- Total grants
- Total funders
- Total recipients
- Earliest award date
- 30 Jan 2009
- Latest award date
- 07 Dec 2009
- Total GBP grants
- Total GBP awarded
- Largest GBP award
- Smallest GBP award
- Total Non-GBP grants
Infection and Immunity The origin and function of theand uninfected wounds FAP + cels in infected and uninfected wounds 18 Feb 2009
The aim of the project is to define the origin of the FAP _ cells in the tumour stroma. The first question to be asked is whether they are composed of haematopoietic cells, mesenchymal cells or a mixture of both. If they are haematopoietic in origin they could represent a case of a haematopoietic to mesenchymal transition as has been proposed for the stellate cells in the liver which are c045 but come from c045 progenitors (Miyata E et al. 2008). Transdifferentiation has also been proposed to occur from mesenchymal stem cells (MSCs) (Uccelli A et al. 2008) and it could be the case that these contribute c045 cells to the stroma. To answer this question OVAGFP BAC transgenic and non-transgenic mice would be sublethally irradiated and then would be reconstituted with sorted haematopoietic stem cells from non-transgenic or transgenic mice, respectively, differing at the thy1.1 locus. These mice would then be injected with LL2 or 816 tumour cells and the resulting tumours analysed for the presence of CD45+ and CD4s EGFP+ cells in the tumour stroma. This would allow the contribution of both non HSCs and HSCs to the FAP compartment to be ascertained. - Along with this lineage analysis further characterisation of the FAP cells would also be carried out. FACS-sorted GFP+ stromal cells could be further analysed to examine their surface markers. Recently it has been suggested that FAP _ marks human MSCs from the bone marrow (Bae S et al. 2008). This study was limited by the fact that it used commercially obtained MSCs and so it is unsure how well it applies to in vivo but it would be worth looking for GFP+ cells in the bone marrow. If these were found, they could be analysed for expression of CD45, indicating HSCs, or for several other markers as CD45.CD73.CD9<rCD105+ cells would indicate an MSC origin. Cells from the tumours would also be analysed. To do this tumours would be obtained and single cell isolates prepared this could allow the phenotypes of the CD45+ and c045 populations to be better defined by other markers. An example would be looking for alpha smooth muscle actin which would define 'reactive' myofibroblasts in the stroma on CD45- cells. Finally functional studies would be performed to see what these cells were doing. They could function as APCs despite being CD45 , as has been reported for the stellate cells in the liver (Miyata E at al. 2008; Winau F et al. 2007). It would also be worth checking if they were anti-inflammatory for example through production of heme-oxygenase 1, or whether they produce retinoic acid which has been shown to be involved in inducing regulatory T-cells (Xiao S et al. 2008). This latter possibility again ties in with the hepatic stellate cells in the liver which also store a large amount of vitamin A (Reuben A, 2002).
Body weight and fat mass are highly heritable traits. I plan to investigate copy number variants (CNVs) that have been identified in a cohort of patients with severe early onset obesity established by Dr Farooqi at the University of Cambridge. My first goal is to identify CNVs of interest by prioritizing those present in multiple affected individuals and enriched in cases versus controls. Bioinformatic databases will be used to identify any prior information known about whether the genes may be associated with obesity. Multiplex ligation-dependent probe amplification (MLPA) will validate the CNVs and a fine-mapping custom array will attempt to identify the breakpoints. Co-segregation in families may suggest potential pathogenicity. In addition, working with Dr Barroso's team at the WTSI, I will sequence potential genes contained within these CNVs to identify whether rare variants are also associated with severe early onset obesity. I intend to establish whether the zebra-fish can be used as a model organism to study new candidate genes for obesity. Mutations in the melanocortin 4 receptor (MC4R) gene can cause severe obesity in humans. In collaboration with Dr Stemple's group at the WTSI, I plan to characterise the phenotype of an MC4R mutant fish that has been identified and in doing so, develop the techniques for studying other obesity candidate genes in this model organism.
In infant humans and small mammals, brown adipose tissue (BAT) functions as a tissue for thermoregulation. This is achieved through its ability to uncouple oxidative metabolism from ATP synthesis; thus facilitating the conversion of caloric energy, especially fats, into heat. With the recent discovery that BAT is present in biologically relevant amounts in both lean and obese adults, the possibility of activating BAT as a means of increasing caloric loss in obese states are being investigated. When BAT is activated, there is a simultaneous increase in both fatty acid oxidation and synthesis. In addition, the synthesized fatty acids are extensively modified, including acyl-chain elongation and desaturation. We postulate that the ability of BAT to modify fatty acids is crucial to its function. Specifically, fatty acid chain length and saturation helps determine its intracellular fate; either towards oxidation, storage or other cellular compartments. The aim of this study is to investigate the role of fatty acid composition on BAT metabolism and asses the impact of this alteration on whole body energy balance and thermoregulation. Fatty acid elongation will be disrupted by altering the expression of Elovl6 (Elongase of very long chain fatty acid-6) in both in vitro and in vivo models. Supporting this, increasing amounts of published data supports the role of fatty acid chain length in obesity and insulin resistance. In addition, our preliminary data suggests that Elovl6 is highly expressed in BAT and its expression increases with BAT activation.
Genetic investigation of life course phenotypes of mental health and illness in the 1946 British birth cohort. 02 Jul 2009
Using repeated measures collected between ages 13 and 53 years in the British 1946 birth cohort we will test associations between hypothesis-driven candidate genes and novel, empirically derived life-course phenotypes of depression and anxiety, cognition and personality. Our new phenotypes are longitudinal latent classes and latent quantitative traits derived from innovative psychometric approaches that we have piloted (Colman et al, 2007). Goal 1 confirms and 2 extends the phenotypes from m ental health to personality (neuroticism) and cognition ( g ). Goal 3 involves conducting single and multi-marker genetic association studies exploring the impact of sets of genes related to brain development, neurotransmission, cognitive ageing, and endocrine function on these summary measures of life-course risk for health and psychiatric symptoms, personality and cognition. Our overarching hypothesis is that these higher-fidelity, longitudinal measures will represent more valid targets for genetic research than conventional cross-sectional, binary diagnoses. Goal 4 considers the impact of three specified effect modifiers (G x E interaction). These include an early environmental adverse event (childhood separation), pubertal stage (in early teenage years) and a protective factor, educational/occupational attainment. This research will serve as a platform for thematically and methodologically linked collaborative work in other British cohorts, and other relevant samples.
What is the mechanism by which initiation factor eIF4GI, and its paralogue, p97 (DAP5), selectively promote translation of different subsets of mRNAs?. 25 Feb 2009
We have previously proposed that resumption of scanning and reinitiation following translation of a short uORF requires that the eIF4G/eIF3/40S subunit interactions (which were instrumental in promoting initiation at the uORF AUG) persist throughout the time taken for uORF translation; and that, by implication, the canonical initiation factors are sufficient, with no additional protein factor(s) required specifically for rescanning/reinitiation. We will use the defined in vitro system, consistin g of purified initiation, elongation and termination factors, defined aminoacyl-tRNAs, and ribosomal subunits to test these two propositions. In addition, working with this purified system and also unfractionated reticulocyte lysate (RRL) we will use formaldehyde fixation and sucrose gradient centrifugation to test directly for short-term persistence of the eIF4G/eIF3/ribosome interactions following 80S initiation complex formation. For p97, a priority will be to identify targets of p97 action (by using micro-array analysis of polysomal mRNA from cells in which p97 has been silenced). This will allow the design of appropriate dedicated reporters. The effects of recombinant p97, and its caspase cleavage product, p86, on translation of these dedicated reporters (compared with standard reporters) will then be studied in RRL or HeLa cell extracts, eIF4G-depleted versions of these systems, and the fractionated system.
Reading and writing the prolongation of life. 26 Mar 2009
This research examines the role of writing in seventeenth-century works on the prolongation of life. It examines the relationship between knowledge and the extension of experience presented by authors, and situates such projects within a wider natural philosophical endeavour to use artifice to overcome man s natural limitations. It proposes that what appear to be practical schemes for prolonging life are in fact about the feat of longevity that is made possible only by the artifice of writing. This research will offer a new understanding of the relationship between natural philosophy and medicine by providing a new reading of these works. It will show that these works conceive knowledge and long life to be fundamentally connected, and reads them as an intervention in contemporary debates about progress. These authors present prodigious old age as a feature of the experience accumulated in the library, rather than chronological precedence. In doing so, they make literary and i ntellectual endeavour rather than the arts the site of progress through experience. The key goal of this research is a monograph on feats of the library, bringing together my thesis research on extending experience through space with the proposed research on extending experience through time.
Chronic hepatitis (CH) in canines is frequently diagnosed and usually idiopathic although histological and clinical features are classical for a viral aetiology. Breed predilections indicate a genetic predisposition to disease, and some breeds show a particularly severe form of CH. CH in humans has multiple defined aetiologies (most commonly viral). Some idiopathic cases remain for which one or more novel viral agents are suspected. Histopathological lesions between a subset of these human cases an d idiopathic canine CH are strikingly similar. Hypothesising that canine CH is of viral aetiology, the key goals of the project are: 1. To identify a virus causing CH. 2. Investigate the MHC haplotypes. I will use state of the art PCR and high-throughput sequencing techniques to identify viruses present in severely affected tissues. Using serological techniques and specific PCR assays I will determine the correlation of infection and disease. Pedigree analysis and MHC haplotyping will be used to investigate the genetic correlates of disease susceptibility and severity.
Genetic and Cellular mechanisms underlying novel inherited disorders of insulin and IGF-1 action. 01 Apr 2009
Insulin and insulin-like growth factor 1 (IGF1) act via closely similar receptors and signalling pathways, yet exert distinct effects on whole body physiology, predominantly modulating fuel metabolism and growth respectively. Understanding this discrepancy is critical in view of the current pandemic of insulin resistance, and the importance of aberrant IGF1 signalling in oncogenesis. Our analytic approach to this question is to identify patients with rare single gene defects in insulin/IGF1 ac tion and to compare the physiological effects of the defined single gene defect with those seen in common forms of disease where the defect is unknown. Identifying the genetic defect in rare patients is the rate limiting step, and previous candidate gene approaches have had only limited success. This project now proposes to use a hybrid approach to identifying genetic defects in well characterised patients with extreme defects in both insulin action and growth. Positional cloning using high d ensity genotyping microarrays will be combined with signalling studies in primary cells, with adjunctive use of microarray transcriptomics and highly parallel sequencing technologies as required. When causative mutations are found they will be recreated and studied in vitro in cell culture models of insulin action.
Investigation of the role if transriptional repression in cell fate decisions during mouse development. 29 May 2009
Embryonic stem (ES) cells have two qualities that make them developmentally and clinically important: the ability to self-renew and the ability to differentiate into any embryonic cell type (pluripotency). A lot of effort has gone into defining molecular requirements of self-renewal, but little is known about how cells commit to differentiation and even less about how ES cells are derived. We have shown that the NuRD co-repressor complex required for the development of pluripotent cells in vivo and in potency of ES cells. This proposal builds on these findings to define the role of two major co-repressor complexes in derivation, potency and lineage commitment of pluripotent cells. We will identify epigenetic silencing events required for lineage commitment of pluripotent cells and define how silenced transcripts influence self-renewal, lineage commitment, and pluripotency. We will define molecularly the differences between ES cells and the embryonic cells from which they are derived and define the role played by epigenetic silencing in this process. We will biochemically define these complexes and identify their partners and targets in embryonic cells. Together this proposal comprises a molecular, biochemical, genetic and functional dissection of epigenetic silencing and cell fate decisions in pluripotent cells.
This program will address some of the fundamental unanswered questions about L-cell biology. L-cells produce hormones affecting glucose homeostasis and appetite, and are the target of current drug development programs for type 2 diabetes. The prediction is that stimulating GLP-1 release from L-cells will provide oral therapies that could be used alongside recently-licensed DPPIV inhibitors. Understanding L-cell physiology is critical to the success of this type of approach, but has been hindered by difficulties in identifying and culturing intestinal endocrine cells. We made transgenic mice with fluorescently labelled L-cells, and developed methodologies to characterise the cells by expression analysis, electrophysiology and imaging, providing a first opportunity to study primary L-cells at a single cell level. This technology will be used to probe the physiology of the L-cell using a range of electrophysiological and optical recording techniques, and to address key questions and hypot heses about L-cell function, such as that electrogenic transporters act as sensors of the luminal constituents, that L-cells from different gut regions are not identical and that the cells exhibit plasticity in number/function in an altered metabolic environment. In parallel, we will develop new technologies to study L-cells from humans, and downstream targets of GLP-1 in the intestine.
Interdisciplinary Training Programme for Clinicians in Translational Medicine and Therapeutics at the University of Cambridge: Support for an additional 2009 MPhil appointment. 31 Aug 2009
We propose an innovative training scheme for Translational Medicine and Therapeutics (TMAT) which builds on the exceptional conjunction on the Cambridge campus of leading scientists and clinical specialists, with an industrial research environment embraced both by international pharmaceutical and local biotech companies. Much of this is found under the same roof, the Addenbrookes Centre for Clinical Investigation (ACCI), with a track record of integrated training: academic with industrial, clinical with scientific, pharmacology & therapeutics with patient-based specialties. The novel TMAT programme will attract the brightest candidates at several levels of seniority, ranging from MB PhD students to clinical lecturers, some wishing translational skills in their chosen specialty, others not yet differentiated who may become future leaders and teachers of TMAT. Each trainee will have a customised programme. Part of this will be a bespoke, modular MSc modelled on the well-known small-group lectures and supervisions of the Cambridge final year undergraduate courses. However the centrepiece for most candidates will be a PhD including formal teaching in a wide range of translational and pharmacological skills, and a project which takes proof-of-concept studies in cell or animal systems forward to proof-of-concept studies in humans. We have assembled an outstanding faculty of PhD supervisors spanning a wide choice of skills and experience in basic and clinical science. All trainees will have the opportunity for hands-on exposure to the design and conduct of experimental medicine studies investigating the therapeutic potential of new drugs, in collaboration with our industrial partner, GlaxoSmithKline (GSK). Our product will be a new generation of clinician scientists with 360-degree vision of the complex landscape of modern therapeutic medicine, who can rise to the challenges and opportunities of 21st century drug development.
Core support for Wellcome Trust Centre for Stem Cell Research-award for supplementary funds. 22 Sep 2009
The new Institute for Stem Cell Biology in Cambridge will be an international centre of excellence in fundamental stem cell research. The Institute will focus on definition of the genetic and biochemical mechanisms that control stem cell fate, providing foundations for applications in disease modelling, drug discovery and regenerative medicine. This proposal is for provision of core resources for embryonic stem cell manipulation and transgenesis. A central resource of skilled personnel will maximise research productivity and continuity, promote cooperation and synergy, and accelerate technological innovation. Timely and efficient production of customised gene-modified stem cells and mice is essential underpinning. Specialised expertise will support advanced genetic engineering of mouse and human stem cells, and operation of robotic platforms to develop screening methodologies for isolating genetic, protein and chemical regulators. A dedicated PdD programme in stem cell biology will capitalise on the opportunity for high level research training provided by the intellectual environment and core facilities in the Institute. A Strategic Award will immediately establish the Institute for Stem Cell Biology amongst the best-resourced and most attractive environments for stem cell research world-wide, providing a magnet for recruitment, and a much-needed focus for UK and European stem cell biology.
British Society for the History of Science Postgraduate Conference 2010 to be held on 5-7th January 2010 30 Nov 2009
Network building, development of presentation skills and exchange of feedback are the central objectives of this meeting, to be supported by sessions encouraging early career scholars to consider their professional options. This meeting provides a supportive environment in which postgraduate students in the history of science, technology and medicine can convene. Delegates can practise their presentation skills outside their home institution, gain conference experience and meet peers from across the world. This not only creates a forum for feedback on work in progress, but also allows graduate students to develop their own professional network. Two skills sessions allow delegates to consult senior academics on questions relating to career development. These sessions focus on academic publishing and the museums sector, and will encourage participation from all delegates, including those not presenting a formal paper. A third postdoc-led session raises awareness of the outreach and education activities headed by the BSHS. These sessions encourage delegates to consider and investigate a range of career options. While the focus of the meeting is on encouraging interaction between graduate-level scholars, these three sessions and a.drinks reception will provide opportunity to discuss experiences with more experienced academics who may be able to offer advice.
From Generation to Reproduction. 14 May 2009
To reassess change and continuity in practices and meanings of 'generation' and 'reproduction', we will combine expertise in periods from antiquity to the present, and approaches from historical demography to history of science. Four teams will work in complementary research strands: 1. 'Patients and practitioners' will study the 'reproductive patient' to recover the wider patterns of intervention and concern. 2. 'Reproducing generations: conception and survival' will consider how matern al, fetal, infant and childhood health affected adult health and fertility, and the reproductive impact of sexual behaviour and venereal disease. 3. 'Representation and communication' will show how changing understandings of sex, development and evolution were produced, debated and used. 4. 'C20 transformations: technologies, experiences and regulation' will make a concerted attempt to account for the recent reproductive revolutions. Using research leave, research assistance and events, we w ill tackle the big questions, and produce high-quality publications (essay collections and single-authored books:Q14) and funding applications. An accessible, co-edited volume will provide a fresh vision for the whole field. We will continue to innovate in teaching and training, and expand a critical mass of expertise. We also propose enhanced public engagement, including 'The book of generation', a major exhibition at Cambridge University Library, and regular outreach activities (Q16).
Maintenance of the cell surface of trypanosomes. 01 Oct 2009
To investigate the roles of critical trafficking pathways underpinning the host-pathogen interface of trypanosomes, and specifically, aspects of the biosynthesis, maintenance and turnover of cell surface components. 1. To extend understanding of the role of ubiquitylation and peptide-based signals in targeting, internalization and turnover of membrane proteins, using TbAT1, the melarsoprol P2 transporter, as a model. 2. The examine the roles of ENTH/ANTH-domain adaptor protein family, AP18 0 and EpsinR, in sorting of the variant surface glycoprotein (VSG), invariant surface glycoproteins (ISGs) and other proteins. 3. To assess contributions of three Ras-like GTPases, TbRab21, TbRab23 and TbRab28, to endocytic trafficking and sorting, and address how these molecules interact with established endosomal and intraflagellar transport (IFT) pathways. 4. To assess the contributions of the exocyst complex to exocytosis of VSG, ISG and other proteins, and to explore the relationship bet ween exocyst function in vesicle trafficking and IFT.
The proposed Programme will investigate the neural and neurochemical substrates of compulsive behaviour in animals and in patients with obsessive-compulsive disorder (OCD). We argue that this cross-species approach will enable an analysis of the nature of the compulsivity (the tendency to perseverate in responding in the face of adverse consequences) and how it is mediated by fronto-striatal circuitry, modulated by monoaminergic systems. We hypothesise that compulsivity arises from core c ontributions from specific forms of executive dyscontrol, aberrant habit learning and anxiety, leading to such typical OCD symptoms as checking, illusory control, failure to complete , and harm avoidance. We will assess the particular role of two major fronto-striatal circuits in compulsive behaviour, and their modulation by 5-hydroxytryptamine (5-HT), dopamine and noradrenaline neurotransmitter systems, with implications for understanding current and future drug therapies. These circuits are o rbitofrontal cortex-medial striatum, and also a lateral prefrontal cortex-associative striatal circuit. This translational approach will allow us to gain further insight into the therapeutic effects of selective 5-HT agents in OCD. We maintain that OCD and its comparison with other disorders, such as adult Attention Deficit Hyperactivity Disorder, will also afford us an enhanced understanding of the executive functions of the frontal lobes.
Protein interactions and functional properties of P2Y1 nucleotide receptors in neurones. 08 Oct 2009
In previous and current work we have shown that the G-protein-coupled P2Y1 receptors (P2Y1Rs) for ATP are located in some CA1-3 pyramidal neurons, as is the scaffolding protein, NHERF-2, to which they can bind, and that they mediate inhibition of the neuronal M(Kv7)-current therein to increase neuronal firing. This is likely to be important because ATP has recently been reported to be constantly released onto central neurons by neighbouring astrocytes. We now propose: (1) to apply transgenic tec hnology to reveal the cellular location of P2Y1 and selected other identified P2Y subtypes in mouse brain in situ by pre-attached fluorophores; (2) to use these (with other approaches) to study (a) the neuronal function of P2Y1Rs by electrophysiology on identified P2YR-expressing neurons, (b) the sites of scaffolding protein NHERF-2 interaction with P2Y1Rs, (c) possible P2YR heterodimerization, and (d) to test the hypothesis (based on observations on hippocampal neurons and sympathetic neurons, which are devoid of NHERF-2) that NHERF-2 serves to confine P2Y1Rs (and possibly some mGluRs) to neuronal microdomains which facilitate coupling to M(Kv7)-channels and preclude coupling to Ca2+-channels and Kir(GIRK) K+-channels. If true, this will have major implications for neuronal receptor-ion channel regulation and pharmacology through membrane-linked microdomains.
Insulator elements are proposed to play key roles in the organisation of the eukaryotic genome and in gene regulation. In this project we will investigate the association between insulator proteins and the regulation of gene expression. Using ChIP-array to map the binding profiles of insulator components in specific tissues we will examine insulator protein binding to genes in different states of activity. This will identify which insulator complexes, and which specific insulator components, are constant, contributing to the fixed architecture of the genome, and which vary with gene expression state, highlighting a role in the regulation of gene expression. We will use functional assays to test these roles and we will link insulator binding to changes in chromatin configuration in different activity states. We will focus on the Ubx Hox gene and will use the 4C chromatin conformation assay to investigate the configuration of the Ubx gene both the ON and OFF states. Having both a h igh resolution map of chromatin configuration and a detailed profile of insulator protein binding in differing states of gene expression will give us an unprecedented view of the association between insulator complexes and the regulation of gene expression.
Fragment-based approaches to the design of candidate drugs that interrupt protein-protein interactions involved in cell regulation 06 Nov 2009
The use of fragment-based drug discovery to identify novel drug candidates that modulate the BRCA2-RAD51 interaction for the treatment of cancerThe Trust has awarded over £2.4 million to Chris Abell, Tom Blundell, Marko Hyvonen, Grahame McKenzie and Ashok Venkitaraman at the University of Cambridge to use fragment-based approaches to design and make molecules that disrupt the interaction of two important proteins in human cells, the recombinase RAD51 and the product of the breast cancer-associated gene BRCA2.These proteins are involved in the repair of DNA breakage, and blocking their interaction should result in sensitization of cancer cells to DNA damage, e.g. by radiation, or directly induce cancer cell death during proliferation. Potent compounds will be developed by synergistic use of X-ray crystallography and synthetic organic chemistry, and improved in a highly focused way to make sure they are safe and suitable for development into possible drug candidates. The lead compounds will be tested against different cancer cell lines to identify susceptible cancers, the most probable therapeutic target being lung cancer. This funding follows on directly from a Translation Award to pioneer the use of fragment-based approaches against protein-protein interactions. That project established the use of biophysical methods, especially NMR spectroscopy and X-ray crystallography to identify fragments that bound at specific sites on a protein interface and to iteratively grow these fragments into successively more potent compounds.
My studies of Lrig1, a negative regulator of receptor tyrosine kinases, have demonstrated a link between stem cell self-renewal and receptor activity. I will establish whether the effect of Lrig1 on stem cell behaviour is a general feature of receptor activation and Lrig family members. Lrig3 will be assessed for its role in vivo in epidermal homeostasis and the effect of receptor tyrosine kinase inhibition and activation will be investigated in vitro. Regulation of receptor kinase activity is e ssential for homeostasis and these aims will help us understand disease progression. I have demonstrated that Lrig1 defines murine interfollicular epidermal stem cells. I will use expression profiling to define the unique properties of Lrig1 expressing cells and perform lineage analysis to establish their role in homeostasis and repair. This will require the generation of a Lrig1::dsRed-IRES-CreERT2 reporter mouse. My studies demonstrate that Lrig1 governs stem cell quiescence. To address whe ther hyperplasia in Lrig1 KO mice is caused by increased stem cell contribution, I will perform in vivo clone assays. The combination of these aims will define the role of Lrig1 and interfollicular epidermal stem cells. This will provide a unique tool to understand how these cells participate in epidermal disorders.