NIHRDataInsights/HRCSData huggingface dataset:
Title: Optic nerve microstructure: a significant indicator of disease severity.
Funder: Health and Care Research Wales (Welsh Government)

Abstract: Glaucoma is the second most common cause of irreversible blindness and in 50% of cases is undetected. Primary open-angle glaucoma results in retinal cell and axon death leading to bilateral blindness in 4-6.3% of patients. This research aims to characterise microstructural features within optic nerve heads that can be used to identify those optic nerves that are at risk of either development of the disease or of rapid progression. Early detection of disease onset of glaucoma is critical to prevent irreversible axon and vision loss. This study will be achieved using a novel imaging device called optical coherence tomography. Our device uses a longer wavelength of light than many commercial devices, which enables us to visualise the structures within the retina and optic nerve. There is now substantial evidence that changes in the lamina cribrosa, a supportive structure to axons within the optic nerve as it leaves the eye on its journey to the brain, initiates retinal ganglion cell (RGC) death in glaucoma. Thus the principal outcome measure of this study is anticipated to be the characterisation of the lamina cribrosa configuration at different stages of disease, in relation to cell and axon loss in the eye blinding disease, glaucoma. In addition, as there is variation in eyes in the normal population, a number of other ocular features will be taken into account when evaluating statistical significance to remove any biase with respect to influencers such as ocular size and age. The outcomes of this study will be of great importance for the discrimination of patients who are at risk of developing glaucoma (e.g. ocular hypertensive patients) or glaucomatous disease progression. In turn, this will enable earlier intervention, prior to irreversible vision loss.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: Disorders of human brain development: genetics and function
Funder: Medical Research Council

Abstract: Purpose: Identify genes important in brain development from the study of human neurogenetic disorders. Characterise these genes cellular and developmental functions utilising cell biology and model organisms, to gain insights into the pathogenesis of human disease.||Background: Determination of brain size is an important component of neurodevelopment and has experienced significant evolutionary change. Brain size has markedly increased during mammalian evolution, and in the last 2 million years the human brain has more than doubled in size. In this respect, the human disorder of primary microcephaly appears to recapitulate an earlier evolutionary state, with a brain size comparable with that of early hominids. Microcephalin is a protein that I have identified as a cause of primary microcephaly and its further characterisation currently forms the major focus of my work. This protein has been subsequently found to have roles in cell cycle regulation, chromosome organisation and DNA repair.||The gene has also experienced Darwinian adaptive evolution in primates, indicating that the protein may well play a role in the evolutionary expansion of brain size.||Current work||- Identify the molecular mechanisms associated with aberrant regulation of chromosome condensation in microcephalin deficient cells.||- Characterise patient cell lines from individuals with primary microcephaly and related conditions for DNA repair and cell cycle defects.||- Positionally clone novel microcephaly genes (Seckel Syndrome) to identify novel components of the ATR signalling pathway.||- Utilise the Drosophila microcephalin model that we have generated to investigate developmental mechanisms and identify genetic interactions||- Investigate the pathogenic basic of Aicardi-Goutieres syndrome (SCF Fellowship)


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: EYE
Title: The ultrastructural basis of corneal dysfunction and the development and optimization of novel therapeutic strategies
Funder: Medical Research Council

Abstract: The cornea of the eye is a uniquely transparent, exquisitely shaped tissue whose functionality depends heavily on the hierarchical ultrastructures and complex microanatomy of its cells and extracellular matrix. Despite the importance of the cornea for vision, the basis of corneal transparency and shape, and their compromise due to injury or disease, are only now starting to be understood in any detail. Our ultimate objective is to build upon the success of our previous studies and relate loss of transparency and changes in corneal shape/astigmatism to tissue micro- and ultra-structure. This will be achieved by taking advantage of recent advances in microfocus synchrotron x-ray scattering, electron tomography, multiphoton laser scanning microscopy and volume scanning electron microscopy to study human foetal corneas, normal and pathological adult corneas, post-surgical corneas and artificial corneal constructs. Our aims are to (i) model corneal transparency at the cellular and fibrillar level, and to extend this to explain the loss of transparency in corneal wounds and diseases; (ii) characterise the full three-dimensional structure of the cornea and explain the structural basis of astigmatism, and demonstrate how surgical treatments can be modeled to predict the cornea's macroscopic behaviour by finite element analysis; (iii) develop and better understand new methodologies for treating or stabilizing corneas, such as cell-based therapies for improving transparency and mechanical strength in corneal wounds and crosslinking methods to stabilize corneas to prevent ectasia; (iv) use the new three-dimensional structural information to assess and improve different approaches being used to develop biological artificial corneal replacements.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: Investigating the role of complement factor H-like protein-1 and factor H related proteins in age-related macular degeneration
Funder: Medical Research Council

Abstract: Age-related macular degeneration (AMD) is the most common form of blindness in the Western world and is characterised by the presence of extracellular debris including drusen at the interface between the retinal pigment epithelium (RPE) and Bruch's membrane. Drusen contain complement proteins suggesting a central role for the complement cascade in their pathogenesis. The common Y402H polymorphism in the gene encoding complement factor H (CFH) has been associated with an increased risk of developing AMD. Recently, I have obtained preliminary evidence showing that a protein resulting from a splice variation of the CFH gene, factor H-like protein 1 (FHL-1) that is also subject to the Y402H polymorphism, is the main regulator on the Bruch's membrane. I hypothesise that FHL-1 confers protection to the Bruch's membrane and that interplay exists between this protein and proteins encoded by related, but separate complement genes, referred to as the factor H-related proteins. My aim is to test this hypothesis by examining the macula of genotyped human eye tissues: donor tissue genotype will be mapped for all the disease-associated genes. Donor eyes will be analysed histologically (including using Picro-Mallory staining) and grouped into unaffected, early or late AMD. Fluorescent immunohistochemistry will show the endogenous localisation of the FHL/FHR proteins in the various disease states. Immuno-EM will elucidate the macromolecular nature of protein/Bruch's interaction and how this changes during the disease process. Functional assays will determine the effect of the Y402H polymorphism on FHL-1 regulatory activity and how this alters with disease. This project is focused on understanding the biochemistry of the early stages of AMD progression, and as such may help guide improvements in treating this disease early in it pathogenesis, before the devastating, life-changing, symptoms occur.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: EYE
Title: Morphogenesis and growth of the eye in health and disease
Funder: Medical Research Council

Abstract: This programme of research will elucidate the genetic pathways, cellular behaviours and developmental mechanisms underlying the specification and morphogenesis of the vertebrate eye. Our studies will help reveal the mechanistic bases of ocular phenotypes such as anophthalmia and coloboma. The knowledge we acquire from genetic analyses of mutant zebrafish will be used to screen for comparable genetic defects in patients with ocular malformations. One goal will be to identify the genetic lesions in a novel collection of fish lines carrying mutations that alone, or in combination lead to defective eye specification or morphogenesis. We will use next generation sequencing approaches to identify the genetic lesions underlying the phenotypes. Once the mutated genes are identified, we will use a transcript counting approach which involves 3' capture of mRNAs in mutants to identify the changes in gene expression that precede the appearance of morphological phenotypes and/or sensitise embryos to eye defects. We will resolve the cellular behaviours and developmental mechanisms that underlie optic vesicle evagination and choroid fissure closure in normal conditions and in embryos carrying genetic lesions. This will be achieved through the use of transgenes that label eye cells or subcellular structures in vivo, coupled with high-resolution multi-photon imaging of wildtype and mutant eyes. Finally, we will use results from the studies above to help to identify genetic lesions that contribute to human ocular phenotypes. We will use an amplicon based approach to sequence exons from candidate genes in patients but may switch to exome sequencing should costs reduce significantly. In parallel, we will expand our collection of lines of fish that carry mutations in genes that render carriers susceptible to ocular phenotypes. These genetically sensitised lines will be used to assess the consequence of mutations in genes that are candidates for causing human disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: EYE
Title: Optimisation of human ESC-derived photoreceptor cell differentiation
Funder: Medical Research Council

Abstract: Retinal degenerations with loss of photoreceptors are the leading cause of blindness in Europe. As there are no treatments that restore lost photoreceptors and vision, there is a clear need for new therapeutic approaches. In 2006 we showed that photoreceptor can be transplanted into the adult retina and recently we have been able to provide the first definitive evidence of restoration of rod-mediated vision, following transplantation of rods into a model of stationary night blindness. Importantly, we have also recently demonstrated that following transplantation, mESC-derived rod photoreceptors are able to integrate, mature & connect with the adult host retinal circuitry. Thus our research to date has established principles of effective transplantation using donor and mESC-derived photoreceptors. Whilst we demonstrated that retinal repair by photoreceptor transplantation is feasible, for clinical application we need to develop protocols for generating & transplanting human photoreceptors from a renewable source. In this project we aim to develop a defined and xeno-free differentiation system to generate and store photoreceptors from clinical grade hESC lines that will support the development of a clinical trial in retinal dystrophy patients. We will (i) develop xeno-free protocols for hESC lines (ii) grow EUCTD-grade hESC lines in the optimal conditions established in (i) (iii) modify our hESC differentiation protocols to be compatible with clinical application (iv) characterize the development of xeno-free EUCTD-grade hESC-derived photoreceptors & utilise viral vectors and CD markers to purify photoreceptor precursors for transplantation (v) transplant photoreceptor precursors generated from EUCTD-grade hESC lines; (vi) develop cryopreservation protocols for photoreceptor & determine their survival after thawing (vi) determine which EUCTD-grade hESC lines are the most effective for generating photoreceptor that can be transplanted following cryopreservation


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: Using cell surface antigens for the isolation of human photoreceptor precursor cells for retinal stem cell therapy
Funder: Medical Research Council

Abstract: Retinal degenerations with loss of photoreceptors are the leading cause of untreatable blindness in Europe. In 2006 we showed that photoreceptors can be transplanted into the adult retina and recently we have been able to provide the first definitive evidence of restoration of rod-mediated vision, following transplantation of rods into a model of stationary night blindness. Importantly we have also recently proved in the mouse model the concept of developing and utilising a cell surface biomarker panel to isolate transplantation-competent photoreceptor precursor cells from 3D embryonic stem cell retinal differentiation cultures. Whilst we have demonstrated that retinal repair by photoreceptor transplantation is feasible, for clinical application we need to develop protocols for purifying human photoreceptors from a renewable source. In this project we aim to identify a human cell surface biomarker panel and to develop robust methodologies for identification and purification of human rod and cone photoreceptor precursors from clinical grade human pluripotent stem cell cultures that will support the development of a clinical trial in retinal dystrophy patients. To achieve this goal we will: (1) Test the biomarkers we have identified in the mouse for labelling and isolation of human fetal photoreceptor precursor cells and immunophenotype human fetal photoreceptor cells; (2) Define the biomarker profiles of human ESC-derived rod and cone photoreceptors by transcriptome analysis at sequential stages of maturation in human ESC-derived retinal cultures, and compare these profiles to transplantation competent mouse photoreceptor precursor cells; (3) Evaluate combinations of identified biomarkers for purification of hESC-derived rod and cone photoreceptors; (4) Evaluate biomarker selected hESC-derived cells in transplantation experiments; (5) Develop robust biomarker panel-based isolation protocols and quality control assays for human photoreceptor precursors.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: Development of an opticin-based anti-angiogenic therapeutic for the treatment of ocular disease.
Funder: Medical Research Council

Abstract: Several blinding eye conditions, including retinopathy of prematurity and proliferative diabetic retinopathy, are characterised by preretinal neovascularisation, i.e. pathological angiogenesis with new blood vessels growing from the retina into the vitreous. The new blood vessels cause visual loss through bleeding and/or tractional retinal detachment. Current management includes panretinal photocoagulation and surgery. Both are associated with predictable side-effects and better treatments are needed. Our solution is to develop recombinant human opticin as a therapeutic that is delivered by intravitreal injection. Opticin is a glycoprotein that occurs naturally in vitreous. We have shown that opticin inhibits preretinal neovascularisation in a dose-dependant manner, with excess opticin having a greater anti-angiogenic effect than endogenous levels. We have elucidated its mechanism of action: it binds to collagen and thereby inhibits endothelial cell adhesion and collagen-mediated pro-angiogenic signalling that is essential for angiogenesis. Aims of this proposal are to: - Investigate whether intravitreal opticin injection induces vascular regression, - Perform ocular PK studies to investigate the half-life of opticin in the vitreous cavity and to determine whether any injected opticin escapes into the circulation or brain, - Determine whether therapeutic levels of opticin affect retinal vascular development, morphology or function, - Undertake additional mechanistic studies to confirm results with fibrillar rather than monomeric collagen. The above experiments are key to determining the potential of opticin as a therapeutic for preretinal neovascularisation, and for attracting funding for further preclinical development and subsequent clinical trials.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: EYE
Title: Optical coherence tomography in infants and children: A normative database and assessment of clinical use
Funder: Medical Research Council

Abstract: Optical coherence tomography (OCT) provides high-resolution scans of the retina and optic nerve and has contributed to significant progress in the diagnosis and treatment of retinal and optic nerve diseases in adults. However, infants and young children have been deprived of the diagnostic progress as they cannot cooperate with conventional OCT. The advent of a new hand-held OCT (HH-OCT) allows scanning infants and small children. Over the last three years, we have used the HH-OCT successfully to establish progression of normal human retinal development and develop a diagnostic algorithm for nystagmus. However the lack of normative data and knowledge about presentation of diseases prevents wider clinical use. The normal optic nerve is not mature at birth and is likely to undergo significant changes until adulthood. However, to date little is known about the normal postnatal development. Developmental abnormalities of the optic nerve and childhood optic nerve diseases such as glaucoma are leading causes of child blindness. First, we plan to optimise and acquisition protocols to enable fast and reliable volumetric scans. We then aim to establish a normative database for optic nerve and retina with OCT in infants and children. We will investigate to what extent OCT can help in improving diagnosis of childhood optic nerve disease. We also will explore whether visual acuity can be predicted from OCT scans. We will investigate to what extent OCT can help improve monitoring of childhood glaucoma and optic nerve glioma and reduce the number of examinations under anaesthesia. Software for automatic segmentation will be developed for easier and faster interpretation of OCT scans. We anticipate that our study will be an important step in validating and providing access of OCT in young children with optic nerve diseases. The normative database will provide the groundwork for using OCT in young children.


Research activity codes: 2.6 RESOURCES AND INFRASTRUCTURE (AETIOLOGY);4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: EYE
Title: A Phase I/IIa Clinical Trial of a Humanised Monoclonal Antibody Against LRG1
Funder: Medical Research Council

Abstract: This is a proposal to conduct a Phase I/IIa clinical trial (LABINA) of Magacizumab, a humanised monoclonal antibody against LRG1, in patients with neovascular age-related macular degeneration (AMD). The methodology follows a conventional series of preclinical steps including the development of a master cell line and preparation of clinical grade antibody, then safety and toxicology testing in pigs and human Lrg1 knock-in mice specifically generated for this purpose as part of our current DPFS project. We will then secure ethical and regulatory approval before progressing to Phase I of the clinical trial. This will take the form of a dose-escalation study in patients who respond poorly to standard of care (SOC), and will be followed by Phase IIa in which Magacizumab is tested in combination with SOC in naive or newly diagnosed patients. The primary endpoint of the LABINA trial will be safety but in Phase IIa we will also look for indications of biological activity using techniques such as optical coherence tomography, fluorescein angiography and visual acuity testing. A successful outcome to this project will mark a major step towards improving therapeutic outcomes for patients with neovascular AMD, among whom there are many who either do not respond to SOC or become refractory. In addition, we will generate new intellectual property that will add to the patents we already hold around LRG1. This will place us in a strong position for onward exploitation of the research through licensing or creation of a spinout.


Research activity codes: 6.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: Restoring vision in mouse models of retinal degeneration using human rod opsin.
Funder: Medical Research Council

Abstract: Degeneration of rod and cone photoreceptors is a major cause of blindness. In patients with this condition, inner retinal neurones survive and we know from work with electronic prostheses that if they can be presented with visual information the machinery exists to transmit it to the brain where it can be interpreted. This proposal is based around a great deal of preliminary data showing that intra-vitreal injection of AAV2/2 driving expression of human rod opsin in surviving ON bipolar cells in a mouse model of advanced retinal degeneration (rd1) can render them photoreceptive and support visual discrimination. The quality of vision produced by this simple approach compares very favourably with that provided by alternative strategies. This proposal builds upon that breakthrough, and comprises a thorough investigation of the safety and efficacy of the rod opsin based intervention in mice using behavioural and electrophysiological assays established in the applicants' laboratories. We aim to determine the best visual performance achievable with this approach by measuring its spatial and temporal resolution over a range of background light intensities and visual contrasts; ask how stable the restored vision is over time; whether it is effective for other forms of retinal dystrophy; and whether it would augment or interfere with low vision at early stages of degeneration. In preparation for clinical application we also ask whether the restored vision can be assayed using clinically relevant electrophysiology (electroretinograms and visual evoked potentials) and can be produced by sub-retinal delivery of virus. Our experiments will also address the question of safety by revealing what impact (if any) exogenous expression of rod opsin has on function or viability of the inner retina. We propose making a transgenic mouse line in which human rod opsin is expressed in all ON bipolar cells to augment our established viral gene delivery methods in addressing these questions.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: EYE