HIGH-THROUGHPUT HIGH-CONTENT PLATFORM FOR IMAGE-BASED SCREENS

描述(申请人提供)：许多人类疾病，包括神经疾病、糖尿病和与衰老相关的疾病，本质上是分子和遗传的。为了从根本上了解这些疾病并发现药物干预，使用模型遗传系统(包括蠕虫、苍蝇和鱼类)中的分析来筛选大型试剂库不仅是一种可行的方法，而且是一种富有成效的方法。然而，目前的瓶颈是手动和半定量技术，如视觉屏幕或基于图像的屏幕，这往往限制了实验的吞吐量；此外，将大型试剂库与这些分析结合使用实际上是不可能的或非常困难的。我们的长期目标是开发和使用工程化的高通量和高含量的方法，以加快使用模式生物的发现过程。本项目的目标是开发一种新的高通量定量方法，结合微流控和自动化技术来研究与突触传递功能有关的基因。从这项研究中发现的基因和途径可能成为神经疾病治疗的靶点。这种方法是创新的，因为这里开发的技术极大地提高了现有筛选工具的能力，并广泛适用于不同实验系统中的各种问题。这项拟议的研究意义重大，因为它填补了高通量和高内容屏幕的技术空白，以及突触传递基因和途径的知识空白。 公共卫生相关性：突触传递是将基因与突触和神经系统的功能联系起来的一个重要和活跃的研究领域。它直接应用于许多人类疾病，如运动控制功能障碍和精神疾病。

DESCRIPTION (provided by applicant): Many human diseases including neurological diseases, diabetes, and diseases associated with aging are molecular and genetic in nature. To fundamentally understand these diseases and to discover pharmaceutical interventions, screening large libraries of reagents using assays in model genetic systems (including worms, flies, and fish) is not only a viable but fruitful approach. The current bottlenecks, however, are in the manual and semi-quantitative techniques such as visual screens or image-based screens, often limiting both the throughput of the experiments; in addition, it is practically impossible or very difficult to use large reagent libraries in conjunction with these assays. Our long- term objective is to develop and use engineering-enabled high-throughput and high-content methods to speed up the discovery processes using model organisms. The objective of this project is to develop a novel high-throughput quantitative method combining microfluidics and automation for studying genes involved in synaptic transmission functions. Genes and pathways emerging from this study could potentially become targets of therapeutics in neurological disorders. The approach is innovative because the technology developed here dramatically increases the capabilities of existing screening tools, and is widely applicable to a variety of problems in different experimental systems. The proposed research is significant because it fills a technology gap in high-throughput and high-content screens, and a knowledge gap in genes and pathways in synaptic transmission. PUBLIC HEALTH RELEVANCE: Synapse transmission is an important and active area of research linking genes to the functions of synapses and the nervous system. It has direct applications to many human diseases such as dysfunctional motor control and mental illnesses.