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

描述（由申请人提供）：阿尔茨海默病、帕金森病、亨廷顿病、ALS等重大神经系统疾病引起的行为和认知方面的具体问题是由于整个大脑中选定区域的神经元进行性变性和功能障碍引起的。与自然衰老相关的行为和认知能力的普遍下降也有类似的原因。然而，通过自然衰老而逐渐畸形和功能失调的整个神经元补体仍是未知的。发现通过自然衰老和疾病模型来预防神经退行性变的药物的一种范式转换方法是，以高通量的方式研究每种化合物在一个定义良好的模式生物的整个神经系统中的作用。我们建议使用光学和微流体（光流体）开发一种新的高通量筛选平台，该平台将在遗传模型秀丽隐杆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。超快速筛选方法与新型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筛选提供可能性。我们之所以选择秀丽隐杆线虫，是因为它是唯一一种具有完全特征的神经系统的动物，适合用微流体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。所提出的光流体平台将能够在几毫秒内快速量化动物神经系统中每个神经元通过微流体通道时的形态完整性。单个神经元可以很容易地通过在单个动物中多种荧光报告基因的组合表达来识别。除了高速定量功能外，它还能自动连接96孔板或384孔板，从而可以将大量经过不同化合物处理的蠕虫装载到光流体芯片中。从这些研究中发现的原则将对理解行为表现如何在疾病和衰老中下降的神经元基础以及如何防止人类这种下降产生深远的影响。

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.