Insights into the effects of α-synuclein expression and proteasome inhibition on glutathione metabolism through a dynamic in silico model of Parkinsonʹs disease: validation by cell culture data

Dopaminergic neurodegeneration during Parkinson disease (PD) involves several pathways including proteasome inhibition, α-synuclein (α-syn) aggregation, mitochondrial dysfunction, and glutathione (GSH) depletion. We have utilized a systems biology approach and built a dynamic model to understand and link the various events related to PD pathophysiology. We have corroborated the modeling data by examining the effects of α-syn expression in the absence and presence of proteasome inhibition on GSH metabolism in dopaminergic neuronal cultures. We report here that the expression of the mutant A53T form of α-syn is neurotoxic and causes GSH depletion in cells after proteasome inhibition, compared to wild-type α-syn-expressing cells and vector control. Modeling data predicted that GSH depletion in these cells was due to ATP loss associated with mitochondrial dysfunction. ATP depletion elicited by combined A53T expression and proteasome inhibition results in decreased de novo synthesis of GSH via the rate-limiting enzyme γ-glutamyl cysteine ligase. Based on these data and other recent reports, we propose a novel dynamic model to explain how the presence of mutated α-syn protein or proteasome inhibition may individually impact on mitochondrial function and in combination result in alterations in GSH metabolism via enhanced mitochondrial dysfunction.