An automated high throughput phenotypic screen for schistosomiasis drug discovery

描述（由申请人提供）：血吸虫病是一种热带寄生虫病，感染超过2亿人。治疗依赖于一种药物，吡喹酮（PZQ）。在没有PZQ治疗谱的备用药物的情况下，对PZQ的耐药性和最终药物失败的风险是一个主要问题。传统的表型筛选，使用成虫阶段的S。mansoni）是低通量的并且与现代高通量筛选（HTS）系统不兼容。为了与NIAID的使命保持一致，本提案旨在将新开发的中等通量表型筛选（MTS）转变为全自动定量HTS，以加速针对该传染病的药物发现。该提案涉及三名PI，他们正在进行合作，并分别拥有生物学，筛选技术和生物计算技能，他们专注于这一目标。作为该提案的第一个研究轨道，我们将利用内部自动化和高内容筛选（HCS）系统，以显着提高吞吐量超过我们公布的MTS方法。该提案将涉及;扩大寄生虫的机器人接种，制定明场和荧光显微镜检查方案，并调整商业图像分析软件，以识别（分段）、定量描述和跟踪寄生虫的运动，以期优先考虑进一步临床前开发的化合物。由于商业HCS分析工具不太可能优化记录这种多细胞寄生虫显示的复杂和动态表型，我们还将进行第二次平行研究。具体来说，我们将重新开发一种自动图像分析筛选技术，以定义，识别和量化这种寄生虫可能的表型反应（形态和行为）范围。最终，实验和计算轨道将共同产生一个标准化的HTS协议和一个全面的，定量的图像分析程序套件，以分类寄生虫表型。这种严格性将有助于筛选大量潜在的化合物，并将其优先纳入内部可用的二级和三级筛选测定中。我们还打算公开算法框架，包括其方法和实现。 公共卫生相关性：该项目的主要目标是将血吸虫病的中等通量表型筛选（MTS）系统转变为全自动的定量高通量筛选（HTS）。通过这样做，治疗这种全球性热带疾病的药物的发现率将提高。

DESCRIPTION (provided by applicant): Schistosomiasis is a tropical parasitic disease infecting over 200 million people. Treatment relies on a single drug, praziquantel (PZQ). In the absence of back-up drugs with PZQ's therapeutic spectrum, the risk of resistance to PZQ and eventual drug failure is a major concern. Traditional phenotypic screens, using adult- stage S. mansoni, are low-throughput and incompatible with modern high-throughput screen (HTS) systems. In keeping with the NIAID's mission, the present proposal aims to turn a newly developed, moderate- throughput phenotypic screen (MTS), into a fully automated, quantitative HTS to accelerate drug discovery for this infectious disease. The proposal involves three PIs with ongoing collaborations and respective biological, screening-technology and bio-computational skills who are focused on just this goal. As a first research track for this proposal, we will utilize in-house automation and a high-content screening (HCS) system to significantly increase throughput over our published MTS approach. The proposal will involve; expanding robotic plating of the parasite, developing protocols for bright-field and fluorescence-based microscopy and adapting commercial image-analysis software to identify (segment), quantitatively describe and track the motion of parasites with a view to prioritizing compounds for further pre-clinical development. Because commercial HCS analysis tools are not likely optimized for recording the complex and dynamic phenotypes displayed by this multicellular parasite, we will also pursue a second and parallel track of research. Specifically, we will develop de novo an automated image-analysis screening technology to define, identify, and quantify the range of phenotypic responses (morphological and behavioral) possible in this parasite. Ultimately, both the experimental and computation tracks will together produce a standardized HTS protocol and a comprehensive, quantitative suite of image-analysis programs to categorize parasite phenotypes. Such rigor will facilitate the screening of large numbers of potential compounds and their prioritization into the secondary and tertiary screening assays available in-house. We also intend to make the algorithmic framework including its methods and implementations, publicly available. PUBLIC HEALTH RELEVANCE: The major goal of the project is to turn a moderate-throughput phenotypic screen (MTS) system for schistosomiasis, into a fully automated, quantitative high-throughput screen (HTS). By so doing, the rate of discovery of drugs to treat this global tropical disease will be increased.