Alison Hardcastle – Research Summary

 

The complexity of genetic eye diseases is unprecedented. The research focus of our lab is inherited eye disease, from gene discovery to defining cellular function through to development of potential therapies. Our molecular genetic, functional and phenotypic studies have continually resulted in new discoveries that influence patient care. We have defined genetic mechanisms of disease and function of disease proteins in the retina, lens, cornea and developing eye, and have harnessed technological advances in next generation sequencing and stem cell biology to address our research questions.

One of our specialties is X-linked forms of retinal degeneration. Some notable examples include identification of the cause of RP23 as a deep intronic mutation resulting in introduction of cryptic exon in a ciliopathy gene. We also described progressive cone dystrophy associated with a novel cone opsin misfolding mutation, and subsequently identified SNP haplotypes that result in splicing defects, leading to molecular understanding of cone opsin array mutational mechanisms and clinical outcome.

In collaboration with Prof Mike Cheetham, we have defined the function of the X-linked retinitis pigmentosa protein RP2. Following on from our important functional studies, we have reprogrammed patient derived iPSCs into RPE cells, in collaboration with Prof Pete Coffey, and successfully shown that readthrough drugs can restore protein function and hence may be a viable treatment for patients with nonsense mutations.

Applying a similar research pathway from genetic mechanism to cellular function, our studies have shown that X-linked cataract is allelic with Nance-Horan Syndrome (NHS), and we defined the function of NHS as a regulator of actin cytoskeletal dynamics and cell size.

We were the first to describe identification of the X-linked megalocornea gene, with consequent effects on brain morphology, influencing our understanding of eye development.

I have developed a successful research program on corneal dystrophies in collaboration with Mr Stephen Tuft, including complex and monogenic diseases, from gene discovery through to developing therapies. This includes a planned GWAS, RNA-seq and whole genome sequencing approach to study keratoconus, and to translate any significant associations into meaningful biological and pathological context.