Assay of chemicals for Parkinson’s toxicity in human iPSC-derived neurons

DESCRIPTION provided by applicant Parkinsonandapos s disease PD is the nd most common neurodegenerative affliction affecting approx of the human population The symptoms of PD include tremors in motor function of limbs due to progressive death of dopaminergic neurons in the brain While certain forms of PD are inherited it is clear that environmental toxins including pesticides and likely industrial solvents contribute to PD but the pathways via which this occurs are incompletely understood Synuclein Syn a protein of unknown function participates in PD pathology likely due to mis folding aggregation and toxicity to mitochondrial function Further environmental toxins linked to PD also inhibit mitochondrial function A mutation in Syn A T has been identified which causes an early onset of PD Induced pluripotent stem cells iPSCs have been prepared from a subject harboring the Syn A T mutation and dopaminergic neurons derived from these iPSCs display increased PD related responses to pesticides compared to isogenic control iPSC cells The goal of this Phase I SBIR project will be to develop a high throughput screen to test environmental toxins for their ability to increase the risk of PD using iPSC derived dopaminergic neurons harboring the A T mutation To identify the biomarkers that will provide the best indication of PD related pathologies we will test pesticides on these neurons and quantify effects on synaptic structures mitochondria and the generation of reactive oxygen and nitrogen species Customers for the assay include government agencies such as the US EPA Vala Sciences Inc specializes in developing cell based assays relevant to environmental toxicity and is an assay provider for the EPA ToxCast program The Phase II goals will be to further refine and expand the capability of the assays that can be done with these neurons PUBLIC HEALTH RELEVANCE Parkinsonandapos s disease PD is the nd most common neurodegenerative affliction affecting approx of the human population The symptoms of PD include tremors in motor function of limbs due to progressive death of certain neurons in the brain While certain forms of PD are inherited it is clear that environmental toxins including pesticides and likely industrial solvents contribute to PD Recently a patient with an early onset form of PD was identified and found to have a mutation in a certain protein and researchers were able to prepare stem cells from a sample of the patientandapos s skin that can be used to form neurons in a dish Furthermore these neurons have proven to be sensitive to environmental toxins We propose to develop methods so that the stem cell derived neurons from the PD patient can be used to screen chemicals that might escape into the environment for their potential to cause PD using the stem cell derived neurons The methods will involve the use of automated robotic microscopes