Retinal dystrophies (RD) are a group of rare diseases that cause photoreceptor degeneration and subsequent loss of vision; in which Retinitis pigmentosa is the most common. These diseases are genetically heterogeneous, with mutations associated in specific retinal genes, but also in ubiquitous expression genes, like PRPF splicing factors, that cause autosomal dominant retinitis pigmentosa (adRP). A panel of 107 genes associated with RD was designed for the molecular diagnosis of DR by next generation sequencing (NGS). This panel was validated and is used in the routine of the laboratory. The Whole Exome Sequence (WES) technique was used in order to characterise new candidate genes to be responsible of adRP in families that were not resolved through the DR panel. By WES one of the four analysed families was resolved. Induced Pluripotent Stem Cells (iPSCs) were obtained from skin fibroblasts of an adRP patient with a mutation in the PRPF8 splicing factor. These cells were differentiated into retinal pigment epithelium (RPE) cells, allowing to generate an in vitro cellular model to study the molecular mechanism that cause the disease. Using RNA. seq technology, it has been possible to observe for the first time significant differences in the expression of genes and isoforms (especially intron retention forms) in specific retinal cells (RPE) with mutated PRPF8. These differences were not observed in other cell types, such as skin fibroblasts. For the first time, a differential molecular mechanism induced by a mutation in the PRPF8, is shown in retinal cells. This mechanism could be related to the development of adRP in the patient.
|Date of Award||30 Nov 2018|
|Supervisor||Adolfo Diez Perez (Director), Francesc Xavier Nogues Solan (Director) & Mónica Rodríguez Carballeira (Director)|