Investigating DNA double-strand break repair mechanisms in mammalian cells

研究哺乳动物细胞中 DNA 双链断裂修复机制

• 批准号: 10207031
• 负责人: Xiaohua Wu
• 金额: $ 44.38万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10207031
• 关键词: Address; Biological; Chromosomal Breaks; Chromosomal Rearrangement; DNA Damage; DNA Double Strand Break; Double Strand Break Repair; Equilibrium; Genetic Diseases; Genetic Recombination; Genomic Instability; Human Genome; Knowledge; Light; Malignant Neoplasms; Mammalian Cell; Mediating; Oncogenic; Other Genetics; Pathway interactions; Principal Investigator; Process; Repetitive Sequence; Reporter; Research; Role; Source; Stress; Structure; System; Variant; base; cancer cell; cancer genetics; coping; human disease; insight; prevent; repaired

Principal Investigator: Wu, Xiaohua Project Summary Gross chromosomal rearrangement (GCR) is a hallmark of cancer cells and the cause of structural variant formation in genetic disorders. DNA double-strand breaks (DSBs) are a major source to induce GCR, but the details on how the DSB repair process leads to genome instability in cancers and other genetic disorders remains elusive. Microhomologies are prevalent at chromosomal break junctions in cancer and other human diseases, suggesting that microhomology-mediated end joining (MMEJ) is one major player to mediate GCRs. Repetitive sequences are abundant, constituting about half of the human genome, which raises the question of how recombination between repetitive sequences is suppressed to prevent genome instability. In this proposal, we aim to investigate the mechanisms underlying MMEJ and repeat-mediated recombination in mammalian cells. We have established multiple EGFP-based DSB repair reporter systems, which enable us to study different DSB repair pathways in mammalian cells. By using newly established MMEJ reporters, we identified new players in the MMEJ pathway. We will investigate how these MMEJ players regulate MMEJ to balance its usage and its mutagenic consequences. We will also study the biological role of MMEJ in repairing replication- associated DSBs and in coping with replication and oncogenic stress, and address how MMEJ contributes to genome instability and the oncogenic process. Additionally, we will determine the mechanisms of repeat-mediated recombination in causing deletions and translocations, and study the role of heteroduplex rejection in preventing recombination among divergent repeats. These studies will generate much-needed new knowledge to fill the gap in our understanding of DSB repair mechanisms and the cause of genome instability that is associated with the oncogenic process and other genetic disorders.

首席研究员：吴，小乌亚 项目摘要 总染色体重排（GCR）是癌细胞和 遗传疾病中结构变异形成的原因。 DNA双链断裂（DSB） 是诱导GCR的主要来源，但有关DSB维修过程如何导致的细节 癌症和其他遗传疾病中的基因组不稳定性仍然难以捉摸。 在癌症和其他 人类疾病，表明微型学介导的结束（MMEJ）是一个主要 介导GCR的玩家。重复序列很丰富，约占一半的一半 人类基因组，提出了一个问题，即重复性之间的重组 序列被抑制以防止基因组不稳定性。在此提案中，我们的目标是 研究MMEJ的基础机制和重复介导的重组 哺乳动物细胞。 我们已经建立了多个基于EGFP的DSB维修记者系统， 使我们能够研究哺乳动物细胞中不同的DSB修复途径。通过新的 既定的MMEJ记者，我们在MMEJ Pathway中确定了新玩家。我们将 调查这些MMEJ播放器如何调节MMEJ以平衡其用法和诱变 结果。我们还将研究MMEJ在修复复制中的生物学作用 - 相关的DSB并应对复制和致癌应力，并解决 MMEJ有助于基因组不稳定性和致癌过程。此外，我们会的 确定重复介导的重组在引起缺失和 易位，并研究杂化排斥反应在预防重组中的作用 在不同的重复序列中。这些研究将产生急需的新知识以填补 我们对DSB修复机制的理解和基因组原因的差距 与致癌过程和其他遗传疾病相关的不稳定性。