Genome instability in cancer: telomeres and DNA repair

癌症中的基因组不稳定性：端粒和 DNA 修复

• 批准号: 9186655
• 负责人: Titia de Lange
• 金额: $ 90.27万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-02 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9186655
• 关键词: Biological Models; CRISPR/Cas technology; Cells; Chromatin Structure; Clinic; DNA Repair; Data; Development; Double Strand Break Repair; Epithelial Cells; Excision; Functional disorder; Genes; Genome; Genomic Instability; Human; Image; Maintenance; Malignant Neoplasms; Microscopy; Molecular; Pathway interactions; Poly(ADP-ribose) Polymerases; Positioning Attribute; Resolution; Structure; TP53 gene; Tumor Suppressor Proteins; XCL1 gene; base; cancer genome; cancer therapy; genome editing; genome integrity; genome sequencing; in vitro Model; innovation; insight; mouse model; new technology; p53-binding protein 1; public health relevance; research study; telomere; tumor; whole genome

PROJECT SUMMARY/ABSTRACT Recent sequencing data has revealed an unanticipated level of genomic instability in cancer. The origin of these changes is largely unknown. This proposal focuses on two aspects of the maintenance of genome integrity relevant to cancer: telomere dysfunction and DNA repair. We propose to examine the molecular basis of the telomere tumor suppressor pathway whereby telomere attrition leads to dysfunctional telomeres and proliferative arrest. These experiments will make use of normal human epithelial cells in which critical genes have been removed with CRISPR/Cas9 genome editing and will apply innovative imaging of telomere structure using super-resolution STORM imaging in combination with expansion microscopy. When the telomere tumor suppressor pathway fails because of the loss of p53 and Rb, telomeres continue to shorten during early tumor development, leading to large numbers of dysfunctional telomeres and genome instability (referred to as telomere crisis). We propose to determine the changes in the genome that result from progression through telomere crisis. These experiments will involve a new in vitro model for telomere crisis in epithelial cells combined with whole genome sequencing. Finally, we propose to study the mechanism of double-strand break (DSB) repair, which is carefully regulated to maintain genome integrity. In particular, we will focus on the mechanism by which 53BP1 represses the resection of DSBs and promotes their mobility using the mouse model systems we have developed and new technologies for the analysis of chromatin structure (ATAC sequencing) and locus position (DamID-LaminB1 marking). These two attributes of 53BP1 are highly relevant to the induction of lethal genome instability resulting from inhibition of the PARP1 poly(ADP-ribose) polymerase in BRCA-deficient cells. Collectively, the proposed experiments will illuminate new aspects of genome instability in cancer. The objective of these studies is to gain insights that will ultimately inform and guide decisions in the cancer clinic.

项目摘要/摘要 最近的测序数据揭示了癌症中基因组不稳定性的意外水平。的起源 这些变化在很大程度上是未知的。这项建议侧重于基因组维护的两个方面 与癌症相关的完整性：端粒功能障碍和DNA修复。我们建议研究分子基础 端粒肿瘤抑制通路，端粒磨损导致端粒功能障碍和 增殖性逮捕。这些实验将利用正常的人类上皮细胞，其中关键基因 已通过CRISPR/Cas9基因组编辑移除，并将应用端粒结构的创新成像 使用超分辨率风暴成像与扩张显微镜相结合。当端粒肿瘤 抑癌通路因P53和Rb丢失而失效，肿瘤早期端粒持续缩短 发育，导致大量端粒功能障碍和基因组不稳定(称为 端粒危机)。我们建议确定基因组在进化过程中产生的变化 端粒危机。这些实验将涉及一种新的上皮细胞端粒危机的体外模型。 结合全基因组测序。最后，我们提出了研究双链断裂的机理。 (DSB)修复，它受到仔细的监管，以保持基因组的完整性。我们将特别关注 利用小鼠研究53BP1抑制DSB切除并促进其移动的机制 我们开发的模型系统和染色质结构分析的新技术(ATAC 测序)和位点位置(DamID-LaminB1标记)。53BP1的这两个属性高度相关 抑制PARP1多聚(ADP-核糖)聚合酶导致致死性基因组不稳定性 在BRCA缺乏的细胞中。总的来说，拟议中的实验将阐明基因组的新方面。 癌症中的不稳定性。这些研究的目标是获得最终将提供信息和指导的见解 癌症诊所的决定。