Fuchs endothelial corneal dystrophy (FECD) is a common vision-impairing disease that affects 4.5% of the population over 50 years of age. Both eyes are generally affected, and this is because the cornea, the transparent window at the front of the eye, loses its ability to remove excess fluid from its outer layers and becomes swollen and cloudy. The only effective treatment for advanced FECD is corneal transplantation. In the UK, 80% of the FECD cases are caused by a common genetic disorder called a 'CTG18.1 repeat expansion', where this short sequence of genetic code repeats and expands to a threshold that causes disease. FECD is the most common reason for corneal transplant in the UK. However, the surgery is costly, and there is a global shortage of donor tissue. In an effort to reduce the demand for corneal tissue, multiple innovative treatment options are emerging, including a gene therapy developed at our lab. The long-term success of these therapies will rely on identifying individuals at risk of developing FECD and initiating early treatment before irreversible damage occurs. Having a better understanding of the genetic mechanism in FECD would help to achieve these. In other better-understood diseases caused by abnormally repeated genetic codes (eg. myotonic dystrophy), the more the disease-causing genetic code is repeated, the earlier the onset of disease and the more severe symptoms patients will have. Moreover, the number of these repeated genetic codes is larger in the affected tissues. We also know that in these conditions, the number of the disease-causing genetic code increases when the disease is passed down from one generation to the next, resulting in earlier and more debilitating disease in each succeeding generation. This project aims to find out whether these clinically important patterns also occur in FECD, which is currently a poorly understood disease. Our study will take place in the laboratory setting at UCL Institute of Ophthalmology, where we have already performed extensive genetic analysis of a large group of FECD patients. Aim 1. I will generate new genetic data and use statistical models to examine the relationship between the number of CTG18.1 repeats of our recruited patients and their age when they require a corneal transplant. Following my preliminary work, I expect to see a trend where patients with larger CTG18.1 repeats require corneal transplants earlier. This knowledge will enable us to predict how fast disease will progress based on genetic information. Aim 2. I will use an innovative technique called Single Molecule (SM)-PCR to analyse the DNA in the affected corneal tissues removed from FECD patients during corneal transplant surgery. Due to technical limitations, analysis of diseased corneal cells has not been successfully performed before. As the cornea is the only tissue affected by FECD, I predict the number of CTG18.1 repeats in corneal cells may be a lot larger than in the blood cells of the same patient. Aim 3. I will analyse the DNA of FECD patients' siblings to assess the proportion of individuals carrying the CTG18.1 expansion that also develop FECD. Adult children of FECD patients will also be recruited to discover how the repeats behave (i.e. if it gets bigger or smaller) when passed onto successive generations. This will enable us to have a better understanding of how CTG18.1 changes within families and its risk associated with FECD. Currently, it is not known how the disease-causing gene of FECD modifies its clinical features. This important research will help us gain more in-depth knowledge about FECD, and potentially allow doctors to give more accurate genetic counselling on the progression of FECD and its risk implication to the next generation of FECD patients. Knowledge of CTG18.1 repeats in affected corneal cells may also open up research for new treatments options.

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

The cornea on the front surface of the eye is our window to the world. If transparency is compromised, visual impairment and even blindness can occur. Aniridia is a rare blinding disease (1:100,000 incidence) caused by a mutation in PAX6 which is one of the genes responsible for development and healthy function of the eyes. Aniridia-related keratopathy (ARK) manifests as persistent, chronically painful defects of the outer epithelial layer of the cornea called the epithelium. Blood vessels grow into the cornea and scarring also occurs, both of which obstruct vision. ARK induced light sensitivity can lead to social exclusion. The cause of these symptoms is related to the inability of the mutated limbal epithelial stem cells (LESC), stromal cells and tissue to maintain normal corneal epithelium. Current treatments include whole donor tissue transplantation and cultured LESC therapy which have both shown poor long-term outcomes. Since stem cells require support from neighbouring cells and proteins in their environment, transplanting cultured LESC alone may not be sufficient to preserve / improve patient vision. An optimal ARK treatment would include transplantation of both healthy LESC and a stroma (corneal support tissue) populated with stromal cells. Our proposed solution utilises our patented technology known as RAFT (Real Architecture for 3D Tissues) in which LESC and stromal cells are cultured together in a transplantable collagen matrix (artificial tissue). We are able to make this tissue in compliance with the required regulations which are known as Good manufacturing practise, or GMP for short, in our clinical cell therapy production laboratory. Pre-clinical safety studies supports us moving forward to a first in human study. In this project we will undertake 1) full GMP protocol validation for RAFT in our Cells for Sight licensed manufacturing facility, 2) write and submit all of the documentation required to obtain regulatory approval for a clinical trial and 3) proceed to first in human RAFT transplantation studies in patients with ARK. In this phase I/II clinical trial we will perform RAFT transplantation in one eye of 21 ARK patients to assess RAFT safety and preliminary efficacy. The outcome measures will include restoration of a normal corneal epithelium without any defects, blood vessel ingrowth or scarring. If successful, this therapy could provide a much-needed solution for patients with ARK and also act as a springboard for the development of RAFT for other blinding eye diseases.

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

Glaucoma is a condition that affects the human eye and is an umbrella term for a family of related eye conditions. A common trait of these conditions is a functional abnormality of the retina and optic nerve, leading to loss of visual field. This vision loss is usually only part of the visual field, although, untreated, glaucoma often leads to blindness. It is thought that by 2010, there will be over 60 million people worldwide suffering from the various forms of Glaucoma. Visual field tests are crucial to the diagnosis and management of all types of glaucoma. Such tests require the level of retinal sensitivity to light to be sampled at a number of points typically between 50 and 100, depending on the type of test; the eye is then assigned a numerical value in the range 'no perception' (lowest) to 'perfect perception' (highest). A specialised machine is used to conduct these tests - a typical clinical test can take between six and seven minutes per eye. Once diagnosed with glaucoma or suspected glaucoma, a patient is monitored and recommended to undergo the same tests every six months (or more frequently in some cases). However due to the psychophysical nature of the glaucoma test, the results can therefore vary in quality dramatically. For example, they can be affected by patient fatigue (as the test can last for long periods) and attention span deficits (particularly in the elderly and children).This proposal aims to use data quality metrics (such as false positive and negative rates) to incorporate uncertainty into computational models that will also take into account the spatial and temporal nature of visual field data. The proposed research will use probabilistic Cellular Automata (CA) with an appropriate rule learning approach as the technique to model the visualfield deterioration of glaucoma sufferes. The aim is to accurately model visual field progression and to provide an aid to the clinical practitioners.This project is in collaboration with Moorfields Eye Hospital, London, UK.