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于细胞在基因毒性应激后生存和增殖至关重要。细胞含有复杂的分子机制，可在各种外在或内在损伤（例如暴露于紫外线和电离辐射、环境毒素、正常代谢产物以及基因转录和 DNA 复制过程中形成的不稳定 DNA 结构）后停止细胞周期并修复受损的 DNA。尽管 DNA 损伤在正常组织中具有明显的诱变和肿瘤诱导潜力，但由于其细胞毒性作用，它却成为现代癌症治疗的支柱。通过 DNA 损伤治疗（例如放射治疗）选择性杀死肿瘤依赖于正常组织和肿瘤组织之间对基因毒性应激的差异反应。不幸的是，相当多的肿瘤类型对放射治疗有抵抗力。能够调节细胞对电离辐射反应的小分子的鉴定将在破译DNA损伤反应的分子细节方面具有巨大的实用性，为探索环境引起的遗传损伤的基本细胞生物学提供化学工具，并鉴定用于开发临床有用的辐射增敏剂或辐射防护剂的先导化合物。在本提案中，我们改进、调整和优化了基于自动显微镜的测定，以同时询问人类癌细胞中 DNA 损伤反应的多个成分，用于 NIH 分子库生产中心网络的化合物高通量筛选，作为 NIH 分子库和成像路线图计划的一部分。该测定使用新颖的数字图像分析技术来创建对单个细胞进行多次定量测量的独特数据集，从而优化后续高通量小分子筛选的成功机会。叙述在本提案中，我们改进、调整和优化了一种基于显微镜的自动化测定，以同时询问人类癌细胞中 DNA 损伤反应的多个成分，用于 NIH 分子库生产中心网络对化合物的高通量筛选，作为 NIH 分子库和成像路线图计划的一部分。该项目的结果应有助于开发化学工具，用于研究细胞对环境辐射和损害基因组的毒素的反应，并有助于识别可用作放射防护剂和放射增敏剂的化合物，以增强当前的癌症治疗形式。