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

 描述（由申请人提供）：帕金森氏病（PD）是第二大最常见的神经退行性疾病，影响约。1%的人口。PD的症状包括由于大脑中多巴胺能神经元的进行性死亡而导致的肢体运动功能的震颤。虽然某些形式的PD是遗传的，但很明显，环境毒素，包括农药，以及可能的工业溶剂，有助于PD，但发生这种情况的途径尚不完全清楚。α -突触核蛋白（α-Syn）是一种功能未知的蛋白质，参与PD病理学，可能是由于错误折叠、聚集和对线粒体功能的毒性。此外，与PD相关的环境毒素也抑制线粒体功能。已发现α-Syn（A53 T）突变导致PD早发。诱导多能干细胞（iPSC）已经从携带α-Syn A53 T突变的受试者制备，并且与同基因对照iPSC细胞相比，源自这些iPSC的多巴胺能神经元显示出对农药的增加的PD相关反应。该I期SBIR项目的目标是开发一种高通量筛选，以测试环境毒素增加PD风险的能力，使用含有A53 T突变的iPSC衍生的多巴胺能神经元。为了确定将提供PD相关病理的最佳指示的生物标志物，我们将在这些神经元上测试农药，并量化对突触结构，线粒体以及活性氧和氮物质产生的影响。该分析的客户包括政府机构，如美国环保署。瓦拉科学公司专门从事开发与环境毒性相关的基于细胞的测定，是EPA ToxCast计划的测定提供商。第二阶段的目标将是进一步完善和扩大可以用这些神经元进行的测定的能力。

 DESCRIPTION (provided by applicant): Parkinson's disease (PD) is the 2nd most common neurodegenerative affliction, affecting approx. 1 % 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. α -Synuclein (α-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 α-Syn (A53T) has been identified which causes an early-onset of PD. Induced pluripotent stem cells (iPSCs) have been prepared from a subject harboring the α-Syn A53T 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 A53T 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.