NIHRDataInsights/HRCSData huggingface dataset:
Title: Magnetic nanoparticle mediated delivery of neurotherapeutic genes to multipotent neural stem cell transplant populations
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Transplanted neural stem cells (NSCs) offer key dual advantages for neural regeneration and delivery of therapeutic biomolecules to injury foci. Their features include self-renewal and multipotentiality, ease of derivation from embryonic stem cells, migratory capacity (notably towards pathology sites and across the blood brain barrier), amenability to genetic engineering, ability to differentiate and integrate in injury foci and suppression of injury mechanisms (eg. astrocytosis and inflammation). From a technical perspective, most studies in transplantation neurobiology use viral vectors for gene delivery. While efficient, such vectors have several drawbacks including safety issues and large scale production limitations highlighting a major need for development/optimisation of alternative, nonviral approaches for basic research and translational applications. We recently provided 'proof-of-concept' that MNPs safely mediate single/combinatorial gene delivery (including that of plasmids encoding therapeutic growth factors) to NSCs without adverse effects on stem cell viability, migration, proliferation and differentiation. Gene transfer was dramatically enhanced using 'multifection' (repeat transfections) and 'magnetofection' (application of static/oscillating magnetic fields to assist transfection, the latter method using a patented oscillating field system manufactured by Keele spin-off company nanoTherics Ltd). Transfection levels were found to be similar or superior to many viral/nonviral methods currently used in neurobiology with the critical advantage of cell safety. Transfected cells could survive, migrate and differentiate normally post-transplantation. We will optimise MNP based delivery of neurotherapeutic genes to NSCs derived for transplantation and investigate the cellular mechanisms for oscillating field enhanced transfection. The repair potential of 'engineered' NSCs will be tested in a novel 3D injury model in organotypic spinal cord slice cultures.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: The role of presynaptic calcium at ageing synapses
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Ageing is associated with a loss of memory. Subtle changes in synaptic structure and function in the prefrontal cortex and hippocampus may account for such age-related deficits in cognitive function. Ageing is accompanied by alterations in the homeostatic mechanisms that control calcium signalling. These can have profound affects on synaptic transmission and the induction of synaptic plasticity, the process that underpins memory formation. Here we wish to examine how ageing affects calcium signalling, specifically in presynaptic terminals of the hippocampus. We have developed a transgenic mouse that expresses a novel, genetically encoded, ratiometric calcium sensor selectively at presynaptic terminals, with particularly high expression in the hippocampus. The sensor consists of a form of GCaMP linked to mCherry, which is in turn fused to the vesicular protein synaptophsin. Driven by the thy1.2 promotor, the sensor is highly expressed in the hippocampus and precisely targeted to presynaptic terminals. The mCherry signal serves two purposes. First it identifies the presynaptic terminals where the sensor is located, and second, it can be used to quantify the calcium signal from the calcium sensor through ratiometric methods. In the CA1 region, stimulation of Schaffer collateral fibres leads to a robust calcium signal. Similar responses are observed in dentate gyrus and CA3. Sensor expression increases with development allowing measurement of presynaptic calcium from ageing animals. We wish to use this unique mouse model to characterise the contribution of presynaptic calcium to synaptic transmission at each of the main relays of the hippocampal circuit. We will establish how the calcium signals are modified by pharmacological intervention at inhibitory and excitatory circuits and how intracellular stores influence presynaptic calcium. We will then examine whether presynaptic calcium signalling is modified in mice with age-related deficits in hippocampal function.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Role for glia-neuron interaction in the brain ageing and longevity
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Brain function depends on the interaction between two cellular circuits, the electrically excitable neuronal network and glial cells which control cerebral circulation and homeostasis. Nowadays glial cells, especially brain astrocytes, are seen as competent participants of brain signaling. Astrocytes receive signals mediated by variety of neurotransmitters, integrate them and provide feedback to neighbouring cells by releasing gliotransmitter molecules. Important role for astrocytes in regulation of neuronal excitability, synaptic plasticity and neuro-vascular coupling has been demonstrated recently. It has also been shown that astrocytic dysfunction is implicated in the course of various age-related neurodegenerative diseases. Our recent work highlighted the importance of glutamate and ATP-mediated signalling for neuron-glial communications. We have shown for the first time the age-related decline in the level of expression of glutamate and ATP receptors and Ca2+-signalling in the astrocytes of brain cortex. One can expect that age-related decline in astroglial control of synaptic transmission and neuro-vascular coupling may strongly affect many brain functions, in particular learning and memory. Thus, glia-neuron communication may be of crucial importance for brain longevity and healthy ageing. However, age-related changes in astroglial signalling and their impact on brain longevity remain almost unexplored. The overall aim of the proposed project is to characterize the integrative role for astrocytes in aging brain. In particular, we shall test the hypothesis that age-related changes in glial modulation of neuronal signalling contribute to the decline in synaptic plasticity and memory. We shall also elucidate whether enriched environment and enhanced physical activity have a positive impact on glia-neuronal signalling and vascular-neuronal coupling in the aged brain.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Commercial scale manufacture of adult allogeneic cell therapy for regenerative medicine
Funder: Engineering and Physical Sciences Research Council

Abstract: The proposal aims to demonstrate the generic potential of a regulator-approved cell immortalization approach pioneered in the UK as a platform technology for scalable expansion of cells for therapy. It will enable 20 years R&D into neuronal regeneration strategies by Prof Raisman's group to be translated into clinical practice. We will adDr.ess robust scalable expansion and co-culture bioprocessing of olfactory ensheathing cells (OECs) and fibroblasts essential for therapy. Multiple animal studies have demonstrated safety and efficacy of OECs. However, a significant hurdle is that the present method uses autologous cells but treatment needs to occur within 14 days of injury. This is not adequate time to take a biopsy from the patient, process the cells to a therapeutic dose range, characterize and implant into the injury site. For routine therapies available in the limited time window, an off-the-shelf product is essential. We will therefore make a step change to universal (allogeneic) cell lines and explore parallel single cell cultures and direct co-culture options. Using a technique that is already approved by the MHRA we will produce conditional inducible OEC lines. The technology (c-MycER) employed to achieve conditional growth control and genetic stability is a fusion protein comprising a growth promoting gene, c-Myc, and a hormone receptor that is regulated by a synthetic Dr.ug, 4-hyDr.oxy-tamoxifen. These modified OECs will proliferate due to activation of c-myc. Cells successfully transduced will be selected and expanded on the basis of resistance to geneticin (only successfully transduced cells will express the geneticin-resistance gene). The resulting OEC lines will then be combined with fibroblasts at various ratios to determine the optimum blend for cell therapy application. The outputs will be tested in vitro, using a rapid, high-throughput screening method established in our lab. We will then proceed to an animal model to determine efficacy and safety.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: NEUROLOGICAL
Title: Manufacturing of iPS cell derived neurons from patients with familial Alzhiemer's disease for drug discovery and stratified medicine applications
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Progress in understanding many diseases has been hindered by a lack of suitable in vitro models, particularly for diseases in which affected cell types are difficult to access or obtain. Induced pluripotent stem cells (iPSCs) are an ideal alternative source as they can be derived from patients' skin cells and have the potential to form any cell type in the body. However, current technologies are hindered by the inability to reliably produce mature cell types. We have developed a method that enables iPSCs to be reliably differentiated at low cost and high purity into neurons. This offers the ability to utilise these cells for understanding Alzheimer's disease (AD) in the petri dish and developing new treatments and diagnostics for such patients. Ultimately, we aim to produce a technology capable of diagnosing AD and informing treatment options for patients as well as providing industry with the ability to screen and develop novel drugs for this incurable disease.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;5.1 PHARMACEUTICALS
Health categories: NEUROLOGICAL
Title: Gustotopic mapping in humans: a high resolution fmri study to assess detailed topography and modulations
Funder: Biotechnology and Biological Sciences Research Council

Abstract: There are five basic tastes: sweet, salt, bitter, acid, umami. Animal studies using extracellular recordings suggest a chemotopic organization of the primary gustatory cortex (PGC), and a recent optical imaging study supports the preferential response to tastants in defined spatial areas of the PGC. A number of functional MRI (fMRI) studies have assessed the brain's response to tastants, however no human studies to date have achieved the high spatial resolution required to assess responses at the level of a chemosensory gustotopic map. Our research will use experimental designs and analyses of ultra-high field (UHF) MRI data to map the representation of tastants in the PGC at very high spatial resolution. We have pilot data to show that bitter and sweet tastants are separated in the PGC. This proposal builds on this, forming a gustotopic map of primary tastants and examining other putative 'metallic' and 'fat' tastes. Taste perception varies depending on PROP phenotype/gustin genotype, and a new thermal phenotype called 'thermal taster status', for whom thermal stimulation to the tongue elicits a phantom taste. We will investigate variation in gustotopic maps across these phenotypes. We will also directly examine the phenomenon of thermal taster status, studying how phantom taste modulates responses in the PGC. We will determine how cortical response is modulated by increasing tastant concentration to establish whether this is a linear response or saturates. We will then address the question of intra-modal responses, assessing combinations of tastants that lead to enhancement (umami/salt) or suppression (bitter/sweet), and how they modulate activity in the PGC. Taste stimuli for fMRI studies will be well characterised using a sensory panel, and assessed by all subjects to link the brain and perceptual responses. These neuroscience questions fit squarely in the BBSRC remit of understanding normal human function and have a strong interdisciplinary component.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: The genetic mechanisms underlying the regenerative potential of ensheathing glial cells in drosophila
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The aim is to discover the molecular mechanisms by which the transcription factor Prospero (Pros) confers regenerative potential upon ensheathing glial cells. Central nervous system (CNS) injury induces a Glial Regenerative Response (GRR) across species that triggers ensheathing glial proliferation and leads to spontaneous remyelination and functional recovery of locomotion. In mammals, the GRR is implemented by NG2+ oligodendrocyte progenitor cells (OPCs). However, it is not known how to induce NG2+ cell differentiation to enable remyelination. NG2+ cells can be both permissive and inhibitory to axonal growth, and how this occurs is not understood either. With previous BBSRC funding, we established a novel injury paradigm to investigate the GRR in the fruit-fly drosophila, we discovered a gene network underlying the GRR, we found that Pros controls ensheathing glial differentiation, and that manipulating Notch and Pros levels in glia induced glial regeneration and axonal neuropile repair. We also found that the mammalian homologue of pros, Prox1, is expressed in NG2+ OPCs. Pros is the key factor that controls glial differentiation and influences neuropile repair. Here, the aim is to find out what genes are regulated by Pros in glia conferring this regenerative potential. We have identified 39 candidate Pros target genes. The experimental objectives are: (1) Using a combination of RT-PCRs, in situ hybridizations and genetics, we will validate these genes in relation to Pros and the GRR gene network and select five. (2) We will analyse the functions of five genes in glial differentiation and repair using genetics, immunostainings, confocal and TEM microscopy, and Optical Projection Tomography. (3) We will test whether the top candidate genes can influence neuronal regeneration, with a combination of DeadEasy software, time-lapse confocal microscopy and Flybow, glial manipulation with the LexA system, and locomotion recordings with FlyTracker and Trikinetics monitors.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Rising to the Challenge: Understanding causes and consequences of neural overactivation in young adult APOE-e4 carriers
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The e4 variant of apolipoprotein E (APOE-e4) has attracted attention as the single most important known genetic risk factor for late onset Alzheimer's disease. It is also associated with steeper age-related decline in cognitively intact older adults. Paradoxically, experiments in my laboratory and others have established that young adult carriers of APOE e4 significantly out perform non-carriers on tasks placing demands on sustained attention, achieving this through a distinct pattern of overrecruitment of brain regions in response to cognitive challenge. We suggest that this early life overactivation has two consequences: 1. it places unusual metabolic stress on key brain regions, 2. it exhausts the capacity for compensatory activations normally recruited as part of healthy aging. Building on this work, and using a unique combination of behavioural, brain imaging, neuropharmacology and pupilometry techniques, this proposal will establish whether e4 overactivation is limited to particular modes of processing, whether it is related to the amount of effort engaged by the task, and whether the overactivity can be 'switched off' under defined conditions. In addition, we will test the relationship between brain overactivity and the release of cortisol, thereby linking the brain activity directly to a potential mechanism through which long-term brain changes occur. Research outcomes will be: 1) an improved understanding of the early life consequences of carrying the e4 variant of APOE, 2) the identification of factors that modulate brain overactivations of in e4 carriers, and 3) an understanding of the mechanisms that link chronic overactivity to metabolic stress, as indexed by cortisol release. Together these advances will directly inform the development of behavioural and cognitive strategies and techniques which can be recommended to young carriers of e4 in order to mitigate the effects of brain overactivation and enhance the prospects for healthy cognitive ageing.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: microRNA-mediated Regulating of Axonal Structural Integrity and Nerve Injury Response
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The architecture of the nervous system is dependent on sophisticated connections established by axons and dendrites. The ability of neurons to maintain the integrity of long axonal projections throughout the life span of an organism is a fascinating biological phenomenon. Elucidating the molecular mechanisms controlling this process under physiological conditions and pathological stress remains a fundamental scientific question. miRNAs are ~22 nucleotide non-coding RNAs that emerged as essential post-transcriptional regulators of gene expression programs in development and disease. Interestingly, miRNAs are abundantly expressed in the CNS, and have been implicated in modulating neurogenesis, axonogenesis, as well as synaptic morphogenesis and plasticity. Directly relevant to this proposal, both acute and prolonged alterations in the levels of several miRNAs following traumatic never injury have been reported, suggesting they may play important roles in neuronal injury response. We propose to explore the role of miRNAs in controlling axonal structural integrity and the response to traumatic nerve injury, using a multi-pronged and multi-disciplinary experimental strategy in the genetically tractable organism drosophila melanogaster. Acute changes in miRNA homeostasis following traumatic nerve injury will be detected using state of the art miRNA profiling tools. High-throughput transgenic technologies will be used to discover miRNAs functionally required for axonal structural integrity and nerve injury response. Deciphering the regulatory pathways controlled by candidate miRNAs will be attained using cutting edge technologies for genome-wide miRNA target identification (Ago-HITS-CLIP). Finally, recently pioneered genome editing tools (TALEN/CRISPR) and epistasis experiments will be used to refine and validate direct effector genes. These studies promise to engender new insights into the molecular mechanisms underlying axonal structural integrity and pathophysiology.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: NEUROLOGICAL
Title: Lemur tyrosine kinase-2 and axonal transport of cdk5/p35 and protein phosphatase-1
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Cyclin dependent kinase-5/p35 (cdk5/p35) and protein phosphatase-1 (PP1) are two signaling proteins that function in a number of key processes within the synapse. We and others have shown that both cdk5/p35 and the catalytic subunit of PP1 (PP1C) bind to the serine/threonine kinase lemur tyrosine kinase-2 (LMTK2). In common with most neuronal proteins, cdk5/p35, PP1C and LMTK2 are all believed to be synthesized in cell bodies and then transported through axons to presynaptic termini. However, the mechanisms that mediate this transport are largely unknown. Kinesin-1 is a major neuronal molecular motor that drives anterograde axonal transport and we have obtained evidence that LMTK2 binds to the kinesin-1 light chains (KLC) so as to mediate attachment of LMTK2, cdk5/p35 and PP1C to kinesin-1 motors. The aim of this project is therefore to test the hypothesis that binding of LMTK2 to KLCs mediates kinesin-1 driven axonal transport of a complex containing LMTK2 and associated cdk5/p35 and PP1C to presynaptic termini. The objectives are:- 1. To finalise our studies demonstrating that LMTK2 scaffolds cdk5/p35 and PP1C to KLCs. 2. To properly characterise axonal transport of LMTK2 in transfected living rat neurons using time-lapse microscopy. 3. To study the role of LMTK2 on axonal transport of cdk5/p35 and PP1C. 4. To investigate the mechanisms that regulate axonal transport of the LMTK2-cdk5/p35-PP1C complex and in particular, how phosphorylation of LMTK2 affects its binding to cdk5/p35 and PP1C, and how phosphorylation of KLC1 affects its binding to LMTK2.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: NEUROLOGICAL
Title: Functional role(s) of oestrogen signalling on neuronal progenitor cell development and fate in the brain
Funder: Biotechnology and Biological Sciences Research Council

Abstract: We will apply an oestrogen responsive transgenic zebrafish model (developed at Exeter) and develop new transgenic models to investigate the roles of oestrogen in neurogenesis and brain development, investigating for sex related differences, including in intact animals in real time using advanced imaging methods. Specifically we will: 1.) Apply our current transgenic zebrafish model (ERE- GFP) to establish the role of oestrogens in telencephalic radial glial cell (RGC) differentiation and function, identifying the different cell types in the brain responsive to oestrogens, their ontogeny, sensitivity and behaviours. The work will include ablation of oestrogen responsive neurones during early life using two-photon microscopy. 2.) Develop a new transgenic zebrafish model [ERE_Gal4ff x UAS_GFP_KalTA4] and use this to follow the fate and behaviour of oestrogen responsive RGCs and other neural cells in juvenile and adult fish brains, investigating also for differences between the sexes. 3.) Create a further transgenic zebrafish [ERE_Gal4ff x UAS_ mCherry x UAS _GCaMP3] to identify neuronal activity in oestrogen responsive neurons. We will investigate the spatio-temporal activation of individual oestrogen responding neurons in the telecephalon associated with olfaction, including assessing for sex related differences. Oestrogen receptor morpholino knockdown will be applied to assess how oestrogen signal interference during early life affects subsequent telecephalon oestrogen responsive cell development and neuronal activity for olfactory stimuli. 4.) Apply our ERE_Gal4ff x UAS_GFP_KalTA4 transgenic model to investigate effects of exposure to a series of endocrine disrupting chemicals (EDCs) on oestrogen responding neuronal progenitors and their subsequent fate. Also examine for effects of these EDCs on the neuronal activity of oestrogen sensitive telencephalic cells using the ERE_Gal4ff x UAS_mCherry x UAS _GCaMP3 transgenic line.


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