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Recipients:
University of Cambridge
Amounts:
£500 - £1,000

Results

The role of FAPa-expressing stromal cells in therapeutic vaccination in a mouse model of pancreatic ductal adenocarcinoma. 15 Feb 2010

Mesothelin as a potential target for immunotherapy- project aim 1 The fir.;t objective of this project is defining an antigen that could be used in the vaccine. Several TAA have already been described to be uniquely expressed in human pancreatic cancer and not in the normal tissue. On~ of the most promising candidates is mesothelin - GPI anchored differentiation antigen that was found in all mesotheliomas an~ pancreatic aden9carcinomas (22), and approximately 70% of ovarian cancer.; and 50% of lung adenocarcinomas [23]. Additionally, mesothelin expression was restricted to pancreatic adenocarcinomas, whereas adjacent normal pancreas did not stain for mesothelin. Mesothelin as an antigen has already been used in allogeneic granulocyte-macrophage colony-stimulating factor (GM-CSF)-secreting tumour vaccines to treat patients with surgically resected adenocarcinoma of the pancreas (24). 3 out of 14 patients developed post-vaccination delayed-type hypersensitivity (OTH) response to autologous tumour, which was associated with prolonged survival. To find whether mesothelin is also expressed in KPC mouse model we isolated RNA from lung, pancreas from healthy mice and POA from tumour-bearing animals. Our preliminary qPCR data suggest that 3 out of 5 tumour.; that were examined showed high mesothelin RNA level (Fig. 3). In order to verify this result on protein level tumour sections will be stained with anti-mesothelin antibody. However, as there are few antibodies (Ab) directed against mouse mesothelin, it is required to screen commercially available Ab and validate then on western blot. Preliminarily, cancer cell lines isolated from PDA tumours can be used for those tests. Cancer/Testis antigens as potential targets for immunotherapy- project aim la (contingency plan) Mesothelin is not the only potential PDA-associated antigen. Cancer/testis antigens (CTA) are immunogenic molecules expressed in normal tissues, but restricted to placenta, testis, fetal ovary, germ cells and placenta, as well as in a wide variety of tumours. There are approximately 150 CT As identified to date (www.cta.lncc.br) strictly regulated by epigenetic mechanisms. Vaccines containing the CT antigens- MAGEA and NY-ES0-1/CTAGIB- are utilized in patients with melanoma as well as lung and ovary cancers. Generation of viral vectors for immunization- Vaccinia-project aim 2 In this project, to mount an effective anti-tumour T cell response, a vaccine strategy utilizing ~ poxvirus as TAA delivery vehicles in combination with T -cell costimulatory molecules will be introduced. The attenuated strain of Modified Vaccinia virus Ankara (MV A) is widely used. as a safe non-replicating recombinant vaccine vector in humans and other mammals. Among the first TAA successfully used for immunization with VV are carcinoembrionic antigen (25), prostate-specific antigen [26] and many melanoma-associated antigens. Generation of viral vectors for immunization-gamma-herpesvirus- project aim 2 Herpesviruses persist latently in the infected host indefinitely effectively evading the immune response. Sporadic reactivations lead to release of infectious virions and facilitate spreading to new host. As y-herpesvirus (?HV) reactivation is only observed in immunosuppressive conditions, it suggests that immune system is able to control the infection. It was demonstrated that more potent antiviral response mediated by antigen-specific COS+ T cells was found in persistent ?HV infection [27)[28). Moreover, during latent infection with ?HV -68 increased turnover rate of CD8+ T cells indicates that these cells are restimulated with antigen. Using recombinant methodology sequence of the antigen of interest will be inserted in place of ovalbumin in both vectors. Infection of LL20VA-bearing mice with ?HV -OVA maintained high level of SIINFEKL-specific CD8+ T cells (data unpublished), which controlled tumour growth when F APa ablation was performed. Assessment of immune response to vaccines project aim 3 Before performing therapeutic vaccination on tumour-bearing mice it is necessary to evaluate the effect of immunization. Cellular response will be assessed by performing killing assay or by studying the profile of major cytokines (IL-2, IL-4, IL-10 and IFN-y). In order to evaluate CDS+ T cell killing antigen-expressing cell line with matching MHC will be used and chromium release will be monitored to detect lysis. EVALUATION OF TUMOUR GROWTH- PROJECT AIM 4 For the purpose of measuring tumour growth, tumour volume will be quantified by 30 ultrasonography [ 17]. 30 ultrasound provides a new method for the objective quantitative assessment of stromal echogenicity and vascularity as well as blood flow within the tumour. Moreover, the biggest advantage of this technique is its non-invasiveness, which enables assessment of the effects of therapy without sacrificing animals. This will be done in collaboration with Tuveson's lab.

Amount: £21,929
Funder: The Wellcome Trust
Recipient: University of Cambridge

Transgenic animals: Genetically modified mice in Britain and the United States, 1970-2000 10 Jun 2010

To explore the history of GM mice, I will begin by studying technical innovations in mammalian embryology and reproductive physiology in the 1970s. In particular, I will consider how the techniques travelled, through multiple exchanges between laboratories in Britain and the US, and how this research was variously combined with recombinant DNA technologies developed in the 1970s to produce multiple transgenic mice in 1980-1. Then I will explore the parallel lines of research on embryonic carcinoma (EC) and embryonic stem (ES) cells in the 1970s and 1980s. To assess the effect of the new kinds of animals on the infrastructure of their breeding and supply, and the stabilisation of their production, I will use as a case study the MRC Radiobiology Unit (Mammalian Genetics Unit since 1995) in Harwell, a stock centre and research facility for mouse experimentation, in the 1980-1990s. I will pay special attention to criteria for genetic modification and negotiations over the status of the mice. Finally, to analyse the (contested) use of GM mice as models of human disease, I will consider the induction of breast cancer through oncogene insertion.

Amount: £85,708
Funder: The Wellcome Trust
Recipient: University of Cambridge

Clinical PhD Programme at the University of Cambridge: 'Functional characterisation of responses to DNA damage in human cells.' 21 Sep 2010

Failure of accurate DNA damage sensing and repairing mechanisms manifest as a variety of human diseases including cancer. Cytotoxic chemotherapy and radiotherapy remain the most beneficial in terms of cancer treatments and largely act through the generation of DNA damage. Accuracy and efficiency of DNA damage detection and repair requires the recruitment and subsequent post-translational modification of various proteins. Ubiquitylation and sumoylation are highly dynamic and reversible enzymatic processes that play crucial roles during the execution of DNA damage sensing and repair. Further characterization of the enzymes involved in these processes is required in order to fully understand how the DNA damage response (DDR) is regulated and has the potential to provide new therapeutic targets. PIAS SUMO E3 ligase enzymes are necessary for human cells to respond appropriately to and effectively repair DNA double-strand breaks. ZIMP7 and ZIMP10 are two PIAS -like proteins that also contain a SP-RING/Miz zinc finger domain that potentially confers E3 ligase activity. Very little previous research has been focused on these proteins. I have demonstrated that both ZIMP7 and ZIMP10 are recruited to sites of DNA damage induced by laser micro-irradiation in cultured human cell lines. My goal now is to characterize the precise functions of ZIMP7 and ZIMP10 in the DDR.

Amount: £221,618
Funder: The Wellcome Trust
Recipient: University of Cambridge

Clinical PhD Programme at the University of Cambridge: 'Defining the origin and fate of Fibroblast Activation Protein (FAP) expressing cells in health and disease'. 21 Sep 2010

A population of tumour stromal cells, identified by Fibroblast Activation Protein (FAP) expression, are essential for suppressing the immune response directed toward the tumour. FAP+ cells are present in inflamed tissues, in healing wounds, during embryological development and recent work from our laboratory has found them in a wide range of normal adult tissue, including muscle, bone marrow and the kidney. Depleting FAP+ cells in mice leads to significant weight loss and skeletal muscle loss. Clearly FAP+ cells have a fundamental role in both health and disease. Their function in different tissues is not clearly defined but raises the question of whether they are all related. Are all FAP+ cells derived from a common lineage? Understanding the origins and fates of these cells will have significant implications for developing effective cancer therapies but may also provide evidence of a cell linking inflammation, healing and tissue specific regeneration. Key goals 1. Generate a BAC Transgenic (Tg) mouse in which FAP+ cells, and all their progeny, can be conditionally marked to express yellow fluorescent protein in vivo. 2. Use the Tg mouse to determine whether FAP+ cells in the tissues are derived from a common embryonic precursor and whether FAP+ cells can become FAP- in vivo. 3. Define the origins of FAP+ cells in acutely and chronically inflamed tissues.

Amount: £240,390
Funder: The Wellcome Trust
Recipient: University of Cambridge

Bayesian inference using a norm based face coding scheme in humans 21 Sep 2010

Recently large amounts of evidence have accrued suggesting that the human brain often supports optimal Bayesian inference, but it has not been shown that this is so in face recognition. In parallel a leading hypothesis for the encoding of faces is the so-called norm coding scheme, whereby each face is defined by its position in face-space. In face-space the average face sits at the origin, and, in a Bayesian interpretation of inference in this context, the peak of the prior probability distribution should correspond to this rather than another face. By using a multivariate analysis, sensitive to the fine structure of fMRI responses, we hope to characterise these implicit probability distributions inthe aIT (anterior inferotemporal cortex) of human subjects, and show that whensubjects hallucinate a face in noisy stimuli this is the average face ratherthan another. This supports a Bayesian interpretation of the norm based scheme, since otherwise any face might be hallucinated . Finally, we aim to show that while face and anti-face (its opposite in face space) can be differentiated in aIT (the face identifier), this cannot be donein the fusiform face area (the face detector).

Amount: £259,443
Funder: The Wellcome Trust
Recipient: University of Cambridge

The epidemiology of glaucoma. 21 Sep 2010

Glaucoma is one of the leading causes of blindness worldwide, second only to cataract. However, unlike cataract, visual loss from glaucoma is irreversible.Uncorrectable loss of vision is associated with threats to independent living,safety and emotional well-being, resulting in increased dependence on social and community services. It has been estimated that if 10% of glaucoma sufferers received earlier treatment that arrested significant visual loss, the United Kingdom economy would benefit by £555 million. Currently, intraocular pressure (IOP) is the only known modifiable risk factor for glaucoma, and all treatment modalities are aimed at reducing IOP. Identification of lifestyle factors associated with glaucoma may enable publichealth strategies as well as new treatments to help prevent this disabling condition. EPIC-Norfolk is an established population study which this project will form part of. Extensive lifestyle data from multiple time points are available and the last health check included a comprehensive ophthalmic examination. The primary goals of this project are to: - Update the definition of glaucoma- Describe IOP, optic nerve measures and glaucoma in the EPIC-Norfolk cohort -Determine nutritional, lifestyle and biological measures associated with the presence of glaucoma and associated physiological measures.

Amount: £361,924
Funder: The Wellcome Trust
Recipient: University of Cambridge

Clinical PhD Programme at the University of Cambridge: 'Early life immune and metabolic function and psychotic symptoms in the ALSPAC birth cohort.' 21 Sep 2010

Schizophrenia is associated with proinflammatory changes (increased cytokines in serum and CSF). Recent evidence suggests important effects of hormone leptin on immune system and cognition. Cognitive dysfunction is also prominent in patients with schizophrenia. However, prospective studies are required to establish importance of inflammatory cytokines and their modulators, such as leptin in the pathophysiology of schizophrenia. Increased risk of schizophrenia is also observed among individuals exposed to immune activation or infection during early life. There is evidence, inflammatory cytokines can increase fetal exposure to maternal glucocorticoids leading to long term changes in hypothalamic-pituitary-adrenal (HPA) axis functioning. Alterations of HPA axis functioning by early life environmental factors may be a common link between chronic diseases of adult life, such as hypertension, type 2 diabetes and schizophrenia. Besides, schizophrenia is associated with subtle alterations in motor and cognitive development early in life. Early environmental factors such as immune activation may interfere with neurodevelopment, and thus contribute to disease risk. However, effects of early life immune dysfunction on human neurodevelopment are yet to be explored. We propose a study of early life (pre and post natal) immune activation and risk of psychotic symptoms, and motor and neurocognitive development in ALCPAC birth cohort.

Amount: £224,505
Funder: The Wellcome Trust
Recipient: University of Cambridge

Wellcome Trust PhD Programme for Clinicians at the University of Cambridge: Investigating rare copy number variants (CNVs) associated with severe early-onset obesity. 14 Jun 2010

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.

Amount: £4,373
Funder: The Wellcome Trust
Recipient: University of Cambridge

Wellcome Trust PhD Programme for Clinicians at the University of Cambridge: 'Role of fatty acid chain length on energy balance and adaptive thermogenesis.' 14 Jun 2010

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.

Amount: £38,568
Funder: The Wellcome Trust
Recipient: University of Cambridge

Wellcome Trust PhD Programme for Clinicians at the University of Cambridge: 'Natural and modified variations in human induced pluripotent stem cells'. 14 Jun 2010

Human induced pluripotent stem cells (hiPSCs) have revolutionized stem cell biology and have renewed aspirations of moving the field towards clinical therapies. By genetic manipulation, fibroblasts have been shown to re-program to a pluripotent state similar to human embryonic stem cells (hESCs). This would allow for patient specific hiPSCs, generating models for disease as well as immune compatible cell therapies. A major problem in the field is heterogeneity in the derived hiPSCs both in derivation as well as differentiation. Additionally it is unknown whether hiPSCs may be made using cells other than fibroblasts and if these hiPSCs retain an ?epigenetic? memory following differentiation. The knowledge gained from the basis of hiPSC variability will be used to optimise strategies towards clinically relevant therapies. Key goals: 1) Generate hiPSCs from different human tissues 2) Compare hiPSC lines derived from the same individual and so same genetic background 3) Identify new lineage-specific markers through gene expression arrays 4) Explore the differentiation capacity and bias of the different hiPSC lines 5) Investigate epigenetic ?memory? in derivation of hiPSCs and subsequent differentiation 6) Use knowledge of the basis of variability to optimise differentiation strategies 7) Gain robust scientific training in stem cell biology and genetics

Amount: £30,392
Funder: The Wellcome Trust
Recipient: University of Cambridge

Wellcome Trust PhD Programme for Clinicians at the University of Cambridge: 'The cell biological and developmental roles of microcephaly-associated genes within the developing zebrafish central nervous system'. 14 Jun 2010

My project aims to investigate the cell-biological basis of abnormal neurogenesis and reduced brain size caused by mutations in genes associated with microcephaly including ASPM, CDK5RAP2, CENPJ, STIL, MCPH1 and PCNT. These genes all encode centrosomal proteins and they are thought to be important in the control of neural proliferation, although their precise roles in central nervous system (CNS) development and regulation of brain size are not known. To investigate this question I am working on embryonic Zebrafish retina as a model for the developing CNS due to its experimental accessibility and amenability to genetic manipulation as well as in vivo confocal time-lapse imaging. FolIowing morpholino-induced gene knockdown or expression of dominant negative constructs I will examine in detail their cell biological roles. These may include regulation of the symmetry of cell divisions and/or cell cycle dynamics. These studies will contribute to our understanding of human microcephaly and nervous system development.

Amount: £8,385
Funder: The Wellcome Trust
Recipient: University of Cambridge

Developing models of thalamocortical unsupervised attentional selection and competitive learning 15 Jul 2010

The project will aim to establish the following. (1) The characteristics of a modular neural network architecture for unsupervised learning in which each module receives weighted predictions (expectations) and bottom-up activation, calculating a 'performance' output and a mismatch error. (2) The impact of introducing global competition for learning based on locally derived prediction error. (3) The subsequent effects on learning rates of introducing biologically based synaptic autostabilization mechanisms. (4) The networks' performance in simple learning-theory paradigms and in perceptual identification with prior information. (5) The unification of such models with methods for implementing high-speed dynamic binding. (6) The effects of network-wide parameter manipulations, including manipulations relevant for psychopathological states such as hallucinations. (7) The simulation of a large-scale version of the computational network and analysis of its topological and statistical properties, for comparison with existing MRI analyses. (8) The simulation of a three-dimensional version of the network and the derivation of predictions relating to surprise in learning, for comparison of the model's predictions with MEG experiments in normal humans and then patient groups.

Amount: £194,487
Funder: The Wellcome Trust
Recipient: University of Cambridge

Defining the impact of inflammatory bowel disease associated SNPs on the immunobiology of human mucosal T lymphocytes 15 Jul 2010

AIM: The phenotypic analysis of intestinal T lymphocytes including functional studies of genetic polymorphisms associated with Crohn's disease. Lymphocytes will be extracted from blood and intestinal biopsies from healthy individuals stratified for two key SNPs identified from GWA studies as associated with IBD susceptibility. One SNP is a coding variant in the IL23 receptor, involved in differentiation of T lymphocytes to IL17 production, and one a non-coding variant associated with chemokine receptor gene CCR6, involved in T lymphocyte chemotaxis and expressed by IL17 producing cells. Lymphocytes will be characterised and sorted by polychromatic flow cytometry based upon expression of coreceptors defining T lymphocyte sub-populations, including unconventional populations that we suggest play a key immunoregulatory role, and simultaneously analysed for chemokine receptor expression. Functional studies will examine CCR6 driven chemotaxis and IL17 differentiation in sorted mucosal and systemic T lymphocytes. cDNA from sorted lymphocyte subpopulations will be analysed for chemokine receptor and cytokine expression by qPCR and allele specific expression. Key goals include the definition of chemokine receptor and cytokine expression by populations of unconventional mucosal lymphocytes, and assessing the phenotypic and functional impact of the two Crohn's-associated SNPs on mucosal lymphocytes including the homing and cytokine production of unconventional subpopulations.

Amount: £268,900
Funder: The Wellcome Trust
Recipient: University of Cambridge

Structural Cell Biology of transport vesicle and organelle biogenesis. 10 Jun 2010

The identity and function of cellular membranes are largely defined by their transmembrane protein composition. Transmembrane protein cargo of an enormous variety of functions is moved between the cell's organelles and its limiting membrane in coated transport vesicles of which clathrin-coated and COPI-coated vesicles are the most common. Coated vesicle formation requires the complex interplay of many cytoplasmic proteins, the membrane itself and the many types of transmembrane protein cargo th at need to be selected for inclusion into a vesicle. When the cell wants to degrade transmembrane or an extracellular proteins, they are delivered to a late endosome, which subsequently fuses with the cell's degradative enzyme store, the lysosome. The HOPS, AP3 and retromer complexes, VARP and many of their binding partners are important players in the critical processes that produce a fully functional late endosomes and lysosomes. We will use an integrated combination of structural studies ( X-ray crystallography and electron microscopy) combined with in vitro and in vivo studies to try and understand how general cargo and SNAREs are selected for incorporation into clathrin, COPI and retromer-coated vesicles and also how the formation and fusion with target membranes of late endosomes, lysosomes and secretory lysosomes are controlled.

Amount: £3,273,391
Funder: The Wellcome Trust
Recipient: University of Cambridge

Statistical methods for the study of molecular mechanisms of disease 14 Jun 2010

Genome-wide association (GWA) studies have been very successful in pointing to genetic loci associated with risk of type 1 diabetes (T1D) and other diseases. We knew only two or three such loci for T1D when I took up my fellowship at the end of 2000 while now we have 45. The pace at which new data has been generated in the last two to three years has been such that some analysis remains to be done on existing datasets. Examples include pathway?based analyses and meta-analysis of results for multiple autoimmune diseases. But GWA studies cannot identify all disease susceptibility loci. In particular, low frequency variants with larger effects are certain to occur and may prove more valuable for further study of disease mechanisms. Advances in high-throughput sequencing and genotyping arrays promise to allow us to extend the spectrum of frequency and types of disease variants which we can identify. But statistical problems will be faced in the design and analysis of such studies. I would hope to contribute to the solution of these problems.

Amount: £307,306
Funder: The Wellcome Trust
Recipient: University of Cambridge

Signalling in T-cell immunity 10 Jun 2010

An immune response against opportunistic infections, foreign transplants and in autoimmune disorders is critically dependent on the generation of intracellular signal by the antigen-receptor (TcR?/CD3) and the co-receptors (CD28, CTLA-4) on the surface of T-cells. Key mediators of signalling include CD4/CD8-p56lck complexes, the tyrosine kinase ZAP-70 and a novel family of immune specific adaptor proteins. In this application, we propose to extend our studies on the nature of protein-tyrosine kinases and adaptors that regulate cytokine production in T-cells. Specifically, we propose to focus on the adaptors FYB and SKAP-55, their phosphoryation sites, interactions with other proteins, nuclear vs. cytoplasmic function and their regulation of thymic differentiation and peripheral T-cell function. Likewise, we propose to elucidate the molecular mechanisms that regulate CD28 positive vs. CTLA-4 negative signalling, and their connection with the TcR?/CD3 complex. Ultimately, we hope to provide insights that will allow for the development of therapies designed to enhance immune responses against infectious agents, and dampen inappropriate responses in autoimmune disorders.

Amount: £320,175
Funder: The Wellcome Trust
Recipient: University of Cambridge

Signalling in T-cell immunity. 10 Jun 2010

The overall goal of this application is to gain a better understanding of the signalling mechanisms that regulate T-cell immunity. This understanding is needed ultimately to understand the molecular basis of T-cell function and to develop new strategies for the treatment of immune deficiencies, graft vs. host responses and autoimmune disorders. This project is divided into four aims: Aim #1: ADAP-SKAP1 regulation of inside-out pathway and immune function; Aim #2: Define the basis of ADAP-M12 b lockade of HIV-1 transcription; Aim #3: SLP-76-RanGAP and the nuclear pore complex and Aim #4: The reverse-stop signal model and chemokine reversal of CTLA-4 anergy.

Amount: £2,101,730
Funder: The Wellcome Trust
Recipient: University of Cambridge

Regulation of self-renewal and differentiation in the Drosophila CNS. 01 Jul 2010

The Drosophila nervous system is an excellent model system in which to analyse the molecular mechanisms controlling stem cell proliferation and differentiation, often at single cell resolution. We will use classical genetic and reverse genetic approaches, in combination with transcriptional profiling of small groups of stem cells and genome-wide DNA-binding analysis, to identify the molecular switches involved in the transition from symmetric to asymmetric division, and from self-renewal to diff erentiation. We will: 1) characterise the genetic switches that convert a symmetrically dividing neural stem cell into an asymmetrically dividing cell 2) identify the genetic networks leading from a neural stem cell to a differentiated neuron 2) test the role, in vivo, of key members of this network by targeted RNAi and screen for brain tumours (neuroblast overproliferation) or small brains (premature differentiation) 2) identify co-factors that modify the activity of the cell fate determi nant, Prospero, enabling it to repress stem cell genes and activate differentiation genes 4) identify the regulatory loops controlling the temporal progression of neurogenesis.

Amount: £1,183,145
Funder: The Wellcome Trust
Recipient: University of Cambridge

The Road from Pluripotency to Lineage Determination. 22 Apr 2010

Pluripotency is the capacity of individual cells to initiate all lineages of the mature organism in response to signals from the embryo or cell culture environment. Pluripotency has no predetermined programme; it is a tabula rasa. This is the foundation of mammalian development and of embryonic stem (ES) cell biology. Genetic and cell biological studies point to transcription factor command rather than epigenetic governance of the pluripotent state. Persuasive support for this view comes from th e demonstration that pluripotency can be recreated from somatic cells through transcription factor induced reprogramming. We now wish to investigate the other side of this coin; how pluripotent cells exit from a naive ground state and become committed to different lineages. Our hypothesis is that fibroblast growth factor stimulation of the mitogen activated protein kinase cascade perturbs the ground state and creates a metastable condition in which cells are poised for multilineage commitment. W e aim to isolate cells in this transition state and interrogate their regulatory circuitry at both population and single cell levels using quantitative transcriptomics, proteomics and biochemistry, complemented by genetic manipulation and live cell tracking. Through these studies we hope to deepen understanding of the underlying design principles of pluripotency lineage choice.

Amount: £1,736,222
Funder: The Wellcome Trust
Recipient: University of Cambridge

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.

Amount: £1,009,035
Funder: The Wellcome Trust
Recipient: University of Cambridge