Genome maintenance by the mouse Hus1 checkpoint gene

小鼠 Hus1 检查点基因的基因组维护

• 批准号: 7876855
• 负责人: Robert S Weiss
• 金额: $ 27.75万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-15 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7876855
• 关键词: Address; Adult; Affect; Alleles; Animals; Apoptosis; Biological Process; Bone Marrow; Cell Cycle; Cell Cycle Arrest; Cell Proliferation; Cell Survival; Cells; Clinical; Collection; Complement; Complex; Cultured Cells; DNA; DNA Damage; DNA Repair; DNA damage checkpoint; DNA lesion; Defect; Development; Disease; Embryonic Development; Functional disorder; Genes; Genetic; Genetic Complementation Test; Genetic Polymorphism; Genetically Engineered Mouse; Genome; Genome Stability; Genomic Instability; Genomics; Genotoxic Stress; Goals; Growth; Head; Hus1 protein; Hypersensitivity; Impairment; Investigation; Ionizing radiation; Knowledge; Light; Maintenance; Malignant Neoplasms; Mammalian Cell; Mammals; Measures; Mediating; Mediator of activation protein; Mitomycins; Modeling; Molecular; Mouse Strains; Mus; Mutagens; Mutation; Oncogenes; Organism; Output; Pathway interactions; Physiological; Post-Translational Modification Site; Predisposition; Premature aging syndrome; Procedures; Process; Protein Kinase; Research; Research Project Grants; Resources; Role; Signal Transduction; Site-Directed Mutagenesis; Skin; Stress; Testing; Tissues; Toxic effect; Tumor Suppression; Tumor Suppressor Proteins; Variant; Vertebrates; anti-cancer therapeutic; base; biological adaptation to stress; cancer cell; cancer prevention; cancer therapy; cell transformation; chemical carcinogenesis; crosslink; design; experience; gastrointestinal; improved; in vivo; insight; lung tumorigenesis; member; neoplastic cell; novel; prevent; protein protein interaction; public health relevance; research study; response; stem; therapeutic target; tool; tumor; tumorigenesis

DESCRIPTION (provided by applicant): Cells continuously experience genome damage that, if not properly attended to, can cause developmental defects, premature aging, and increased cancer predisposition. DNA damage checkpoints defend against these consequences of genomic instability by regulating cell cycle transitions, apoptosis, and DNA repair. In mammals, the checkpoint protein Hus1 is a critical component of a pathway that responds to replication stress and a variety of DNA lesions. The Hus1- dependent pathway is essential for embryonic development, and consequently many of its biological functions in animals are not well-defined. The long-term goals of the research described in this proposal are to understand how the Hus1-dependent checkpoint pathway protects genomic integrity and to determine how defects in this mechanism affect tumorigenesis and physiological DNA damage responses. The proposed studies will advance these important objectives using a unique collection of targeted modifications at the mouse Hus1 genomic locus. Aim one addresses how partial or complete Hus1 inactivation affects tumor development induced by activated oncogenes in order to resolve the opposing roles of Hus1 as a candidate tumor suppressor and a factor required for cancer cell survival and proliferation. In Aim Two, the requirements for Hus1 during in vivo DNA damage responses will be determined by measuring the tissue-specific genotoxic stress responses of mice with reduced Hus1 expression, in part to gain new insights into how checkpoint function impacts sensitivity to DNA damaging anti-cancer therapies. These analyses of the physiological consequences of checkpoint dysfunction in the first two aims will be complemented by the investigation of Hus1 molecular mechanisms in Aim Three, in an effort to define the Hus1 functions that are essential for the survival of genotoxic stress and to establish how Hus1 sequence variations affect genome maintenance. Together, the proposed studies will define how an essential checkpoint mechanism functions during tumorigenesis and tissue-specific DNA damage responses, and will provide insights into the biomedical implications of checkpoint dysfunction stemming from spontaneous mutations, natural polymorphisms, or pharmacological inhibition. PUBLIC HEALTH RELEVANCE: By defending against damage to the genome, DNA damage checkpoints are critical for normal embryonic development, protective responses to environmental stresses and DNA damaging chemotherapeutics, and the tumor-free survival of adult organisms. The experiments of this proposal make use of novel genetically-engineered mouse strains to analyze the biological functions of the essential DNA damage checkpoint gene Hus1. These studies hold great promise for improving the understanding of how cancers develop and how normal and neoplastic cells respond to anti-cancer therapies, as well as how these processes are influenced by Hus1 sequence variations, including spontaneous mutations and natural polymorphisms.

描述（由申请人提供）：细胞持续经历基因组损伤，如果不适当处理，可能导致发育缺陷、过早衰老和癌症易感性增加。DNA损伤检查点通过调节细胞周期转换、凋亡和DNA修复来防御基因组不稳定性的这些后果。在哺乳动物中，检查点蛋白Hus1是响应复制应激和各种DNA损伤的途径的关键组成部分。Hus1依赖性通路对于胚胎发育至关重要，因此其在动物中的许多生物学功能尚未明确。该提案中描述的研究的长期目标是了解Hus1依赖性检查点途径如何保护基因组完整性，并确定该机制中的缺陷如何影响肿瘤发生和生理DNA损伤反应。拟议的研究将使用小鼠Hus1基因组位点的独特靶向修饰集合来推进这些重要目标。目的之一是解决部分或完全的Hus1失活如何影响肿瘤的发展所诱导的癌基因激活，以解决Hus1作为一个候选的肿瘤抑制因子和癌细胞生存和增殖所需的因素的对立作用。在目标二中，体内DNA损伤反应期间对Hus1的需求将通过测量Hus1表达减少的小鼠的组织特异性遗传毒性应激反应来确定，部分原因是为了获得关于检查点功能如何影响对DNA损伤抗癌疗法的敏感性的新见解。在前两个目标中检查点功能障碍的生理后果的这些分析将通过目标三中的Hus1分子机制的调查来补充，以努力定义Hus1功能，这些功能对于遗传毒性应激的生存至关重要，并确定Hus1序列变异如何影响基因组的维持。总之，拟议的研究将定义在肿瘤发生和组织特异性DNA损伤反应期间基本检查点机制如何发挥作用，并将提供对自发突变，自然多态性或药理学抑制引起的检查点功能障碍的生物医学意义的见解。公共卫生相关性：通过防御对基因组的损伤，DNA损伤检查点对于正常胚胎发育、对环境压力和DNA损伤化疗的保护性反应以及成年生物体的无肿瘤生存至关重要。本方案的实验利用新型基因工程小鼠品系来分析必需的DNA损伤检查点基因Hus1的生物学功能。这些研究对于提高对癌症如何发展以及正常和肿瘤细胞如何对抗癌疗法作出反应的理解，以及这些过程如何受到Hus1序列变异（包括自发突变和天然多态性）的影响，具有很大的希望。