in the heart and brain, and test them in animal models of IP and NP. In separate parallel studies they will use a known non-selective blocker to carry out proof-of-principle studies in human NP.

Amount: £25,000
Funder: The Wellcome Trust
Recipient: King's College London