FUNCTIONAL CHARACTERISATION OF THE A30P MUTATION IN ALPHA-SYNUCLEIN GENE IN A PATIENT-DERIVED CELLULAR MODEL OF PARKINSON’S DISEASE

Our study aims to perform detailed phenotyping of the A30P alpha-synuclein familial case of PD, allowing to identify underlying mechanisms of the disease that may translate into novel therapies.
Parkinson’s disease (PD) is the second most common neurodegenerative disease. Approximately 20% of PD cases are known to have a genetic cause. From these, mutations in SNCA, the gene encoding alpha-synuclein, are linked to an autosomal dominant inheritance of the disease. In 1998, our group discovered the second known point mutation within the SNCA gene, causing an A30P exchange of the peptide sequence.
We generated first patient-derived cellular model of the A30P alpha-synuclein mutation carrier, by obtaining fibroblasts from an affected sibling of the index patient, an unaffected sibling of the patient, and an age-matched gender-matched non-PD control. We reprogrammed these fibroblasts into induced pluripotent stem cells (iPSCs), and differentiated them into midbrain dopaminergic neurons.
We obtained enriched cultures of 80% midbrain neurons (FoxA2+/Tuj1+), with approximately 12 % dopaminergic (TH+), for which we observed electrophysiological activity and dopamine release. We detected a significant reduction of the protein level of mitochondria complexes II, IV, and V in the patient lines compared with the controls, additionally we found a significant impairment of mitochondrial respiration and an increased susceptibility of the cells to oxidative stress. Gene-edited isogenic controls were generated to dissect mutation-specific effects. Furthermore, we investigated mitochondrial morphology and dynamics, and how these processes contribute to the dopaminergic neurodegeneration. Additionally, we were implementing previously established readouts on our high-throughput automated screening platform that will allow us to identify FDA approved compounds with potential to be re-purposed and used as PD treatment.
We believe that detailed phenotyping of the A30P alpha-synuclein monogenic case may help to identify underlying mechanisms of the disease that may translate into novel therapies, which would also apply to the more common sporadic forms of PD.