NIHRDataInsights/HRCSData huggingface dataset:
Title: Enhancing ocular gene therapy using glycosidic enzymes
Funder: Medical Research Council

Abstract: The overall aim of this study is to: Investigate strategies to improve retinal transduction efficiency using combinations of intravitreally delivered AAV serotypes and glycosidic enzymes. Specific objectives of this study are to: 1. Optimize combinations of glycosidic enzymes and AAV serotypes for the transduction of inner and outer retina using an intravitreal approach in a mouse model 2. Assess the safety of these treatment combinations in the mouse using functional studies 3. Confirm our ability to target cells in ganglion cell layer and photoreceptor layer in a therapeutically relevant manner using this technique Methodology: AAV containing a cDNA encoding enhanced GFP under the control of a chicken beta-actin promoter will be delivered intravitreally into anaesthetized adult mouse eyes in conjunction with enzymes. Various combinations and doses of glycosidic enzymes including hyaluronan lyase, chondroitin ABC lyase and heparinase III will be co-injected with the AAV2. In addition, I will investigate whether other AAV serotypes (AAV5, AAV8 and AAV9) can achieve retinal transduction following intravitreal injection when used in combination with these enzymes. Two weeks later retina flat mounts will be examined by confocal microscopy to analyze the amount and depth of penetration of the GFP fluorescence (using ImageJ software). Other retinas will be snap frozen, cryostat sectioned and co-stained with antibodies to GFP and to proteins expressed in specific types of retinal neurons to establish which particular cell types are transfected. At two weeks post treatment retinal function will be evaluated with dark- and light- adapted electroretinograms (ERG) and visually evoked potentials (VEPs), which will be compared to control eyes. Next, I will generate triple-knockout (Opn4-/-, Rho-/-, Cnga3-/-) mice lacking rod, cone and melanopsin transduction. I will then be able to use AAV containing cDNA for melanopsin (Opn4), rod opsin (Rho) or the cone cyclic GMP-gated channel A-subunit 3 (Cnga3) to restore phototransduction in melanopsin ganglion cells, rods or cones respectively. The readouts for these experiments will be circadian photo-entrainment (melanopsin photoreceptor activity), pupil reflex activity, ERGs and VEPs. The retinas will also be examined using immunohistochemistry. Scientific and medical opportunities of the study: If successful, this work will validate an intravitreal injection as a route for ocular gene therapy. As the current the approach of sub-retinal injection has many practical limitations, my work will meet an immediate clinical need, which is only likely to grow as the range of ocular disorders amenable to gene therapy increases.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE
Title: Incidence and progression of posterior segment eye disease in a population-based cohort in Kenya
Funder: Medical Research Council

Abstract: Background There has been limited focus on posterior segment eye diseases (PSED) by ‘prevention of blindness‘ programmes in Africa. Diagnostic challenges surround these conditions and assumptions have been made that they are rare. However, recent surveys indicate that PSED are now responsible for up to one third of visual impairment and blindness in Africa. There is currently a great lack of data on the incidence and progression of PSED in Africa. Aim To assess the five-year incidence and progression of diabetic retinopathy (dr), age related macular degeneration (AMD) and glaucoma in a Kenyan population aged ?50 as a follow up of the Nakuru PSED study. Methods Recruitment: In 2007/8, 4381 participants aged ?50 years were recruited in a population-based survey in Nakuru, Kenya. They underwent comprehensive ophthalmic examination and fundus image capture. These participants will be re-invited to attend for examination at a local centre, tracked through GPS points taken of their homes. A pilot follow-up study undertaken in January 2009 suggests that 5-years post baseline, at least 65% of the original participants (n=2,848) would be retraced and examined, approximately 2,205 of whom will have gradable images from baseline. Data Collection: Study participants will each undergo: 1) Visual acuity testing; 2) Visual field assessment; 3) Anterior segment assessment with a slit lamp and measurement of intra-ocular pressure; 4) Retinal photography using a Topcon non-mydriatic camera (dilated); 5) Clinical assessment of posterior segment (dilated) with slitlamp biomicroscopy and OCT); 6) Random blood glucose; and 7) Interview on eye conditions and treatment. Analysis: The Moorfields Grading Centre will grade retinal images for signs and stages of dr, AMD and glaucoma, using fundus photos. Assessment of disease progression and independent predictors of progression will be undertaken on participants with signs of dr, AMD or glaucoma at baseline. The incidence and predictors of individual PSED development will be analysed on those participants without PSED at baseline. Scientific and medical Opportunities These data will contribute to establishing diagnostic and prevention strategies, advocacy for provision of resources and identification of high-risk groups. Specifically, Africa is on the cusp of a diabetes epidemic and the impact of dr needs to be assessed. People from African populations appear to be more vulnerable to glaucoma, however, currently incidence and progression data are lacking. The baseline data on AMD suggests this disease presents differently in the Kenyan population, and it is important to assess how it progresses.


Research activity codes: 2.4 SURVEILLANCE AND DISTRIBUTION
Health categories: EYE
Title: To investigate the performance of scanning laser polarimetry and scanning laser ophthalmoscopy in glaucoma case finding
Funder: Medical Research Council

Abstract: Aims: To investigate if optic disc/ retinal nerve fibre layer imaging is a viable method for glaucoma case finding. Objectives: 1. To investigate the performance of scanning laser polarimetry (GDx) and scanning laser ophthalmoscopy (HRT) in glaucoma case finding in a community setting in the UK 2. To assess whether combination of GDx and HRT measures may enhance case finding performance 3. To explore whether additional demographic, socio-economic or ophthalmic features may be used to further enhance the performance of case finding. Design: The European Prospective Investigation of Cancer (EPIC) study is a large multi-centre cohort study looking at the connection between diet and cancer. One of the cohorts, EPIC-Norfolk, consisting of 30,446 men & women, was established in 1993. Since 2004, detailed eye examination data on 6,277 subjects aged 49 years and older have been gathered, and examination on 8,000 subjects is expected to be complete before the start of this project. The data includes among others, parameters for glaucoma assessment, such as visual acuity, intraocular pressure, retinal nerve fibre layer imaging with scanning laser polarimetry, optic nerve head tomography with Heidelberg HRT2, as well as disc & macular digital photographs. Participants with abnormal test results have been referred to a local NHS consultant ophthalmologist for a full examination and, if necessary, further management. Preliminary analysis indicates that only 29% of normal pressure glaucoma has been identified in the population of Norfolk. Methods: This project will examine the performance of HRT2, GDX and combinations of test using sensitivity, specificity, false positive and false negative rates, positive and negative predictive values, and receiver operating characteristic analyses, with 95% confidence intervals. We will explore the feasibility of individual tests and combinations of tests in data from 4,000 participants, and test the performance most successful model(s) using data from a different 4,000 participants. To quantify the false negative rates of these tests, optic disc photographs of all people aged 70 years and older and a proportion (5%) of those aged 49 to 69 years who were tested negative will be examined to detect undiagnosed glaucoma based on optic nerve structure.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: EYE
Title: Genetic Analysis of Corneal Endothelial Dystrophies
Funder: Medical Research Council

Abstract: Loss of corneal clarity causes 2% of blind registrations in UK children and is primarily caused by rare Mendelian forms of corneal endothelial dystrophy. More common but genetically complex Fuchs endothelial corneal dystrophy (FECD) is a disease of older age, with a prevalence of 4% in those over 40 years of age, accounting for up to 20% of corneal transplants. This proposal aims to estimate the frequency of rare Mendelian corneal endothelial dystrophies, correlate genotypes with phenotypes, determine aetiological overlap with FECD and find new genes/mutations involved in these conditions. Frequency will be determined by commissioning a nationwide surveillance project. Endothelial status in SLC4A11 mutation carriers said to be at risk of developing FECD will be determined, and audiology testing will determine whether homozygotes develop hearing loss. An existing cohort of endothelial dystrophy cases and families will be expanded and screened for mutations in five genes implicated in these conditions. The remaining patients/families will be tested for linkage to new or existing loci then subjected to Next Generation sequencing to identify the genes involved. Transcriptome analysis will determine which of the genes at the uncharacterised loci are expressed in corneal endothelium and whether their expression is altered in FECD, highlighting new candidate genes for these conditions, thus enhancing our knowledge of the pathogenesis of the condition. This may have future implications for targeted gene therapy for FECD and other endothelial dystrophies.


Research activity codes: 2.4 SURVEILLANCE AND DISTRIBUTION;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: EYE
Title: Functional Imaging in the Zebrafish Visual System
Funder: Medical Research Council

Abstract: Understanding how the brain processes sensory information is a major challenge in neuroscience. The aim of this proposal is to use the optic tectum of larval zebrafish as a system to investigate how a defined visual stimulus is processed at defined stages in the visual pathway. Specifically we will characterize the circuit components that underlie the perception of motion. A major focus of the application is to also investigate how such circuitry develops. Firstly we will characterise direction-selective retinal ganglion cells (DSGCs). We have established a system to provide precisely controlled visual stimuli to zebrafish while performing in vivo imaging in the tectum. In order to visualise neural activity in RGCs specifically we have fused the genetically encoded calcium sensor, GCaMP3 to the presynaptic vesicle protein, synaptophysin to generate SyGCaMP3. We have generated a transgenic line of zebrafish that expresses SyGCaMP3 specifically in RGCs, thus allowing us to visualise motion-induced activity at the presynaptic terminals of RGCs in the tectum. Using this approach we will characterise the functional development of DSGCs and also map the laminar location of different types of DSGC axon in the tectum. This functional map will then be used as a point of reference to ask how information from DSGCs is processed and integrated in the postsynaptic tectal neurons. This will be achieved using bulk loading of the fluorescent calcium indicator, Oregon Green BAPAT 488 AM into tectal cells. We will determine how tectal neurons are tuned to motion in different directions and the location of each characterised cell within the tectum will also be mapped. We will also examine how the tuning properties and functional organisation of tectal cells develops. We will also characterise the functional properties of morphologically identified tectal neurons. This will be achieved through expression of a membrane targeted version of GCaMP3 which will allow morphological and functional characterisation of neurons. By describing each of the circuit components in this way we can begin to understand how the tectum as a whole processes visual stimuli. As part of this last aim we hope to isolate genomic regulatory regions that will permit expression of transgenes in subsets of tectal neuron. With these molecular tools in hand we can selectively manipulate different cell types and in the long-term ask how they contribute to the perception of motion and the generation of behaviours.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: EYE
Title: Gene therapy for blindness caused by choroideraemia - a Phase I clinical trial.
Funder: Department of Health (England)

Abstract: We are seeking support to start a clinical trial using gene therapy to treat a disease that causes blindness known as choroideraemia. This condition is currently incurable and affects thousands of people worldwide. The principle of gene therapy is to use the shell of a virus (known as a vector) to carry a segment of DNA into the cells of affected patients where it can have a beneficial effect. In the case of choroideraemia there is deficiency of a gene known as REP1. We have put this gene into a viral vector and shown in our laboratory that it can correct the choroideraemia defect. We have now reached the point where we are ready to assess the potential benefit of this treatment in patients. We have designed a study involving 12 patients across four NHS sites that would represent the world's first ever clinical trial for this disease. We will assess the effects of the gene therapy two years after treating each patient and if it is shown to be successful we will set up regionally located follow-on studies.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: Accurate and patient-friendly measurement of binocular visual function using a 3D smartphone
Funder: Department of Health (England)

Abstract: Stereo vision, often called 3D vision, is the ability to use both eyes together to see depth. Clinicians use specialised vision tests, called stereotests, to measure patients' 3D vision in disorders like squint. This helps them monitor progress, assess whether treatment is helping, and can guide decisions like when to operate. However, existing stereotests are not very reliable. Young children may not understand the test or be willing to cooperate, e.g. by wearing 3D glasses. This limits the usefulness of current stereotests. Recently, the first glasses-free 3D tablet computers have come on the market. We want to use these to produce a better stereotest. For children, our stereotest will be embedded in a fun, colourful game. Patients will give their answers by touching the screen. The device will use these responses to interactively adapt the test for that particular patient, customising it for each individual. The device will monitor patients while they do the test and automatically correct for any changes in viewing distance. As a result, the device will provide clinicians with more accurate data on 3D vision, especially in small children. This will help healthcare professionals to track the progress of treatment and make the best clinical decisions.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: EYE
Title: A New Minimally Invasive Surgery for the Treatment of Corneal Endothelial Disease.
Funder: Department of Health (England)

Abstract: The cornea is the transparent tissue at the front of the eye which allows light to reach the retina. If the cornea becomes cloudy, vision is impaired, a bit like looking through a frosted glass window. Often the only treatment is a cornea transplant in which a surgeon removes the damaged cornea and replaces it with a donor cornea or part of a donor cornea. Often this has a good outcome, but the number of donor corneas available for surgery is limited, and risks such as infection or rejection remain. We plan to develop a new surgery for corneal cloudiness which is caused by diseased or damaged cells that line the inside of the cornea. Basically, under local anaesthetic, we will very gently touch the front of the cornea for a few seconds with a new surgical device which has the same curvature as the cornea and which very quickly freezes cells. In doing so the diseased endothelial cells at the back of the cornea are destroyed. Then we will apply eye drops which will encourage healthy endothelial cells to grow and repopulate the inside of the cornea. In this way the cornea can become healthy and clear again.


Research activity codes: 5.4 SURGERY
Health categories: EYE
Title: Scalable production of RPE cells from induced pluripotent stem cell under GMP conditions for cellular replacement therapy of the dry form of AMD
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Stem cells can be differentiated from somatic cells by reprogramming with multiple factors which have the capability to be pluripotent. Forced expression of four transcription factors, Oct4, Sox2, Myc and Klf4, can reprogramme these cells to ESC-like cells with pluripotent qualities, termed induced pluripotent stem cells (iPSCs). This can be achieved using different combinations of transcriptions factors, such as Oct4, Sox2, Nanog and Lin28, which avoids the use of Myc, a known proto-oncogene. The goal of this project is to produce iPSC's which could be used therapeutically and manufactured under cGMP processes. We will then use clinically approved protocols to generate eye-like cells from iPSCs. We will manufacture retinal pigmented (RPE) cells for therapeutic replacement of these cells for the dry form of age-related macular degeneration. The iPSC-derived RPE cells form monolayers that exhibit polarity, form tight junctions, express genes vital to the visual cycle and are able to phagocytose. Transplantation of iPSC-derived RPE into animal models of retinal degeneration has been shown to improve visual function. This will produce the data to enable the submission of a clinical trial application.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE
Title: PRISSM (Perfecting Refraction in India with Superior Service Models): A cluster-randomized controlled trial of three models of school-based spectacle service delivery in India
Funder: British Council for Prevention of Blindness

Abstract: Uncorrected refractive error (URE) causes 85% of vision impairment among Indian children and worldwide. School glasses programs improve children’s learning, as targeted in Sustainable Development Goal (SDG) #4, and help redress Indian girls’ unequal healthcare access (SDG#5). Indian government policy and numerous institutions support school glasses programs, but to expand them, cost-effectiveness must be improved. The most important questions are where refraction is done, and whether glasses are free. Refractive programs fall into the “School Model” (screening, refraction and spectacle delivery in schools) and “Referral Model” (children failing screening are refracted at outside facilities). The former model covers most children, but few countries have sufficient refractionists. The latter requires fewer refractionists, but often few families accept referral. Reviews by applicants Reddy/Bassett show: involving school teachers in vision screening and referral support significantly increases uptake. We propose a randomized trial in India involving partners in the Orbis REACH network. Our hypothesis: An “Optimized Referral Model” with strong teacher involvement will double “Referral Model” acceptance to two-thirds, making it more cost-effective than the “School Model.” We will also study impact of selling glasses in the “Referral Model.” A successful trial will benefit countless children in India and globally.


Research activity codes: 8.1 ORGANISATION AND DELIVERY OF SERVICES;8.2 HEALTH AND WELFARE ECONOMICS
Health categories: EYE
Title: Training in the use of smartphones and tablets to improve quality of life in visual impairment
Funder: Fight for Sight

Abstract: Mobile electronic devices (MED) including, smartphones and tablets, offer a new type of assistive technology for visually-impaired people (VIP). They offer the possibility to replace optical magnifiers for those with mild impairment, and braille or auditory for those with severe visual loss, using standard consumer devices, which are relatively cheap and convenient. However not all VIP and rehabilitation professionals are familiar with the devices and their potential. This study aims to determine usage of MED by VI users; how information about them is disseminated; and barriers to use (Aim 1). Having determined the typical format of training courses designed for VIP to learn how to use MED (Aim 2), we will recruit and train VIP who are interested in purchasing a MED. We will determine the effectiveness of this training (Aim 3) by assessing the usage of devices by the participants from completion of training to 6 months, using questionnaires, and by remote monitoring of their MED. We will use validated questionnaires to measure changes in quality of life, depression, adaptation to vision loss, and ability to carry out everyday tasks, before and after training, and 6 months later. Ongoing support has been found to be important when introducing users to new technology, in the form of volunteers to provide assistance and advice. We will therefore compare the standard “training course only” model to a scheme in which each participant is paired with a “buddy” (a university student) who can provide continuing support by visiting the VIP regularly at home.


Research activity codes: 6.6 PSYCHOLOGICAL AND BEHAVIOURAL;7.1 INDIVIDUAL CARE NEEDS
Health categories: EYE
Title: Developing rhodopsin gene therapy to treat dominant retinitis pigmentosa
Funder: Fight for Sight

Abstract: Retinitis pigmentosa (RP) is the most common genetic cause of irreversible sight loss in the developed world with the dominant form being the most frequent. Around 25% of cases of dominant RP are caused by mutations in the rhodopsin gene. As a single gene disorder involving a small protein, gene therapy using an adeno-associated virus (AAV) vector (which has been used successfully in a number of mouse models of retinal degenerations and Phase 1 human clinical trials) may represent a viable treatment option for RP. Recent evidence has shown that gene augmentation may be of benefit in treating dominant conditions- the assumption being that augmentation of the wild-type allele may increase the ratio of wild-type to mutant gene at the mRNA level, leading to reduced mutant protein. Rescue of dominantly inherited RP by over-expression of the wild-type gene (but without direct suppression of the mutant gene) has been demonstrated in the P23H-rhodopsin mouse, in which one allele contains a human rhodopsin variant that causes RP (Mao et al., 2011). The purpose of this proposed PhD (DPhil) project is to develop an efficient AAV vector that expresses the human rhodopsin gene to a sufficient level that might be applicable for a follow-on clinical trial in patients. The first phase will involve the generation of an AAV rhodopsin vector by assessing the optimal AAV capsid combinations and DNA cloning sequence to produce highly efficient rhodopsin expression. This will be achieved through the use of self-complementary double stranded DNA. In the second phase, I will assess the effect of this vector in wild type and rhodopsin knock-out mice to ascertain the optimal dosage. The final third phase will consist of an analysis of gene silencing activity of the optimized vector in the P23H or P347S rhodopsin mouse model of dominant RP and in human retinas in vitro. It is expected that all data generated from this project would be submitted for future Phase I clinical trial approval.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: EYE