Cell Division Errors as a Mechanism Driving Massive Genomic Rearrangements

细胞分裂错误作为驱动大规模基因组重排的机制

• 批准号: 9371237
• 负责人: Peter Ly
• 金额: $ 11.81万
• 依托单位: LUDWIG INSTITUTE FOR CANCER RES LTD
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9371237
• 关键词: Abnormal Cell; Address; Aneuploidy; Automobile Driving; Award; Biogenesis; Biological; CRISPR/Cas technology; California; Cancer Biology; Categories; Cell Cycle; Cell Cycle Stage; Cell Division Process; Cell division; Cells; Cellular biology; Centromere; Characteristics; Chromosomal Instability; Chromosomal Rearrangement; Chromosome Segregation; Chromosome abnormality; Chromosomes; Coin; Communities; Complex; Coupled; DNA; DNA Damage; DNA Sequence Rearrangement; DNA sequencing; Derivative Chromosome; Development; Disease; Encapsulated; Engineering; Epigenetic Process; Event; Evolution; Faculty; Foundations; Generations; Genes; Genetic Transcription; Genome Stability; Genomic Instability; Genomics; Goals; Health; Hematologic Neoplasms; Heritability; Human; Human Cell Line; Inherited; Institutes; Interphase; Maintenance; Malignant Neoplasms; Mediating; Membrane; Mentors; Methodology; Mitosis; Mitotic; Modeling; Molecular; Nuclear Envelope; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Positioning Attribute; Process; Recruitment Activity; Research; Research Training; Resolution; Science; Series; Shapes; Site; Solid Neoplasm; Specific qualifier value; Structure; System; Technology; Training; United States National Institutes of Health; Universities; anticancer research; base; cancer cell; cancer genetics; cancer genome; career; career development; chromothripsis; deep sequencing; genome-wide; genomic signature; human tissue; innovation; insight; interdisciplinary approach; live cell imaging; multidisciplinary; next generation; nuclease; pressure; prevent; programs; public health relevance; repaired; scientific atmosphere; spatiotemporal; telomere; tenure track; tissue culture; tissue/cell culture; tumor; tumorigenesis

PROJECT SUMMARY/ABSTRACT (DESCRIPTION) Advances in DNA sequencing technologies have uncovered remarkable structural complexities within the human cancer genome, including a new category of massive and localized intra-chromosomal rearrangements coined chromothripsis. Since its discovery in 2011, the signatures of chromothripsis have now been detected in a broad spectrum of solid and hematological tumors. These alterations are thought to occur during a single catastrophic event; however, their underlying mechanistic origins are not well understood. Errors in mitotic cell division can provoke chromothripsis through entrapment of missegregated chromosomes into aberrant structures called micronuclei. Dr. Ly previously identified that chromosomes in micronuclei are subjected to extensive shattering in mitosis, and the resulting DNA fragments are reassembled by canonical end-joining repair in the subsequent interphase. This proposal for an NIH Pathway to Independence Award seeks to understand how chromosome segregation errors during mitosis initiate a cascade of downstream genomic instability events to directly shape or contribute to the cancer genome. In the first aim, examples of fully functional chromosomes with de novo chromothripsis will be generated through development of a powerful and reversible centromere-specific inactivation approach coupled to a chromosome-specific selection strategy in human tissue culture cells. Genome-wide sequencing will be used to identify the hallmark features of chromothripsis from unique clonal derivatives following chromosome missegregation into micronuclei. In the second aim, the precise mechanisms and spatiotemporal dynamics of chromosome shattering and reassembly events will be explored through multidisciplinary cell biological approaches, including gene disruption and live- cell imaging. The third aim will focus on how chromosomes with gross rearrangements that lack a functional alphoid centromere are able to propagate indefinitely through the epigenetic formation of a stable, new centromere at non-alphoid loci. By leveraging his expertise in engineering sophisticated tissue culture models combined with his background in cancer biology, these collective efforts by Dr. Ly will establish the mechanisms and consequences of mitotic errors in triggering genomic instability – insights that are critical for understanding the biogenesis of complex genomic features commonly manifested in patient tumors. During the mentored K99 phase of the Award, Dr. Ly will receive additional and needed training at the Ludwig Institute for Cancer Research under the guidance of Dr. Don Cleveland – a widely recognized and established leader in the fields of cell division and aneuploidy. The Ludwig Institute for Cancer Research, the University of California at San Diego, and the surrounding La Jolla scientific community serves as an exceptional atmosphere for research training, collaborative science, and career development. An excellent team of collaborators (Drs. David Page and Andrew Shiau) and consultants (Drs. Richard Kolodner, Paul Mischel, and Karen Oegema) has been assembled to provide Dr. Ly with additional scientific training and career support before, during, and after transitioning toward an independent tenure-track faculty position. The R00 Award phase will set the foundation for Dr. Ly's long-term career goals of establishing a rigorous research program that seeks to address exciting, unanswered questions in the fields of cell biology, cancer genetics, and genome stability through use of cutting-edge, interdisciplinary methodologies.

项目摘要/摘要（描述） DNA测序技术的进步已经发现 人类癌症基因组，包括新的大规模和局部染色体内重排的新类别 创造的铬壳。自2011年发现以来，现在已经发现了Chromothripsis的签名 广泛的固体和血液学肿瘤。这些变化被认为是在单个过程中发生的 灾难性事件；但是，他们的基本机理起源尚不清楚。有丝分裂细胞的错误 分裂可以通过将错误分析的染色体陷入异常而引起染色体 结构称为微核。 Ly博士先前识别出微核中的染色体受到 有丝分裂的大量破碎，由典型的最终连接重新组装了所得的DNA片段 在随后的相间进行修复。这项针对NIH独立奖的提案旨在 了解有丝分裂过程中染色体分离误差如何引发下游基因组级联 直接塑造或有助于癌症基因组的不稳定性事件。在第一个目标中，完全 具有从头染色体的功能性染色体将通过开发强大的和 可逆的丝粒特异性灭活方法与特定于染色体的选择策略相结合 人组织培养细胞。全基因组测序将用于识别 染色体错误地分析后，来自独特的克隆衍生物的染色体术。在 第二个目的，染色体破碎和重新组装的精确机制和空间时间动态 事件将通过多学科细胞生物学方法进行探讨，包括基因破坏和生命 细胞成像。第三个目标将集中于缺乏功能性的总重排的染色体 字母的丝粒能够通过稳定的新的表观遗传形成无限期地传播 在非αNophoidlotai的Centromere。通过利用他的工程精致组织培养模型的专业知识 结合他的癌症生物学背景，Ly博士的这些集体努力将建立 有丝分裂错误在触发基因组不稳定性中的机制和后果 - 对洞察力至关重要 了解通常在患者肿瘤中表现出的复杂基因组特征的生物发生。在 指导了奖项的K99阶段，Ly博士将在路德维希研究所接受额外且需要的培训 在唐·克利夫兰博士的指导下，癌症研究是一位公认和既定的领导者 细胞分裂和非整倍性的领域。加利福尼亚大学路德维希癌症研究所 在圣地亚哥和周围的拉霍​​亚科学界是一种非凡的氛围 研究培训，协作科学和职业发展。一支优秀的合作者团队（Drs。 David Page和Andrew Shiau）和顾问（Richard Kolodner博士，Paul Mischel和Karen Oegema） 已集会以向Ly博士提供其他科学培训和职业支持 在过渡到独立的终身教师职位之后。 R00奖励阶段将设置 Ly博士的长期职业目标的基础是建立一项严格的研究计划 在细胞生物学，癌症遗传学和基因组稳定性领域中解决令人兴奋的，未解决的问题 通过使用尖端的跨学科方法。