NIHRDataInsights/HRCSData huggingface dataset:
Title: The impact of network remodelling on outcomes for regenerative medicine in the retina - defining the therapeutic window
Funder: Medical Research Council

Abstract: Some of the most encouraging data in regenerative medicine have come from attempts to restore photosensitivity in retinal degeneration. Introduction of stem cell-derived photoreceptors, optogenetic actuators, or electronic prosthetics to the degenerate retina have all been shown to restore light responses in human patients or animal models of the disease. Clinical development of these therapies currently concentrates on subjects with end stage degeneration and profound visual loss. However, retinal degeneration is accompanied by profound neuroanatomical reorganisation of the surviving visual circuits, raising the possibility that early therapeutic intervention could be advantageous. Here we have devised a program that determines the impact of the time of intervention upon the level of visual restoration using the rd1 mouse. We will first define the impact of remodelling on the capacity of retinal/central circuits to transmit and process visual signals. We will then determine the extent to which therapy at early or late stages of degeneration alters the progress of remodelling to improve the functional capacity of visual circuits, before finally asking whether time of intervention impacts on the quality of restored vision. We propose an innovative 'double-optogenetic' approach, using expression of a red-shifted channelrhodopsin in ON bipolar or ganglion cells as a circuit interrogator in combination with electrophysiological recordings to probe the transfer of information through visual circuits. Our candidate therapeutic will be an optogenetic intervention (ectopic expression of rhodopsin in ON bipolar cells) shown previously to restore vision in rd1 mice. By defining the impact of remodelling on circuit function and its interaction with therapy our study addresses issues of substantial concern to regenerative medicine, while answering the specific practical question of whether intervention at the early stages of retinal degeneration improves efficacy.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE;NEUROLOGICAL
Title: Neuroglobin in the Retina. Use of a Knockout Mouse for Functional assessment / Phenotyping and examining Human relevance, towards Neuroprotection.
Funder: Medical Research Council

Abstract: 1. Is there loss of function and structure time-course for retinal ganglion cell and amacrine cells in an Ngb KO mouse retina? 2. Are there changes in Ngb KO brain distribution of Ngb, structure, Apoptosis over an extended time? A longitudinal assessment of retinal function and retina / brain characteristics, will be performed on a cohort of mice from each genotype. Retinal structure by Optical Coherence Tomography (OCT scan), retinal function (ERG) in these animals will be assessed at 1, 3, 6, 12 and 18 months, at which point the animals will be used for immunohistochemical and apoptosis marker (TUNEL) analysis of the retina and the brain. Extra cohorts of mice will be used for immunohistochemical analysis of the retina and the brain at the intermediate time points described above. 18 month experimental study with completion of research questions by 30 months. 3. Are there similarities between human and mouse retina and brain Ngb distribution, tissue structure and Apoptosis markers? In collaboration with the UCL/ Moorfields histopathology service we will carry out immunostaining on selected library research human tissue sections. 18 month study based on 1. (above) 4. Is there any association between variations of human Ngb gene (or determinants of transcription) and markers for glaucoma/ ganglion cell/ optic nerve disease? I will work with experts in Bio-informatics/ UK Biobank experience, to look at glaucoma / optic nerve dysfunction, using gene-based statistical techniques to look up whether genetic variation in Neuroglobin in humans is associated with Ganglion cell variates in 40,000 participants in UK Biobank with the potential to search other databases. 12 months data gathering/ mining followed by 12 months analysis.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE;NEUROLOGICAL
Title: Elucidating splicing factor function and retinal splicing programmes: developing new therapeutic strategies for splicing factor retinitis pigmentosa
Funder: Medical Research Council

Abstract: Alternative splicing is a pre-mRNA processing step that removes or includes specific exons/introns enabling generation of multiple isoforms from a single gene. Mutations in pre-mRNA splicing factors (PRPFs) cause 15-20% of autosomal dominant retinitis pigmentosa (adRP). It is unknown how mutations in ubiquitously expressed splicing factors cause retinal disease. In collaborative work, we have used patient-specific induced pluripotent stem cells (iPSC) differentiated to retinal cell-types to show that PRPF31 haploinsufficiency results in reduced spliceosome activity, defective splicing and impaired ciliogenesis in retinal pigment epithelium (RPE) and photoreceptors. Building upon these multidisciplinary skills and expertise, we aim to focus on a key essential structural component of the activated spliceosome, PRPF8, in order to identify genes and functional pathways that are affected by splicing factor dysfunction and their contribution to adRP. Mechanistic insights into splicing regulation of key candidate genes will inform the development of proof-of-concept splice-correcting gene therapies for RP. The specific aims of this proposal are to: 1) use RNA-seq, quantitative proteomics and cellular assays to determine the effect of PRPF8 mutations on retinal cell structure and function, and identify mis-spliced genes and dysregulated splicing programmes; 2) use biochemical & in vivo splicing assays to assess the effect of PRPF8 mutations on splicing kinetics, and spliceosome assembly, stability, activity and function; 3) use RNA-Seq/iCLIP-Seq to identify the physiological & disease relevant RNA-binding sites of PRPF8 in the nucleus and cytoplasm; 4) use splice-switching morpholino oligonucleotides to validate the candidate mis-spliced transcripts in retinal cells; 5) gain insights into functional & splicing defects in PRPF8-related RP to develop new targeted therapeutic interventions, leading to translation into Phase I clinical trials for PRPF8-RP.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE;NEUROLOGICAL
Title: Dynamic decisions: Integrating dynamic gene expression with mechanical signals to understand retinogenesis and ocular disorders.
Funder: Medical Research Council

Abstract: Eye development is a dynamic process, but we do not understand how retinal cells undergo dynamic transitions between progenitor and differentiated states. I have developed live imaging techniques which have significantly contributed to the understanding of pulsatile and oscillatory expression of the cell fate determinants HES1 and HES5 (HES1/5) in cell fate decisions in neural tissue, but the relevance in the retina and functions of oscillatory expression are unclear. Furthermore, although we know that extrinsic factors regulate intrinsic cell fate determinants, the extrinsic control of ultradian oscillations in HES1/5 is unknown. This is important as mis-regulation of cell fate can lead to developmental disorders and disease. Critically, the co-occurrence in many patients of coloboma, a structural disorder, and microphthalmia, a cell fate disorder, suggests a mechanistic link between the mechanical environment and cell fate in the developing eye. I will investigate how mechanical properties of the cellular environment regulate the intrinsic expression dynamics of cell fate determinants HES1/5 in the retina. To address this, I propose a novel analysis integrating cutting-edge live imaging of protein expression dynamics, measurements of physical forces and human organoid modelling of ocular developmental disorders with the following aims: 1. Determine functional importance of dynamic cell fate factor expression in retinal progenitor cells. 2. Characterise mechanical properties of the retina and determine the impact of mechanical signals on expression dynamics of fate determinants and cell fate decisions. 3. Determine the impact of mutations linked to microphthalmia and coloboma on cell fate. These mechanistic insights would be clinically valuable by helping to explain why coloboma and microphthalmia can co-occur.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE;NEUROLOGICAL
Title: New genetic therapy approaches for inherited retinal diseases
Funder: Medical Research Council

Abstract: Inherited retinal diseases, including retinitis pigmentosa and Leber congenital optic neuropathy (LHON) are the leading cause of blindness in the younger population in developed countries. Retinitis pigmentosa is caused my mutations in genes in outer retinal cells that ultimately lead to the loss of photoreceptors but the inner retinal cells, such as retinal ganglion cells (RGCs) remain intact. LHON is caused by mutations in mitochondrial genes that primarily affect the ganglion cells. There is no cure for these diseases. After loss of photoreceptors, optogenetic therapy aims to introduce light sensitive proteins to surviving cells of the retina, including RGCs, and confer ability to detect light. For patients with LHON, a promising strategy for the treatment of mitochondrial mutations in RGCs is allotopic expression-based gene therapy via intravitreal delivery of adeno-associated viral (AAV) vectors. However, efficient targeting of RGCs with therapeutic vectors has not been achieved to date. Herein, we aim to develop a surgical procedure using a robot to more effectively target the cells of the inner retina. The procedure will involve robot-assisted direct infusion into the optic nerve of non-human primates which is currently not possible to perform manually in patients. Following this, we aim to optimise AAV gene delivery platforms by optimising mitochondrial delivery for LHON applications and by restricting transgene expression to soma and dendrites of RGCs, for optogenetic applications. Lastly, we aim to explore the possibility of activating endogenous opsins in RGCs using a gene editing tool, CRISPRa, as an alternative novel strategy for optogenetic therapy. The approaches have potential to lead to treatment of optic neuropathies and any late stage retinal degeneration.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.1 PHARMACEUTICALS;5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE;NEUROLOGICAL
Title: Targeting residual visual pathways to develop effective rehabilitation for patients with hemianopia
Funder: Wellcome Trust

Abstract: Hemianopia is common following acquired brain injury, and is estimated to affect over 230,000 people living in the UK. Despite its high prevalence and well-described impact on society, it is a neglected clinical problem with no treatments to recover vision. My research shows that patients with hemianopia can show residual vision in their blind field, a phenomenon called 'blindsight'. Patterns of blindsight and one's capacity for visual retraining appear to vary according to the presence of preserved 'secondary' visual pathways. Where pathways remain intact, they represent an important potential target for rehabilitation. I will use cutting-edge neuroimaging techniques in patients and healthy controls to build a normative connectivity atlas of the central visual pathways. I will quantify the relationship between secondary visual pathway integrity and function in patients and controls, to learn how to optimally drive their response. I will use results to develop a novel visual-training protocol targeting the ventral visual stream, and test whether the integrity of ventral and dorsal visual pathways can predict improvement with visual training protocols targeting those two domains in patients with hemianopia. Understanding the secondary visual pathways will expand neural targets and enhance precision through a customised approach to treat hemianopia.

Lay abstract: Sight loss is extremely common after a stroke or brain injury, caused by damage to the vision centre at the back of the brain. This condition, known as 'hemianopia', can cause significant distress through loss of a driving license and social isolation. Unlike other parts of the body, there are no treatments to recover sight loss after a stroke. However, my research shows that the brain can still respond to images, even though you may not be aware of seeing anything at all. This is called 'blindsight', and it relies on previously unknown connections between the eyes and spared areas in the damaged brain that bypass the main vision centre. My proposal uses state-of-the-art brain imaging and vision tests to measure and target these pathways using new visual training techniques. The purpose will be to select the right training tool for the right person, to maximise recovery of sight loss.

Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: EYE;NEUROLOGICAL
Title: Purpose and Mechanisms of Behavioural Modulation in Retina and Superior Colliculus
Funder: Wellcome Trust

Abstract: The project aims to uncover why and how behavioural states like arousal affect neural processing in the early visual system by recording from the retina and superior colliculus in awake mice using techniques I have previously developed. Behavioural states affect task performance but their effect on early sensation is unclear; understanding their impact on early visual processing will reveal fundamental computational principles of adaptation. Our objectives are: 1. determine the effects of behavioural states on different cell types in the early visual system by recording activity of retinal and collicular neurons classified by function, genetics and projection targets in awake mice. 2. determine the purpose of behavioural state modulation in terms of information processing in the early visual system by recording activity of retinal and collicular cell types in awake mice viewing visual stimuli that elicit nonlinear responses or are behaviourally relevant. 3. determine the mechanisms by which behavioural states influence early visual processing by measuring and manipulating the activity of neuromodulators, serotonin and noradrenaline, controlling behavioural states while recording from retinal and collicular neurons. The results of this project impact on basic neurosciences and other fields including the development of artificial intelligent systems and the treatment and diagnosis of psychiatric disorders.

Lay abstract: Our state of mind largely determines our cognitive performance, which is significantly compromised in mental disorders like attention deficit disorder. Although we know that mental states affect neural activity in many brain regions, an open question is how effects on sensory processing lead to drops in cognitive performance. For example, does fatigue limit our reading ability because our sense of sight suffers? My goal is to determine how mental states change visual signals as early as in the eye and how this affects our behaviour. I will investigate how neurons with different roles in vision are affected by mental states, how the processing of behaviourally relevant visual input changes, and how brain messengers like serotonin control the effect of mental state on vision. I will use large-scale imaging and electrical recordings of hundreds of neurons; genetic and neural tracing tools to distinguish cell types; neural manipulation to elucidate functional roles.

Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE;NEUROLOGICAL
Title: Daylight vision beyond cone photoreceptors
Funder: Wellcome Trust

Abstract: The ability of cones to detect small differences in light intensity over fine spatiotemporal scales is the basis of high acuity vision. However, high amplitude and coarse patterns are common in visual scenes. An ‘intelligent designer’ may thus include a separate photoreceptor optimised for detecting such features. Our latest data leads to the striking conclusion that such a function is indeed provided by the inner retinal photopigment, melanopsin. I present an inventive and original programme of work to test this hypothesis, built upon my laboratory’s world-leading technological expertise in multi-primary visual display design (allowing us to produce images visible to melanopsin for the first time), and its multi-disciplinary infrastructure spanning molecular biology and cell culture to electrophysiology and behaviour. I will employ new and existing transgenic mouse lines and our colony of Rhabdomys, a diurnal mouse with strong cone vision. I will define melanopsin’s contribution to vision in healthy animals with fully intact visual systems, and in mouse models of photoreceptor degeneration. I will determine whether enhancing melanopsin’s activity in retinal degeneration can augment residual vision. My project holds the promise of a step-change in our understanding of daytime vision with wide-ranging practical applications.

Lay abstract: Daytime vision is thought to originate with light sensitive cells in the retina called cones. Excitingly, I have recently found that another light detector (melanopsin) can also detect images under daylight conditions. In this project I will determine how melanopsin and cones work together to allow us to see. My work takes advantage of the first visual displays capable of presenting images visible to melanopsin (a breakthrough analogous to the appearance of colour displays for colour vision research), and the latest genetic tools, to study melanopsin vision in laboratory mice and in an African diurnal mouse with strong daytime vision. The project promises a step-change in our understanding of vision, and has realistic translational potential. My findings could establish ‘melanopsin displays’ as a technology for improving image quality in commercial visual displays, and establish a new range of therapies targeting melanopsin to enhance vision in retinal degeneration.

Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE;NEUROLOGICAL
Title: How to connect an eye to a brain
Funder: Wellcome Trust

Abstract: Retinal ganglion cells (RGCs) are the vertebrate eye’s only projection neurons to the brain. Through RGCs, species transmit ~50 visuo-ecologically relevant image features in parallel. However, what these features are, and how their information is used by central circuits to inform visual decisions remains unclear in any species. A key difficulty has been to link the activity of RGCs in the in vivo eye with behaviour. To address this major knowledge gap, I will capitalise on the visual system of the larval zebrafish where the activity of RGCs in the eye and their presynaptic terminals in the brain can be non-invasively monitored and manipulated in the live animal. I will ask: 1. What information does the fish’s eye send to the fish’s brain? 2. Does the brain “tune” its own input from the eye? 3. How does the brain use information from RGCs to guide behaviour? Taking reference of our and others’ data on RGC processing from mice and primates, we ultimately aim to arrive at a more general theory of how an eye can communicate with its brain, that encompasses coding strategies employed by diverse species with distinct visual abilities and requirements. Key-words: Vision, projection neuron, zebrafish, 2-photon imaging

Lay abstract: Background. Our eYES communicate with the brain via functionally diverse nerve cells that transmit information about specific image aspects such as the orientation of an edge or the direction of an object’s movement. However, what all these nerve cells tell the brain, how these messages differ between animals that use their eYES for very different things, and how they are used to guide behaviour remains unclear in any species. Approach. Working on the transparent and experimentally accessible larvae of zebrafish, a central model in vision research, I will non-invasively monitor and manipulate the activity of 1,000s of visual nerve cells to address these major gaps in knowledge in the live, behaving animal. Impact. I aim to establish crucial information that will ultimately inform a general theory of how an eye can communicate with its brain.

Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE;NEUROLOGICAL
Title: Investigating mechanisms involved in retinal signalling in health and disease through detailed in vivo human electrophysiological investigation
Funder: Wellcome Trust

Abstract: Retinal imaging investigates structure, but not cellular function. Electrophysiology allows direct assessment of the latter. The electroretinogram (retinal electrical response to light) can be recorded non-invasively. As understanding of retinal processing improves, with refinement of mathematic models of current flows, light and dark adaptation, there is scope to probe retinal signalling in great detail. Findings from animal and in vitro studies can be specific to species or laboratory conditions. This project uses in vivo human recording. Novel stimulus protocols will be used in patients with largely monogenic disease (including natural “knock-outs”), and in genetically characterised twins (both using the classic twin study and by exploring further genetic associations including the retinal functional significance of myopia-associated genetic loci). Project goals include the following: 1. A more integrated model of sensory signalling pathways in the retina: elucidating roles of specific proteins (in the photoreceptor-RPE complex and in signal transmission to bipolar cells) 2. Elucidation of a myopia signalling cascade: functional effect of variants that confer susceptibility to myopia These will yield insights into neural processing (with wider implications for neuroscience), retinal disease and myopia. As new gene and stem cell therapies emerge, novel functional assessments can provide objective outcome measures.

Lay abstract: The retina is the structure at the back of the eye that changes light into electrical signals. These signals travel to the brain allowing us to see. Retinal diseases cause blindness. Also, short-sightedness (which is becoming extremely common and which has complications that can cause blindness), is driven by abnormal signals in the retina. Technology looking at retinal structure has advanced; we can see single cells in the eye. Measuring retinal electrical function is not as developed, and this project will address this. By making safe, non-invasive recordings of retinal electrical responses from large numbers of human subjects (patients with retinal diseases and thousands of twin volunteers), this project will help us understand how the retina works, and why alterations in retinal function cause blindness or short-sightedness, opening the door to new approaches for developing treatments. As the retina has similarities with the brain, this can also help us understand some brain diseases. We are now starting to try out some very new treatments (gene and stem cell therapies) to try to restore function to parts of the retina. This project will provide new ways of precisely measuring retinal function, so we know whether these treatments actually work.

Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: EYE;NEUROLOGICAL;MENTAL HEALTH