IGF::OT::IGF MULTIPLEXED HIGH-CONTENT ASSAY TO MEASURE CELLULAR STRESS MECHANISMS FOR PREDICTIVE TOXICOLOGY STUDIES

IGF::OT::IGF 多重高内涵检测可测量细胞应激机制以进行预测毒理学研究

• 批准号: 9355770
• 负责人: MARY LUDLAM
• 金额: $ 150万
• 依托单位: CAIRN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-15 至 2018-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9355770
• 关键词: Automation; Biological Assay; Biology; Cell Line; Cells; Cellular Stress; Contractor; Contracts; Development; Engineering; Evaluation; Foundations; Generations; Homeostasis; Libraries; Life; Measures; Monitor; Organelles; Pharmacologic Substance; Phase; Public Health; Reporter; Reporting; Running; Sampling; Small Business Innovation Research Grant; Testing; Therapeutic; Time; Toxic Environmental Substances; Toxic effect; Toxicology; assay development; cost; environmental chemical; high throughput screening; improved; novel; response; screening; tool

The Contractor currently provides a high-throughput assay platform that rapidly pinpoints compound mechanism of action and toxicity liabilities through its unique ability to monitor multiple facets of previously inaccessible biology in live cells in real-time. Through this SBIR Phase II contract, the Contractor's existing platform will be applied to the development of a novel live-cell multiplexed fluorescent assay that reports an aggregate of at least 10 distinct readouts of cellular stress and organelle homeostasis per well. A panel of multicolor cell stress reporters will be generated and stably integrated into cell lines relevant for compound toxicity evaluation. The resulting panel of multicolor stable reporter cell lines will be used as the foundation for our multiplexed assay. Our assay will be validated by testing its response to tool compounds known to induce cellular stress. Our assay's suitability for high-throughput screening will be evaluated by running a 10,000-sample pilot screen, followed by scaling to at least 100,000 samples/day throughput in 384-well plate format with full assay automation. The Contractor's proposed assay will benefit public health by reducing the time and cost to pinpoint toxicities of pharmaceutical and environmental chemicals, thus improving our ability to identify beneficial therapeutics, and to proactively manage harmful environmental toxins.

承包商目前提供一个高通量检测平台，通过其独特的能力来快速查明化合物的作用机制和毒性责任，实时监测活细胞中以前无法访问的生物学的多个方面。 通过这份 SBIR II 期合同，承包商的现有平台将用于开发一种新型活细胞多重荧光测定，该测定报告每孔至少 10 个不同的细胞应激和细胞器稳态读数。将生成一组多色细胞应激报告基因，并将其稳定地整合到与化合物毒性评估相关的细胞系中。由此产生的多色稳定报告细胞系组将用作我们多重测定的基础。我们的测定将通过测试其对已知诱导细胞应激的工具化合物的反应来验证。我们的检测对高通量筛选的适用性将通过运行 10,000 个样本的试点筛选进行评估，然后在 384 孔板格式中扩展到至少 100,000 个样本/天的通量，并具有完全的检测自动化。 承包商提出的检测方法将通过减少查明药物和环境化学品毒性的时间和成本来有益于公众健康，从而提高我们识别有益疗法和主动管理有害环境毒素的能力。