Integrated ImageXpress Micro Confocal High Content Screening System

项目摘要 在过去的20年里，学术界和工业界都从基于靶向的药物发现转向了 表型筛选长期以来，生物医学研究企业的一个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。培养来源于患者或动物的类器官或3D组织的最新进展 祖细胞提供了应对这一挑战的机会。人们普遍认为，药物发现使用 这种体外模型代表了候选药物临床前测试的加速途径 与培养的细胞系相比， 或生化分析。该领域的一个持续挑战是仪器、技术和 可视化和测量响应分子治疗的组分和特征（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- 建立了学术药物发现中心，利用创新技术推进科学和临床 治疗发现目前，VHTS设施有一个自动荧光显微镜，用于高- 内容物成像（HCI）和表型筛选，其与机器人臂和培养箱集成， 用于微量滴定板中的自动活细胞和固定细胞成像。利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、新治疗开发的药物筛选、药物作用机制和耐药性，以及药物 组合研究。然而，由于缺乏共焦成像能力，这一系统是严重不足的 用于成像和测量类器官和3D结构内的特征。此外，目前的供应商不再 支持机器人手臂和培养箱的维修。因此，获得新的自动化HCI系统至关重要， 配有机器人设备，以实现最先进的高通量表型筛选 使用类器官和3D结构的疾病生物学和药物发现方法。推荐的HCI药物 发现系统将集中在分子设备（MD）ImageXpress微共聚焦HCI系统上， Precise Automation PreciseFlex PF 400机械臂和Liconic自动化培养箱的集成。这 仪器的组合将允许在每个期望的水平进行高通量表型筛选， 容量我们注意到，MD致力于设计最强大的自动化显微镜仪器， 旨在针对3D结构的HCI实现速度和精度的优化， 类器官拟议中的系统将为已经建立的范德比尔特客户增加重要价值，因为 以及VICB和VHTS设施在药物发现方面的使命， 确定具有临床相关性的治疗策略。

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.