Suite of high content assays for mitochondrial dynamics in neurons

项目总结/摘要 迫切需要开发用于治疗精神疾病和精神分裂症的新类别的治疗剂。 神经系统疾病尽管有这种需要，但在过去两年中，在实现这一目标方面取得的进展甚微。 几十年这种失败的一种解释是，旨在识别新铅的细胞筛选 化合物传统上利用永生化的非神经元细胞系。这是可以理解的，在 回顾过去，为什么过去的屏幕没有产生希望的过多的新的和重要的类， 与其他细胞类型相比，神经元具有非凡的结构和生理学特性。一 这种失败的第二个可能原因是大多数基于细胞的筛选已经利用了基于目标的方法， 其中鉴定了特异性蛋白质靶点，这可能为新疗法的开发带来希望。 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。这一事实表明， 基于表型的筛查可能会带来更多的希望。表型筛选搜索 影响特定的细胞生物学表型，而不知道特定的分子靶点， 影响表型。 我们已经开发了一种新的筛选平台，采用原代培养的神经元和细胞生物学， 表型读数，以确定小分子的影响。该方法灵活、可扩展， 提供了许多优于传统方法的优点。在这个项目中，我们建议使用 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， 分支和损伤。我们将使用线粒体的荧光标记来跟踪线粒体， 分析，并进行三个筛选的小分子对线粒体动力学的影响，一旦 优化了测定。线粒体功能障碍与多种精神疾病如情绪有关 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或基因过表达对 神经元中的线粒体动力学

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.