NIHRDataInsights/HRCSData huggingface dataset:
Title: Blocking chemotherapy-induced peripheral neuropathy by preserving axons
Funder: Medical Research Council

Abstract: Axon degeneration plays a prominent, early role in many neurodegenerative conditions. Despite substantial progress in preserving axons in mice, translation remains a key challenge. This project focuses on chemotherapy-induced peripheral neuropathy (CIPN) as a particularly tractable and important disorder, and has wider implications for other peripheral neuropathies such as diabetic neuropathy, and for other disorders with axonal transport disruption such as multiple sclerosis and glaucoma. CIPN is a neuropathic pain disorder experienced by 50-90% of cancer chemotherapy patients and a dose-limiting side effect. It is caused by axon loss during a treatment period lasting only days or weeks, but becomes a lifelong condition for around half of survivors. Quality of life is greatly reduced by chronic pain, burning, tingling and sleep disturbance and current therapies are palliative and inadequate. A prophylactic treatment is urgently needed to protect axons and thereby prevent or minimise CIPN. The underlying mechanism in CIPN is believed to involve axonal transport impairment. Other mechanisms have been proposed but this model fits with the ability of the slow Wallerian degeneration protein (WldS) to preserve axons in two CIPN models. However, important questions remain. We have made significant advances in understanding why axons degenerate when axonal transport is impaired. We have developed tools for quantitative live imaging of axonal transport in peripheral nerves, for investigating axon degeneration mechanisms and for blocking axon degeneration genetically and pharmacologically when transport is impaired. Here, we will apply these to CIPN models to test whether axons can be permanently rescued, whether this is effective in vivo for a wider range of CIPN-inducing drugs, whether neuropathic pain is reduced, what this tells us about the degeneration mechanism and whether we can block this axon degeneration pharmacologically.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Machinery and Mechanisms of Endosomal Protein Sorting - A key Pathway Associated with Alzheimer Disease
Funder: Medical Research Council

Abstract: The goal of the proposed research is to identify and characterise genes encoding endosomal protein sorting machinery. The genes that function at endosomes will be tested for a role in regulating the localisation and processing of amyloid precursor protein (APP). Those genes are likely to be most relevant for Alzheimer disease (AD). The endosome-to-Golgi retrieval pathway has been shown to play a key role in mediating the localisation and processing of APP. This occurs through the association of APP with the Vps10-domain containing protein SorL1. The SorL1 protein is a cargo protein for the retromer complex and requires retromer for its retrieval from endosomes to the Golgi. Loss of SorL1 or retromer function increases processing of APP to the neurotoxic Abeta peptide that is causal in AD. A genome-wide siRNA screen for genes required for endosome-to-Golgi retrieval has been performed in the SEAMAN lab and has identified 88 genes required for endosome-to-Golgi retrieval including SFT2D2, ZDHHC5 and GRINA. The specific functions of the STF2D2, ZDHHC5 and GRINA proteins will be investigated and proteins that function with them identified by native immunoprecipitation and GST-pulldown experiments. The requirement for the 88 endosome-to-Golgi retrieval genes in regulating APP localisation and processing will be determined and the role of these genes in regulating SorL1 localisation investigated in neuronal cells. Genes identified through the siRNA screen as enhancing endosome-to-Golgi retrieval when silenced will be validated and their role in regulating APP processing determined. Additionally a small molecule screen to identify modulators of endosome-to-Golgi retrieval, both positive and negative, will be performed using an antibody-uptake assay in a high throughput format. Those compounds that affect endosomal protein sorting will be characterised with respect to APP processing and effects on the endo-lysosomal system.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Microglial phagocytic receptors P2Y6 & VNR in brain pathology and therapy
Funder: Medical Research Council

Abstract: Many brain pathologies, such as brain ischaemia (stroke), trauma, infections, epilepsy and neurodegeneration, in addition to causing direct damage to neurons, also cause inflammation, which induces a delayed loss of neurons and synapses. Microglia are brain macrophages that regulate inflammation, and when inflamed can phagocytose neurons and synapses. We and others have found that microglia can phagocytose live neurons in models of Alzheimer's disease, Parkinson's disease, Frontotemporal dementia, AIDS dementia, brain trauma, brain infection, epilepsy, stroke and brain development. If we block the P2Y6 or VNR receptors then we can prevent neuronal and synaptic loss in models of brain pathology in culture and in vivo. These are novel and surprising findings, opening new perspectives on brain pathology and therapy, and in this project we want: 1. To determine whether blocking (by drugs or genetics) the phagocytic receptors P2Y6 or VNR is beneficial in a variety of brain disease models. 2. To find novel ways of blocking P2Y6 or VNR that may used as therapy for brain pathology. 3. To determine whether blocking P2Y6 or VNR is beneficial by blocking phagocytosis of neurons and synapses. 4. To generate genetic knockout mice for P2Y6 and VNR and test whether they are protected against a variety of brain pathologies. If this work is successful, it would potentially lead on to the development of novel treatments for stoke, trauma, epilepsy, AIDS dementia, Alzheimer's disease, Parkinson's disease and/or other brain pathologies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Molecular mechanisms of TDP-43 mediated neurodegeneration
Funder: Medical Research Council

Abstract: Tar DNA binding protein of 43 kDa (TDP-43) has been identified as the major disease protein present in cytoplasmic inclusions in Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Lobar Degeneration (FTLD), as well as in other neurodegenerative disorders including Alzheimer's, Parkinson's and Huntington's disease, thereby defining a novel proteinopathy. Dominant missense mutations in the encoding TARDBP gene characterise both familial and sporadic cases of ALS and FTLD, and recent studies in drosophila, zebrafish and mice reveal that both loss and toxic gain of TDP-43 function contribute to disease onset and progression. TDP-43 proteinopathy is associated with synaptic defects which precede behavioural abnormalities and progressive age-related neurodegeneration. However, the molecular mechanisms underlying neurodegeneration remain elusive. Our hypothesis is, and our preliminary data suggest, that TDP-43 regulates dSarm/Sarm1 and Hiw/mycBP2 that mediate an active autodestruction program causing dying back-like neurodegeneration. Here we propose to use animal and cell culture models to test this hypothesis and to dissect the molecular mechanisms by which TDP-43 regulate dSarm/Sarm1 and Hiw/mycBP2. We also aim to identify ways to rescue disease formation in a drosophila model of TDP-43 related neurodegeneration, and to translate these findings into the human disease condition by detecting RNA/protein changes for human homologues of the autodestruction pathway. Given that TDP-43 proteinopathy characterises several neurodegenerative disorders, including dementia, the expected findings will be of general relevance and application as they likely identify novel biomarkers and therapeutic targets for TDP-43 related neurodegeneration.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: TRPM3 regulation of chronic pain
Funder: Medical Research Council

Abstract: The treatment of chronic pain is an area of significant unmet medical need. The current treatments are associated either with low efficacy in a significant percentage of patients or with undesirable side effects. The identification of a mechanism that can be exploited to develop novel analgesic drugs would be a major advance. We have discovered that genetically modified mice lacking functional TRPM3 ion channels throughout the body have normal sensitivities to mechanical and cold stimuli but fail to develop any neuropathic pain behaviours (von Frey filament, paw pressure, hot plate or cold plate) in a nerve injury model. Furthermore Trpm3-/- mice, do not develop heat or cold hypersensitivities in an inflammatory pain model. As TRPM3 is expressed in some sensory neurons and also in the spinal cord, we will use tissue specific deletion of TRPM3 using the Cre-Lox system to determine the important site(s) of action. We will cross LoxP flankedTRPM3 mice with tissue specific Cre expressing mice to delete TRPM3 in specific cells. The effects of Sensory neuron specific deletion (Advillin-Cre mice) will be compared with sensory and spinal cord deletion (Hoxb8-Cre mice) using behavioural measures and histochemical markers of neuronal activation (e.g. p-ERK, p-p38). The importance of TRPM3 for the maintenance of chronic pain will be determined. We will use tamoxifen-inducible Cre expression to delete TRPM3 in sensory nerves or throughout the nervous system and determine if established hypersensitivities are reversed. We will also use RNA interference, administration of inhibitory monoclonal antibodies directed against TRPM3 and small molecule TRPM3 antagonists to examine the reversal of chronic pain. One or more of these approaches could offer downstream therapeutic opportunities. We will study the possible role of TRPM3 in the relay and processing of nociceptive signals using electrophysiological approaches using sensory neurons and spinal cord slices.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Dissecting the role of LRRK2 in the regulation of autophagy
Funder: Medical Research Council

Abstract: Mutations in Leucine Rich Repeat Kinase 2 (LRRK2) are the most common genetic cause of Parkinson's disease (PD). Recent genome wide association studies have identified variation at the LRRK2 locus as a significant risk factor for idiopathic PD, linking the Mendelian and sporadic forms of Parkinson's. In combination with the presence in the LRRK2 open reading frame of two enzymatic active sites (a kinase and a GTPase), these facts make LRRK2 one of the most attractive drug targets for PD. This proposal investigates the role of LRRK2 in the regulation of autophagy. Data from my group and from other laboratories around the world have revealed that altering LRRK2 function, either by inhibition, knockout, knockdown or mutation, impacts on autophagy although the precise mechanism remains obscure. We have preliminary data that localizes LRRK2 downstream of mTORC1 in the regulation of autophagy, as chemical inhibition of LRRK2 kinase activity results in an induction of autophagy in the absence of any impact of translational targets of mTORC1 (S6 and 4E-BP1). In this new investigator research grant, I will use a combination of cellular and chemical approaches to manipulate LRRK2 in order to establish the exact point at which LRRK2 intersects with the induction of autophagy, specifically examining the phosphorylation of ULK1 and ATG13 as potential effectors for LRRK2s inhibitory impact on autophagy. In parallel to this, I will examine the impact of mutations in LRRK2 upon the regulation of autophagy at the endogenous level using fibroblasts isolated from individuals harbouring mutations in LRRK2 and age/sex matched controls. Preliminary data suggests that mutations result in a decreased ability to enter autophagy upon starvation, and so in this proposal I will carry out a detailed examination of autophagic flux, along with an examination of the impact of LRRK2 mutations on key regulators of the induction of autophagy using in vitro and cellular models.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Neuronal vulnerability to ischaemia: the role of AMPA receptor trafficking.
Funder: Medical Research Council

Abstract: Transient global brain ischaemia induces delayed neuronal death in hippocampal CA1 region via GluA2-lacking, Ca2+-permeable AMPA receptors (CP-AMPARs). This occurs via two mechanisms: a delayed downregulation of GluA2 mRNA expression, and a rapid trafficking event during the insult whereby a fraction of the normal complement of GluA2-containing Ca2+-impermeable AMPA receptors are replaced by CP-AMPARs. Certain populations of pyramidal neurons show a greater vulnerability to ischaemia than others. Typically, in the mature brain, hippocampal CA1 neurons are considerably more vulnerable than their counterparts in CA3 or cortical regions. Our data suggest that oxygen/glucose deprivation (OGD) causes distinct AMPAR trafficking events in CA1, CA3 and cortical neurons. We propose that these differences contribute to the differential vulnerability to ischaemia, and that manipulating these processes could influence the vulnerability of neurons in response to OGD. We will use electrophysiology, biochemical and cell imaging techniques to define OGD-induced AMPAR trafficking events in neuronal populations that exhibit different vulnerabilities to ischaemia. We will focus on the differences between cortical and hippocampal CA1 neurons, because OGD induces a rapid expression of CP-AMPARs in both neuronal populations, but the precise mechanisms are different. In particular, we will investigate the role of subunit-specific endocytosis, recycling and degradation, and the function of subunit interacting proteins in controlling the expression of CP-AMPARs at the synapse. In addition, we will apply genetic or peptide-based tools that prevent or reduce synaptic CP-AMPAR expression to cell death assays, with the aim of improving neuronal viability in response to OGD.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Imaging the assembly, disruption and restoration of nodes of Ranvier in myelinated nerves
Funder: Medical Research Council

Abstract: The purpose of this project is to use advanced optical microscopy and live imaging to reveal how fluorescent proteins associated with voltage-gated sodium channels (Nav), including Neurofascin186, are trafficked to the node of Ranvier during myelination, how they behave once there and how their localization and dynamics are affected by demyelination and remyelination in the CNS. We have generated transgenic mouse lines shown to be suitable for live fluorescence imaging in collaborative pilot experiments with Prof. T. Misgeld, Technical University Munich. A fluorescently-labelled derivative of saxitoxin developed for live imaging of Nav by Dr Justin Du Bois, Stanford University will be supplied to us which enables real-time imaging of native Navs in live cells at the single-molecule level. Nodes are approximately 1 micron wide but spatial resolution in conventional optical microscopy is ultimately limited by the diffraction of visible light (250 nm). Several techniques have been invented to achieve higher spatial resolution to <50 nm collectively termed super-resolution microscopy which offer the opportunity not only of visualizing much finer detail within nodes but also of providing quantitative data on mobilities, dynamic behaviours, interactions and spatial patterning of individual molecules during myelination, demyelination and remyelination. Two recent awards by MRC for optical imaging infrastructure in Edinburgh now make this work feasible (£2M, ESRIC, 2013, MR/K01563X/1; £1.7M, 2013: Ref. MR/K015710/1). Prof. Rory Duncan is PI and Director of ESRIC and will be our collaborator in super-resolution microscopy. We believe that at the conclusion of this project we will have a much clearer view of nodal protein dynamics as they occur in living tissue; further, by applying these approaches to a genetically tractable model we expect to gain new insights into how the targeting and dynamics of these proteins are influenced by demyelination and repair in the CNS.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: How expectations become reality: The neurobiology underlying the influence of expectations on pain
Funder: Medical Research Council

Abstract: Pain is a major health care problem worldwide. It affects the well-being of millions of individuals, and its financial burden upon our societies is considerable. Per annum, it is estimated that the cost of chronic pain to Europe is 200 billion euros. Improvements in the ability to prevent and treat chronic pain are therefore urgently needed to reduce its socioeconomic and personal impact. Pain is not solely determined by sensory input but is critically influenced by expectations and beliefs individuals hold. The influence of expectations on pain has been shown in the exacerbation of symptoms when patients catastrophize about pain, in placebo analgesia and expectancy-related modulations of pharmacologically induced analgesia (Tracey, 2010). Here, I will capitalize (i) on the formalization of the influence of expectations as an inferential process ('perception as inference') and (ii) the investigation of this inferential process using a combined approach of recent advances in computational modeling (Friston & Dolan, 2010) and neuroimaging techniques. This allows for the characterization of neural mechanisms underlying the different steps involved in the inferential process and the investigation of aberrant processing. In four cross-sectional studies using functional neuroimaging in combination with computational modelling in healthy volunteers I will focus on the following questions: - How can expectations lead to a biased perception of pain? (Study 1) - What triggers expectations of pain in the brain? (Study 2) - When does a discrepancy between expectation and sensory input lead to updating of the expectation? (Study 3) - How do expectations affect the perception of change in pain during pain treatment? (Study 4) References: 1. Tracey, I. (2010). Nature Medicine, 16(11), 1277-1283; 2. Friston, K.J., Dolan, R.J. (2010). NeuroImage, 52(3), 752-765.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Early identification of Alzheimer's disease: dynamic biomarkers for enrichment of trials
Funder: Medical Research Council

Abstract: Aim: To use integrated biostatistics and bioinformatics to relate genetic, peripheral and cognitive biomarkers to central Alzhiemer's disease (AD) pathology, in order to reduce barriers to clinical trials in the prodromal phase of the disease. Objectives 1. Characterise genetic and environmental influences on candidate protein markers. 2. Identify tasks from cognitive tests that reflect amyloid pathology. 3. Relate markers to central pathology using a systems biology approach applied to post-mortem brains. 4. Build a causal model of genetic factors, brain Abeta, cognitive test items and candidate protein markers. Methods Objective 1 Sample: 106 twin pairs (for main analysis) Analyses: An established quantitative-genetic model to test for genetic and environmental control of plasma protein levels and cognitive ability. Analysis of association of protein levels with other AD-related phenotypes, and with genetic data. Objective 2 Sample: 144 + 273 + 56 = 473 subjects combined (across cohorts) Analyses: Meta-analysis of association of cognitive task scores with brain amyloid beta levels. Multivariate regression and machine learning to find optimal combination of cognitive task scores. Objective 3 Sample: ~200 case and ~200 control brains (for main analysis) Analyses: Candidate markers will be assessed for differential expression in AD brains, taking into account genetic differences that affect their levels. Common genetic control of these traits will be further explored using 'co-localisation' analysis. Finally, genome-wide analyses on multiple genomic levels will be performed in an integrated fashion. Objective 4 Sample: 116 + 273 + 170 = 559 combined subjects (across cohorts) Analyses: The most promising markers from Objectives 1-3 will be studied further, with the causal relationship between genetics, brain amyloid levels and these markers investigated using Structural Equation Modelling.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: NEUROLOGICAL
Title: MICA: [11C]BU99008 - A novel Positron Emission Tomography ligand for the Imidazoline2 Binding Site, First in human and initial patient group
Funder: Medical Research Council

Abstract: The Imidazoline2 binding sites (I2BS) are located on astrocytes and seem to be involved in the regulation of glial fibrillary acidic protein (GFAP). In addition the densities of the I2BS are altered in a range of psychiatric and neurological disorders (e.g. depression, glial tumours, Alzheimer's and Parkinson's Disease). A positron emission tomography (PET) ligand for these sites would be invaluable in studying these binding sites and disorders in vivo, in humans. Due to the association of I2BS with GFAP such a PET ligand could also be an in vivo marker for astrocytes and their role in these and other psychiatric and neurological disorders. We have developed and evaluated, such a PET ligand, [11C]BU99008, in a range of pre-clinical species. The results of these investigations have indicated that this is an excellent ligand for the I2BS showing high specificity and selectivity for these sites in vitro and in vivo. The next step is to evaluate this ligand in humans. This project aims to do this. The ligand will be initially evaluated in healthy subjects and its pharmacological specificity, selectivity and practicality as a PET ligand in humans, will be determined. On successful completion of these investigations we will then study [11C]BU99008 in a patient population. We plan to use people diagnosed with mild cognitive impairment (MCI) as a group potentially suffering from the early stages of dementia as an initial study group. We will compare the signal obtained in this group with that of the healthy subjects. As the I2BS is correlated with age, care will be taken to ensure that the healthy subjects and the MCI patients are as closely matched for age as possible.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: NEUROLOGICAL