High-content screening for modifiers of the DNA damage response

DNA 损伤反应调节剂的高内涵筛选

• 批准号: 7559378
• 负责人: Scott R Floyd
• 金额: $ 16.32万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-15 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7559378
• 关键词: Biological Assay; Cell Cycle; Cells; Cellular biology; Chemicals; DNA Damage; DNA Repair; DNA Structure; DNA biosynthesis; Data Set; Development; Exposure to; Genetic; Genetic Transcription; Genome; Genotoxic Stress; Human; Image; Image Analysis; Individual; Ionizing radiation; Lead; Measurement; Metabolism; Microscopy; Molecular; Molecular Bank; Mutagenesis; Normal tissue morphology; Numbers; Production; Proliferating; Radiation; Radiation therapy; Radiation-Protective Agents; Radiation-Sensitizing Agents; Radio; Resistance; Screening procedure; Signal Transduction; Techniques; Tissues; Toxic Environmental Substances; Toxin; United States National Institutes of Health; base; cancer cell; cancer therapy; cytotoxic; digital imaging; environmental radiation; high throughput screening; killings; neoplastic; novel; response; small molecule; success; tool; tumor; ultraviolet

DESCRIPTION (provided by applicant): Project Summary: A coordinated and integrated response to DNA damage is critical for cells to survive and proliferate after genotoxic stress. Cells contain an elaborate molecular machinery that halts the cell cycle and repairs damaged DNA after a wide variety of extrinsic or intrinsic insults such as exposure to ultraviolet and ionizing radiation, environmental toxins, products of normal metabolism, and unstable DNA structures formed during gene transcription and DNA replication. Although DNA damage has clear potential for mutagenesis and tumor induction in normal tissues, it paradoxically forms a mainstay of modern cancer treatment due to its cytotoxic effect. Selective tumor killing by DNA damaging treatments such as radiation therapy relies on a differential response to genotoxic stress between normal and neoplastic tissue. Unfortunately, a significant number of tumor types are resistant to radiation treatment. The identification of small molecules capable of modulating the response of cells to ionizing radiation would be of tremendous utility in deciphering the molecular details of the DNA damage response, providing chemical tools for exploring the basic cell biology of environmentally-induced genetic damage, and identifying lead compounds for the development of clinically useful radiation sensitizers or radioprotective agents. In this proposal we refine, adapt and optimize an automated microscopy-based assay to simultaneously interrogate multiple components of the DNA damage response in human cancer cells for use in high- throughput screening of compounds by NIH Molecular Libraries Production Centers Network as part of the NIH Molecular Libraries and Imaging Roadmap Initiative. This assay uses novel digital image analysis techniques to create a unique dataset of multiple quantitative measurements on individual cells that optimizes the chances of success in subsequent high-throughput small molecule screening. Narrative In this proposal we refine, adapt and optimize an automated microscopy-based assay to simultaneously interrogate multiple components of the DNA damage response in human cancer cells for use in high-throughput screening of compounds by NIH Molecular Libraries Production Centers Network as part of the NIH Molecular Libraries and Imaging Roadmap Initiative. The results of this project should facilitate the development of chemical tools for studying the response of cells to environmental radiation and toxins that damage the genome, and help identify compounds that could be used as radio- protective and radio-sensitizing agents to enhance current forms of cancer treatment.

描述(由申请人提供)：项目摘要：对DNA损伤的协调和综合反应对细胞在遗传毒性应激后的生存和增殖至关重要。细胞包含一种复杂的分子机制，在受到各种外部或内在的伤害后，如暴露在紫外线和电离辐射、环境毒素、正常代谢的产物以及在基因转录和DNA复制过程中形成的不稳定的DNA结构，细胞可以停止细胞周期并修复受损的DNA。虽然DNA损伤在正常组织中具有明显的诱变和肿瘤诱导的潜力，但由于其细胞毒性作用，它却矛盾地形成了现代癌症治疗的主流。通过DNA损伤治疗(如放射治疗)选择性杀伤肿瘤依赖于正常组织和肿瘤组织对遗传毒性应激的不同反应。不幸的是，相当数量的肿瘤类型对放射治疗具有抵抗力。识别能够调节细胞对电离辐射的反应的小分子将在破译DNA损伤反应的分子细节、为探索环境诱导的遗传损伤的基本细胞生物学提供化学工具以及为开发临床上有用的辐射敏感剂或辐射防护剂识别先导化合物方面具有巨大的实用价值。在这项提案中，我们提炼、调整和优化了一种基于显微镜的自动分析方法，以同时询问人类癌细胞中DNA损伤反应的多个组件，以用于NIH分子文库生产中心网络作为NIH分子文库和成像路线图计划的一部分的高通量化合物筛选。该分析使用新的数字图像分析技术来创建单个细胞的多个定量测量的独特数据集，从而优化后续高通量小分子筛查的成功机会。在这项提案中，我们改进、调整和优化了一种基于显微镜的自动分析方法，以同时询问人类癌细胞中DNA损伤反应的多个组件，以用于NIH分子文库生产中心网络作为NIH分子文库和成像路线图计划的一部分进行的高通量化合物筛选。该项目的成果应有助于开发化学工具，研究细胞对环境辐射和破坏基因组的毒素的反应，并有助于确定可用作辐射防护剂和辐射增敏剂的化合物，以加强目前的癌症治疗形式。