NIHRDataInsights/HRCSData huggingface dataset:
Title: Identifying the early biochemical signature of amyotrophic lateral sclerosis
Funder: Medical Research Council

Abstract: Aims - Characterise the pre-symptomatic cerebrospinal fluid (CSF) and CSF extracellular vesible (EV) proteome in ALS - Identify key biochemical pathways reflecting pathological or compensatory mechanisms amenable to therapeutic intervention in both presymptomatic and symptomatic ALS Objectives - Establish a well-characterised longitudinal cohort of asymptomatic individuals carrying the highly-penetrant ALS-causing C9orf72 hexanucleotide repeat expansion - Perform deep proteomic analysis of individual whole CSF and CSF EV samples from asymptomatic gene carriers, healthy controls and ALS patients - Identify alterations in pathways implicated in ALS that vary longitudinally in gene carriers and predate clinical symptoms - Explore mechanistic role of these pathways using in vitro models and post mortem tissue Methodology High-depth liquid chromatography tandem mass spectrometric analysis. Univariate and multivariate analysis to identify key proteins and pathways, including use of gene ontology, pathway and protein-protein interaction network analysis. Comparative studies of whole CSF and EV results. Subsequent validation using targeted proteomics of whole CSF, serum and urine as well as CSF and blood EVs. Further study of implicated pathways by quantitative immunohistochemistry of post mortem tissue and in C9orf72 iPSC-derived motor neuron model of ALS. Scientific and medical opportunities - Broadening the treatable window of ALS to include the period before the onset of symptoms - Identification of previously unexplored markers of the earliest events in ALS pathogenesis and compensatory pathways might yield potential avenues for therapy - Potential diagnostic value and as biochemical measure of response to intervention - Broader value, through data-sharing, through contribution to global efforts to define the human CSF and EV proteome and validation of disease models


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Modelling neuronal dysfunction in early onset epilepsies; a patient-centric approach
Funder: Medical Research Council

Abstract: Aim: to create a patient-derived 3D neuronal model of EIMFS which will give insight into the temporal development of the disease phenotype and act as a platform for novel therapy discovery and testing. Objectives: 1. Establish patient-derived induced pluripotent stem cell (iPSC) and isogenic control lines 2. Create patient-derived 3D neuronal models 3. Investigate the neuronal phenotype of EIMFS 4. Rescue neuronal and network phenotype with novel therapeutic approaches Methodology: 1. Fibroblasts from EIMFS patients with mutations in SLC12A5, KCNT1 or SCN2A will be reprogrammed into iPSCs. Isogenic controls will be created by CRISPR/Cas9 genome editing. 2. iPSC lines from patients, isogenic controls and age-matched controls will be differentiated into 3D cerebral organoids. Using patterning factors, medial ganglionic eminence-like organoids will be generated and fused with cerebral organoids. 3. Cell surface expression will be studied with immunoblotting, immunohistochemistry and confocal microscopy. Patch clamping and multielectrode arrays will assess cellular and network electrophysiological phenotype. Synaptic integrity will be assessed with confocal microscopy for synaptic markers. scRNAseq will examine gene expression differences. 4. SLC12A5 lentiviral gene therapy and KCNT1 antisense oligonucleotides will be assessed using the assays described above. Elucidation of neuronal dysfunction in EIMFS organoids will provide insights into disease mechanisms and allow testing of novel treatments.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: MICA: Neuronal Arginine Metabolism in Health and Disease
Funder: Medical Research Council

Abstract: Arginine downstream metabolites play key-roles in neurotransmission, cellular bioenergetics, gene regulation. These metabolites (i.e. nitric oxide (NO), creatine, polyamines) are involved in various pathophysiological processes e.g. immunology, inflammation, tumorigenesis, but also in schizophrenia and neurodegenerative diseases, highlighting the importance of this pathway in brain physiology and suggesting the relevance of arginine-targeted therapy for some neurodevelopmental or neurodegenerative diseases. Argininosuccinate lyase (ASL) is the only mammalian enzyme enabling endogenous arginine synthesis. The ubiquitously expressed ASL enzyme belongs to the urea cycle, detoxifying nitrogen waste, and the citrulline-NO cycle, essential for NO synthesis from NO synthase. Inherited ASL deficiency (ASLD) causes hyperammonaemia, arginine deprivation and NO deficiency, features of argininosuccinic aciduria. Compared to other urea cycle defects, ASLD shows rarer hyperammonaemic decompensations and a systemic phenotype with developmental delay unrelated to hyperammonaemia. In previous work, I showed that, similarly to humans, ASLD mice present memory deficit, independent of hyperammonaemia and neuroinflammation, and neuronal oxidative/nitrosative stress. This project aims to unravel the role of arginine in neuronal biology using ASLD as a model to interrogate the effect of arginine deprivation on neuronal biology and to identify therapeutic targets. I will study the impact of arginine deprivation on (i) neuronal bioenergetics and antioxidant pathways and mitochondrial function, (ii) neuronal plasticity and neurotransmission, and (iii) assess if ASLD can be a reliable disease model to better understand features of neurodegenerative diseases. On completion, this work will provide a clear understanding of the impact of arginine metabolism on key neuronal functions and its putative role in neurodegeneration.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Dissecting molecular mechanisms of Parkinson's disease
Funder: Medical Research Council

Abstract: Parkinson's disease (PD) is a common neurodegenerative disease that remains defined by pathological hallmarks of end-stage disease, neuronal loss and protein aggregation. The aim of this research program is to understand the earliest molecular events that drive pathogenesis. Powerful clues to pathogenesis are provided by mendelian genes that cause familial PD, and genetic loci that confer risk to sporadic PD. Importantly, both approaches highlight selective convergent processes to be causal in PD, namely protein aggregation, mitochondrial homeostasis, lysosomal biology and inflammation. This provides the rationale for the overall approach of this program: monogenic forms of PD can be harnessed to delineate causative mechanisms in human neurons and astrocytes, and these mechanisms will be relevant in the sporadic PD population. This proposal first addresses when and where protein oligomerization occurs in human neurons, and how oligomerisation is modulated by mitochondrial function, or lysosomal function. Second, we address what is the earliest mitochondrial defect in PD, and the underlying mitochondrial mechanisms that lead to neuronal loss. Single molecule and single cell methods in human iPSC derived monogenic models are used to characterize protein aggregation and mitochondrial function at high resolution, determine their intersection, and causality in monogenic PD. Third, we investigate whether PD specific astrogliopathy is due to reactive astrocytosis and inflammation, or intrinsic molecular changes in astrocytes causing loss of neuronal homeostasis. Co-culture paradigms of astrocytes and neurons are combined with imaging and transcriptomics to dissect the role of astrocytic biology and inflammation in PD. Fourth, the proposal defines a pipeline to harness information from monogenic PD to validate its relevance in sporadic PD, with the ultimate aim of critically refining our classification of the molecular origins of sporadic disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Do adult human oligodendrocytes remyelinate poorly and can we change this to better treat progressive multiple sclerosis?
Funder: Medical Research Council

Abstract: Multiple sclerosis (MS) is a common chronic demyelinating, neurodegenerative disease of the CNS, with major societal impact. There are now effective treatments to reduce relapses in early disease, but not to slow, stop or reverse disability progression in later disease. As disability reflects neurodegeneration, one focus of research into therapies to limit progressive disease is to enhance repair of demyelinated lesions (remyelination), restoring saltatory conduction and metabolic support to the underlying axon. Three such therapies have now been tested in clinical trials but with only minor effects. This may be due to two problems: 1) we have shown that aged adult rat oligodendrocytes do not respond to these pro-remyelinating drugs compared to young adult cells, but these drugs become effective again after metformin treatment, 2) our snRNAseq from human brain shows six distinct human oligodendrocyte 'states', different from rat, the proportion of which are different in MS, suggesting it may be best to enhance particular oligodendrocyte states e.g. those better at remyelination. We hypothesise that pro-remyelinating drug efficacy will be improved if preclinical testing is performed on aged human oligodendroglia with relevant functional states similar to those in MS. In this project, we will use adult human oligodendrocytes to determine whether remyelination declines with age and is rejuvenated after metformin treatment. Thus we need in vitro adult human oligodendrocytes, preferably from a stem cell source for a limitless supply. We will obtain some from human brain biopsies, but we will test strategies to induce 'aged' cells from stem cells. We will assess oligodendrocyte states formed under these conditions. These drugs and metformin are being considered for use in the MS Society UK's 'Efficient Clinical Trials Platform'. This project will provide a relevant testing platform for current and future drugs, accelerating movement of effective drugs into MS patients.


Research activity codes: 6.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: Susceptibility to Chronic Pain: role of Cerebellar - Periaqueductal Gray Communication
Funder: Medical Research Council

Abstract: There is a failure in translation from pre-clinical studies of chronic pain to effective therapeutic interventions in humans. A confounding factor is that experimental animals of the same strain are often assumed to be a homogeneous group which overlooks inherent variability in outbred strains, including susceptibility to chronic pain. We want to exploit individual variability, with the overall aim of gaining a better understanding of the neural mechanisms that determine why some individuals are more susceptible to transition from acute to chronic pain, with the goal of determining the underlying neural network dynamics responsible for this transition. Risk factors that predispose an individual to the development of chronic pain include changes in the functioning of pain networks in the CNS resulting in an enhancement of the pain signal as it ascends from the spinal cord to the brain. This involves a shift from inhibitory to facilitatory control of the transmission of the pain signal by the actions of so-called descending pain modulatory systems (DPMS) that originate in brainstem regions, including an area called the periaqueductal grey (PAG). DPMS controls pain transmission at the spinal level and a shift from inhibitory to facilitatory actions of DPMS are believed to underpin the transition from acute to chronic pain, but why some individuals are more susceptible than others is a key unknown. The hypothesis to be tested is the new concept that the cerebellum plays an important role in regulating the DPMS and changes in this regulation underlie susceptibility. In an out bred strain of rats, chemogenetic silencing of specific pathways between the cerebellum and the PAG in behavioural and electrophysiological studies will focus on two previously neglected questions; (i) the effect of individual variability (measured by rates of fear extinction) on DPMS and (ii) the role of cerebellar-PAG-DPMS axis in the susceptibility to chronic pain.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Defining the primary afferent circuitry that drives neuropathic pain
Funder: Medical Research Council

Abstract: Neuropathic pain arises as a consequence of a lesion or disease of the somatosensory nervous system. It is common, affecting circa 8% of the general population. Unfortunately, current treatments are inadequate due to both poor efficacy and tolerability. We will focus on hyper-excitability in primary sensory neurons as a target because this has been shown to be critical in the maintenance of neuropathic pain. Indiscriminate nerve fibre conduction blockade (for instance with local anaesthetic) is not clinically viable due to its non-specific nature and other safety concerns. Importantly, sensory neurons consist of heterogenous sub-populations categorised by molecular profile, connectivity and stimulus-response function. We will determine which sensory neuron sub-populations are critical in the development and maintenance of neuropathic pain in order to develop means to suppress aberrant activity in these circuits. We will use chemogenetic technologies to reversibly silence specific sensory neuron sub-populations following induction of traumatic or chemotherapy induced painful neuropathy in mice. We will determine how these sub-populations contribute to specific aspects of neuropathic pain related behaviour such as brush evoked allodynia versus assays of spontaneous pain. We will investigate the pathophysiological changes such as transcriptional dysregulation and altered connectivity to dorsal horn neurons within these sub-populations in order to enhance the precision of treatment. To assess the translational potential of this chemogenetic approach we will compare the sensory neuron sub-populations that we have identified in mouse to human DRG and use a newly developed fully humanised chemogenetic system in both animal models and human iPSC derived sensory neurons. As an outcome we hope to have defined the critical circuit changes which drive neuropathic pain but also provide proof of concept that a chemogenetic approach could be applied to neuropathic pain patients.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Elucidating the consequences of aberrant mGluR1 signalling in cerebellar ataxia
Funder: Medical Research Council

Abstract: The spinocerebellar ataxias (SCAs) are a complex group of neurodegenerative disorders characterized by the progressive dysfunction of the cerebellum resulting in a loss of motor coordination. No effective treatments exist for the SCAs, and there is thus a pressing need for better models in which to study the underlying disease-causing mechanisms and to identify potential therapies. Aberrant mGluR1-TRPC3 signalling and downstream dysregulation of calcium homeostasis has been hypothesized as a key pathological event in several distinct genetic forms of cerebellar ataxia, but the underlying mechanisms remain unclear. Recently, we were the first to identify dominant gain-of-function mutations in the GRM1 gene - encoding mGlur1 - in patients with SCA44. Here, we will take a multi-facetted approach to elucidate the molecular and cellular mechanisms by which mutant mGluR1 causes cerebellar ataxia and to identify small molecule compounds that could be used as potential therapeutics. We will functionally characterize the novel mutations that we have identified in cellular assays including in primary Purkinje cells. We have already generated the first mouse model for SCA44 that carries one of the patients' gain-of-function mutation in Grm1. We propose to carry out a detailed investigation of the disease phenotypes in this mouse related to both motor and cognitive dysfunction. We will subsequently exploit the identified in vitro and in vivo SCA44 phenotypes to identify compounds that modulate the disease. We have already identified lead compounds that might have therapeutic benefits including the FDA-approved drug Nitazoxanide and a novel potent TRPC3 antagonist that we propose to further explore. Together, our research will provide a platform for drug discovery in SCA44 and related ataxias and identify promising molecules that could be of therapeutic benefit for these disorders.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: Transplantation of genetically modified haematopoietic stem cells for Friedreich's Ataxia
Funder: Medical Research Council

Abstract: Background Friedreich's ataxia (FA) is a neurodegenerative disease currently lacking any proven treatment. Experimental findings have provided robust evidence that allogeneic haematopoietic stem cell (HSC) transplantation may offer an effective treatment for the disease. When used in clinical practice, however, allogeneic transplantation is associated with significant morbidity and mortality. To mitigate this limitation, we propose to investigate whether genetic modification of endogenous HSCs (insertion of functional frataxin gene (FXN) ex vivo) and subsequent autologous HSC transplantation, is of therapeutic benefit. This approach offers the prospect of a significantly safer and rapidly translatable therapy which may have significant clinical impact on both symptoms and long-term disability for people with FA. Aims This project aims to produce a clinically relevant lentiviral transduction protocol for stable FXN delivery to isolated HSCs; to determine the efficacy of autologous FXN-corrected HSC transplantation to reverse neurological features of Friedreich's ataxia; and define mechanisms by which corrected cells prevent atrophy of large sensory dorsal root ganglion (DRG) neurons. Methods Lentivirus vector-mediated genome editing and transplantation techniques will be employed to demonstrate the efficacy of HSC gene therapy in models of FA. Neurobehavioural, neurophysiological and biochemical changes will be assessed to measure the impact treatment has on protecting or repairing nervous system function. Single cell RNA-sequencing will be employed to explore mechanisms of the DRG neuroprotection. Super-resolution confocal and transmission electron microscopy will provide further in-depth histological assessment. Importance Developing autologous FXN-corrected HSC transplantation as a disease-modifying treatment for people with FA may lead to major therapeutic advances in the foreseeable future.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: All-optical interrogation of the hippocampal neural code underlying episodic memory
Funder: Medical Research Council

Abstract: The hippocampus is essential for the formation and retrieval of episodic memories. The discovery of hippocampal place cells suggested a cellular mechanism for spatial memory. However, a causal role of place cell firing has yet to be established; and the general neural code employed by hippocampal neurons to form and recall episodic memories is unknown. We will address these problems by leveraging a novel "all-optical" approach for interrogating neural circuits in the intact brain. By combining simultaneous 2-photon calcium imaging and 2-photon targeted optogenetic stimulation, the activity of thousands of neurons can be recorded and simultaneously manipulated at cellular and millisecond resolution in the behaving animal. We will use this approach to determine the hippocampal neural code underlying episodic memory across both space and time. First, we will test the functional role of place cells in mice performing a spatial memory task in virtual reality. If specific optogenetic activation of place cells representing a rewarded location biases the behaviour towards that displayed in the rewarded location, this provides a direct causal link between place cell activity and spatial memory. We will also assess the role of stereotypical patterns in spatial learning by artificially increasing place cell sequence reliability in naive mice. Second, complementary experiments will be performed during an olfactory paired-associates task with a delay to test whether hippocampal neural sequences support memory across time. We will compare the efficacy of consistent versus scrambled sequences of activity in boosting learning rates and memory retrieval. Our experiments will provide unprecedented causal links between specific patterns of activity in hippocampal neurons and memory. This information will guide diagnosis and novel treatments in dementias and neurodegenerative diseases, which cause memory deficits due to disruptions of cellular function and coding in the hippocampus.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Defining and manipulating the neural basis of hypoactive and hyperactive behaviours in Parkinson's disease.
Funder: Medical Research Council

Abstract: People with Parkinson's disease (PD) often experience neuropsychiatric symptoms ranging from hypoactive behaviours such as apathy and depression to hyperactive behaviours such as impulsive compulsive behaviours. Although these symptoms are common and disabling, their physiopathology is unclear and there is a lack of effective treatment. It has been suggested that behavioural hypo- and hyper-activation might be linked to hypo and hyper motoric states in PD and that the subthalamic nucleus (STN) might play a key role in their pathophysiology, as for motor symptoms. Deep Brain Stimulation (DBS) of STN is a powerful treatment for PD motor complications but its role in behavioural disorders is unclear. Our study aims to record the local field potential (LFP) in PD patients with STN-DBS and to identify common neural biomarkers of hypo and hyperactive behaviours. We have 3 specific hypotheses that we will test: 1. Different spatio-spectral patterns of STN LFP activity and of LFP coupling with frontal EEG are associated with different aspects of hypo- and hyperactive behaviours in PD. We will record STN LFP with high spatial resolution using directional electrodes, simultaneously with EEG, during tasks challenging key aspects of behavioural control e.g. the Stop-Signal Reaction Time task, Temporal Binding task and Effort-based decision-making task. 2: Different spatio-spectral patterns of LFP activity are causally important in aspects of hypo-and hyperactive behaviours in PD. We will steer electrical currents to target the topography of relevant relevant LFP patterns in the STN to manipulate task performance with DBS. 3: The amplitude of spatio-spectral patterns relates to the severity of hypo- and hyperactive behaviours in PD. We will contrast task performance during random bursts of DBS with bursts triggered by activity changes in the STN. Our findings will help clarify the pathophysiology of hypo and hyperactive behaviours and help us adapt DBS for the their treatment


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL