Integrated ImageXpress Micro Confocal High Content Screening System

项目总结 在过去的20年里，学术界和工业界都从以靶向为基础的药物发现显著转变为 表型筛选。长期以来，生物医学研究企业中一个尚未满足的需求是人类疾病 具有可靠翻译内容的模型，可用于筛选小分子或RNAi/gRNA/ORF文库 以获得理想的表型变化。从患者体内培养器官或3D组织的研究进展 祖细胞为迎接这一挑战提供了机会。人们普遍认为，药物发现使用 这种体外模型加速了对候选药物的临床前测试，因为有机化合物 与培养的细胞系相比，3D结构更接近于疾病组织的复杂生理病理 或者生化分析。该领域的一个持续挑战是仪器、技术和 可视化和测量分子治疗反应中的成分和特征(表型)的方法 在这些3D模型中。 范德比尔特化学生物学研究所和范德比尔特高通量筛选(VHTS)设施是一个很好的- 建立学术药物发现中心，利用创新技术推进科学和临床 治疗方面的发现。目前，VHTS设施拥有一台基于自动荧光的显微镜，可用于高分辨率的 与机械臂和孵化器集成的内容成像(HCI)和表型筛选，从而实现 用于微滴定板中的自动活细胞和固定细胞成像。使用该系统的研究包括IPSC- 衍生细胞筛选、功能基因组学筛选(siRNA、ORF)、病毒复制机制、药物毒性 研究、用于新治疗开发的药物筛选、药物作用机制和耐药性以及药物 联合研究。然而，由于缺乏共焦成像能力，该系统严重不足 用于成像和测量有机物体和3D结构中的特征。此外，目前的供应商不再 支持机械臂和孵化器的服务。因此，获得一个新的自动化人机交互系统是至关重要的， 配备机器人设备，以实现最先进的高通量表型筛选 利用有机化合物和三维结构进行疾病生物学和药物发现的方法。建议的HCI药物 Discovery系统将以分子设备(MD)ImageXpress微共焦人机界面系统为中心，具有 集成了Precision Automation PreciseFlex PF400机械臂和利康自动孵化器。这 仪器的组合将允许在每个所需的水平上进行高通量的表型筛选和 容量。我们注意到，MD致力于设计最强大的自动显微镜仪器， 实施速度和精度优化的意图是专门针对3D结构的人机交互和 有机化合物。拟议的系统将为已经建立的范德比尔特客户增加重大价值，因为 以及VICB和VHTS设施使用具有很强潜力的疾病模型进行药物发现的使命 确定与临床相关的治疗策略。

PROJECT SUMMARY In the last 20 years, both academia and industry have significantly shifted from target-based drug discovery to phenotypic screening. An unmet need in the biomedical research enterprise has long been human disease models with reliable translational content that can be used to screen small molecule or RNAi/gRNA/ORF libraries for desirable phenotype changes. Recent advances in culturing organoids or 3D tissue derived from patient or progenitor cells provide opportunities for meeting this challenge. It is widely accepted that drug discovery using such in vitro models represents an accelerated route to pre-clinical testing of drug candidates because organoids and 3D structures more closely resemble the complex physiopathology of diseased tissue than cultured cell lines or biochemical assays. An ongoing challenge in the field is the upgrading of instrumentation, technology, and methods to visualize and measure components and features (phenotypes) in responses to molecular treatment within these 3D models. The Vanderbilt Institute of Chemical Biology and Vanderbilt High-throughput Screening (VHTS) facility is a well- established academic drug discovery center that uses innovative technologies to advance scientific and clinical therapeutic discoveries. Currently, the VHTS facility has an automated fluorescence-based microscope for high- content imaging (HCI) and phenotypic screening that is integrated with a robot arm and incubator, which allow for automated live and fixed cell imaging in micro-titer plates. Studies that have utilized this system include iPSC- derived cell screens, functional genomics screens (siRNA, ORF), mechanisms of viral replication, drug toxicity studies, drug screens for novel therapeutic development, drug mechanisms of action and resistance, and drug combination studies. However, due to the lack of confocal imaging capability, this system is severely inadequate for imaging and measuring features within organoids and 3D structures. In addition, the current vendor no longer supports service of the robot arm and incubator. Therefore, it is critical to acquire a new automated HCI system, complete with robotic equipment, in order to enable state-of-the-art high-throughput phenotypic screening approaches to disease biology and drug discovery using organoids and 3D structures. The proposed HCI drug discovery system will center on a Molecular Devices (MD) ImageXpress Micro Confocal HCI system, with integration of a Precise Automation PreciseFlex PF400 robot arm and Liconic automated incubator. This combination of instruments will allow for high-throughput phenotypic screening at every desired level and capacity. We note that MD has committed to designing the most robust automated microscope instrument with the intention of implementing optimization for both speed and accuracy specifically for HCI of 3D structures and organoids. The proposed system would add significant value to the already established Vanderbilt clientele, as well as, to the mission of VICB and VHTS facility in drug discovery using disease models with strong potential to identify therapeutic strategies with clinical relevance.