NIHRDataInsights/HRCSData huggingface dataset:
Title: The Role of the Alzheimer's Disease Risk Gene PICALM in Blood Brain Barrier Transport, Alzheimer's Disease and Amyloid Angiopathy
Funder: Medical Research Council

Abstract: We have developed exciting data suggesting that: a) in human brain, PICALM is expressed at highest levels in Cerebral Microvascular Endothelial Cells (CMECs); b) non-coding variants in PICALM are associated with increased risk for AD and Cerebral Amyloid Angiopathy (CAA); c) the AD/CAA risk allele promotes increased PICALM expression in CMECs from human brain; d) equivalent overexpression of PICALM in cultured CMECs modulates several aspects of Abeta biology, but the strongest effect is to enhance uptake and subsequent re-secretion of Abeta as amyloid seeds. To explore the underlying biology we will: 1) Deploy well-characterised models (immortalised human CMECs and polarised, multicellular trans-well culture systems) that possess several advantages over other models; 2) Use super resolution microscopy to characterize the effect of changes in PICALM expression on the quantity and direction of secretion of Abeta seeds (brainvessel/vessel-brain); 3) define the subcellular localization of PICALM in models expressing normal, reduced or increased PICALM; 4) map changes in intracellular distribution of endocytosed Abeta in models expressing normal, reduced or increased PICALM; 5) investigate how changing PICALM expression alters the protein interactome of PICALM; 6) validate which of the changes in the PICALM interactome are necessary and sufficient to alter Abeta uptake and secretion. Our proposal builds upon our expertise in the biology of the BBB (Abbott), bioinformatics (Oliver), intracellular vesicular trafficking (CIMR collaborators), advanced imaging (Kaminski), and human functional genomics (St George-Hyslop). This work will clarify the role of PICALM and may identify potential therapeutic targets to ameliorate vascular clearance of Abeta in AD.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Dopamine release and blood flow changes during induced epileptic seizures assessed with simultaneous PET-MR-EEG
Funder: Medical Research Council

Abstract: The PET Centre at St Thomas' Hospital hosts the UK's second simultaneous PET-MR. We have established and validated EEG inside the scanner with phantom and volunteer studies to obtain simultaneous PET-MR-EEG. We will use PET-MR-EEG to investigate dopaminergic neurotransmission during hyperventilation-induced absence seizures, a relatively easily accessible model of epileptic seizures generally. Twenty controls, and 20 patients with idiopathic generalised epilepsy and hyperventilation-inducible seizures, most already identified in specialist clinics, will undergo a single-injection scan with [18F]fallypride, a high-affinity slowly equilibrating dopamine D2/D3 receptor ligand. The long scanning time (3h) will be split into three sessions for subject comfort. The tracer allows visualisation of endogenous dopamine release through receptor blockade (e.g. pilot data in Case for Support). We will use the two early PET-MR sessions to determine subject-specific early [18F]fallypride kinetics and use the "free" simultaneous MR time to acquire structural (3DT1, DESPOT, DTI, FLAIR) and functional information (blood flow - ASL, rsfMRI) as well as sequences necessary for MR-based attenuation correction (MRAC). Until full validation of MRAC (including our own technique), a low-dose one-minute CT scan for attenuation correction will also be acquired at the Centre. During PET-MR-EEG in the third session, subjects will be asked to hyperventilate, thus provoking absence seizures which are harmless and compatible with remaining still in the scanner. We will simultaneously record dynamic [18F]fallypride data (timing resolution in the minute range thanks to its high k2 constant), ASL (every 16s) and EEG (ms resolution) to test the hypotheses that dopamine release occurs, and occurs in response to seizures. Volunteers scanned with and without hyperventilation will allow disentangling the EEG, blood flow, and dopaminergic neurotransmission effects of seizures and hyperventilation.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Dementias Platform UK Stem Cell Partnership
Funder: Medical Research Council

Abstract: A major hurdle to developing treatments for dementia is the lack of human models to study disease mechanisms. However, induced pluripotent stem cell (iPSC) technology and methods to derive neurons and glia now allow study of the pathogenic processes affected in dementia. This proposal partners six major UK Dementia Stem Cell Centres supported by the MRC Dementias Platform UK [DPUK] (Cambridge, Cardiff, Edinburgh, Manchester, Oxford, University College London). Our Partnership capitalises on the £8M DPUK investment in technology platforms to significantly increase the capacity, competitiveness and effective resource use of UK dementia stem cell research by promoting a highly collaborative research network to feed into drug discovery initiatives. The Partnership will create reference iPSC lines from all the major forms of dementia as well as deeply phenotyped healthy aged controls, to form a global highly phenotyped stem cell resource. Critically, we will define standard operating procedures to allow collaborative phenotyping using the DPUK technology platforms including state-of-the-art high-throughput imaging, electrophysiology, metabolomics and proteomics. Development of robust assays / phenotypes for dementia will accelerate the identification and validation of drug targets by industry and academic partners. Four appointed Network Technical Scientists will work across the six sites. Each will be dedicated to specific techniques and goals and will disseminate expertise across the Partnership and beyond. The Partnership will also support PhD students working across Centres. The programmes will leverage existing MRC investment by equipping the UK with researchers highly skilled in interdisciplinary stem cell technology and engage new UK collaborators through regular symposia and training. In summary, the Partnership will create unique stem cell resources and expertise to drive joined-up dementia research to identify new disease-modifying treatments.


Research activity codes: 2.6 RESOURCES AND INFRASTRUCTURE (AETIOLOGY)
Health categories: NEUROLOGICAL
Title: Revealing the dynamic mechanisms of seizures: An integrated mathematical and clinical approach
Funder: Medical Research Council

Abstract: Epileptic seizures are characterised by periods of abnormal synchronous activity distributed over large areas of the brain. This has lead to the current understanding of epilepsy as a dynamic disorder of brain networks, where patients have an enduring susceptibility to seizures, which occur unpredictably from otherwise healthy brain function. By understanding how seizures emerge from healthy background activity from clinical recordings, we can start to understand how networks of people with epilepsy differ from the networks of people with non-epileptic seizures (e.g. syncope), and how AEDs might alter the network structures to decrease the seizure-likelihood. This will put us in a better position to support clinicians in making decisions with respect to diagnosis and prognosis of people with epilepsy. I have used a combination of mathematical modelling and data-analysis to show that connectivity structures within resting-state functional brain networks of people with idiopathic generalised epilepsy (IGE) and their first-degree relative have altered properties to those of healthy controls. This implies that the underlying mechanisms responsible for seizures are reflected in resting state EEG, but that additional mechanisms play a role as well because the abnormal features are characteristic of both people with IGE as well as their unaffected relatives. In order to identify a sufficient set of dynamic biomarkers characteristic of people with epilepsy, I will develop a model framework that combines connectivity structures inferred from resting state EEG with mathematical descriptions of the dynamics of individual brain regions (e.g. neural mass). By using a novel parameter estimation technique, I will then find optimal model frameworks that allows me to infer a set of dynamic biomarkers generating the best discrimination between people with epilepsy and people without epilepsy, as well as seizure-free people and people with ongoing seizures using follow-up data.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Glycosphingolipid recycling in health and disease: the GALC-SapA lipid processing complex
Funder: Medical Research Council

Abstract: This proposal describes structural and functional studies to determine how sphingolipid processing occurs in cells. Lysosomal hydrolases require activating saposin proteins to process substrates. Defects in these proteins cause a range of diseases including early-onset neurodegenerative disorders. Specific enzyme:saposin complexes are essential for lipid hydrolysis but it is not understood how these proteins interact or how specificity is determined. Our current work is focused on GALC, a lysosomal enzyme that when defective causes Krabbe Disease. Our preliminary data reveals a direct interaction between GALC and SapA that occurs at low pH, consistent with formation of this complex in the acidic environment of the lysosome. We have determined the structure of a GALC:SapA:detergent complex identifying how these two proteins interact and where detergent molecules bind. We now intend to exploit the insights from this structure to test how bona fide lipid substrates are bound in the complex and processed in cells. We will determine structures of the complex in the presence of lipid substrates as well as custom-made, high-affinity lipidated inhibitors. The composition of these complexes will be verified using biophysical methods. Protein interaction studies will be used to test whether specific mutations at the interaction interface reduce or abolish binding. This will be done using structurally-informed mutations and Krabbe disease variants that may cause disease by interfering with SapA binding. We will transfer these insights into functional assays monitoring lipid-processing in cells by generating GALC deficient cell lines and re-introducing wild-type and mutant GALC. To monitor defective lipid processing in these cells we will determine their lipid composition using mass spectrometry, monitor the processing of lipid substrates by immunofluorescence and electron microscopy and carry out cell-based activity assays when the GALC-SapA interaction is disrupted.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: MICA: Defining G protein- and arrestin-dependent signalling pathways of biased DOPr agonists and the relevance to their in vivo effects
Funder: Medical Research Council

Abstract: The drug discovery company Pharmnovo AB and Pharmnovo UK Ltd, the industrial partner in this application, has synthesised and developed a number of very selective DOPr agonists with "biased signalling" which can couple efficiently to G protein but partially or not at all to arrestins. These ligands provide analgesia in experimental models of chronic pain. The best way to determine bias and assess its functional significance must be to monitor as many signalling outputs as possible following activation of the endogenous receptor in the organism or relevant part of an organism, and then correlate the pattern of signalling with the observed in vivo effects of the biased ligand. Hence if the global signalling fingerprint (phosphoproteome) in a neuron following DOPr activation can be determined, then by comparing functional effects, such as the ability of DOPr agonists to induce analgesia or an antidepressant effect, with the neuronal phosphoproteome produced by each ligand, we can begin to identify signalling pathways associated with desirable effects and those associated with undesirable effects of DOPr ligands. Consequently the objectives of this present proposal are to: - determine precisely the G protein versus arrestin bias of novel DOPr agonist ligands - apply advanced phosphoproteomics and bioinformatics to native DOPr in neurons to determine and compare the global signalling response of novel DOPr agonists with variable G protein/arrestin bias - correlate the neuronal signalling profiles identified from the phosphoproteome with the in vivo functional profiles of the tested DOPr agonists (e.g. their relative abilities to induce analgesia, and to potentially promote seizure activity, tolerance and dependence) - use these data to identify the optimal neuronal signalling profile for future DOPr agonist ligands for the treatment of pain, depression and anxiety


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: The Pathogenesis of Spinal and Bulbar Muscular Atrophy
Funder: Medical Research Council

Abstract: All the experimental protocols described are established in my host labs 0-18m: Characterise iPSCMNs Initially 2 patient, 2 healthy control lines. More lines are available Differentiate MN: Spinal: Developmentally rationalised cues used including retinoic acid (RA) and Sonic Hedghog (SHH). Bulbar: WNT agonist combined with RA for rostrocaudal patterning. MN creation: Immunocytochemistry (HB9, ISL1, SMI32, ChAT), electrophysiology SBMA: karyotype analysis, CAG using Sanger sequencing, expression and localisation of AR Cell death: cell counts +/- dihydrotestosterone by immunofluorescent staining using Dapi, B-III tubulin and actived caspase 3 (quantified with Western Blot) Axonal transport: Live cell imaging and a fluorescent atoxic tetanus toxin fragment for retrograde AT, Mitotracker for anterograde/retrograde mitochondrial AT Protein mishandling: WB and immunostaining for components of heat shock response (HSR) at baseline and following exposure to cell stressors ER stress: Changes in cytosolic Ca2+ levels using ER stressors (thapsigargin, ionomycin) and fura-2. Markers (PDI, CHOP, BiP ATF4) with WB and immunostaining Mitochondria: Live cell imaging with fluorophores (TMRM, FCCP, Rhod 5N, Indo-1 AM) to measure: membrane potential, redox state, and Ca2+ levels. Electron microscopy to assess morphology 0-24m: Isogenic controls CRISPR spacers to replace expanded CAG repeat with normal length sequence in a patient line and insert an expanded repeat into a control line. Confirm genetic correction with PCR 18- 36m: Astrocyte-neuronal interaction Differentiate astrocytes: Precursors cycle in FGF2 for >80d then terminal differentiation with BMP4 and LIF Phenotype: Reactive state using process length, GDNF and GFAP upregulation. Co-cultures of iPSC-astrocytes and MNs, to examine Ca2+ regulation and mitochondria function 28-36m: Therapeutics Test potential therapeutics which may ameliorate MN dysfunction as described in the experiments above


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Parenteral interventions to support families of children with neurodisability in low resource settings
Funder: Medical Research Council

Abstract: With the reduction in childhood survival in Africa, many more children are living with neurological disabilities. There are few services to support these children or their families in resource poor settings, such as in sub-Saharan Africa, since there are few therapists. The parents and carers report that the two main features that impair their child's and family's quality of life are difficulties with behaviour and communication. Recent reviews of the literature have supported that parent interventions may be the most acceptable and feasible way to improve communication and behaviour. In particular, the Stepping Stones parenting programme, which has been developed for parents of children with disabilities was suggested as an appropriate intervention, since it does not require specialised therapists, does not involve hours of therapy and empowers the parents and carers. This programme has been used in over 23 countries, shown to be effective in improving behaviour and communication; but has not been tested in low-resource settings. The World Health Organization has also recognised these interventions, and promotes them through the mental health treatment gap interventions for children with developmental disorders. In addition the WHO has developed a a parent based intervention i.e. Parent Skills Training to improve communication and behaviour, but this intervention has not been tested in the field. We plan to examine the cultural acceptability and feasibility of administering two interventions i.e. Stepping Stones Programme and Patent Skills Training delivered through parents or carers to improve the communication, behaviour and quality of life of children with neurological disabilities. These studies will be conducted in two poor areas of Kenya ie informal settlements in Nairobi and rural Kilifi.


Research activity codes: 6.6 PSYCHOLOGICAL AND BEHAVIOURAL
Health categories: NEUROLOGICAL
Title: Does kisspeptin in the amygdala control the timing of puberty?
Funder: Medical Research Council

Abstract: Although it is recognised that hypothalamic kisspeptin plays an integral role in the gonadotrophin-releasing hormone (GnRH) pulse generating neural network, and is obligatory for puberty, the upstream regulatory mechanisms that drive the GnRH pulse generator to trigger the onset of puberty have remained stubbornly elusive, as have those underlying its delay by stress. We have recently discovered that the posterodorsal subnucleus of the medial amygdala (MePD) plays a key role in pubertal timing, involving intrinsic GABA and glutamate receptor mechanisms, and that kisspeptin signalling in this extra-hypothalamic structure surprisingly exerts a profound influence on GnRH pulse generator frequency. Additionally, activation of MePD corticotrophin-releasing factor type 2 receptor (CRF-R2) by the endogenous ligand urocortin3 delays puberty. We aim to test the hypothesis that in the amygdala the kisspeptin and GABA/glutamate neural circuits are interdependent and form a higher-order regulator of the GnRH pulse generator that is critical for triggering puberty. Our multidisciplinary approach will involve a combination of neuropharmacological, optogenetic, neuroanatomical and molecular biological techniques applied to well established animal models. The project will focus on (i) determining the anatomical and functional relationship between the kisspeptin and GABA/glutamate systems in the MePD and hypothalamic kisspeptin-GnRH neuronal signalling to regulate pubertal timing and, (ii) determine the neural mechanisms by which the urocortin3/CRF-R2 system alters MePD kisspeptin and/or GABAergic signalling to delay puberty. This project will not only provide novel insight to key upstream regulatory mechanisms that activate the GnRH pulse generator to trigger puberty onset, but may provide new insights into the control and treatment of early and late puberty and hence lessen the burden of many adverse outcomes on health and wellbeing that persist and emerge in adulthood.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Astrocyte-secreted chaperones as a non-cell autonomous form of neuronal protection in Alzheimer's disease
Funder: Medical Research Council

Abstract: A major challenge in biomedical research is to design effective therapeutic strategies for Alzheimer's disease (AD). Currently, the molecular mechanisms underlying AD remains to be elucidated, in part due to a traditional neurocentric view of the disease that has hampered the study of other brain cells such as astrocytes. The objective of this proposal is to shed light into the mechanisms of intercellular signalling between neurons and astrocytes and how these are modulated in disease. I aim to identify endogenous factors that are employed by astrocytes to protect neurons and that could represent the target of novel therapies for AD. In preliminary data I have shown that astrocytes secrete molecular chaperones, intracellular proteins that prevent protein misfolding and that are protective in neurodegenerative diseases. However, the function of chaperones as extracellular proteins is not understood and is a completely novel notion in the context of astrocyte-neuron communication. I hypothesize that molecular chaperones secreted from astrocytes provide a non-cell autonomous form of protection to neurons in AD. We will study the profile of chaperones secreted from astrocytes in AD-like conditions and characterize the impact that these extracellular chaperones have for relevant aspects of AD, such as tau aggregation, synapse and neuronal health. We will also explore their mechanisms of neuroprotection, such as internalization into neurons to provide a novel mechanism to increase neuronal chaperone load. As a proof of concept to validate their therapeutic use, we will use organotypic brain slice cultures from AD transgenic mice to test the protective effects of astrocyte-secreted chaperones in a relevant disease model. Understanding how astrocytes and neurons interact and whether chaperones are intermediaries will help identify 1) important molecular mechanisms underlying AD, as well as 2) novel protective factors to exploit as disease-modifying therapie


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;2.3 PSYCHOLOGICAL, SOCIAL AND ECONOMIC FACTORS
Health categories: NEUROLOGICAL
Title: Regulatory and functional pathways mediating the control of central osmotic defences by hypothalamic transcription factor CREB3L1
Funder: Medical Research Council

Abstract: The brain reflexes regulating water balance are centred on hypothalamic neurones that synthesise the antidiuretic hormone vasopressin (AVP). These neurones send axonal projections to the posterior pituitary, from where AVP is released into the blood. As a consequence of the depletion of pituitary stores that accompanies chronic osmotic stimulation, there is a need to synthesise more AVP. We showed 25 years ago that this starts with an increase in transcription, which leads to more abundant mature mRNAs, but the molecular mechanisms responsible remained elusive until we described transcriptome changes in the rat hypothalamus following osmotic stimulation. We identified basic leucine zipper transcription factor CREB3L1 as being robustly up-regulated in AVP neurones by hypersomostic cues, and showed binding to the AVP gene promoter and activation of AVP gene transcription. Further, we have now shown that CREB3L1 mRNA expression is up-regulated by cAMP pathways, and that shRNA knockdown of CREB3L1 blocks the cAMP-mediated activation of AVP expression. Interestingly, inhibition of protein synthesis blocks the cAMP-mediated up-regulation of CREB3L1 expression, suggesting the requirement for an intermediary "Mystery Transcription Factor". We will now we now address 6 questions, the answers to which will reveal the detailed mechanisms CREB3L1 regulation and function, within the context of the brain mechanisms that regulate water balance: 1. What are the molecular mechanisms that mediate the regulation of CREB3L1 by physiological cues? What is the identity of the "Mystery Transcription Factor". 2. What are the molecular interactions in the hypothalamus that facilitate CREB3L1 actions? 3. What genes does CREB3L1 regulate in the SON and PVN? 4. How does CREB3L1 affect neuropeptide biosynthesis in the HNS? 5. Can we construct and test in silico models of the integrated mechanisms of CREB3L1 function? 6. What are the physiological roles of CREB3L1 and interacting genes.


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