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

描述（由申请人提供）：由包括阿尔茨海默氏病、帕金森氏病、亨廷顿病和ALS在内的主要神经系统疾病引起的行为和认知方面的特定问题是由于整个大脑中选定区域的神经元的进行性变性和功能障碍而产生的。类似的原因也被假设为与自然衰老相关的行为和认知的常见下降。然而，通过自然衰老而逐渐变形和功能失调的整个神经元仍然未知。通过自然衰老和疾病模型发现预防神经变性的药物的范式转变方法将是以高通量方式研究每种化合物在明确定义的模型生物体的整个神经系统中的作用。我们建议开发一种新的高通量筛选平台，使用光学和微流体（光流体），这将使每个神经元在整个神经系统的表征在毫秒内与亚细胞分辨率的遗传模型秀丽隐杆线虫。超快速筛选方法与新型体内模型的结合将为无偏倚筛选提供可能性，这些筛选不需要任何潜在药物靶点和途径的先验知识。我们选择了C。因为它是唯一具有完全表征的神经系统的动物，适合于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经验证的模型。所提出的光流体平台将能够在动物神经系统中的每个神经元通过微流体通道时在毫秒内快速量化其形态完整性。单个神经元可以很容易地通过在单个动物中组合表达不同的荧光报告基因来识别。除了高速定量能力之外，与96孔或384孔板自动连接的能力将使得能够将大量的蠕虫种群加载到光流控芯片中，每个蠕虫种群用不同的化合物处理。从这些研究中发现的原则将对理解疾病和衰老中行为表现如何下降的神经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.