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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.

Structure and mechanism of the Ino80 chromatin remodelling complex 28 Nov 2017

Compaction of the genome into chromatin helps to protect the genetic material but also causes problems in regard to access for essential processes such as transcription, replication and repair. Chromatin remodelling complexes alter the state of chromatin through a number of processes that includes chemical modifications of nucleosomes and sliding their position on DNA. Nucleosome sliding is catalysed by a number of protein complexes, one of which is the multi-subunit INO80 complex. INO80 contains an ATP-dependent translocase motor, that is common to all nucleosome sliders, but also a variety of other subunits, most of which have unknown roles. Furthermore, not only does it require two INO80 complexes interacting with a single nucleosome to promote sliding, but the complex also has an ability to "sense" the presence of other nucleosomes to space them evenly on DNA indicating interactions with multiple nucleosomes. The mechanism for this process is poorly understood, particularly at a molecular and structural level. INO80 is highly regulated in several distinct ways, including chemical modifications, small molecule effectors and subunit interactions but none of these are well understood. Finally, how the various subunits, many of which are ATPases in their own right, contribute to INO80 activities is also unclear.

Amount: £1,701,911
Funder: The Wellcome Trust
Recipient: Imperial College London

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

Suppression of adaptive immunity by Salmonella 28 Nov 2017

Dendritic cells (DCs) have a crucial role in the development of adaptive immunity to bacteria. DCs transport the intracellular pathogen Salmonella from intestinal Peyer’s Patches to mesenteric lymph nodes where they present bacterial antigens to CD4+ T cells using MHCII molecules. DCs also secrete cytokines that stimulate recruitment and activation of T and NK cells. Salmonella is a globally important intracellular pathogen that survives in DCs and interferes with the processes of DC migration, cytokine production/sensing and T cell activation. The overall goal of this application is to understand mechanisms by which Salmonella interferes with these processes. Recently we identified an effector of the SPI-2 type III secretion system (SteD) that reduces the number of mature MHCII molecules on the surface of DCs. A significant component of the planned work is to understand its mechanism of action in detail. We will use candidate-based and unbiased screens, along with molecular cell biological approaches to characterize mechanisms involved in suppressing DC migration, production of IL-12 and IFN-gamma-stimulated host cell signaling. Collectively, this research will advance the field by providing novel insights into different mechanisms by which a bacterial pathogen subverts the development of adaptive immunity.

Amount: £2,170,670
Funder: The Wellcome Trust
Recipient: Imperial College London

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

The molecular mechanisms of structural conversion and toxicity in amyloid disease. 14 Oct 2010

Despite the importance of amyloid disorders in today s population, attempts to inhibit the progress of amyloidosis have met with limited success. New therapeutic strategies require the structure, stability and dynamics of every species populated during assembly to be determined and the effects of individual species on cellular function deduced. Here we propose to combine biophysical, biochemical and cell biological approaches to address three questions that lie at the heart of our quest to u nderstand amyloidosis at a molecular level: (i) how does molecular self-recognition occur in the earliest stages of amyloid assembly; (ii) which species nucleate fibril formation and what is the structure of higher order oligomers and amyloid fibrils; and (iii) how do amyloid fibrils and fibril-associated species exert their toxic effects? Using beta2-microglobulin as a paradigm, and embracing other assembling proteins/peptides, our aim is characterise all species possible on an as sembly landscape in order to define the entire molecular assembly pathway from monomer to fibril. In parallel, by combining different strategies we aim to derive new understandings of the origins of amyloid-associated cytotoxicity. Together the programme will provide the much-needed structural, biophysical and cellular insights required for therapeutic intervention in the years ahead.

Amount: £925,927
Funder: The Wellcome Trust
Recipient: University of Leeds

The relationship of the menopausal transition to healthy aging and chronic disease risk. 20 Oct 2010

To complement an existing profile of work concerned with the relationship of women s reproductive heath and her healthy ageing, in the current proposal we will determine the role of the menopausal transition on healthy ageing and chronic disease risk. Data from the mothers in ALSPAC, together with other collaborating cohorts, will be used. Key goals that will be achieved are: 1. Determine patterns of change and inter-relationships in sex hormones, physical and cognitive capability, mental hea lth, cortical and trabecular bone density, fat and lean mass, blood pressure, fasting glucose, insulin and lipids as women go through the menopause. 2. Identify temporal and inter-individual variation in DNA methylation as women go through the menopause. 3. Determine the associations of earlier markers of women s reproductive health and their earlier lifestyle characteristics with menopausal age and menopausal transition changes. 4. Identify genetic variants that are robustly associated wi th variation in age at menopause and sex hormone levels, and use these to explore the extent to which these are causally related to cardiovascular and musculoskeletal health and physical and cognitive function. 5. Provide a detailed intergenerational resource that can be used by a wide-range of scientists to improve understanding of women s health.

Amount: £1,289,829
Funder: The Wellcome Trust
Recipient: University of Bristol

The structural and functional diversity of anti-glycolipid antibody repertoires and their nerve binding domains in human autoimmune neuropathy. 07 Oct 2010

The concept that formation of heteromeric complexes between two different glycolipids might influence antibody binding has long been suspected, but has never been systematically investigated in a clinically relevant context. In autoimmune neuropathy driven by anti-glycolipid antibodies, this is becoming of major importance as we uncover countless examples of new autoantibody specificities whose antigen binding capability is entirely dependent upon the formation of complexes between different gly colipids. This expands the extensive existing data on single glycolipids as the key antigens in these disorders. Three patterns of neuropathy-associated anti-complex autoantibody binding have emerged to date: complex enhanced, complex inhibited and complex independent, all of which are likely to profoundly influence pathogenic capability, and thus clinical classification and prognostics. New analytical methods and transgenic mouse models we have developed now provide a tractable route into disco vering complex-dependent autoantibody specificities and their neuropathological significance. Our key goals are to map the relationships between clinical phenotypes and anti-complex antibody patterns and to explore pathological effects of clinically-relevant complexes in mouse models. In doing so we will build a comprehensive understanding of complex recognition by autoantibodies, and how these antibodies underlie the undiscovered pathogenic pathways in human autoimmune neuropathy.

Amount: £1,557,397
Funder: The Wellcome Trust
Recipient: University of Glasgow

Extension to A record- funds for sequencing: Measuring the impact of naturally acquired immunity on the expression of Plasmodium falciparum variant surface antigens. 31 Aug 2011

Plasmodium falciparum infected erythrocytes express on their surface, members of a diverse family of parasite molecules called PfEMP1 that are encoded by a family of 60 var genes. These molecules interact with the surface of host cells and mediate parasite sequestration in tissues including the brain, an important step in the pathogenesis of cerebral malaria. Although naturally acquired antibodies to PfEMP1 provide specific protection against the molecular variants that they recognise, PfEMP1 ar e often considered too diverse to be vaccine candidates. However, we and others have shown that parasites infecting children with severe malaria tend to express serotypes that are more broadly recognised suggesting they may be antigenically restricted. Studies of a laboratory isolate suggest that the most broadly recognised PfEMP1 types are encoded by a genetically distinct subset of groupA var genes. By sequencing short var sequence tags expressed in parasites from Kenya, we have shown that expression of sequences with groupA-like features are associated with younger children and that a smaller subset of these sequences are associated with severe malaria. We propose to identify new targets of malaria intervention by characterizing these specific subsets of PfEMP1 that are associated with infections of children with low natural immunity.

Amount: £34,024
Funder: The Wellcome Trust
Recipient: University of Oxford

Molecular biology of bunyavirus host cell interactions. 12 Jul 2011

The Bunyaviridae is a family of mainly arthropod transmitted viruses that contain a tripartite single-stranded RNA genome. Some members of the family are of medical importance causing encephalitis or haemorrhagic fever in man. My laboratory has used Bunyamwera virus, the prototype of the family, as the representative on which to perform molecular biological studies such as functional analyses of recombinant expressed viral proteins, and the development of a reverse genetic system to recover infe ctious virus entirely from cDNAs. This proposal exploits the materials and reagents we have developed to continue and expand our work analysing the replication processes of bunyaviruses at the molecular level, with the overall goal of understanding how the virus interacts with host cells of mammalian and insect origin. Our underlying hypothesis is that differences in virus:cell protein:protein interactions between these cell types determine the outcome of infection. Specifically, using a combina tion of molecular, biochemical and cell biological approaches we will address the following interrelated areas: (i) interactions of viral proteins with cellular proteins during bunyavirus replication; (ii) real-time microscopic analysis of bunyavirus infection using fluorescently tagged viruses; and (iii) functional analysis of the bunyavirus NSs protein in mammalian and insect cells.

Amount: £283,910
Funder: The Wellcome Trust
Recipient: University of St Andrews

Regulation of N type calcium channel expression, trafficking and modulation. 31 Aug 2011

Voltage-gated calcium channels are essential to numerous aspects of the function of excitable cells. In neurons they are involved in the release of neurotransmitters, and in a number of other processes in which a rise in intracellular Ca2+ is the primary signal. Two very important neuronal subtypes of calcium channels are N-type (CaV2.2) and P/Q-type (CaV2.1), with the former being particularly important at peripheral synapses and neuro-effector junctions, in the autonomic and sensory nervous systems. In nociceptive dorsal root ganglion neurons they are a therapeutic target for anti-nociceptive drugs. The function of these voltage-gated calcium channels may be modulated by several means, for example their expression is controlled by the auxiliary ß as well as a2d subunits, and in the short term they are inhibited by the activation of G-proteins via a variety of G-protein coupled receptors. Over a number of years, my group has examined the properties of cloned calcium channels, expressed both in non-neuronal heterologous expression systems and in neurons. The key goals of the current proposed programme of work relate to four interlinked themes involving CaV2.2 calcium channels: (i) examination of CaV2.2 calcium channel mRNA trafficking and stability, since the mRNA for these channels has particular trafficking motifs in it, (ii) examination of CaV2.2 calcium channel protein trafficking and the role of the auxiliary ß subunits and (iii) study of the mechanism of G-protein modulation of plasma membrane CaV2.2 calcium channels. Finally (iv) we will study the pathophysiological consequences of mis-assembly of CaV2.2 calcium channels. The last theme could also generalise to CaV2.1 calcium channels and may relate to the calcium channelopathy, episodic ataxia-type 2, which is caused by mutations in this channel that are often truncating mutations

Amount: £333,308
Funder: The Wellcome Trust
Recipient: University College London

Ethics, Policy and Practice of Poliomyelitis Vaccination in Orissa: A case study in community, professional and governmental attitudes to mass public health programmes in India. 02 Jun 2011

This interdisciplinary project brings together two experienced academics from different disciplines (history and philosophy) to work together on a project that naturally develops out of their individual past interests. It has three key aims. First, to gather relevant empirical evidence about attitudes to poliomyelitis vaccination campaigns in Orissa from three key participant groups: parents, community workers and those involved in the planning and implementation of the campaign (e.g. government officials). Second, to outlineand systematically explore the ethical issues that will arise in relation to the empirical material gathered as part of this project. Third, to develop research capacity in ethics, particularly public health ethics, in a state in India (Orissa) where there is at the moment almost none. This project is only the first of an expected continuing collaboration between the applicants and their institutions.

Amount: £63,698
Funder: The Wellcome Trust
Recipient: University of Birmingham

Ethics, payments and maternal survival in Tanzania. 02 Mar 2011

In the context of Tanzania's high maternal mortality and low levels of access to life-saving interventions, the limited existing research on charging levelsfor emergency obstetric and post-natal care confirms their potentially exclusionary levels, and facility surveys identify problems of poor quality and staff morale. Both the charging practices and the other quality problems are widely seen as unethical, but there is a gap in the research literature ontheir interactions at facility level. This project aims to add to knowledge about the ethical content and implications of individual payment systems for maternal care, and to generate policy-relevant evidence as to how health management can achieve more ethical and inclusive care through breaking interactive linkages between individual payments, abuse, exclusion and collapse in staff morale. The project methodology is largely qualitative, to be undertaken in four districts of Tanzania including districts which have, and have not, been the subject of policy intervention to improve quality and access to maternal care. The results will be fed into policy and health management debate through strong links of Economic and Social Research Foundation (ESRF) to local policy processes and through international dissemination of results.

Amount: £106,669
Funder: The Wellcome Trust
Recipient: Research on Poverty Alleviation

Perceptions of scientific collaboration in transnational HIV research -- an emergent bioethical issue. 04 Nov 2010

The growth of collaborative global health research networks presents a number of ethical issues not previously encountered such as community engagement, global ethics governance, and the ethics of global research collaboration. Recently, researchers and bioethicists have observed that the protections that medical and research ethics aim for are modeled on a western tradition which often prove limited in effect and inadequate in scope especially when cast against a wider backdrop of global healt h, economic inequalities and cultural diversity. These discussions have not been a preserve for researchers and bioethicists. They do take place amongst the public through debates in the media, by politicians and policy makers as well as the general public, including the communities where research is conducted (discussing power relations among scientists, control of research agenda, trusting or mistrusting certain agencies). The proposed project will focus on the experiences of scientists workin g in collaborative global health research initiatives and the public perception of collaboration between local and international scientists as a measure of the way in which the public engages with health research in the broadest sense. The role of the media in informing and shaping this public opinion on collaborations is investigated as part of this inquiry.

Eugenics and Genetics: dissemination project. 19 Oct 2010

There is a lot of confusion about what exactly eugenics is and about some of the ethical arguments closely associated with eugenics. With this is mind, the main aim ofthis Dissemination Project is to inform ethical discussion both within and between the following key groups: Activist, Campaigning, or Educational Organisations with specialist interests in genetic and reproductive technologies Health Care Professionals involved in Reproductive Medicine Policy and Regulatory Bodies with remits or interests in genetics or reproduction Stakeholder Organisations representing people with disabilities, people with genetic disorders, or actual/prospective recipients of infertility treatment services The proposed dissemination methods are: (a) three or four accessible briefing papers; (b) at least two papers for health care professional journals; (c) a series of (three) stakeholder engagement seminars. One important conclusion of my past research is that we ought to be cautious about using the term eugenics', because its meaning is disputed and unclear, and because it can beused as a powerful emotive term. Therefore, much of the proposed dissemination activity will not use, or at least will not foreground, the term eugenics' since that would be counterproductive and may well damage attempts to enhance understanding of the underlying arguments and concepts.

Amount: £34,009
Funder: The Wellcome Trust
Recipient: Keele University

Consent to and community engagement in global health research - reviewing and developing research and practice. 18 Oct 2010

This is a proposal for funding to organize a four-day international collaborative workshop bringing together leading groups engaged in research on community engagement and consent in collaborative global health research. The workshop will bring together individuals responsible for introducing or sustaining community engagement activities in developing countries, and for designing and implementing appropriate consent practices in these settings, and those conducting social science research on these topics. The aim will be to: 1. share ideas, experiences and needs regarding community engagement and consent interventions, including monitoring and evaluation, and related social sciencemethodologies 2. review ethical theories and approaches, and recent community engagement andconsent guidelines, and to consider how these frameworks can be incorporated into implementation activities and social science studies. 3. develop proposals for future research, including comparative studies, and to consider transferability of findings of studies conducted to date. 4. discuss the possibility and practicalities of establishing an internationalnetwork of groups working on community engagement and consent which might, for example, agree todevelop online collaborative resources for the sharing of experiences and of training materials.

Amount: £29,300
Funder: The Wellcome Trust
Recipient: University of Oxford

Assessing voluntariness of consent to research in South African HIV prevention and treatment trials. 18 Oct 2010

Consent to research participation is only valid if voluntarily given. Voluntariness in research generally implies that a person is able to act of their own free will, free from coercive influences and undue pressure, when deciding whether or not to participate in research. The high rates of HIV/AIDS in South Africa make HIV prevention and treatment research essential in this country. Studies on the voluntariness of consent conducted in South Africa suggest that participants may believe they are not free to make decisions to participate or to withdraw from research. However, voluntariness and its impairments have been poorly conceptualised and studied to date (Appelbaum, Lidz & Klitzman, 2009a).This study aims to develop a method to assess voluntariness of consent and to provide empirical data to contribute to this debate and inform future studies of voluntariness in the context of HIV prevention and treatment research. This will be achieved by identifying, reviewing and comparing appro aches to the assessment of voluntariness; developing a method to assess voluntariness; and piloting the method to assess voluntariness of consent to research to explore the degree of voluntariness exercised by research participants in their decision to enrol in HIV prevention and treatment trials.

Amount: £25,488
Funder: The Wellcome Trust
Recipient: University of Kwazulu Natal