Human centromere variation and function

人类着丝粒变异和功能

• 批准号: 10506033
• 负责人: Glennis Amelia Logsdon
• 金额: $ 10万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10506033
• 关键词: Address; Advisory Committees; Affect; Aneuploidy; Architecture; Biological Assay; Biological Process; Biological Sciences; Biology; Cancer Etiology; Cell division; Cells; Centromere; ChIP-seq; Chromatin; Chromatin Fiber; Chromosome Segregation; Chromosome abnormality; Chromosomes; Collaborations; Computing Methodologies; Congenital Abnormality; DNA Methylation; Data; Defect; Disease; Ensure; Environment; Epigenetic Process; Event; Fiber; Foundations; Genetic; Genetic Transcription; Genetic Variation; Genomics; Goals; Haplotypes; Health; Human; Individual; Kinetochores; Knowledge; Lead; Life; Location; Malignant Neoplasms; Maps; Mentors; Methods; Microtubules; Modeling; Mutation; NCI Center for Cancer Research; Nature; Phase; Population; Positioning Attribute; Process; Protein Isoforms; Research; Research Training; Resources; Rest; Role; Scientist; Spontaneous abortion; Structure; Tandem Repeat Sequences; Technology; Testing; Training; Transcript; Universities; Untranslated RNA; Variant; Washington; Work; base; carcinogenesis; career development; contig; epigenetic variation; genetic information; human model; innovation; multiple omics; nanopore; novel; programs; segregation; skills; tool

Project Summary/Abstract The equal segregation of chromosomes during cell division ensures the accurate inheritance of genetic information. Aberrant chromosome segregation can cause an imbalance in chromosome number, or aneuploidy, which can result in spontaneous abortion and birth defects and is a major cause of cancer. The locus required for the equal segregation of chromosomes is the centromere. In humans, centromeres are comprised of repetitive α-satellite sequences that span several megabases on each chromosome. The repetitive nature of these regions has challenged efforts to determine their sequence, structure, and variation using short-read sequencing data. As a result, we have a very limited understanding of the natural variation of centromeres and the impact of this variation on essential cell biological process critical for life. In this proposal, I aim to address this gap in knowledge by sequencing and assembling centromeres from diverse humans using a combination of long-read sequencing technologies and novel computational assembly tools (Aim 1; K99 phase). Additionally, I propose to assess the natural variation of human centromeres at the genetic, epigenetic, and transcriptional level using innovative computational methods and multiomic sequencing approaches, ultimately building a model of human centromere variation (Aim 2; K99 phase). Finally, I propose to determine how variation among centromeres affects the accurate segregation of chromosomes during cell division using cell-based assays, long- read sequencing, and multiomic sequencing approaches (Aim 3; R00 phase). Together, this work will provide the first comprehensive assessment of human centromere variation and reveal how this variation affects centromere function in cells. The tools, resources, and skills developed in the K99 phase will be applied in the R00 phase to determine the functional consequences of centromere variation and its role in human health and disease. My goal is to build an independent research program that spans the gap between genomics and centromere biology. I will receive the necessary interdisciplinary training from my mentor (Dr. Evan Eichler), co- mentor (Dr. Sue Biggins), and the rest of my postdoctoral advisory committee (Drs. Deborah Nickerson, Andrew Stergachis, and Kelley Harris). In addition, I will participate in career development activities offered through the University of Washington and the Fred Hutchison Cancer Research Center. Together, my research training, mentors, advisory committee, and academic environment will prepare me well as I transition to an independent position as an academic scientist.

项目摘要/摘要 细胞分裂过程中染色体的平等分离确保了基因的准确遗传 信息。异常的染色体分离会导致染色体数量的不平衡，或非整倍体， 这会导致自然流产和出生缺陷，也是癌症的主要原因。所需的轨迹 因为染色体的平等分离就是着丝粒。在人类中，着丝粒由以下组成 重复的α-卫星序列，跨越每条染色体上的几个百万碱基。的重复性 这些区域挑战了使用短阅读来确定它们的序列、结构和变异的努力 测序数据。因此，我们对着丝粒和着丝粒的自然变异的了解非常有限 这种变异对生命至关重要的基本细胞生物学过程的影响。在这项提案中，我的目标是解决 通过对来自不同人类的着丝粒进行测序和组装，利用组合的 长读测序技术和新的计算组装工具(目标1；K99阶段)。另外， 我建议从遗传、表观遗传和转录三个方面评估人类着丝粒的自然变异 使用创新的计算方法和多基因组测序方法，最终建立一个模型 人类着丝粒变异(目标2；K99期)。最后，我建议确定 着丝粒影响着丝粒在细胞分裂过程中的准确分离，使用基于细胞的分析，长- 阅读测序和多组体测序方法(目标3；R00阶段)。共同努力，这项工作将提供 首次全面评估人类着丝粒变异并揭示该变异如何影响着丝粒 着丝粒在细胞中起作用。在K99阶段开发的工具、资源和技能将应用于 R00阶段，以确定着丝粒变异的功能后果及其在人类健康和 疾病。我的目标是建立一个独立的研究计划，跨越基因组学和 着丝粒生物学。我将从我的导师(Evan Eichler博士)那里接受必要的跨学科培训， 导师(Sue Biggins博士)和我的博士后咨询委员会的其他成员(Deborah Nickerson博士，Andrew 斯特加奇斯和凯利·哈里斯)。此外，我还将参加通过 华盛顿大学和弗雷德·哈奇森癌症研究中心。一起，我的研究训练， 导师、顾问委员会和学术环境将为我过渡到独立的 学术科学家的职位。