The structural function of ATR in development, oncogenesis and cancer therapy

ATR 在发育、肿瘤发生和癌症治疗中的结构功能

• 批准号: 9886205
• 负责人: Shan Zha
• 金额: $ 42.18万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9886205
• 关键词: ATR gene; Affect; Aging; Alleles; Appearance; CHEK1 gene; Cell Cycle Checkpoint; Cells; Cessation of life; Chromosomes; Combined Modality Therapy; DNA Damage; DNA Double Strand Break; DNA Repair; DNA biosynthesis; DNA replication fork; DNA-dependent protein kinase; Defect; Development; Dominant-Negative Mutation; Embryo; Embryonic Development; Enterobacteria phage P1 Cre recombinase; Estrogen receptor positive; Evolution; Female; Genetic Models; Genetic Recombination; Genomic Instability; Genotoxic Stress; Growth; Hematopoiesis; Human; Hypersensitivity; Impairment; In Vitro; Infertility; Knock-in; Lymphocyte; Malignant Neoplasms; Mediating; Meiosis; Microcephaly; Mitotic; Modeling; Mus; Mutagens; Mutation; Oncogenes; Oncogenic; Pathway interactions; Phosphorylation; Phosphotransferases; Process; Proliferating; Proteins; RNA Splicing; Radiation; Replication Origin; Resected; Role; Seckel syndrome; Sex Chromosomes; Spermatogenesis; Sterility; Structure; Structure of thyroid parafollicular cell; Symptoms; TP53 gene; Tamoxifen; Testing; Therapeutic; Therapeutic Effect; Tumor Suppression; base; cancer cell; cancer initiation; cancer therapy; chemotherapy; exhaust; genotoxicity; in vivo; in vivo regeneration; inhibitor/antagonist; kinase inhibitor; leukemia; male; mouse model; overexpression; preclinical trial; premature; prevent; replication stress; response; side effect; stem; stem cells; tissue culture; tissue regeneration; tumorigenesis

PROJECT SUMMARY/ABSTRACT Genomic instability drives cancer initiation, progression and treatment. The ATR kinase is a master regulator of DNA damage responses (DDRs) during DNA replication and strand breaks. Although complete loss of ATR (Atr-/-) abrogates embryonic development and cellular viability, ATR kinase inhibitors are well-tolerated in preclinical trials and have generated promising therapeutic effects when used in combination with genotoxic treatments including radiation. To determine whether ATR protein has a structural function in DNA replication and repair (i.e., independent of its kinase activity or regulated by auto-phosphorylation), we generated a mouse model expressing kinase-dead ATR protein (AtrKD). Atr+/KD mice are born at expected Mendelian ratio and of normal size. Moreover somatic inactivation of the Atr conditional allele (AtrC) using Tamoxifen inducible Cre recombinase is very well tolerated in AtrC/KD mice but not AtrC/- mice. While female Atr+/KD mice are fertile, Atr+/KD male mice are sterile due to meiosis defects during spermatogenesis that were not seen in Atr+/- mice. Here we propose to use the Atr null (Atr-), conditional (AtrC), and kinase-dead (AtrKD) mouse models to investigate kinase-independent structural function during normal DNA replication and aging (aim 1), auto-phosphorylation dependent dominant negative function of AtrKD protein during spermatogenesis(aim 2) and the oncogenic potential as well as the chemo-sensitivity profile of the AtrKD mutation. Together, the results will elucidate the structural functions of ATR during DNA repair and oncogenesis, and the mechanisms underlying the therapeutic as well as side effects of ATR inhibition.

项目摘要/摘要 基因组不稳定性驱动癌症的开始，进展和治疗。 ATR激酶是 DNA损伤反应（DDR）在DNA复制和链断裂中断裂。虽然完全失去了ATR （ATR - / - ）废除了胚胎发育和细胞活力，ATR激酶抑制剂在 临床前试验并与遗传毒性结合使用时产生了有希望的治疗作用 包括辐射在内的治疗。确定ATR蛋白在DNA复制中是否具有结构功能 和修复（即独立于其激酶活性或通过自磷酸化调节），我们产生了一只小鼠 表达激酶已故ATR蛋白（ATRKD）的模型。 ATR+/KD小鼠以预期的Mendelian比率出生 正常大小。此外，使用他莫昔芬诱导CRE的ATR条件等位基因（ATRC）的体细胞失活（ATRC） 重组酶在ATRC/KD小鼠中的耐受性良好，而不是ATRC/ - 小鼠。雌性ATR+/KD小鼠肥沃，ATR+/KD 雄性小鼠由于在ATR +/-小鼠中未见的精子发生过程中的减数分裂缺陷而导致无菌。我们在这里 建议使用ATR NULL（ATR-），条件（ATRC）和激酶已故（ATRKD）鼠标模型来研究 在正常DNA复制和衰老期间，非激酶无关的结构功能（AIM 1），自磷酸化 精子发生过程中ATRKD蛋白的依赖性显性负功能（AIM 2）和致癌 电位以及ATRKD突变的化学敏感性。结果一起，结果将阐明 ATR在DNA修复和肿瘤发生过程中的结构功能以及依据的机制 ATR抑制的治疗和副作用。