NIHRDataInsights/HRCSData huggingface dataset:
Title: Dkk1-Wnt signalling pathway in synapse degeneration: implication for early stages of Alzheimer's disease
Funder: Medical Research Council

Abstract: Deficient Wnt signalling has been shown to contribute to Alzheimer's disease (AD). Given the role of Wnts at the synapse, we propose that the secreted Wnt antagonist Dkk1, which is elevated by A-Beta, contributes to synapse failure in AD. Here we will examine the mechanisms responsible for synapse vulnerability and their impact to AD. We address the following specific aims: 1) To investigate the contribution of downstream components of the Wnt pathway in Dkk1-mediated synapse degeneration. Using a candidate approach, we will study key synaptic components of the Wnt pathway, the LRP6 receptor and the scaffold protein Dishevelled-1 (Dvl1). LRP6 is blocked by Dkk1 and has been linked to late onset AD in a genome wide association study. Moreover, Dvl1 is required for excitatory synapse formation and function. Here, we will evaluate the role of these proteins in synapse vulnerability. 2) To identify novel mechanisms contributing to Dkk1-mediated synapse vulnerability. Proteome and transcriptome analyses will be used to identify proteins that contribute to Dkk1-mediated synapse degeneration. Candidate molecules will be selected for functional studies in primary neuronal cultures. In addition, these molecules will be tested for their contribution to Dkk1-mediated synapse vulnerability. 3) To examine the in vivo role of Dkk1 target molecules. The most highly selected candidates identified in our screens will be chosen for in vivo studies. Gain and loss of function of selected candidates will be examined using viral vectors injected into the adult hippocampus of wild-type and transgenic mice expressing Dkk1. 4) To investigate the role of the Dkk1 target molecules in AD models. Behavioural studies will be performed using the most highly selected candidates from our screen to investigate the ability of these new molecules to modulate synapse degeneration in AD animal model.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: R Idro, Makerere University - The pathogenesis and treatment of nodding syndrome
Funder: Medical Research Council

Abstract: Nodding syndrome (NS) is a devastating epileptic encephalopathy of an unknown cause affecting thousands of children in Africa. Our goal is to understand the pathogenesis and develop specific interventions. The objective is to examine if NS is a neuro-inflammatory disorder induced by Onchocerca volvulus or its symbiotic bacteria, Wolbachia and whether doxycycline may be used as treatment. The specific aims are: Aim 1: Determine the relationship between NS, antibodies to host neuron surface proteins (NSPs) or leiomodin and O.volvulus. I hypothesise that NS is an O.volvulus or Wolbachia-induced epileptic encephalopathy with antibodies against O.volvulus or specific sero-groups and variant species of Wolbachia cross-reacting with host NSPs/leiomodin. We will recruit 165 cases and 165 sibling controls, obtain blood and CSF (cases only) and examine: a) the relationship between NS and antibodies to leiomodin, the voltage-gated potassium channel complex and other surface proteins; b) in cases, examine for neuro-inflammation in CSF; c) determine the relationship between these antibodies and inflammatory markers, microfilaria density, Wolbachia sero-groups and disease severity, epileptiform discharges on EEG and structural changes on brain MRI. Aim 2: Conduct a pilot study to examine the role of Onchocerca by targeting the adult worm. There is no routine treatment for adult Onchocerca. However, antibiotic depletion of Wolbachia results in marked reduction in microfilaria density, sterilisation and premature death of the adult worm. I hypothesise that this will lead to reduced markers of inflammation and potential subsequent clinical improvement. In a proof of principle pilot study I will randomise 152 patients to either doxycycline or placebo and determine the effect on levels of antibodies to NSPs or leiomodin, disease severity, inflammatory markers, microfilaria density and Wolbachia load at 6 and 24 months.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;6.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: Investigating the role of Myosin VI in quality control of mitochondria linked to Parkinson's disease pathology
Funder: Medical Research Council

Abstract: In this project we will investigate the molecular function of the actin-based motor protein myosin VI in mitochondrial quality control. Defects in mitochondrial turnover are closely linked to Parkinson's disease (PD), the second most common neurodegenerative disorder. A hereditary form of PD is caused by mutations in the E3 ubiquitin ligase Parkin; this protein has been shown to mediate selective autophagy of damaged mitochondria, called mitophagy. Several lines of evidence and data from published large-scale whole genome studies suggest a functional link between Parkin and myosin VI and its cargo adaptors. To test this hypothesis and analyse the link between myosin VI and Parkin in more detail we will use in situ proximity labeling and a combination of confocal fluorescence microscopy, live cell and super-resolution microscopy as well as electron microscopy. To determine the requirement of myosin VI and its adaptor proteins for Parkin-mediated mitophagy, we will use siRNA KD cells, KO cells created by CRISPR/Cas9 genome editing, primary fibroblasts and neurons from our myosin VI KO mouse as well as iPS cells generated from human PD patient fibroblasts. In addition we will employ multiple approaches to assess whether loss of myosin VI or its adaptor proteins affects mitophagy and thereby leads to an accumulation of damaged mitochondria with reduced function.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: How do astrotactin-1 and astrotactin-2 act in the determination of mammalian cell polarity?
Funder: Medical Research Council

Abstract: This grant application focuses on the use of structural biology, biophysics and live cell imaging to study the structures of astrotactins (ASTNs) 1 and 2 and the interactions they undergo both with each other and with other proteins within cells and tissues. Recent data indicate that the role of ASTN1 in controlling cerebellar granule neuron migration depends on ASTN2 which drives its recycling via endocytosis between successive interstitial junctions with glial fibres. In other data, our collaboratoing Project Partners in the Nathans lab (Johns Hopkins) have found a link between ASTN2 and planar cell polarity (PCP) determination. This makes our proposed study extremely timely and its medical importance can be gauged from the involvement of ASTNs - as assessing by largescale clinical and genetic studies - in conditions as diverse as ADHD, schizophrenia and Alzheimer's, beside non-neuronal effects. Having already crystallised and partially solved the structure of the ASTN2 ectodomain we will complete the X-ray structure determination at neutral and then at acidic pH and then make use of biophysical methods to study its self-interaction. We will also solve the structure of the cytoplasmic ASTN2 N-terminal domain, using crystallography or NMR. The absence of exons 4 and 5 from the NTD has been noted in normal tissue and a PCP variant, respectively. The structure of ASTN1 will then be solved, again using crystallography for the ectodomain and crystallography or NMR for the NTD, also studying its interactions with ASTN2 biophysically. We will use live cell studies with FRET measurement incorporating lifetime imaging to look at the interactions undergone by ASTN1 and ASTN2 in a cellular context, including how this is affected by their cycling through the endosomal system, and also aim to extend our work into investigation of the interaction of ASTNs with proteins which are components of PCP signalling.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: How does the cell distinguish between coding and non-coding RNAs in the cytoplasm?
Funder: Medical Research Council

Abstract: This project will elucidate the mechanisms by which cells distinguish between non-coding and coding RNAs in the cytoplasm. Whilst the majority of our genome is transcribed, only a small fraction is protein-coding. This implies that many non-coding RNAs exist, some of which are similarly processed to mRNAs, termed long non-coding RNAs (lncRNAs). The majority of characterised lncRNAs are nuclear but recent ribosome profiling results have revealed that many are translated. Although controversial, it is clear that numerous lncRNAs are cytoplasmic, polysome-associated and translated. The line between coding and non-coding has become blurred and some lncRNAs are really mRNAs. I hypothesise that cells can differentiate between coding and non-coding RNA, by virtue of the different proteins bound. I will identify what these molecular markers are and investigate their biological importance. LncRNAs are enriched in neuronal tissues and several have been found associated with neurological conditions e.g. Alzheimer's disease. An inability to differentiate between coding and non-coding RNAs will have profound effects on cellular health. To better understand the molecular basis of neuronal disease it is vital that we appreciate how coding and non-coding RNAs are distinguished by the cell and the role of cytoplasmic lncRNAs in regulating gene expression. I developed Poly-Ribo-Seq to identify regions of active translation with an improvement to ribosome profiling and discovered which lncRNAs are translated in Drosophila S2 cells. Using cultured human neurons, I will now employ Poly-Ribo-Seq to identify which RNAs are translated and which are not in human neurons. I will purify lncRNPs and mRNPs from Drosophila and human neurons to identify which proteins bind to the different RNA pools. This will demonstrate the molecular differences between lncRNAs and mRNAs and provide novel and timely insight into how mis-differentiation of RNA populations can promote neurological disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: NEUROLOGICAL
Title: Understanding the Molecular Origins of the Toxicity of Alpha-synuclein in Parkinson's Disease
Funder: Medical Research Council

Abstract: In this project we assembled a cohort of complementary skills and combined our longterm research programmes to carry out an interdisciplinary study of the structural bases of the toxicity of prefibrilar alpha-synuclein oligomers in view of the crucial interaction with the membrane, which represents the key step in the aetiology of Parkinson's disease. Our research proposal comes at a most opportune time, since we now have all the tools, background research and materials to perform such an ambitious study. The proposal is articulated on three independent and complementary streams of increasing scale of complexity (i.e. from oligomers structure to membrane interaction and to the effects of pathological mutations). To fulfil this ambitious goal, we will carry out an investigation of solid-state NMR (ssNMR), solution NMR and computational biology that will provide an accurate structural characterisation of toxic and non-toxic oligomers, previously individuated by a part of the team (Cremades et al, Cell, 2012 149:1048-59). Moreover, to probe the selective interaction of the two types of oligomers with a variety of lipid membranes, we will adapt our recent approach that successfully unveiled the nature of the interaction between monomeric alpha-synuclein and lipid membranes (Fusco et al, Nature Communications, 2014 5:3827). The structural investigation will be complemented biophysical and cellular toxicity measurements to define the underlying structural bases of cellular toxicity of alpha-synuclein.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: The Genetic Architecture of Carpal Tunnel Syndrome
Funder: Medical Research Council

Abstract: The aim of this study is to understand the molecular genetic basis of carpal tunnel syndrome (CTS). The specific aims of the research are divided into overlapping themes of genetic analysis and functional characterisation: Theme 1 - Genetics (Months 1-16) Genotypes generated by UK Biobank will be used to compare CTS cases with controls. We estimate that there will be approximately 7500 cases following quality control, and the control group will be matched using age and gender, outnumbering the cases 10:1. Analysis will be conducted mainly using PLINK and R. Fine-mapping of the associated loci will be performed using imputed data released from UK Biobank, expected in early 2016. Theme 2 - Functional Characterisation (Months 17-32) By the end of 2015, the Bennett Group will have developed a cohort of >150 patients with electrophysiologically and clinically diagnosed CTS, and age- and gender-matched controls, with detailed phenotypic information including electrophysiology, skin biopsies from the median nerve distribution, DNA and RNA samples. Patients recruited from late 2014 onwards will also undergo 7T MRI imaging of the carpal tunnel, and biopsies from fibrotic synovial tissue will be taken during decompression surgery. Functional studies will be driven by the results of the genetic analysis. First, the entire CTS cohort will be genotyped using Sequenom MassArray at all associated SNPs identified in the GWAS, and genotype will be correlated with clinical phenotype. A thorough analysis of the potentially most functionally interesting variants will then be undertaken, and the location of these variants will be analysed in relation to genes of interest. CTS-patient derived tissues will be used to determine the effects of variants on RNA and protein expression. If there is evidence for a functional effect of a variant on a particular gene in CTS-derived cells, gene knockdown or over-expression will be used to affect the function of this gene.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Neuroanatomical and functional characterization of hindbrain leucine-sensing circuits implicated in the regulation of feeding behaviour
Funder: Medical Research Council

Abstract: The brain, through the function of specialized cell populations, receives and integrates convergent neuronal, hormonal and nutritional signals of energy availability, and orchestrates the behavioural, endocrine and metabolic feedback control necessary for the maintenance of energy homeostasis. The hypothalamus is established as one of the major brain regions containing these metabolic sensing cell populations, but there is expanding body of knowledge highlighting the role of extra-hypothalamic centres in the co-ordination of energy balance. These include the nucleus of the solitary tract in the caudo-medial brainstem, which combines nutrient and hormonal sensing properties with a unique neuro-anatomical connection to both gustatory and visceral nutrient sensors, and forebrain homeostatic and hedonic circuits. Our previous research supports the idea that circulating L-leucine concentrations represent a signal of energy availability, detected by hindbrain nutrient-sensing cells to modulate central feeding-regulatory circuits. The aim of the proposed research is to use a combination of state-of -the-art neurosurgical, histological, imaging, viral tract-tracing and molecular genetic tools to characterize the neurocircuits engaged downstream from hindbrain L-leucine sensing to regulate food intake. Results from this research will increase our understanding of the connectomics of the central regulation of energy balance, may challenge the hypothalamic-centric view of central metabolic sensing that is currently predominant in the field, and promote the idea that the brainstem is a major center of metabolic integration and effector coordination in the control of energy homeostasis.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Non-invasive brain stimulation to suppress pathological tremors
Funder: Medical Research Council

Abstract: Tremor is a disabling symptom in many movement disorders, estimated to affect over 3.4 million people in the UK alone. The lack of any clear pathophysiology means that there have been no new interventional tools developed for the treatment of tremor in the last 20 years. Recently, I pioneered an approach in which non-invasive transcranial alternating current stimulation (TACS) was delivered during bouts of tremor over sensorimotor cortex at the patients' individual tremor frequency. When delivered open-loop a slow precession occurred between stimulation and tremor waveforms (as measured by accelerometry), permitting those alignments that induced the greatest suppression in tremor to be identified. By fixing this relationship in a closed-loop arrangement I was able to induce a 50% reduction in resting tremor amplitude for Parkinson's disease. Yet there is a growing body of evidence that the effect-size can be dramatically increased. It also remains unclear whether this approach could be adopted to provide a generic treatment for all forms of tremor. My objective is the development of a non-invasive form of electrical stimulation as a realisable therapy for those afflicted with disabling tremors. Tremor itself is a complex, evolving entity with an unclear mechanism of emergence. But tremors are distinct - in Parkinson's disease the tremor emerges at rest, while it emerges in Essential Tremor during use of the affected limbs. I aim to employ an integrated approach encompassing contemporary brain stimulation techniques, neuroimaging and electric field modelling to identify those neural substrates responsive to electrical stimulation, and tune that stimulation to maximal effect through cutting-edge electric field steering technology. By evaluating these techniques in the two most common tremor syndromes (Parkinson's disease and Essential Tremor), the widespread efficacy and applicability of the approach will be assessed.


Research activity codes: 6.7 PHYSICAL
Health categories: NEUROLOGICAL
Title: CHARACTERISING KYNURENINE 3-MONOOXYGENASE (KMO) AS A THERAPEUTIC TARGET FOR HUNTINGTON'S DISEASE
Funder: Medical Research Council

Abstract: Huntington's disease (HD) is an incurable, fatal neurodegenerative disorder caused by the expansion of a polyglutamine tract in the huntingtin protein. A hallmark of HD is the perturbation of the kynurenine pathway (KP), such that there is an increase in neurotoxic metabolites (quinolinic acid; 3-hydroxykynurenine) relative to the neuroprotective metabolite (KYNA). The KP enzyme kynurenine 3-monooxygenase (KMO) plays a central role in the regulation of this pathway; indeed its inhibition leads to normalization of the pathway in HD models, and a shift towards neuroprotection. We have found that pharmacological KMO inhibition also ameliorates neurodegeneration and other disease-relevant phenotypes in models of HD, including fruit flies and mice, making KMO inhibition a promising candidate therapeutic strategy. Notably, KMO inhibition specifically in the blood of HD mice is efficacious, but it is not yet known whether inhibition in the CNS yields similar or enhanced levels of neuroprotection. Here we propose to validate KMO inhibition as a therapeutic strategy for HD by a number of approaches. We will employ our recently developed KMO conditional knockout mice, which will be crossed to R6/2 mice to test the efficacy of KMO inhibition by genetic means. We will delete KMO either globally, or specifically in the periphery or CNS in order to dissect the differential effects of KMO inhibition in these regions. We will expand upon this work by testing KMO inhibition in myeloid cells derived from HD patients. In parallel with this work, we will employ genomics approaches to explore the biology underlying KMO inhibition in control and HD mice, as well as in myeloid cells from HD patients. Finally, we will explore the biological relevance of our preliminary findings that KMO and huntingtin interact physically at mitochondria. In total, this work will clarify the therapeutic relevance of KMO inhibition in HD, which - if promising - will help translation to the clinic.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: MICA: Optogenetic dissection of homeostatic and Hebbian components of cortical plasticity
Funder: Medical Research Council

Abstract: Synaptic plasticity is thought to endow the organism with vital functions such as circuit formation during development, learning and memory in adulthood, and recovery of function following trauma. Defective synaptic plasticity either during development or adulthood is thought to occur in many common forms of psychosis and dementia. Our long term goal is to understand synaptic plasticity, not only for fundamental scientific reasons, but also for understanding and ultimately treating debilitating neurological and mental health conditions. Two factors are bottlenecks to progress in this respect (1) understanding the normal circuitry of the cerebral cortex and (2) understanding the diversity of plasticity among cellular subsystems. Recently we found that cortical layer 5 RS and IB cells have fundamentally different plasticity properties. RS cells exhibit little potentiation per se but do show homeostatic rebound from depression. Conversely, IB cells show little depression but do show CaMKII dependent potentiation. Our medium term goal is therefore to determine the pathways and mechanisms underlying these differences and to see if they are general features of cortical architecture. To achieve these objectives we have devised the following 8 experiments: 1 Decomposition of projection pathways to L5 RS cells for sensory transmission 2 Identification of homeostatic pathway(s) operating in RS cells 3 Decomposition of projection pathways to L5 IB cells for sensory transmission 4 Identification of Hebbian pathway(s) operating in IB cells 5 Determination of the dependence of functional plasticity in L5 IB cells on structural plasticity in L5 apical dendrites or L2/3 cells 6 Determination of whether L5 RS and IB plasticity distinctions generalise to visual cortex 7 Determination of the dependence of visual cortex plasticity on structural plasticity 8 Identification of structural differences between homeostatic and Hebbian plasticity


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL