Frequency and consequences of chromosome missegregation in breast cancer

项目总结 研究。染色体不稳定(CIN)-100多年前被观察到的第一个癌症标志-是 以通过不正常的细胞分裂使整个染色体持续丢失和获得为特征的。这一过程 结果在非整倍体中，染色体数目不正确的状态出现在超过70%的 实体瘤，其中一些表现为非整倍体的复发模式。持续的“错误隔离” 染色体与较差的患者预后和较高的临床特征有关。这归因于 通过不同非整倍体的广泛景观增加了基因组多样性的肿瘤的适应性 克隆人。在较高的错分率下，它似乎会导致细胞死亡和肿瘤抑制。事件的发生 理论上，CIN还可以使肿瘤对紫杉烷等CIN诱导的化疗药物敏感。尽管CIN很长 历史上，它作为紫杉烷疗效的预后标记物和生物标记物的临床应用目前还无法获得。 量化染色体错选率的方法要么在组织中不可行，要么不够 信息量大和/或劳动密集型。这项提议的一个关键目标是将随机计算 利用肿瘤的单细胞测序对细胞分裂进行建模，以允许量化 肿瘤内染色体错选率。此外，染色体后的细胞过程 作为核型选择基础的错误分离还没有被探索。我们假设- 错误的分离转录过程排除或允许克隆的繁殖 特定的非整倍体染色体组合。我们将随机生成许多组合 转化细胞系和未转化细胞系染色体拷贝数的变化及急性 单细胞分辨率下的转录改变和克隆组成。总体而言，这一目标的长期目标 建议解决染色体错误分离和乳腺癌之间的复杂关系 其在人类乳腺肿瘤及其下游转录中的研究进展 作为肿瘤进化基础的后果。这项工作在方法上是创新的，并将显著 提高我们对肿瘤演变和如何评估患者CIN的理解。 训练。通过完成拟议的研究，我将发展我在设计和实施方面的技能 高影响力和严谨的科学研究。通过课程作业和与实验成员的互动以及 合作者，我将在细胞生物学和基因组学/转录组学方面发展多学科专业知识 实验方法和分析方法。我将在科学交流、公共事务方面磨练专业技能 通过提供给我的许多机会与我互动并展示我的 向合作者、领域专家、立法者和社区成员进行研究。独特的、多学科的 这里建议的培训将使我在职业生涯中成为一名独立的调查员 在学术机构或联邦研究机构从事癌症基因组学研究。

PROJECT SUMMARY Research. Chromosomal instability (CIN) – observed as the first cancer hallmark over 100 years ago – is characterized by the persistent loss and gain of whole chromosomes through abnormal cell division. This process results in aneuploidy, the state of having an incorrect number of chromosomes, which is present in over 70% of solid tumors, some of which display recurring patterns of aneuploidy. Persistent ‘missegregation’ of chromosomes is associated with worse patient prognosis and advanced clinical features. This is attributed to the increased adaptability of a tumor having increased genomic diversity via a broad landscape of different aneuploid clones. At higher rates of missegregation it appears to cause cell death and tumor inhibition. The occurrence of CIN is also theorized to sensitize tumors to CIN-inducing chemotherapies like taxanes. Despite CIN’s long history, its clinical use as a prognostic marker and biomarker for taxane efficacy is inaccessible as current methods of quantifying rates of chromosome missegregation are either infeasible in tissue, insufficiently informative and/or labor intensive. A critical goal for this proposal to combine stochastic computational modeling of cell division with single cell sequencing of tumors in order to allow for the quantification of intratumoral rates of chromosome missegregation. Additionally, cellular processes after chromosome missegregation that underlie karyotypic selection have not been explored. We hypothesize that post- missegregation transcriptional processes either preclude or permit the propagation of clones with specific aneuploid chromosome combinations. We will stochastically generate many combinations of chromosome copy number alterations in transformed and non-transformed cell lines and analyze the acute transcriptional alterations and clonal composition at single cell resolution. Overall, the long-term goal for this proposal is to resolve the complex relationship between chromosome missegregation and breast cancer progression with respect to its incidence in human breast tumors and downstream transcriptional consequences that underlie tumor evolution. This work is innovative in its approach and will significantly improve our understanding of tumor evolution and how to evaluate CIN in patients. Training. Through completion of the proposed research, I will develop my skills in the design and implementation of high impact and rigorous scientific studies. Through coursework and interactions with lab members and collaborators, I will develop multi-disciplinary expertise in cell biological and genomics/transcriptomics experimental and analytical methods. I will hone professional skills in scientific communication, public engagement, and networking through the many opportunities afforded to me to interact with and present my research to collaborators, field-experts, legislators, and community members. The unique, multi-disciplinary training proposed here will position me well for a career as an independent investigator in the field of cancer genomics at an academic or federal research institution.