Integrated ImageXpress Micro Confocal High Content Screening System

集成 ImageXpress 微型共焦高内涵筛选系统

• 批准号: 10175498
• 负责人: Joshua A. Bauer
• 金额: $ 79.9万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175498
• 关键词: 3-Dimensional; Academia; Automation; Biochemical; Biological Assay; Biology; Biomedical Research; Cell Line; Cells; Chemicals; Clinical; Complex; Devices; Disease; Disease model; Drug Combinations; Drug Screening; Drug toxicity; Equipment; Fluorescence; Functional disorder; Guide RNA; Image; Incubators; Industry; Institutes; Intention; Libraries; Measures; Methods; Microscope; Mission; Molecular; Open Reading Frames; Organoids; Patients; Phenotype; Preclinical Testing; RNA Interference; Resistance; Robot; Robotics; Route; Scientific Advances and Accomplishments; Services; Small Interfering RNA; Speed; System; Technology; Therapeutic; Tissues; Vendor; Virus Replication; arm; base; cellular imaging; clinically relevant; confocal imaging; design; drug candidate; drug discovery; drug mechanism; functional genomics; high throughput screening; human disease; imaging capabilities; imaging system; in vitro Model; induced pluripotent stem cell; innovative technologies; instrument; instrumentation; meetings; novel therapeutics; response; screening; small molecule; stem cells; therapeutic development; three dimensional structure; three-dimensional modeling

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.

项目概要 在过去的 20 年里，学术界和工业界都已从基于靶点的药物发现显着转向基于靶点的药物发现。 表型筛选。人类疾病长期以来一直是生物医学研究企业未满足的需求 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。培养源自患者或患者的类器官或 3D 组织的最新进展 祖细胞为应对这一挑战提供了机会。人们普遍认为，药物发现使用 这种体外模型代表了候选药物临床前测试的加速途径，因为类器官 与培养细胞系相比，3D 结构更类似于患病组织的复杂生理病理学 或生化测定。该领域持续面临的挑战是仪器、技术和技术的升级 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) 设施是一个良好的 建立学术药物发现中心，利用创新技术推进科学和临床 治疗发现。目前，VHTS 设施拥有一台基于荧光的自动显微镜，可用于高 内容成像 (HCI) 和表型筛选与机械臂和培养箱集成，允许 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、功能基因组筛选（siRNA、ORF）、病毒复制机制、药物毒性 研究、新疗法开发的药物筛选、药物作用和耐药机制以及药物 组合研究。然而，由于缺乏共焦成像能力，该系统严重不足 用于对类器官和 3D 结构内的特征进行成像和测量。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至关重要， 配备机器人设备，以实现最先进的高通量表型筛选 使用类器官和 3D 结构进行疾病生物学和药物发现的方法。拟议的 HCI 药物 发现系统将以 Molecular Devices (MD) ImageXpress Micro Confocal HCI 系统为中心， Precise Automation PreciseFlex PF400 机械臂和 Liconic 自动培养箱的集成。这 仪器组合将允许在每个所需水平进行高通量表型筛选 容量。我们注意到 MD 致力于设计最坚固的自动化显微镜仪器 专门针对 3D 结构的 HCI 实施速度和精度优化的意图，以及 类器官。拟议的系统将为范德比尔特已经建立的客户群增加显着的价值，因为 以及 VICB 和 VHTS 设施利用具有强大潜力的疾病模型进行药物发现的使命 确定具有临床相关性的治疗策略。

项目成果

High-Content Imaging Platform to Discover Chemical Modulators of Plasma Membrane Rafts.

用于发现质膜筏化学调节剂的高内涵成像平台。

• 发表时间: 2022-03-23
• 影响因子: 18.2
• 作者: Fricke, Nico;Raghunathan, Krishnan;Tiwari, Ajit;Stefanski, Katherine M;Balakrishnan, Muthuraj;Waterson, Alex G;Capone, Ricardo;Huang, Hui;Sanders, Charles R;Bauer, Joshua A;Kenworthy, Anne K
• 通讯作者: Kenworthy, Anne K