Centromeres and Ovarian Cancer

着丝粒DNA的组成、组织和功能仍然是 基因组学。这项名为“着丝粒与卵巢癌”的提案将研究着丝粒DNA的作用 在卵巢癌中，序列在着丝粒和染色体分离的正确功能中发挥作用， 以及它对卵巢癌进展的贡献。着丝粒是负责 忠实的染色体分离。未能正确地将染色体分割到子代细胞导致 基因组不稳定和非整倍体(染色体的获得或丢失)，这是癌症的标志。功能着丝粒 结构由着丝粒表观遗传标记组成，这些标记与潜在的DNA序列结合。在牢房期间 在分裂过程中，微管与这些着丝粒结构相互作用，将染色体分开。虽然 着丝粒的特性和功能依赖于表观遗传机制，最近的研究表明着丝粒 序列也有功能作用。然而，人类着丝粒序列还没有组装好 由于它们的重复DNA含量很高，因此其精确的组成、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 序列在卵巢癌的着丝粒功能和染色体分离中发挥作用。要克服虚拟 在人类基因组计划中缺乏着丝粒序列的情况下，P.I.设计了快速、定量的PCR- 基于方法和创新的下一代测序工具来具体研究着丝粒DNA 人类每条染色体的分子水平上的元素。使用这些开创性的技术，我们 发现卵巢癌细胞基因组在特定情况下表现出严重的着丝粒序列丢失 染色体，特别是17号染色体，与健康的匹配组织相比。我们假设 这些特定的着丝粒突变在着丝粒的形成和功能中起着关键作用，并导致 卵巢癌中出现的17号染色体分离错误和基因组不稳定。拟议的研究将使用 使用CRISPR Cas9产生的带有着丝粒缺失的卵巢和输卵管细胞系突变。我们会 研究着丝粒丢失是否改变着丝粒表观遗传标记的形成能力 着丝粒/着丝粒结构。我们将解决这些突变细胞中着丝粒物质的丢失 影响基因组稳定性、染色体分离和转化。这些信息将开始提供一个 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。这些研究 将增强我们对着丝粒和染色体生物学的知识，并可能导致新的 研究癌症遗传缺陷的方法。

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.