SBIR Phase I: A novel multicolor cell line engineering platform that enables high-throughput microscopy-based screening of living cells for drug discovery

SBIR 第一阶段：一种新型多色细胞系工程平台，可实现基于高通量显微镜的活细胞筛选以用于药物发现

• 批准号: 1448764
• 负责人: Mary Ludlam
• 金额: $ 15万
• 依托单位: Cairn Biosciences
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1448764&HistoricalAwards=false
• 关键词: SBIR; Phase; novel; multicolor; cell

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is the development of new tools to study the dynamic behavior of cellular machinery, especially for applications in drug discovery. Unraveling the dynamic aspects of cellular physiology that may be targeted therapeutically requires a new paradigm for screening disease biology in living cells. Physiologically relevant live-cell models that are compatible with visualizing and quantifying the spatiotemporal regulation of disease-relevant signal transduction pathways and cellular machinery will be a key component of this approach. Such models will enable improved prioritization of potential therapeutics by virtue of their ability to deliver enhanced mechanistic insights compared to existing end-point cell-based and biochemical assays, and their increased throughput and reduced cost compared with animal models used in preclinical drug discovery research and toxicology testing. These attributes represent important advances for de novo drug discovery, drug repurposing initiatives and the identification of productive therapeutic combinations, in addition to being valuable capabilities for basic cell biology research.This SBIR Phase I project proposes to develop a novel reporter cell line engineering platform that will enable the rapid generation of multicolor fluorescent cell lines suitable for kinetic live-cell high-content screening (LC-HCS). Stable cell line generation using traditional antibiotic-selection methods is a lengthy and inflexible process ill-suited for the efficient generation of LC-HCS-compatible cells. The aim is to address these shortcomings by implementing a novel strategy to deliver cell lines that incorporate fluorescent reporters of multiple cellular markers under inducible control, and at well-defined, physiologically relevant, and stoichiometrically balanced expression levels at a specified locus within 4-6 weeks of transfection. This will be accomplished through the use of a genome editing strategy that will yield a panel of cell lines that can be quickly configured to report on any desired combination of up to 4 fluorescent markers. The multicolor reporter cell lines generated through this SBIR project will represent valuable new tools for the next generation of drug discovery in clinically relevant living cells and contribute invaluable insights into human disease.

这个小企业创新研究（SBIR）项目的更广泛的影响/商业潜力是开发新的工具来研究细胞机器的动态行为，特别是在药物发现中的应用。 解开细胞生理学的动态方面，可能是有针对性的治疗需要一个新的范例，筛选活细胞中的疾病生物学。与可视化和量化疾病相关信号转导通路和细胞机制的时空调节兼容的生理相关活细胞模型将是这种方法的关键组成部分。 这些模型将能够改善潜在治疗剂的优先级排序，因为与现有的基于终点细胞和生物化学测定相比，它们能够提供增强的机制见解，并且与临床前药物发现研究和毒理学测试中使用的动物模型相比，它们具有更高的通量和更低的成本。 这些属性代表了从头药物发现，药物再利用倡议和生产性治疗组合鉴定的重要进展，SBIR第一阶段项目提出开发一种新型的报告细胞系工程平台，该平台将能够快速生成适用于动态活细胞高含量筛选（LC-HCS）的荧光细胞系。 使用传统的免疫选择方法产生稳定的细胞系是一个冗长且不灵活的过程，不适合有效产生LC-HCS相容性细胞。 目的是通过实施一种新的策略来解决这些缺点，以在转染的4-6周内在指定的基因座处以明确定义的、生理学相关的和化学计量平衡的表达水平递送在诱导型控制下掺入多种细胞标志物的荧光报告物的细胞系。这将通过使用基因组编辑策略来实现，该策略将产生一组细胞系，这些细胞系可以快速配置为报告多达4种荧光标记物的任何所需组合。 通过SBIR项目产生的报告基因细胞系将代表在临床相关活细胞中发现下一代药物的宝贵新工具，并为人类疾病提供宝贵的见解。