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Microfluidic Platform for Investigating the Kinetics of Extracellular Vesicle Induced Metastatic Niche Formation 21 May 2018

Extracellular vesicles (EVs) are believed to be important messengers in the progression of metastatic cancer that prime distant organs for tumour cell colonisation. However, due to an inadequacy of relevant tools, we have a poor understanding of how EVs distribute to, diffuse into and remodel organs into metastatic niches. The goal of this project is to develop novel microfluidic platforms for performing real-time continuous quantification of EV kinetics over multiple days in physiologically-relevant microenvironments. Towards this end, I propose three aims: Develop Microfluidic Metastatic Niche Platforms to explore the interaction of extracellular vesicles with liver tissue and vasculature. Investigate the kinetics of EV distribution, uptake and diffusion in liver and vasculature compartments of Microfluidic Metastatic Niche Platforms. Explore the influence of EV kinetics (distribution, uptake and diffusion) on the ability of cancer cells to attach, invade and proliferate in Microfluidic Metastatic Niche Platforms. The results of this project will enhance our understanding of metastatic cancer progression and will contribute valuable data for numerous follow-up studies aiming to inhibit or even prevent the development of metastatic niches.

Amount: £99,951
Funder: The Wellcome Trust
Recipient: Imperial College London

Mechanisms and pharmacodynamics of antifungal agents for cryptococcal meningitis. 30 Sep 2018

Cryptococcal meningitis (CM) is an infection of the brain and surrounding tissues (the meninges). It is caused by a yeast called Cryptococcus and is responsible for approximately 180,000 deaths annually (26). The most effective drug is amphotericin B (AmB) which needs to be given for 2 weeks and causes dangerous side-effects. A modified formulation, liposomal amphotericin B (LAmB), may be easier to administer to patients because it can be given as a single dose, and appears to be as effective as 2-weeks of conventional AmB (12, 15, 23). This observation raises a number of questions: 1) What is the optimal dosing strategy for LAmB? I will measure drug levels and describe their relationship with reduction in Cryptococcus levels. 2) How does one dose of LAmB exert a prolonged effect? i will image the movement of LAmB in mouse brains and meninges to assess how long LAmB stays in these regions. During treatment for CM, the rate of decline of yeast in spinal fluid is highly variable (24, 25, 27). Therefore, another question is: 3) Do different groups of yeast vary in teir response to treatment? I will collect samples of Cryptococcus and characterise their survival ability in various conditions.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Liverpool

Clinical Characterisation of a Broad Spectrum of Genetic Variation in the General Population 30 Sep 2018

Inborn errors of metabolism (IEM) are severe and extreme changes in metabolism caused by mutations in a single gene. Recent large-scale human studies have shown that genes causal for IEM are associated with nutrients, or ‘metabolites’, in the blood. However, whether these associations cause disease or adverse health outcomes is unknown. In this project, I will use IEM genes identified in these studies to link genetic variation to clinical features in a large human population. To do this, I will assemble a list of IEM genes of interest that were identified in the literature and in large population datasets. I will then test for associations between the variants I find in these genes and a wide range of clinical features found in open-access population datasets. As the IEM genes used in this study have been associated with blood metabolites previously, linking variants in these genes to clinical features will shed light on the molecular mechanisms underlying genes and disease in the general population. Understanding how genetic variation affects disease will help identify novel therapeutic targets and enable health professionals to better manage disease risk.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Cambridge

LMIC Travel Scholarships for Global Climate and Health Forum 30 Sep 2018

The Global Climate and Health Forum is a one-day, high-level convening of global climate and health leaders designed to mobilize stronger health sector engagement in and commitments to climate action. The Forum will bring together 250 leaders from national and local governments, health systems, public health agencies, civil society, and international organizations to build the community of climate and health professionals, strengthen collaboration across sectors, and raise the health voice for climate action. The Forum will be held at the University of California, San Francisco on September 12th, 2018. The Forum is an affiliate event of the Global Climate Action Summit, and co-hosted by the UCSF Global Health Group, Health Care Without Harm, Global Climate and Health Alliance, and US Climate and Health Alliance. In order to make the Forum a truly global event, it is imperative that the Forum includes speakers and participants from low- and middle-income countries who are leading climate mitigation, adaptation, and resilience work in the most vulnerable regions and communities. Funding from the Wellcome Trust will be used to support five travel scholarships for participants from the global South, including all registration, travel, accommodation, and event-related expenses.

Growth and secretion by Campylobacter jejuni under anaerobic conditions 31 May 2018

Campylobacter jejuni is the leading cause of bacterial gastroenteritis and thus poses a significant health risk. The bacteria is part of the natural microbiome of the chicken caecum where it appears to function as a non-invasive commensal but in the human intestine the organism becomes invasive and pathogenic. The Ó Cróinín group have recently reported that DNA supercoiling plays a key role in inducing this invasive phenotype and that relaxation of DNA supercoiling is associated with an increase in invasion and the appearance of an invasion associated secretive. This group have also unpublished data which reveals that DNA supercoiling allows the bacteria to survive and grow under anaerobic conditions which normally do not support growth. Given the anaerobic nature of some areas of the human intestine this could indicate that relaxation of DNA supercoiling could be critical in allowing this bacteria to both secrete virulence factors as well as to survive and grow under anaerobic conditions. The aim of this study is thus to investigate and characterise the effect of DNA supercoiling on the ability of the bacteria to grow under anaerobic conditions as well as to compare the secretion of proteins by microorganisms grown under anaerobic and microaerophilic conditions

Amount: £0
Funder: The Wellcome Trust
Recipient: University College Dublin

Intra-Africa Mobility Initiative on Science, Language and Culture 30 Sep 2018

The Alliance for Accelerating Excellence in Science in Africa (AESA), Wellcome Trust (WT) and the Institut Pasteur International Network propose a researcher mobility program which would support African researchers (who share our ambition) to break down language and cultural barriers that impede greater African research collaboration in the biomedical and health sciences.Over the next 5 years, we aim to support at least 50 researchers to undertake mobility favoring the enhancement of their scientific careers while also increasing the understanding and interconnections of the African research community within different cultural and linguistic environments. Where language is the barrier we will look for flexible ways of providing support and funding language training. The program will aim to; Strengthen scientific collaboration between Anglo and Francophone speaking African scientists Build language skills/capabilities in English and French among African scientists Improve cultural understanding between English and French speaking African scientists The program will build out from the DELTAS and H3Africa researcher communities and the Institut Pasteur network, Our indicator of success will be; By December 2020, 50 African researchers will have experienced the mobility program and report positively on breaking down language and cultural barriers to research.

Amount: £198,662
Funder: The Wellcome Trust
Recipient: African Academy of Sciences

Investigating the Structure of FcγRIIa and its Interactions within the Membrane 30 Sep 2018

Human Fcgamma receptors (FcgammaRs) are proteins found on the surface of immune cells. They bind to antibodies, which are produced by the body, in response to infection. Some antibodies produced recognise their own tissues and are found in many diseases, including rheumatoid arthritis and lupus. It has been shown that genetic changes in the FcgammaRs are found more frequently in rheumatoid arthritis sufferers compared to healthy individuals. This project will focus on FcgammaRIIa, which is present on cells which are responsible for the destruction of many antibody-bound objects. Through a combination of cutting edge techniques, spanning physics, biology, immunology and medicine, we will uncover fundamental information within this field. This information would aim to inform the production of effective therapies to treat diseases such as arthritis, which put a huge strain on the NHS every year.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Leeds

Design and testing of allele specific gene editing tools. 31 May 2018

The Type II Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a programmable RNA guided endonuclease, which is effective at gene editing in mammalian cells. These highly specific and efficient RNA-guided DNA endonucleases may be of therapeutic importance to a range of genetic diseases. The CRISPR/Cas9 system relies on a single catalytic protein, CRISPR associated protein 9 (Cas9), which can be guided to a specific DNA sequence anywhere in the genome by the substitution of a 20-nucleotide sequence, complimentary to the particular target, within a single RNA molecule (sgRNA).A number of computer programs have been developed to predict the sgRNA that cuts the intended target most efficiently with the least off-targetting. I aim to identify, design and characterise sgRNA molecules that could be used to target mutant corneal dystrophy genes by using molecular biology techniques such as in silico identification of mutations in corneal dystrophy genes suitable for CRISPR/Cas9 gene editing with in vitro testing of selected guides, design of sgRNA to target the identified mutation and comparison of sgRNA design and off-target prediction tools.

Amount: £0
Funder: The Wellcome Trust
Recipient: Ulster University

C. elegans – a model system to study causation in the microbiome-gut-brain axis 06 Sep 2018

Metagenomic approaches are generating a long list of health and disease states associated with the microbiome (the microbes inhabiting our digestive tract) but without mechanistic understanding. The challenges arise from the enormous complexity of the microbiome, including genetic, environmental and dietary variations, microbe-microbe interactions, and the cost associated with studying the microbiome using murine models. If the field is to move beyond association, simple, well-controlled and cost-effective models allowing unbiased high-throughput studies are needed. The nematode C. elegans is a genetically tractable model, ideally suited for mechanistic and causative studies of host-microbiome and microbe-microbe interactions shaping host physiology and metabolism. Aims: 1) Establish an experimental microbiome in C. elegans based on strains from the human microbiome associated with health, disease and ageing 2) Generate fluorescent labelled bacterial strains and perform real-time imaging of gut colonisation 3) Characterise the effects of the experimental microbiome on the microbiome-gut-brain axis during ageing The experimental microbiome in C. elegans will be an important contribution to the field, complementing existing models. It will allow me to establish important collaborations, form a basis for my future research and address one of the most important questions of modern biology: the effects of the microbiome on host physiology.

Amount: £99,905
Funder: The Wellcome Trust
Recipient: Queen Mary University of London

A structural investigation into the action of and resistance to ribosome-targeting antibiotics 30 Sep 2018

Antibiotics are crucial to modern medicine, allowing treatment of life-threatening bacterial infections and making many surgeries like transplantations possible. However, pathogenic bacteria are rapidly evolving to resist their effects. Protein synthesis is one of the main antibiotic targets in bacterial cells. I will use structural biology techniques, principally cryoEM and single particle image processing, to understand how both novel natural products and clinical antibiotics bind to the ribosome to bring about their inhibitory effects on protein synthesis. Furthermore, I will investigate the cause of toxicity of certain ribosome-binding antibiotics by examining how they bind to the mammalian mitochondrial ribosome. Finally, I will use a combination of cryoEM and protein X-ray crystallography to elucidate how certain ribosomal-protecting proteins form complexes with the ribosome in order to bring about antibiotic resistance. On an individual level, these studies will allow an assessment of the viability of novel natural products as suitable clinical antibiotics. More generally, they will contribute to our knowledge of how different classes of antibiotics target the ribosomes of pathogenic bacteria, and how these bacteria evolve resistance. This knowledge will help the development of methods to rationally design new ribosome-targeting antibiotics that are able to overcome or circumvent resistance.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Leeds

The role of MINK1 in homoeostasis of the intestinal epithelium 30 Sep 2018

Surgery and chemotherapy, which preferentially kills dividing cells, are the main treatments for colon cancer: a common disease in the developed world. To develop better treatments requires defining the molecular events that cause tumours to grow so that these specific aberrations can be bypassed. Mutations in a gene called Adenomatous polyposis coli (Apc) are the most common molecular change identified in colon tumours. The Näthke lab recently discovered that APC protein (encoded by the Apc gene) binds to and regulates the abundance of another protein (MINK1) with roles in cell division and movement. This raises the possibility that faulty control of MINK1 by mutated APC could explain why cells in colon tumours divide excessively and migrate aberrantly. I will investigate the function and control of MINK1. Using cells grown in dishes and also cells that form mini-gut structures called organoids, experiments will be designed to determine whether and how MINK1 is required for Apc mutations to cause cancer, and how MINK1 affects the structure of the gut lining. These issues will also be addressed in mice whose guts lack the Mink1 gene. Results will help to decide whether human colon cancer can be treated by new drugs that target MINK1.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Dundee

Human Challenge Studies in Endemic Settings: Ethical and Regulatory Issues 30 Sep 2018

This study will investigate the key ethical and regulatory considerations regarding human challenge studies (HCS) in endemic settings—and to examine the ethical processes (e.g., in the design, review, and conduct) involved in such studies to date (with a focus on recent HCS involving diarrheal disease in Thailand and malaria in Kenya and Tanzania). Our approach to this project will include two main data gathering processes: (1) review of relevant literature, and (2) qualitative research, involving in-depth interviews of those involved in the conduct and ethical review of recent HCS in endemic settings (focusing especially on HCS involving diarrheal disease in Thailand and malaria in Kenya and Tanzania) and other relevant stakeholders.

Amount: £99,407
Funder: The Wellcome Trust
Recipient: Monash University

Free Will and Neural Activity in Consequential Action 31 May 2018

Neuroscientific and psychological studies have found that neural activity precedes the intention to act by several seconds, indicating that our actions are predetermined by unconscious neural computation. A major shortcoming of these studies is that the acts tested are trivial (e.g. move a finger at a time of your choosing), having no real-world implications. In a series of EEG experiments, we will assess complex choices that participants make. First, participants will be asked to choose to press one of two buttons. Then, they will be asked to a) ‘reward’ or ‘punish’ a person arbitrarily, b) reward or punish a person in response to a moral act, c) choose how much to reward or punish the individual based on their action, and finally, d) choose reward and punishment allocations within social or economic constraints, to reflect real-world decision-making. This will allow us to test if there is neural activity prior to the intention to act and also to compare between consequential and inconsequential choices. The investigation of neural activity in consequential action, examined in the light of philosophical analyses, addresses an essential unanswered question of past studies, and promises to shed new light on the nature and existence of free will.

Amount: £0
Funder: The Wellcome Trust
Recipient: University of Aberdeen

Complex Urban Systems for Sustainability and Health (London Hub) 30 Sep 2018

The London Hub for Urban Health, Sustainability and Equity aims to be the world’s foremost transdisciplinary hub for research, training and pubic engagement on urban health. It is founded on two constituent projects – Complex Urban Systems for Sustainability and Health (CUSSH) and Pathways to Equitable Healthy Cities (PEHC) – and involves leading London-based institutions and their global network of collaborating institutions. The Hub’s principal objective is to integrate and coordinate research and stakeholder engagement that support evidence-based policies aimed at improving population health, health equity and environmental sustainability in cities around the world. The Hub, and its projects, will achieve this objective through comparative studies that involve participatory research and coproduction of knowledge among academic researchers, policy makers and practitioners, and civil society; developing models for prospective policy evaluation and applying these models to data from our partner cities; and training the next generation of research and policy leaders in urban health, while establishing the foundations for sustaining and expanding the Hub beyond the Wellcome funding period. The CUSSH project focuses on how to transform cities to address vital environmental and population health imperatives, and entails partnership with the cities of London, Beijing, Kisumu, Nairobi, Ningbo and Rennes.

Amount: £5,150,000
Funder: The Wellcome Trust
Recipient: University College London

An Extended Pilot for the Human Cell Atlas: Adult tissues, human development and inflammation-mediated pathologies 30 Sep 2018

The Human Cell Atlas (HCA) is an international, collaborative effort that "...aims to define all human cell types in terms of their distinctive patterns of gene expression, physiological states, developmental trajectories, and location". Here, we will contribute directly to the first phase of the HCA by forming an ‘extended pilot’ to implement UK infrastructure for large-scale, high quality human cell atlas experiments. We will generate a high-level atlas, with spatial resolution, for multiple adult human tissues along with matched data from human fetal material. We will then illustrate the power of a deep and focused investigation of a single tissue (skin) to produce highly-detailed data describing its cellular composition and spatial organisation. Finally, for selected tissues that have been profiled in adults and fetal material, we will analyse samples from immune-mediated disorders as a comparison with our reference data to gain deeper understanding of the pathological mechanisms. This will demonstrate the utility of the HCA as a ‘healthy reference’ for comparison with disease. Throughout, we will generate profound biological insight from primary human cells and lay a foundation of technology development and optimisation with a set of hardened and scalable methods for single-cell RNA-sequencing, spatially-resolved gene expression, and tissue imaging.

Amount: £3,878,823
Funder: The Wellcome Trust
Recipient: Wellcome Trust Sanger Institute

Toxic DNA: a model for all domains of life 17 Jul 2018

The instructions for life are encoded by DNA. As such, it is counter intuitive that certain DNA sequences can be intrinsically toxic to the cell. The problem is acute for DNA where the frequency of adenine and thymine exceeds what is normal for a given organism. Toxic effects of such AT-rich DNA have been reported for species as diverse as E. coli and humans. Working with bacteria, I have shown that misdirection of transcription is closely linked to the toxicity of AT-rich DNA. This happens because promoters, the DNA sequences that instigate transcription, are also AT-rich. Since promoters from all cell types have a high AT-content, I argue this may be a universal phenomenon. If true, the implications are far reaching; AT-rich DNA impacts processes as diverse as antibiotic resistance in bacteria and cancer in humans. My application probes the mechanistic details underlying the toxicity of AT-rich DNA for both prokaryotic and eukaryotic cell types. Hypothesis: The toxicity of AT-rich DNA is a consequence of spurious transcription Aim 1. Understand molecular basis Aim 2. Understand toxic mechanisms Aim 3. Understand evolutionary prevalence

Amount: £1,138,415
Funder: The Wellcome Trust
Recipient: University of Birmingham

Human cell-type specific ion dynamics in neuroinflammation and seizures 06 Sep 2018

Epilepsy is more common in Africa than other parts of the world due to high rates of head injuries and infection, which trigger inflammatory processes in the brain. How neuroinflammation causes epilepsy is not well understood. Disruptions in the transmembrane gradient for two key ions: chloride and hydrogen ions, are prototypic features of hyperexcitable neuronal networks. We hypothesize that neuroinflammation affects the cellular regulation of chloride and hydrogen ions, and hence seizure susceptibility in the brain. To test this hypothesis, we will utilize my expertise for measuring ionic changes in the brain together with rare access to human brain tissue donated by patients who have undergone neurosurgical procedures. Our first aim is to establish human brain slice cultures as a model system. We will then transfect different brain cell types with a genetically-encoded chloride and hydrogen ion sensor I have developed in order to generate the first baseline and activity-dependent measurements of these ions in living human neural tissue. Next we will determine how triggering an innate neuroinflammatory response in human brain slice cultures affects circuit excitability and seizure propensity. Finally, we will observe cell-type specific intracellular chloride and pH dynamics during the acute neuroinflammatory response in the human brain.

Amount: £94,309
Funder: The Wellcome Trust
Recipient: University of Cape Town

How does nonsense-mediated decay contribute to gene regulation in developing neurons? 31 May 2018

Nonsense mediated decay (NMD) is a quality control mechanism used by eukaryotic cells to destroy messenger RNAs containing incorrectly positioned translation termination codons [1]. NMD, in combination with alternative RNA splicing (AS), also provides a potent mechanism for natural changes in gene expression in developing brain [2-5]. The main goal of my summer project will be to test the hypothesis that progressive down-regulation of an RNA-binding protein called PTBP1 during mammalian neurogenesis promotes neuronal identity by changing AS patterns and triggering NMD of multiple neural precursor-specific transcripts. I will first follow up on the RNA sequencing screen carried out in the Makeyev lab and validate bioinformatically predicted AS-NMD targets using Reverse Transcription PCR and quantitative PCR analyses of developing nervous system samples and embryonic stem cells undergoing neuronal differentiation in vitro. I will then analyse AS-MND regulation for one example showing the most robust regulation. This will be achieved by designing minigene and CRISPR-Cas constructs specific for AS-NMD promoting exons and testing these reagents in mouse ES cells undergoing neuronal differentiation or treated with siRNA against PTBP1. The results of this work should improve our understanding of the AS-NMD pathway and evaluate its contribution to neuronal differentiation.

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

The Effect of Acute Caffeine Ingestion on Cognitive Dual Task Performance During Assessment of Static and Dynamic Balance in Older Adults 31 May 2018

The ergogenic benefit of caffeine for sports performance has been reported extensively in younger adults (4) and recent evidence suggests benefits of acute caffeine ingestion for motor performance (5) and cognition (6) in older adults. Given such components are important mediators of postural control, it is proposed that caffeine may be an important nutritional supplement to improve balance in older adults. Studies examining the effect of caffeine on balance are sparse and focus only on static postural control (7). Such assessments give a limited understanding of balance tasks adopted during the completion of tasks of daily living, where falls are more likely during locomotory tasks requiring dynamic balance (8). Furthermore, individuals must maintain balance whilst completing secondary cognitive tasks such as communicating or crossing a road. Previous work had indicated that that completion of a secondary task can impair (9) or improve postural control but at the expense of the secondary task (10). It is unknown weather caffeine can mediate these detrimental effects. The present study aims to assess the effect of caffeine ingestion on cognitive dual performance during assessments of both static and dynamic balance in older adults.

Amount: £0
Funder: The Wellcome Trust
Recipient: Coventry University

Biomaterial-based delivery of neurotrophic factors to the Parkinsonian brain 31 May 2018

The concept of cellular brain repair for Parkinson's disease is relatively simple - if brain cells die in this condition, then it should be possible to replace these cells through transplantation of healthy cells back into the brain. Over the past three decades, numerous animal studies and several clinical trials in human patients have shown that this concept has significant potential for repairing the brain affected by Parkinson's disease. However, poor survival of the healthy cells has limited the widespread roll-out of this cellular reparative approach to patients. Biomaterials, that is, materials that have been engineered to interact safety with living tissue for therapeutic purposes, have the potential to improve such cellular reparative therapies for Parkinson's disease. Specifically, injectable biomaterials gels have the potential to significantly improve cell survival by providing a physical scaffold and pro-survival environment for the implanted cells. Thus the aim of this proposal is to determine if biomaterial hydrogels can be used to improve cellular reparative therapies for Parkinson’s disease using animal models of the condition. We will specifically investigate the beneficial effect of biomaterials on stem cell-derived neuron cell transplantation approaches.