Centromeres and Ovarian Cancer

着丝粒和卵巢癌

基本信息

批准号：
10425088
负责人：
Rafael Alejandro Contreras
金额：
$ 9.12万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-23 至 2023-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10425088
关键词：
Address Affect Aneuploidy Area Binding Binding Proteins Binding Sites Bioinformatics Biological Assay Biology CRISPR/Cas technology Cancer Etiology Cell Line Cell division Cells Centromere Cessation of life Chromatin Chromatin Structure Chromosomal Instability Chromosomal Stability Chromosome 17 Chromosome Segregation Chromosomes Complex Consumption DNA DNA Sequence Data Deposition Development Disease Elements Epigenetic Process Epithelial Cells Exhibits Failure Genetic Genetic study Genome Genome Stability Genomic Instability Genomics Head Human Human Chromosomes Human Genome Human Genome Project Impairment Individual Kinetochores Knowledge Lead Malignant Neoplasms Malignant neoplasm of ovary Mammalian Oviducts Methodology Methods Microtubules Modernization Molecular Mutation Ovarian Play Process Prognosis Proteins Research Research Personnel Role Serous Structure System Tail Technology Time Tissues Woman base cancer cell cancer genetics cell transformation chromatin immunoprecipitation chromosome loss chromosome missegregation chromosome replication contig daughter cell design frontier innovation monomer mortality mutant next generation sequencing novel recruit tool tumor progression virtual

项目摘要

The composition, organization, and function of centromeric DNA remain one of the last frontiers of genomics. This proposal “Centromeres and Ovarian Cancer” will investigate the role centromere DNA sequences play in the correct function of centromeres and chromosome segregation in ovarian cancer, and its contribution to ovarian cancer progression. The centromere is the functional unit responsible for the faithful segregation of chromosomes. Failure to properly partition chromosomes to daughter cells results in genome instability and aneuploidy (gain or loss of chromosomes), a hallmark of cancers. Functional centromere structures are made of centromere epigenetic marks that bind to the underlying DNA sequence. During cell division, the microtubules interact with these centromeric structures and pull chromosomes apart. Although centromere identity and function rely on epigenetic mechanisms, recent studies suggest that centromere sequences have functional roles as well. However, human centromere sequences have not been assembled into linear contigs due to their high content of repetitive DNA, and thus, the precise composition, organization, and function of centromeric DNA remain to be elucidated. We will investigate the role centromere DNA sequences play in centromere function and chromosome segregation in ovarian cancer. To overcome the virtual absence of centromere sequence in the Human Genome Project, the P.I. has devised rapid, quantitative PCR- based methods and innovative Next Generation Sequencing tools to study specifically the centromeric DNA elements of each human chromosome at the molecular level. Using these pioneering technologies, we discovered that the genomes of ovarian cancer cells exhibit severe loss of centromere sequences in specific chromosomes, in particular chromosome 17, when compared to healthy matched-tissue. We hypothesize that these specific centromere mutations play a key role in centromere formation and function, and lead to chromosome 17 missegregation and genome instability seen in ovarian cancer. The proposed research will use mutant ovarian and fallopian tube cell lines with centromere deletions generated using CRISPR Cas9. We will investigate whether centromere loss alters the ability of centromeric epigenetic marks to form centromere/kinetochore structures. We will address whether loss of centromeric material in these mutant cells affects genome stability, chromosome segregation and transformation. This information will begin to provide a better understanding of the role’s centromere sequences play in cell division and cancer genetics. These studies will enhance our knowledge of centromeres and chromosomal biology and could potentially result in novel methodologies to study genetic defects in cancers.

着丝粒 DNA 的组成、组织和功能仍然是人类研究的最后前沿领域之一。该提案“着丝粒与卵巢癌”将研究着丝粒 DNA 的作用。序列在卵巢癌中着丝粒和染色体分离的正确功能中发挥作用，及其对卵巢癌进展的贡献着丝粒是负责卵巢癌进展的功能单位。染色体无法正确分配到子细胞会导致染色体的忠实分离。基因组不稳定和非整倍性（染色体的获得或丢失），是癌症功能性着丝粒的标志。结构由着丝粒表观遗传标记组成，在细胞过程中与底层 DNA 序列结合。分裂时，微管与这些着丝粒结构相互作用并将染色体拉开。着丝粒的身份和功能依赖于表观遗传机制，最近的研究表明着丝粒序列也具有功能作用，然而，人类着丝粒序列尚未组装。由于重复 DNA 含量高，因此可以形成线性重叠群，因此具有精确的组成、组织、着丝粒DNA的功能还有待阐明，我们将研究着丝粒DNA的作用。序列在卵巢癌中发挥着丝粒功能和染色体分离作用。由于人类基因组计划中缺乏着丝粒序列，P.I. 设计了快速定量 PCR- 基于方法和创新的下一代测序工具来专门研究着丝粒 DNA 利用这些开创性技术，我们在分子水平上分析了每个人类染色体的元素。发现卵巢癌细胞的基因组在特定区域表现出着丝粒序列的严重丢失与健康匹配组织相比，染色体，特别是 17 号染色体。这些特定的着丝粒突变在着丝粒的形成和功能中发挥着关键作用，并导致拟议的研究将使用卵巢癌中发现的 17 号染色体错误分离和基因组不稳定。使用 CRISPR Cas9 生成具有着丝粒缺失的突变卵巢和输卵管细胞系。研究着丝粒丢失是否改变着丝粒表观遗传标记形成的能力我们将解决这些突变细胞中着丝粒物质是否丢失的问题。影响基因组稳定性、染色体分离和转化的信息将开始提供。这些研究有助于更好地了解着丝粒序列在细胞分裂和癌症遗传学中的作用。将增强我们对着丝粒和染色体生物学的了解，并可能产生新的结果研究癌症遗传缺陷的方法。