Polyploidy, Aneuploidy and Genome Stability

多倍体、非整倍体和基因组稳定性

• 批准号: 10544815
• 负责人: DAVID S PELLMAN
• 金额: $ 52.87万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-30 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10544815
• 关键词: Address; Adenine; Aneuploidy; Automobile Driving; Base Excision Repairs; Cell Nucleus; Cell Separation; Cell division; Cells; Chromatin; Chromosomes; Coupled; Cytoplasm; DNA; DNA Damage; DNA biosynthesis; DNA glycosylase; DNA replication fork; DNA sequencing; DNA-Directed DNA Polymerase; Data; Deaminase; Defect; Deoxyribonucleases; Development; Evolution; Exposure to; Fluorescence; Funding; Genetic Transcription; Genome; Genome Stability; Goals; Grant; Hybrids; In Vitro; Interphase; Interphase Chromosome; Knock-out; Malignant Neoplasms; Mass Spectrum Analysis; Mitosis; Mitotic; Mitotic Chromosome; Mitotic spindle; Modeling; Mutation; Nuclear Envelope; Nuclear Structure; Oncogenic; Outcome; Pathway interactions; Pattern; Polyploidy; Process; Proteins; Purines; RNA; Reporter; Rupture; Series; Shapes; Single-Stranded DNA; Structure; System; Testing; Transcriptional Regulation; Work; Xenopus; cancer genome; cancer therapy; chromothripsis; deoxyinosine; ds-DNA; egg; endonuclease; experimental study; fitness; genome sequencing; live cell imaging; micronucleus; novel; whole genome

PROJECT SUMMARY/ABSTRACT The goal of this proposal is to determine the mechanistic basis for mutational processes that drive very rapid evolution of cancer genomes. One process causing mutation bursts is chromothripsis, massive rearrangement of one or a few chromosomes occurring within a single cell division. Under this grant, my group established a general mechanism for chromothripsis. This work was enabled by our development of a novel technical approach, the ability to combine live cell imaging, same cell isolation, and single cell whole genome sequencing (Look-Seq). Chromothripsis originates from abnormal structures of the nucleus—micronuclei (MN) or chromosome bridges. We identified a defect in nuclear envelope assembly on chromatin that lags within the mitotic spindle. This results in nuclear envelope (NE) fragility and spontaneous NE rupture. Exposure of MN chromatin to the interphase cytoplasm leads to extensive DNA damage, which is followed by a second wave of DNA damage upon entry into mitosis. The first two aims of this proposal address the mechanisms leading to chromosome fragmentation that is the basis for chromothripsis. First, building on preliminary data we will test the hypothesis that exposure of MN chromatin to the interphase cytoplasm causes DNA damage via an aberrant pathway of DNA base excision repair. Second, we propose hypothesis driven and unbiased approaches to identify the mechanism of MN chromosome fragmentation in mitosis. Finally, we address the mechanism of another poorly understood, common cancer genome catastrophe termed chromoplexy.

项目摘要/摘要 该提案的目的是确定突变过程的机械基础，这些过程驱动很快 癌症基因组的进化。导致突变爆发的一个过程是铬骨，大规模重排 在单个细胞分裂内发生的一个或几个染色体。根据这笔赠款，我的小组建立了 染色体的一般机制。这项工作是通过我们开发新颖的技术来实现的 方法，结合活细胞成像的能力，相同的细胞分离和单细胞全基因组测序 （look-seq）。染色体起源于细胞核的异常结构 - micronuclei（Mn）或 染色体桥。我们确定了在染色质上的核包膜组件中的缺陷，该缺陷滞后于 有丝分裂主轴。这导致核包膜（NE）脆弱性和赞助NE破裂。 Mn的暴露 染色质到相间的细胞质会导致广泛的DNA损伤，然后是第二波 进入有丝分裂后的DNA损伤。该提案的前两个目的涉及导致的机制 染色体碎片化，这是染色体的基础。首先，在初步数据的基础上，我们将测试 MN染色质暴露于相间胞质的假设会通过异常引起DNA损伤 DNA基础惊喜维修的途径。其次，我们提出了假设驱动和公正的方法 确定有丝分裂中MN染色体碎片化的机制。最后，我们解决了 另一个知名度不佳的，常见的癌症基因组灾难称为Chromoplexy。