Suite of high content assays for mitochondrial dynamics in neurons

神经元线粒体动力学的高内涵测定套件

• 批准号: 9236526
• 负责人: Ronald L Davis
• 金额: $ 64.84万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-13 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9236526
• 关键词: Algorithms; Architecture; Autistic Disorder; Axon; Biogenesis; Bioinformatics; Biological; Biological Assay; Biological Response Modifier Therapy; Brain; Brain Diseases; Cell Line; Cell physiology; Cells; Chimeric Proteins; Collection; Complex; Data; Defect; Dendrites; Development; Dose; Effectiveness; Equilibrium; Event; Failure; Family; Functional disorder; Generations; Genes; Genetic; Genetically Engineered Mouse; Goals; Health; Image; Knowledge; Lead; Lesion; Libraries; Liquid substance; Literature; Longevity; Measures; Mediator of activation protein; Membrane Potentials; Mental disorders; Methodology; MicroRNAs; Mitochondria; Molecular; Molecular Target; Mood Disorders; Mus; Nervous System Physiology; Neuraxis; Neurobiology; Neurodegenerative Disorders; Neuroglia; Neurons; Organelles; Pharmaceutical Preparations; Phenotype; Physiology; Porifera; Procedures; Process; Protein Import; Proteins; RNA Interference; Reporter; Research; Resources; Robotics; Schizophrenia; Speed; Surveys; System; Testing; Therapeutic; Time; Toxic effect; assay development; base; biological research; cell type; cheminformatics; design; flexibility; follow-up; high throughput screening; meetings; nervous system disorder; neuropsychiatric disorder; novel; novel strategies; novel therapeutics; overexpression; potency testing; protein degradation; response; scaffold; scale up; screening; small molecule; small molecule libraries; tool

PROJECT SUMMARY / ABSTRACT There is a pressing need to develop new classes of therapeutics for the treatment of psychiatric and neurological disorders. Despite this need, little progress has been made towards this goal over the last two decades. One explanation for this failure is that the cell-based screens designed to identify new lead compounds have traditionally utilized immortalized, non-neuronal cell lines. It becomes understandable, in retrospect, why past screens have failed to yield the hoped-for plethora of new and important classes of compounds given the extraordinary architecture and physiology of neurons compared to other cell types. A second possible reason for this failure is that most cell-based screens have utilized a target-based approach, in which a specific protein target is identified that may hold promise for the development of new therapeutics. However, the brain and how brain disorders influence its function remains a large mystery. This fact argues that the alternative of phenotypic-based screens may offer more promise. Phenotypic screens search for influences on particular cell biological phenotypes without knowledge of the specific molecular targets that influence the phenotype. We have developed a novel screening platform that employs primary cultured neurons and cell biological, phenotypic readouts to identify the influences of small molecules. The methodology is flexible, scalable, and offers numerous advantages over traditional approaches. In this project, we propose to develop assays using this platform for the processes of mitochondrial dynamics, including biogenesis, fission, fusion, protein import, branching, and damage. We will use fluorescent markers for mitochondria to follow mitochondria for such assays, and conduct three screens of small molecules for influences on mitochondrial dynamics once the assays are optimized. Mitochondria dysfunction is associated with multiple psychiatric disorders like mood disorders, schizophrenia and autism; and neurodegenerative disorders as well. Thus, this project promises a rich new resource that can be used to survey small molecules, RNAi, or gene overexpression effects on mitochondrial dynamics in neurons.

项目概要/摘要 迫切需要开发新的治疗方法来治疗精神科和精神科疾病 神经系统疾病。尽管有这种需要，但过去两年在实现这一目标方面进展甚微 几十年。对于这种失败的一种解释是，基于细胞的筛选旨在识别新的先导化合物 化合物传统上利用永生化的非神经元细胞系。就变得可以理解了，在 回想起来，为什么过去的屏幕未能产生人们所希望的大量新的、重要的类别 与其他细胞类型相比，这些化合物赋予神经元非凡的结构和生理学。一个 这种失败的第二个可能原因是大多数基于细胞的筛选都采用了基于目标的方法， 其中确定了一个特定的蛋白质靶标，该靶标可能有望开发新的疗法。 然而，大脑以及大脑疾病如何影响其功能仍然是一个很大的谜团。这一事实论证了 基于表型的筛选的替代方案可能会提供更多希望。表型筛选搜索 在不了解特定分子靶标的情况下对特定细胞生物表型的影响 影响表型。 我们开发了一种新颖的筛选平台，采用原代培养的神经元和细胞生物学， 表型读数可识别小分子的影响。该方法灵活、可扩展且 与传统方法相比具有许多优势。在这个项目中，我们建议开发使用 该平台用于线粒体动力学过程，包括生物发生、裂变、融合、蛋白质输入、 分枝、损伤等。我们将使用线粒体荧光标记来跟踪线粒体，以实现此类目的 测定，并进行三项小分子筛选，以了解对线粒体动力学的影响。 分析得到优化。线粒体功能障碍与情绪等多种精神疾病有关 疾病、精神分裂症和自闭症；以及神经退行性疾病。因此，该项目承诺 丰富的新资源，可用于调查小分子、RNAi 或基因过度表达对 神经元中的线粒体动力学。