To respond to the high attrition rate of current target-based drug discovery, an alternative systems-oriented human cell-based approach has been proposed, in which chemical compounds will be tested in a cell-relevant context at the early stage of the drug development pipeline. For neurological disorders, this is however hampered by the inability to sample live neuronal cells. The seminal discovery that adult somatic cells can be reprogrammed to a pluripotent state could help to circumvent these obstacles. The technology of induced pluripotent stem (iPS) cells might open up a novel paradigm in brain drug development, allowing the employment of live patient neurons for phenotype-based preclinical drug-screenings.
The project aims to generate patient-derived neuronal cell models of two genetic disorders affecting the basal ganglia and lacking effective treatments. Maternally inherited Leigh syndrome (MILS) is an infantile encephalopathy due to point mutations within the MT-ATP6 gene of the mitochondrial DNA (mtDNA). Huntington’s disease (HD) is a neurodegenerative disorder caused by the CAG triplet repeat expansion within the gene encoding the protein Huntingtin. Basal ganglia are highly dependent on mitochondrial-based energy production. Hence, the two diseases represent in a way two paradigmatic examples, one exhibiting a direct mitochondrial implication and one where mitochondrial dysfunctions are considered a secondary insult. Human basal ganglia neurons (GABAergic and dopaminergic) will be obtained from induced pluripotent stem (iPS) cells generated by reprogramming patient and control fibroblasts using footprint-free episomal plasmid-based techniques. The chance to identify a disease phenotype within the reprogramming-derived basal ganglia cells and to develop potential therapeutic strategies might be diminished when using only a conventional reductionist molecular biology approach. In this study, we therefore propose to broaden the levels of investigations by combining standard functional and biochemical assays with global OMICS-driven analyses (systematic transcriptomics, proteomics, and metabolomics) to unravel the disease mechanisms without an a priori knowledge. This would allow the generation of computational models for the two neuronal disease states and to in silico predict the targets of potential interventional strategies. The computational predictions will be validated in follow-up cellular experiments. Finally, based on the identified disease pathways, we will seek to establish scalable assays amenable to high-throughput and focused compound screenings. If successful, this iPS cell-driven systems biology approach may represent an innovative platform for drug discovery of complex genetic brain diseases.
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