Assays to evaluate biological pathways in Parkinsons disease

评估帕金森病生物学途径的测定

• 批准号: 10683012
• 负责人: James Inglese
• 金额: $ 20.9万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683012
• 关键词: Biological; Biological Assay; Biological Models; Caenorhabditis elegans; Cell Culture Techniques; Cell Line; Chemical Exposure; Chemicals; DNA Sequence Alteration; Enhancers; Foundations; Foxes; Funding; Gene Mutation; Genes; Genetic; Genetic Transcription; Goals; Human; Investigation; LRRK2 gene; Laser Scanning Cytometry; Libraries; Link; Methods; Mitochondria; Monitor; Nematoda; Nerve Degeneration; Neurons; Organism; Parkin; Parkinson Disease; Pathway interactions; Pharmaceutical Preparations; System; Therapeutic; alpha synuclein; base; design; dopaminergic neuron; genome editing; neuron loss; neuronal survival; novel therapeutic intervention

Parkin removes damaged mitochondria, which protects neurons from cell death. However, in a form of familial PD, parkin haploinsufficiency cannot fully support neuronal survival. With funding in part from the Michael J. Fox Foundation and using a genome-edited neuronal cell line to monitor the endogenous levels of Parkin, we conducted a qHTS to identify transcriptional enhancers of the parkin locus. In a second strategy, we are employing a Caenorhabditis elegans (C. elegans) PD phenolog based on human PD-linked gene mutations in alpha-synuclein and the leucine-rich repeat kinase 2 (LRRK2). Using laser scanning cytometry methods, we are developing a 384-well qHTS-compatible C. elegans PD model system for evaluating libraries of investigational agents and approved drugs for their ability to inhibit nematode neurodegeneration.

帕金去除受损的线粒体，该线粒体可保护神经元免受细胞死亡的影响。但是，以家族性PD的形式，帕金单倍体不足不能完全支持神经元生存。在迈克尔·J·福克斯基金会（Michael J. Fox Foundation）的部分资金中，并使用基因组编辑的神经元细胞系来监测帕金的内源性水平，我们进行了QHTS，以识别Parkin locus的转录增强子。在第二种策略中，我们采用了基于α-突触核蛋白和富含亮氨酸的重复激酶2（LRRRK2）的人类PD连接基因突变的秀丽隐杆线虫（秀丽隐杆线虫）PD酚。使用激光扫描细胞仪方法，我们正在开发384孔QHTS兼容的秀丽隐杆线虫PD模型系统，用于评估研究剂和批准药物的文库，以抑制线虫神经变性。