NIHRDataInsights/HRCSData huggingface dataset:
Title: Connexin 26-mediated breathing control by the healthy and obese brain
Funder: Medical Research Council

Abstract: Expiration of the CO2 we produce is essential for survival. This is centrally controlled in the medulla. We have shown that CO2 is sensed directly by the brain: It gates the opening of Connexin26, leading to ATP release and increase in breathing volume. We ablated Cx26 from specific tissues of the medulla using recombinase mediated gene ablation. To study mutant breathing responses to hypercapnia we established ATP release assays on medullar slices and whole-body plethysmography in awake animals. We discovered a novel Cx26+ chemosensory cell type derived from the neural crest, which develops in middle age and dies in old age. Ablation of Cx26 inside these cells reduces breathing volume by 40% upon mild hypercapnia. We now wish to learn how epigenesis affects the development of and signalling inside these cells. We will also explore other areas of Cx26's function in central control of respiration: endothelia, leptomeninges and the rostral chemosensing region. Recombinase-mediated conditional gene ablation and lineage labelling, using Cre lines under cell- or region specific control and floxed Cx26 or LeprB alleles. Mutant phenotypes will be assayed as mentioned. As chemosensory cells carry the leptin receptor, a possible powerful modulator of breathing, they might be defective in obesity hypoventilation syndrome. By ablating the leptin-receptor inside these cells we will test how leptin affects their ontogeny and/or function. As chemoreceptors are now genetically tractable we will ablate Cx26 and LepRB inside them & probe transcriptomes at single cell resolution in relation to diets, yielding new pathways and drugs targets. We establish respiration-specific animal models for human conditions with blunted chemosensitivity: Chiari, SIDS, CCHS all leading to serious morbidity or mortality. Understanding these now tractable chemosensors will enable stratification of patient populations and help meet the challenges of an ageing and increasingly obese patient population.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: BRAIN UK: accessing NHS tissue archives for neuroscience research (2015-2019)
Funder: Medical Research Council

Abstract: Formal prospective brain banks mainly focus on neurodegenerative disease and do not cover other common disorders such as stroke, head injury, psychiatric and developmental disorders. There already exists as a result of decades of diagnostic work a vast resource of well-characterised human tissue in NHS Clinical Neuroscience centres. BRAIN UK is a collaborative virtual brain bank, linking together 26 regional centres, covering approx. 90% of the UK population, and the Corsellis Collection, with its large collection of psychiatric disorders such as schizophrenia. Access is provided to tissue from >60,000 post mortem examinations and >400,000 samples from the living (biopsies) permitting very large studies on common disorders, and particularly useful for uncommon disorders. It allows research into the many neurological diseases not covered by dedicated brain banks. Researchers obtaining tissue through BRAIN UK do not need to obtain their own ethical approval, saving approx 6 months per project. 48 applications have been supported (2011-2014) encompassing a wide variety of conditions including stroke, epilepsy, schizophrenia, tumours, disorders of childhood and ageing. In the coming 4 years we aim to (i) continue to supply neuroscience researchers with well characterised human brain tissue samples (ii) add newly obtained tissue samples which are being accumulated at a rate of >16,000/year (iii) establish a system to gather together data generated by researchers on samples provided by BRAIN UK, promoting interactions and collaborations, and (iv) establish links between BRAIN UK and UK Biobank, a resource with rich data on 500,000 participants in the general population to further understanding on the origins of neurological diseases. As it makes use of existing samples BRAIN UK represents excellent value for money. The scale and scope of BRAIN UK is unprecedented and archived tissue that would otherwise be unused has supported important research projects.


Research activity codes: 2.6 RESOURCES AND INFRASTRUCTURE (AETIOLOGY)
Health categories: NEUROLOGICAL
Title: Molecular chaperone function and motor neuron degeneration
Funder: Medical Research Council

Abstract: Many neurodegenerative diseases are characterised by intracellular proteinacious inclusions of aggregated, misfolded protein, suggesting that imbalances of protein homeostasis (proteostasis) are a factor in disease pathogenesis. Molecular chaperones are important regulators of proteostasis and the neuronal chaperone HSJ1 (DnaJB2) can protect against several types of neurodegeneration, including polyglutamine expanded huntingtin and mutant SOD1 in vivo. Furthermore, mutations that cause loss of HSJ1 function lead to distal hereditary motor neuropathy (dHMN) and Charcot-Marie-Tooth type 2 (CMT2), suggesting HSJ1 is particularly important for maintaining motor neuron proteostasis. This project aims to discover the precise molecular mechanisms that underlie the death of motor neurons caused by HSJ1 mutations. We have successfully reprogrammed HSJ1 patient cells to an induced pluripotent stem cell (iPSC) state and have developed a conditional HSJ1 knock-out (KO) mouse. The constitutive KO recapitulates the human phenotype, with death of motor neurons in the spinal cord. Here we will combine these models to understand motor neuron degeneration caused by loss of HSJ1. We will use a multidisciplinary approach to investigate specific functional deficits in iPSC motor neurons. Subsequently, we will rescue the cellular phenotype, by viral delivery of HSJ1 isoforms, to confirm the specificity of these functional defects, in parallel with investigating isoform specific mouse models, to define the HSJ1 isoform requirements for motor neuron survival. The client proteins of HSJ1 in motor neurons will be identified by a combined proteomic approach of identifying co-purifying proteins and monitoring changes in the proteome following loss of HSJ1. The interaction of putative clients with HSJ1 will be defined, and their importance for motor neuron survival tested. Finally, this mechanistic understanding will be used to test potential therapies for HSJ1 mediated dHMN.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: Dissection of voltage-gated ion channel physiology in vivo in the C. elegans oxygen-sensing neural circuit
Funder: Medical Research Council

Abstract: Voltage-gated ion channels are highly conserved in evolution and broadly expressed in nervous systems. However, the neurophysiological function of many of these channel families remains mysterious, as their investigation in nervous systems in vivo has been hindered by a lack of tractable null mutant phenotypes. Voltage-activated channels have been linked to O2 sensing, but their mechanistic role in this is not understood. I have identified several novel, distinct and genetically separable behavioural phenotypes of null mutants of a conserved ion channel in C. elegans, which alter the function of the [O2]-sensing neural circuit and thus oxygen responses, and discovered a novel interaction of the channel with genes that tune acute O2 responses. I will take advantage of these phenotypes to shed light on the mechanism of channel action by 1. performing a structure-function analysis of the channel and its domains; 2. investigating their functional interaction with other sensory proteins in tuning O2 responses; and 3. conducting an EMS mutagenesis screen, using aggregation behaviour as readout, to identify novel interactors / regulators of the channel. We will identify molecular lesions by whole-genome sequencing and characterising the interaction of 2-3 of the hits with the channel. Microfluidics chambers will be used to assay O2-evoked behaviour and neuronal Ca2+ with imaging of YC2.60; confocal microscopy of GFP-tagged channels will be used to assess aberrant localisation of truncated channels. We will complement our C. elegans studies with recording macropatch currents in HEK293 cells expressing the channel or a human homologue. This way, we aim to make significant contributions towards understanding the neurophysiology of O2-linked ion channels.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Variability in human axon survival
Funder: Medical Research Council

Abstract: We study the pathway of axon degeneration after injury: Wallerian degeneration. Data from animal models suggest related mechanisms influence axon survival in some diseases and in ageing, including multiple sclerosis (MS), peripheral neuropathies (PN), glaucoma and some motor neuron diseases. Early axon loss is also prominent in Alzheimer's disease, Parkinson's disease and ALS. There are no disease-modifying therapies. Here, we address the role of this pathway in human axon survival. Mutations of five genes are now known to robustly delay Wallerian degeneration in mice and other model organisms but whether a similar phenotype exists in humans has remained an intriguing but unanswered question. Recent GWAS and exome linkage between Wallerian pathway genes and ALS and a painful neuropathy, and animal studies, suggest that such a phenotype could function as a disease modifier. SNP databases indicate polymorphisms likely to delay Wallerian degeneration and others likely to confer an opposite phenotype of axon vulnerability. Our mouse and preliminary human studies support this notion. We will test which human variants alter axon survival by testing their function in mouse neurons that lack the homologous mouse gene. Using established assays, we will ask whether human variants in a pro-survival protein, NMNAT2, support axon survival and if so how strongly, and whether variants in human SARM1, a pro-degenerative protein, restore rapid Wallerian degeneration when mouse SARM1 is missing. We will then estimate the prevalence of these phenotypes. We will test whether NMNAT2 deficiency is a risk factor for neuropathic pain, and having shown that SARM1 deletion completely rescues nerve growth and early lethality in NMNAT2 null mice, we will test whether it also rescues a premature age-related decline that we find in mice expressing low levels of NMNAT2. These data will complement ongoing GWAS and exome studies to understand the roles of this pathway in human disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Using transcriptomics to transform the diagnosis and understanding of inherited adult neurological disorders
Funder: Medical Research Council

Abstract: The aim of this study is to determine the effect of genetic factors on whole transcriptome expression in the human central nervous system and in brain-relevant cell types with the primary aim of transforming diagnostics for adult inherited neurological disorders. This project is inspired by the growth in whole genome sequencing in patients, which is likely to drive the discovery of an increasing number of potentially pathogenic variants within non-coding regions of the genome. Prioritising such variants for further analysis and understanding the molecular processes and regional/cellular site of action is key to ensuring diagnostic accuracy and driving further research. I aim to address both these important issues. Using control post-mortem human tissue from 12 regions of the human central nervous system, and through analysis of paired nuclear and cytoplasmic RNAseq data, I will define the transcribed portion of the human genome in adult brain. This data will also be analysed using allele-specific expression (ASE) analysis to identify genetic loci that regulate the expression of specific gene transcripts within specific tissues. A similar approach will be taken to the analysis of RNA sequencing data generated from brain-relevant cell-types originating from human induced pluripotent stem cells. This data will be used to explore cell and state-specific regulation of gene expression in brain-relevant cell types. Through collaboration with the neurology domain of the Genomics England Clinical Interpretation Partnership, I hope to test the value of this approach in real clinical scenarios and in some cases through additional RNA sequencing of patient samples. All the data generated by this project will be publicly released as both raw data files for re-analysis and processed information suitable for non-expert users. Thus, I will generate novel disease-relevant findings and provide the neuroscience community with a world-class resource.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: The UK7T Network: developing the ultra-high field MRI platform for biomedical research.
Funder: Medical Research Council

Abstract: We propose to establish a UK7T Network that will bring together the 7T sites in Cambridge, Cardiff, Glasgow, Nottingham and Oxford to share expertise, build capacity, and develop harmonised approaches to image data acquisition, sharing and analysis. Once established, the UK7T Network will serve as a platform for future UK-wide, collaborative research programmes in 7T magnetic resonance (MR), including multi-site, large cohort clinical studies. The Partnership Grant will allow us to: 1) Increase the level of expertise in the UK in UHF magnetic resonance by sharing the experience that has been built up in Nottingham and Oxford, training a cohort of postdoctoral researchers and delivering a programme of targeted training workshops. 2) Ensure that the wider UK MR research community can exploit the UK's investment in 7T infrastructure (via organisation of symposia and workshops, establishment of a Network Advisory Committee, and provision of access to 7T scanning for pilot studies). 3) Produce a set of optimised 7T protocols for brain imaging for use by all of the UK 7T sites, including (i) anatomical and functional brain imaging methods that can be applied quickly; (ii) more advanced methods, such as diffusion imaging and arterial spin labelling, that require additional development work; (iii) standardised quality assurance protocols. 4) Work with the Dementias Platform UK's XNAT-based national image sharing and analysis platform to develop the capability to share, aggregate and analyse UHF data. 5) Demonstrate the UK7T Network's capability as a platform for multi-site studies, by evaluation of two demonstrator cohorts: (i) a core dataset of high-resolution brain images from 1000 healthy adults across the age lifespan; (ii) a pilot clinical study of twenty five early-stage Alzheimer's disease patients. 6) Establish a robust UK7T Network management structure to support the development and exploitation of the UK's UHF infrastructure.


Research activity codes: 2.6 RESOURCES AND INFRASTRUCTURE (AETIOLOGY)
Health categories: NEUROLOGICAL
Title: STRUCTURE AND REGULATION OF THE BLOOD NERVE BARRIER
Funder: Medical Research Council

Abstract: Technical summary A specialised vasculature, the Blood Nerve Barrier (BNB) protects the Peripheral Nervous System (PNS) whereas its breakdown is associated with injury and pathologies such as cancer and diabetic and inflammatory neuropathies. Unlike the Blood Brain Barrier (BBB), it is poorly characterised with little known about its structure, properties and regulation. It is different to the BBB however, in its permeability and it must be different in its structure and how it is regulated, as some of the cell types that confer barrier function in the BBB do not exist in the PNS. Some of the differences in barrier function between the BBB and the BNB may explain why some drugs, such as taxol, preferentially affect the PNS. We have recently developed a mouse model in which signals from Schwann cells in the PNS cause the breakdown of the BNB in a manner related to the normal injury response. This provides a powerful model to study how the BNB can be regulated and to identify the signaling mechanisms responsible. In order to systematically define the BNB and determine how is can be regulated and examine the relevance of these process to disease we will do the following: 1. Using a number of advanced microscopic techniques, including 3D EM, super-resolution microscopy and light-sheet microscopy we will determine the structure, nature and development of the Blood Nerve Barrier (BNB). 2. Determine the BNB transcriptome and characterise molecules important for distinct aspects of BNB function 3. Using our mouse model, we will determine the cell and molecular changes associated with the breakdown and reformation of the barrier and identify the signals responsible for regulating these changes. These findings will be related to pathologies of the PNS including relevant cancers and neuropathies.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Mitochondrial ubiquitin dynamics and apoptotic cell death.
Funder: Medical Research Council

Abstract: Mitochondria play an important role in the orchestration of apoptotic cell death. Synthetic BH3-mimetics promote apoptosis and have shown some success in the clinic. However, additional approaches are required to improve their efficacy and overcome resistance. Mitochondrial dysfunction has been linked to neurodegeneration, in particular to Parkinson's disease (PD), which is characterised by the loss of dopaminergic neurons from the substantia nigra. Two PD-associated genes, the ubiquitin E3-ligase Parkin and PINK1 are involved in the safe disposal of damaged mitochondria by mitophagy. Depletion of the unique mitochondrial deubiquitylase (DUB), USP30, has been shown to promote mitophagy in cells with defective Parkin or PINK1 and rescue motor function and dopamine levels in a corresponding fly model. We have recently shown that USP30 depletion also sensitises cancer cells to BH3-mimetics, expanding the clinical potential of a future USP30 inhibitor. We will generate CRISPR/CAS9 dependent knock-out and/or catalytically inactive knock-in cell lines and use a range of proteomic and cell biological approaches to a) identify new substrates of USP30 and explore its role in Parkin-independent mitophagy, b) explore the synergism between USP30-silencing and a toolkit of BH3-mimetics within a panel of cell lines and c) elucidate the mechanism of action of USP30 in modulating apoptotic cell death. Whilst this proposal is focused on USP30 as the preeminent mitochondrial DUB, we will in parallel evaluate USP9X and USP24, two DUBs which have been proposed to stabilise MCL1, thus contributing to a major mechanism of resistance to BH3-mimetics.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: NEUROLOGICAL
Title: Examining the coupling of small GTPase activation and metabolism of the phosphoinositide lipid PI(3,5)P2 in Charcot Marie Tooth Type 4 Neuropathies
Funder: Medical Research Council

Abstract: This project aims to understand the function of a group of related enzymes in membrane trafficking, and how their loss may cause Charcot Marie Tooth Type 4 neuropathies. We aim to identify critical points in the exo-endocytic cycle which may be open to therapeutic manipulation to arrest or reverse this progressive disorder. We will use proteins produced in human cell suspension cultures as material to identify small GTPase interactors of CMT4 genes using label-free differential mass spectrometry and then characterize these interactions in model cells, human patient fibroblasts (where available) and in in vitro Schwann cell- neuron co-cultures. We will use lentiviral treatments of Schwann cells to establish cell-autonomous effects of CMT4 gene manipulations in myelinating cells in these co-cultures. Our second aim will establish tools to acutely manipulate the lipid PI(3,5)P2 in cell culture and in vitro myelinating culture models using the CRY2-CIBN optogenetic system, so as to establish the location and function of the pool of this lipid in (dis)myelination.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Characterising brain network differences during scene perception and memory in young adult APOE-e4 carriers: multi-modal imaging in ALSPAC
Funder: Medical Research Council

Abstract: Key to reducing the burden of poorer cognitive health in later life (e.g., by delaying disease onset) is the identification of high-risk individuals prior to the onset of cognitive decline. Here, we test the hypothesis that 'vulnerability' to poorer cognitive health in later life may not be exclusively related to 'aging', but instead, to the accumulation of risks across the lifespan. We will investigate whether young healthy individuals, who are at heightened risk of age-related cognitive decline via possession of an allele (APOE-e4) associated with cognitive disorders of ageing (including Alzheimer's disease), show brain and behavioural differences, compared to non-carriers, that are consistent with the early anatomical and cognitive markers of ageing. A multimodal cognitive imaging approach will be applied involving functional and structural imaging at both 3T and 7T (the latter enabling high-resolution segmentation of subfields in the medial temporal lobe). By applying this in individuals from the Avon Longitudinal Study of Parents and Children, who have been studied from birth, and are now aged 26yrs, we will additionally be able to ask how such early life effects may be moderated by cognitive reserve (e.g., childhood IQ), gender and lifestyle factors (e.g., diet, smoking, alcohol and/or exercise). This study will be used to test the overall hypothesis that: APOE-e4 may influence the onset of cognitive decline in later life via its impact on functional and structural connectivity in a brain circuit involving posteromedial cortex and the medial temporal lobe. More specifically, we predict that young healthy APOE-e4 carriers will show specific brain and behavioural differences (compared to non-carriers) linked to scene perception and memory, and that these effects will be moderated by cognitive reserve, gender and lifestyle factors.


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