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

化学物质对人类 iPSC 衍生神经元帕金森毒性的测定

• 批准号: 9020156
• 负责人: PATRICK M MCDONOUGH
• 金额: $ 22.49万
• 依托单位: VALA SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9020156
• 关键词: Accounting; Affect; Age; Alzheimer' s Disease; Apoptotic; Back; Behavior; Biological Assay; Biological Markers; Brain; Calcium; Cell Line; Cell physiology; Cells; Cessation of life; Chemicals; Cognition; Companions; Dementia; Development; Disease; Environment; European; Family history of; Fluo 4; Generations; Goals; Government Agencies; Health; Herbicides; Human; Incidence; Inherited; Insecticides; Label; Leg; Limb structure; Link; Methods; Microscope; Midbrain structure; Mitochondria; Modeling; Moods; Motor; Mutation; National Institute of Environmental Health Sciences; National Toxicology Program; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nitric Oxide; Nitrogen; Nitrogen Oxides; Oxygen; Parkinson Disease; Parkinson' s Disease Pathway; Pathology; Patients; Pesticides; Pharmaceutical Preparations; Phase; Population; Prevalence; Production; Proteins; Protocols documentation; Provider; Research; Research Personnel; Risk; Robotics; Sampling; Science; Skin; Small Business Innovation Research Grant; Solvents; Stem cells; Structure; Substantia nigra structure; Superoxides; Symptoms; Synapses; Testing; Toxic Environmental Substances; Toxic effect; Toxicity Tests; Toxin; Tremor; United States Environmental Protection Agency; Work; Xenobiotics; alpha synuclein; arm; base; beta pleated sheet; dopaminergic neuron; early onset; genome editing; high throughput screening; induced pluripotent stem cell; interest; male; men; mitochondrial dysfunction; pars compacta; programs; protein misfolding; protein structure; public health relevance; response

 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.

 描述(申请人提供)：帕金森病(PD)是第二种最常见的神经退行性疾病，影响大约。占总人口的1%。帕金森病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震颤。虽然某些形式的帕金森病是遗传的，但很明显，环境毒素，包括杀虫剂，可能还有工业溶剂，有助于帕金森病的发生，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，参与了PD的病理过程，可能是由于错误折叠、聚集和对线粒体功能的毒性。此外，与帕金森病有关的环境毒素也会抑制线粒体功能。已发现α-Syn(A53T)突变可导致早发性帕金森病。从携带α-Syn A53T突变的受试者中制备了诱导多能干细胞(IPSC)，与同基因对照IPSC细胞相比，来自这些IPSC的多巴胺能神经元对杀虫剂显示出更高的PD相关反应。这一阶段SBIR项目的目标将是开发一种高通量筛查，以测试环境毒素是否有能力增加帕金森病的风险，使用含有A53T突变的IPSC衍生的多巴胺能神经元。为了确定对帕金森病相关病理提供最佳指示的生物标志物，我们将测试这些神经元上的杀虫剂，并量化对突触结构、线粒体以及活性氧和氮物种产生的影响。该检测的客户包括政府机构，如美国环保局。Vala Science Inc.专门开发与环境毒性相关的基于细胞的检测方法，是EPA ToxCast计划的检测供应商。第二阶段的目标将是进一步改进和扩大可以用这些神经元进行的分析能力。