Investigating the therapeutic potential of newly identified gene targets using stem cell models of neurodegenerative disease

Alzheimer’s, Parkinson’s, and Huntington’s diseases are characterised by the impairment of protein metabolism and mitochondrial function occurring with age. We have used an in vitro model that recapitulates the changes to mitochondria function that occurs during neurodegenerative illnesses to carry out a high-throughput screen of 7500 genes. We also completed a secondary screen and identified thirty druggable genes (can be pharmacologically manipulated) whose role in neurodegenerative disease we wish to investigate further. These genes fall into three broad classes, those that regulate: protein degradation pathways; the generation of reactive oxygen species (ROS); DNA repair and transcription. Targets will be characterised and their therapeutic potential assessed using human neuronal cells derived from the induced pluripotent stem cells (IPSC) of patients. In addition to characterising these target the student will be encouraged to carry out a phenotypic screen based on a novel cellular stress response assay to identify further druggable targets. The student will use a number of measures (e.g. of cellular bioenergetics, ROS, phototoxicity induced mitochondrial depolarisation, mitophagy and measures of protein function and accumulation) and techniques (see below) to assess gene function. There is very strong translational focus, by the end of the PhD genes that can be targeted to mediate neuroprotection would have been characterised. This project offers a unique opportunity for the student to learn molecular (e.g. lentiviral construction, cloning, siRNA/miRNA mediated gene knockdown and genomic techniques to assess the transcriptome) imaging (confocal, fluorescent microscopy), biochemical (measurements of cellular bioenergetics, protein aggregation, assays of mitochondrial function) and stem cell biology (culturing of neurons derived from human IPS cells carrying autosomal dominant mutations in genes implicated in neurodegenerative disease). They will benefit from working with: experienced post-doctoral scientists in both host laboratories; our industrial partner to gain insight into a drug development program and have access to state-of-the-art technologies (e.g. Seahorse Bioanalyser, translating ribosome affinity purification).