High-speed opto-fluidics to screen entire nervous system in aging and disease

描述(申请人提供)：由阿尔茨海默氏症、帕金森氏症、亨廷顿氏症和肌萎缩侧索硬化症等主要神经疾病引起的行为和认知方面的具体问题，是由于大脑中选定区域的神经元进行性退化和功能障碍引起的。类似的原因也被假设为与自然衰老相关的行为和认知能力的普遍下降。然而，由于自然衰老而逐渐变得畸形和功能障碍的神经元的全部组成仍然未知。发现通过自然衰老和疾病模型防止神经退化的药物的一种范式转换方法将是以高通量方式研究明确定义的模型生物体整个神经系统中每种化合物的影响。我们建议开发一种使用光学和微流体(光流体)的新型高通量筛选平台，该平台将能够在遗传模型秀丽线虫中以亚细胞分辨率在毫秒内表征整个神经系统中的每个神经元。超快速筛查方法与新的体内模型的结合将为不需要任何潜在药物靶点和途径的先验知识的无偏见筛查打开可能性。我们之所以选择线虫，是因为它是唯一具有完全特征化神经系统的动物，可以用微流控进行高通量药物筛选，并且是人类衰老和神经系统疾病的有效模型。拟议的光流控平台将能够在动物神经系统中的每个神经元通过微流控通道时，在毫秒内快速量化它们的形态完整性。通过在单个动物中组合表达不同的荧光报告，可以很容易地识别单个神经元。除了高速量化能力外，能够自动与96孔板或384孔板对接的能力将能够将大量的蠕虫种群加载到光流控芯片中，每个蠕虫种群都经过不同的化合物处理。从这些研究中发现的原理将对理解疾病和衰老时行为表现如何下降的神经元基础，以及如何防止人类行为能力下降产生深远影响。

DESCRIPTION (provided by applicant): The specific problems in behaviors and cognition that are caused by major neurological diseases including Alzheimer's, Parkinson's, Huntington's, and ALS arise due to the progressive degeneration and dysfunction of neurons in selected regions throughout the brain. Similar causes are also hypothesized for the common decline in behaviors and cognition associated with natural aging. Yet, the entire complement of neurons that become progressively dysmorphic and dysfunctional through natural aging remains unknown. A paradigm shifting approach for discovering drugs that prevent neurodegeneration through natural aging and disease models would be to study the effect of each chemical compound in the entire nervous system of a well-defined model organism in a high-throughput manner. We propose to develop a novel high-throughput screening platform using optics and microfluidics (opto-fluidics) that will enable characterization of each neuron in the whole nervous system within milliseconds with sub-cellular resolution in the genetic model Caenorhabditis elegans. The marriage of an ultra-rapid screening method with novel in vivo models will open the possibility for unbiased screens that do not require any prior knowledge of potential drug targets and pathways. We have chosen C. elegans because it is the only animal with a completely characterized nervous system, is amenable to high-throughput drug screening with microfluidics, and is a validated model for aging and neurological diseases in humans. The proposed opto-fluidics platform will be able to rapidly quantify the morphological integrity of every neuron in an animal's nervous system in milliseconds as they pass through a microfludic channel. Individual neurons can easily be identified by combinatorial expression of diverse fluorescent reporters in a single animal. Besides high-speed quantification capabilities, the ability to automatically interface with 96- or 384-well plates will enable for loading of a large number of populations of worms each treated with a different chemical compound into the opto-fluidics chip. The principles uncovered from these studies will have a profound impact on understanding the neuronal basis for how behavioral performance declines in disease and aging, and how to prevent this decline in humans.