NIHRDataInsights/HRCSData huggingface dataset:
Title: New humanised mouse models for dissecting the pathobiology of disease, using FUS-ALS as a paradigm
Funder: Medical Research Council

Abstract: Many genes, including the well-known genes FUS, TDP43, SOD1 that are causative for the neurodegenerative disease amyotrophic lateral sclerosis (ALS) are dosage-sensitive. ALS is an incurable and essentially untreatable disease that leads to paralysis and death through the progressive loss of motor neurons. ALS is generally a mid-life disorder but patients as young as 11 years of age (with FUS mutations) have been described. Most familial forms of ALS arise from dominant mutations and are modelled in mice by overexpression of the human mutant transgene. However, phenotypes arise in transgenic mice from overexpression per se, rather than from the effects of the mutation. To create more sophisticated models that overcome issues of dosage-sensitivity and express the biochemically correct human protein at physiological levels, we have developed a new technology that is straight-forward and should be useable by standard-gene targeting laboratories. We have created 'genomically humanised' animals in which the mouse gene is entirely replaced by the human genomic locus. These mice express the mutant protein at physiological levels. We are now applying for funding to achieve two goals, (1) to develop our genomic humanisation technology to create conditional mutants and, (2) to characterise our new mouse lines and shed light on the biology and pathology of our paradigm, FUS, in health and in neurodegeneration. By the end of this program of research we will have informative new FUS models for the community, and a better understanding of how mutant FUS causes neurons to die. The normal biology of FUS is not well understood and FUS mutations can be particularly aggressive, often causing ALS in young teenagers and adults, for as yet entirely unknown reasons. FUS is also involved in other neurological diseases, and is an important target for our attention. This technology is applicable to any organism for which ES cells exist, not just mice.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Rational strategies for neuronal production and maturation from human cells.
Funder: Medical Research Council

Abstract: Generation of human neurons in vitro is prerequisite for development of disease-in-a-dish models of neurodegenerative disorders, and ultimately for producing neurons for transplantation. However, for these applications to be realized, neurons generated must be mature, of defined subtype and with normal electrophysiological function. Recently, neurons have been generated from human fibroblasts and ES cells in vitro (iN cells) using defined transcription factors, with and without additional small molecules. However, the neurons generated have frequently been disappointingly immature. The proneural transcription factor Ascl1, used extensively in these protocols, is a crucial regulator of multiple aspects of neurogenesis, including progenitor maintenance, neuronal differentiation, and neuronal subtype. We find that using a modified form of Ascl1 that cannot be phosphorylated on Ser-Pro sites greatly increases the morphological and functional maturity of iN cells produced from human fibroblasts. Our data indicates that phosphomutant Ascl1 will also enhance generation of mature neurons from ES cells. We aim to manipulate phosphoregulation of Ascl1 to enhance maturation of human neurons generated in vitro, and to characterize the processes involved. Experimentally, we will 1) Compare expression of WT and phosphomutant forms of Ascl1 to characterize effects on differentiation, maturation and neuronal subtype after iN cell generation from human fibroblasts and hES cells, in the presence and absence of signaling modulators. 2) Compare in vitro generated neurons produced by direct fibroblast transdifferentiation and via transcription factor expression in iPS cells from the same familial alzheimers patients to compare modelling methods. 3) Use genome-wide ChIPseq and RNA seq to identify key targets that respond differentially to Ascl1 phosphostatus and to use ChIP proteomics to identify potential differences in co-factor association controlled by phospho-status.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: UK Dementias Platform
Funder: Medical Research Council

Abstract: The UK Dementias Platform (UKDP) is a radically new approach to dementias research, providing a highly efficient and cost-effective translation pipeline from basic discovery through to early phase trials (Fig 1). By combining the analytic and statistical power of >20 clinical and population cohorts representing 2M participants and creating a large readiness cohort, UKDP will create an optimal environment for basic discovery activities relevant to the clinical progression and human impact of dementia. Although UKDP is intended as a strategic and sustainable resource generating increasing scientific benefit over time, it will also deliver short and medium term benefits through the analysis of existing data, the enrichment of strategically selected cohorts, developing new ways of working with industry, and generating interest and further funding for dementias research generally. A single portal informatics hub (funded in stage 1) will be used for integrating data from participating cohorts and triangulating evidence between cohorts. The stage 2 proposal focuses on providing strategic resources for EM by enriching strategically selected cohorts, addressing key methodological issues and pump-priming a programme of EM studies. Further development of the platform is anticipated as the opportunities it provides are more widely understood. For example, predicated on the ability of the platform's ability to provide accurately risk stratification, MRC and NIHR have recently funded a deep and frequent Phenotyping (DFP) feasibility study with a view to providing a further £5M for the full study. The DFP project may be considered a stage 3 development of UKDP and is an example of how UKDP can be used to leverage further resource for dementias research.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;2.6 RESOURCES AND INFRASTRUCTURE (AETIOLOGY)
Health categories: NEUROLOGICAL
Title: Investigating Ionotropic Glutamate Receptor Interfaces as Novel Drug Targets.
Funder: Medical Research Council

Abstract: Ionotropic glutamate receptors (iGluRs) mediate nearly all of the fast neurotransmission in the brain and CNS and consequently are intimately linked with the processes of memory and learning. Given this, it is perhaps unsurprising that they have been implicated in a wide-range of neurological conditions ranging from stroke through to Alzheimer's. Researchers have long since recognized that these receptors could represent routes to potential therapies. However, the field has been hampered considerably by the problem of unwanted side-effects. Therefore there is clear clinical need for better, "cleaner", more-specific compounds that target the correct sub-population of receptors. Previous research has shown that in one subtype of iGluRs, the kainate receptors (KARs), has an allosteric site that binds cations and anions and is directly associated with controlling the receptor moving from the open state into the desensitized state. Recently we also showed how the integrity of this site is controlled at the atomic level using molecular dynamics simulations in conjunction with single-channel experiments and site-directed mutagenesis. Targetting this allosteric region (ie away from the glutamate binding site) opens up new possibilities for the design of new compounds that can take advantage of differences in this region between subtypes and thus will have greater specificity. Here we are proposing to extend our preliminary work on the factors that govern the behavior of this allosteric region, and to explore its potential as a drug-binding site via the use of molecular simulation approaches in conjunction with single-channel experiments and site-directed mutagenesis performed by our collaborators. This approach already has demonstrated proven success and we are hopeful that ultimately, new therapies can arise from this work.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: Understanding axonopathy by defining physiological and pathological functions of the microtubule severing protein spastin at membrane traffic pathways
Funder: Medical Research Council

Abstract: We focus on the microtubule severing protein spastin. Mutations in the spastin gene are the most common cause of hereditary spastic paraplegia, in which axons of motor neurons selectively degenerate. Spastin has also been implicated in the pathogenesis of Alzheimer dementia. Spastin is recruited to endosomes and the endoplasmic reticulum, where it controls functionally important membrane modeling events. This led us to propose a pathological model for spastin-HSP in which lack of spastin-mediated microtubule severing causes failure of microtubule-motor dependent membrane modeling steps, which results in abnormality of membrane functions (e.g. traffic of key membrane receptors) that are critical for axonal health. We will test each part of this hypothesis by answering the following questions: i) Does microtubule severing increase dynein loading on microtubules by generating new microtubule plus ends, so providing power for membrane modeling? We will focus on endosomes. Our results indicate that spastin is required for fission of recycling tubules from the endosomal body. Using TIRF and spinning disc microscopy in living cells, we will examine the spatial and temporal relationship between microtubule severing, microtubule plus-end generation, dynein motor recruitment to the microtubule, and tubule fission. ii) What are the functional consequences of failure of spastin-mediated membrane modeling and how do these cause axonopathy? We will examine specific endosomal trafficking steps that are strong candidates to be regulated by spastin. We will also use an unbiased quantitiative SILAC proteomics approach to identify the full range of cell surface receptors regulated by spastin. We will examine the pathological relevance of functional abnormalities identified, prioritising for study those that are candidates to impact on axonal function, by testing whether they are involved in generating axonal swellings in primary neurons derived from our spastin-HSP mice.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: HSN1 secondary to SPTLC1/2 mutations: Pathogenesis and treatment
Funder: Medical Research Council

Abstract: 1)Determining deoxysphingolipid (DSB) induced neurotoxicity and evaluating L-serine supplementation as a therapeutic option. a)Mammalian in-vitro model (primary motor neuron cultures from wild-type mouse embryos and primary dorsal root cultures from wild type mouse pups): These will be used to determine toxicity by assessing the effects on neuronal survival and neurite outgrowth following treatment with DSBs. Co-treatment with L-serine will then be evaluated to determine if it is neuroprotective. Further studies will look at differential toxicity to DSBs between different sensory subpopulations and evaluate whether there is differential uptake of fluorescently labelled DSBs using confocal microscopy. Cellular pathomechanisms will also be explored, initially looking at whether TRPA1 receptors are activated by DSBs. b)iPSC derived sensory neurons from HSN1 patients' fibroblasts: At first, the iPSCs will be differentiated into sensory neurons and phenotyped using functional calcium imaging following application of nociceptive agents and thermal stimuli and electrophysiological studies. After establishing a lineage that phenotypically resembles HSN1 (for example by assessing DSB synthesis in these neurons), these cultures acting as patient specific in-vitro disease models, will be used to assess the therapeutic effects of L-serine supplementation. Later, this model will be used to explore disease mechanisms. 2)Determining responsive outcome measures. HSN1 patients will undergo a set of investigations which will be repeated after a year's interval. These include: CMT Neuropathy Score, comprehensive neurophysiological assessment, Quantitative Sensory Testing (DFNS protocol), muscle MRI studies of the thighs and calves (T1 weighted, STIR, IDEAL and CPMG sequences), myometry, analysis of plasma DSB levels, upper thigh skin biopsy (epidermal nerve fibre density measurements) and patient questionnaires (SF36 and NPSI)


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: The development of human cortical networks: effects of early life stress and injury
Funder: Medical Research Council

Abstract: Little is known about how and when noxious information is processed in the human infant brain and whether the immature brain can distinguish the effects of clinical injury and physiological stress. This proposal focuses upon the functional maturation of brain networks responsible for discriminating noxious events and creating a state of pain in human infants aged 28 weeks (gestational age) to one year old. The project will be carried out by a group of scientists and clinical investigators who have proven skills, expertise and track record in human infant pain neurophysiology. Using a combination of electroencephalography (EEG) and multi-channel near infrared spectroscopy (NIRS) together with physiological measurements of stress and behaviour we plan to (i) map the developmental trajectory of somatosensory and nociceptive cortical activity patterns and their topographic organisation (ii) discover central brain processes of clinically important, pain hypersensitivity states in preterm infants (iii) measure the interaction between physiological stress and central pain processing in infants, which is an especially important problem in an intensive care environment (iv) investigate the development of individual pain phenotypes in infants at birth and to what extent they are shaped by postnatal experience. These experiments will provide us with novel information about the development of cortical networks in the human infant brain that process nociceptive pain and hyperalgesic pain states and new insights into the developmental regulation of pain and stress and the emergence of individual pain phenotypes in infancy. This data is of fundamental biological importance and will impact substantially upon the treatment of infants in intensive care.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: The importance of precision: a model for movement and movement disorders.
Funder: Medical Research Council

Abstract: Parkinson's disease is the second most common neurodegenerative disorder in the world, affecting ~7 million people, with ~130,000 patients in the UK. Parkinson's patients have pronounced movement tremor and are hypokinetic in their movements, presenting with akinesia, bradykinesia, freezing and rigidity. The aim of the work proposed here is to test a novel theoretical account of movement and movement disorders, investigating whether this account can explain some of the hypokinetic symptoms observed in Parkinson's disease. In particular, this research will test a theory of the functional role of sensorimotor beta oscillations that could explain beta power modulations in healthy subjects and the increase in beta power observed in Parkinson's disease patients. Finally, based on this theoretical framework we propose to test a new non-invasive intervention that could ameliorate some of the hypokinetic symptoms of Parkinson's disease. It has been known for over 50 years that power in the beta frequency range (~15-30 Hz), originating in the sensorimotor cortices in healthy human subjects, is modulated during action execution. However, despite extensive research into these neuronal oscillations their functional role, if any, remains unknown. The importance of understanding their functional role is highlighted by the observation that Parkinson's disease patients have a pathologically higher amplitude of beta oscillations, both in the cortex and sub-cortically in the subthalamic nucleus. In Parkinson's disease patients it has been proposed that this pathologically high amplitude of beta oscillations can lead to some of the motor symptoms of Parkinson's disease. Here, we propose to test a novel theory of the functional role sensorimotor beta oscillations, namely that they are causally linked to the imprecision or demodulation of proprioceptive predictions.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Glucocerebrosidase mutations in Parkinson disease:molecular pathogenesis,and the basis for personalised therapy with small molecule chaperones
Funder: Medical Research Council

Abstract: This application addresses the challenge of slowing the clinical and pathological progression of Parkinson disease (PD) with the use of small molecule chaperones to enhance the activity of glucocerebrosidase enzyme (GCase) activity in turn to reduce alpha-synuclein levels and retard neurodegeneration. The programme will use a large cohort of >200 individuals with GBA mutations already recruited from the National Lysosomal Storage Disorders services at the Royal Free London and Addenbrokes hospitals. We propose to phenotype our expanded cohort for these parameters and to retest our complete group to identify progression of the abnormalities or conversion to clinically diagnostic PD. The objectives of this Pathfinder grant are to: 1. Define the clinical evolution and phenotype of GBA mutation positive individuals as they progress to the development of PD. This will enable these features to be correlated with the underlying biochemical abnormalities. 2. Investigate the mechanisms of glucocerebrosidase enzyme (GCase) deficiency in patient derived primary (fibroblast) cultures obtained from our established cohort of GBA mutation positive individuals, age-matched non-GBA mutant PD patients and controls. 3. Determining the effect of small molecule chaperones to enhance GCase activity and reduce alpha-synuclein levels in patient cells and in animal models of human mutations to establish the validity of pursuing these as the next step as a therapy to slow progression in PD patients.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: Regulation of Wnt and calcium signalling by the Parkinson's disease protein LRRK2
Funder: Medical Research Council

Abstract: Mutations in Leucine Rich Repeat Kinase 2 (LRRK2), a common cause of Parkinson's disease (PD), lead to symptoms indistinguishable from idiopathic late-onset PD. This fact makes LRRK2 and LRRK2 signalling pathways particularly attractive therapeutic targets. Evidence for LRRK2 modulated canonical Wnt-signalling and Ca2+-signalling through NFAT is accumulating. Nonetheless, evidence for LRRK2 effects on additional Wnt- and Ca2+-pathways is also emerging. Our goal, to understand how LRRK2 effects specific Wnt- and Ca2+-pathways, will provide a rationale for disease-modifying treatments for this currently incurable disease. LRRK2 knockout and familial G2019S mutant mouse embryonic fibroblast (MEF) and primary neuronal cell cultures will be utilised in luciferase signalling reporter and biochemical assays to evaluate Wnt- and Ca2+-dependent pathway regulation by wild-type LRRK2 and deregulation by mutant LRRK2. Confocal imaging of dopaminergic SH-SY5Y cell lines stably over-expressing EGFP-tagged wild-type or G2019S LRRK2 will determine the subcellular localisation of LRRK2 signalling complexes in response to different environmental (e.g. toxin) or metabolic (e.g. insulin) stimuli, allowing conclusions on signalling regulation/deregulation. Lentiviral biosensors will provide a luminescence-based, real-time read-out of the physiological and pathological function of wild-type and mutant LRRK2 in canonical Wnt and NFAT/Ca2+-signalling in brain during development and adulthood. We will also assess the potential of Ca2+- and Wnt-signalling modulators on previously observed G2019S phenotypes in MEF and SH-SY5Y cells to generate data on the likely efficacy of these modulators in PD. This research will provide key insights into LRRK2-regulated specific Wnt- and Ca2+-pathways in health and disease, increase our understanding of how mutant LRRK2 affects signalling pathways during early events leading to PD and inform therapeutic strategies to halt or cure PD.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Exosome-based Gene Therapy for Huntington's Disease
Funder: Medical Research Council

Abstract: Huntington's disease (HD) is a monogenic, autosomal dominant disorder affecting 1 in 10,000 people caused by a pathological CAG repeat expansion mutation in exon 1 of the Huntingtin gene (Htt). This is translated into a polyglutamine protein that aggregates into toxic inclusions within neurons leading to neural dysfunction then neurodegeneration, progressive motor dysfunction, cognitive decline and death on average within 20-25 years. Drugs currently only allow symptomatic management. Due to the toxic gain-of-function of mutant Htt, a rational therapeutic strategy is to suppress expression of the mutant protein by RNA interference, an approach that would be APPLICABLE TO ALL HD patients. This has been investigated using antisense oligonucleotides, siRNA and viral vectors expressing shRNA given via local administration into the brain. From a clinical therapeutic perspective, given that Htt is ubiquitously expressed and that HD pathology affects a number of brain regions and has peripheral non-nervous system manifestations, it would be highly advantageous to systemically deliver these silencing agents. However, delivery across the blood brain barrier (BBB) remains a major hurdle. Our PROPOSED SOLUTION is the use biological membrane vesicles termed exosomes for delivery of therapeutic agents systemically to the brain. Exosomes naturally transport RNA and protein between cells and our recent research has shown that they can be targeted to the brain following systemic injection to deliver macromolecular siRNA drugs across the BBB. More recently we have demonstrated that exosome-delivered siRNA significantly decreases alpha-synuclein levels in a transgenic mouse model of Parkinson's disease highlighting the potential of this technology. This project will develop brain-targeted exosome technology for targeted reduction of Htt expression in vivo and to demonstrate efficacy and safety in an HD mouse model.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL