Cellular Responses to Cancer Drug-Induced Genomic Breaks

细胞对癌症药物引起的基因组断裂的反应

基本信息

批准号：
7169561
负责人：
TERRY A BEERMAN
金额：
$ 27.28万
依托单位：
ROSWELL PARK CANCER INSTITUTE CORP
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-14 至 2009-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7169561
关键词：
1-Phosphatidylinositol 3-Kinase Antineoplastic Agents Ataxia-Telangiectasia-Mutated protein kinase C 1027 Cell Cycle Checkpoint Cell Cycle Progression Cells Characteristics Chromosome abnormality Chromosomes Chromosomes, Human, Pair 3 Condition Cytogenetic Analysis DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway DNA-Binding Proteins Disruption Ensure Fluorescent in Situ Hybridization Functional disorder Genetic Recombination Genomics Immunofluorescence Microscopy Interphase Ionizing radiation Ku Protein Measures Metaphase Mitosis Mitotic Modeling Nonhomologous DNA End Joining Oligonucleotides Pathway interactions Pharmaceutical Preparations Phosphotransferases Play Process Property Proteins Regulation Role Spectral Karyotyping Tandem Repeat Sequences Testing Western Blotting antitumor drug cytotoxic homologous recombination insight repaired response telomere

项目摘要

DESCRIPTION (provided by applicant): Cellular responses to ionizing radiation (IR)-induced DNA double strand breaks (DSBs) involve activation, via the phosphatidylinositol 3-kinase-like kinase (PIKK) ATM, of cell cycle checkpoint proteins that delay cell cycle progression to allow DNA repair and to preserve chromosomal integrity. Responses to DNA DSBs induced by the radiomimetic enediyne C-1027 deviate from this model. For example, IR induces ATM-dependent, and C-1027, both ATM- dependent and independent, responses. Furthermore, treatment with equi-cytotoxic levels of IR or C-1027 results in either a very low (2.9%), or an extraordinarily high (92%), percentage, respectively of mitotic cells showing aberrant chromosomal recombination. FISH analysis revealed that C-1027-induced chromosomal aberrations are frequently associated with disruption of telomeres. The mechanism(s) behind these unique responses to the antitumor drug, C-1027 should help extend our understanding of DNA damage responses to DSBs. First, pivotal checkpoint pathway proteins involved in the response to C-1027 will be identified, with emphasis on both ATM-dependent and independent responses. Next the conditions under which C-1027 induces extensive chromosomal fusions, characterized by aberrant end-joining and fragmentation, will be identified. The NHEJ DNA binding protein Ku, which not only is involved in repair responses but also plays a role in regulating chromosomal fusions, will also be examined for its role in C-1027 induced aberrant end-joining. Finally, we will test whether C-1027, which induces DSBs preferentially within a GTTA motif, targets the telomere tandem repeat sequence (GGGTTA) and contributes to aberrant end-joining by inducing telomere dysfunction. Other enediynes with cleavage characteristics different from C-1027, will be tested selectively to provide additional mechanistic insights into which properties of C-1027-induced damage are associated with the DNA damage responses. The specific aims will test the following hypotheses: 1. C-1027 is unique compared to other enediynes and to IR in that it induces both ATM-dependent and independent DNA damage responses. 2. C-1027 is unique compared to other enediynes and to IR in that it causes extraordinarily high levels of rearranged chromosomes. 3. C-1027 is unique compared to other enediynes and IR in that it targets telomeres.

描述（由申请人提供）：细胞对电离辐射（IR）诱导的DNA双链断裂（DSB）的响应涉及通过磷脂酰肌醇3-激酶（PIKK）类似激酶（PIKK）ATM激活，这些细胞周期检查点延迟细胞周期蛋白进展以允许DNA修复并保留染色体完整性。放射性ENEDIYNE C-1027诱导的对DNA DSB的反应与该模型偏离。例如，IR诱导ATM依赖性和ATM依赖性和独立响应的C-1027。此外，用等应毒性水平的IR或C-1027治疗会导致非常低（2.9％）或非常高（92％）的有丝分裂细胞的百分比（92％），表现出异常的染色体重组。鱼类分析表明，C-1027诱导的染色体畸变经常与端粒中断有关。这些对抗肿瘤药物的独特反应背后的机制C-1027应有助于我们对DNA损伤对DSB的响应的理解。首先，将确定参与C-1027响应的关键检查点途径蛋白，重点是ATM依赖性和独立响应。接下来，将确定C-1027诱导广泛的染色体融合（以异常结合和碎片化为特征）的条件。 NHEJ DNA结合蛋白KU不仅参与了修复反应，而且还在调节染色体融合中起作用，还将检查其在C-1027引起的诱导异常连接中的作用。最后，我们将测试C-1027是否靶向端粒串联重复序列（GGGTTTA），该C-1027是否优先诱导DSB，并通过诱导端粒功能障碍而导致异常结合。其他具有与C-1027不同的裂解特性的ENEDIYNES将有选择性测试，以提供其他机械见解，以了解C-1027诱导的损伤的特性与DNA损伤响应有关。具体目的将检验以下假设： 1。C-1027与其他ENEDIYNES相比是独一无二的，与IR相比，它诱导了ATM依赖性和独立的DNA损伤响应。 2。C-1027与其他Enediynes相比是独一无二的，与IR相比，它会引起非常高的重排染色体。 3。C-1027与其他ENEDIYNES和IR相比是唯一的，因为它针对端粒。