Molecular mechanism that suppresses the proliferation of cells with supernumerary centrioles

抑制多余中心粒细胞增殖的分子机制

• 批准号: 10389176
• 负责人: Andrew Jon Holland
• 金额: $ 7.85万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10389176
• 关键词: Address; Aneuploidy; Biochemical; Biogenesis; Biological; Biology; CASP2 gene; Cell Cycle; Cell Proliferation; Cell division; Cells; Centrioles; Centrosome; Chromosome Segregation; Cilia; Data; Development; Distal; Ensure; Genes; Genetic; Goals; Growth; Human; Knowledge; Laboratories; MDM2 gene; Malignant - descriptor; Malignant Neoplasms; Mature Centriole; Meiosis; Microtubules; Mitotic; Molecular; Monitor; Mus; Normal Cell; Oncogenic; Organelles; Pathway interactions; Process; Proliferating; Research; Resolution; Structure; TP53 gene; Translating; appendage; cancer cell; cell transformation; genome integrity; genome wide screen; human disease; in vivo; insight; live cell imaging; mouse model; neoplastic cell; protein complex; tumor

Project Summary The long-term goal of our research is to understand the molecular mechanisms that control centriole biogenesis and how errors in this process contribute to human disease. Centrioles are the structural core of centrosomes, organelles that nucleate microtubules to build mitotic/meiotic spindles and cilia. During a normal cell cycle, centrioles duplicate once to ensure their copy number is precisely maintained. The presence of supernumerary centrioles is a common feature of human tumors and can promote chromosome segregation errors that are sufficient to drive tumor development in mice. To maintain genome integrity, cells have evolved a protective centriole surveillance pathway to restrict the proliferation of cells with extra centrioles. The goal of our application is to unravel the molecular mechanism responsible for ‘sensing’ supernumerary centrioles and evaluate whether inactivation of this pathway facilitates tumor development in cells with extra centrioles. Centriole amplification triggers the activation of the PIDDosome, a trimeric protein complex that acts as an activation platform for Caspase-2. Once activated, Caspase-2 promotes the cleavage of MDM2 and subsequent stabilization of P53. However, there exists a gap in our understanding of how extra centrioles are sensed and how this information is relayed to the PIDDosome to trigger P53 activation. To address this knowledge gap, we developed a genome-wide screening approach to identify genes required to arrest the growth of non-transformed cells with extra centrioles. Our preliminary data show that distal appendages that form on mature centrioles are responsible for activating the PIDDosome following centriole amplification. In Aim 1 of this proposal we will use cell biological, genetic and biochemical approaches to mechanistically dissect how cells ‘sense’ supernumerary centrioles to trigger PIDDosome activation. In Aim 2, we will determine the impact of specifically inactivating the centriole surveillance pathway on the proliferation and oncogenic transformation of cells with extra centrioles in vivo. We are well suited to pursue these studies given our expertise in studying centriole biology; our development of a unique mouse model to study the impact of centriole amplification in vivo; and our collaborative relationship with the Regot and Loncarek laboratories, who are world-experts in high resolution live-cell imaging and correlative light/EM analysis of centriole ultrastructure. Understanding how normal cells detect centriole amplification addresses a fundamental question that will provide insight into how aneuploid tumor cells adapt to proliferate robustly with extra centrioles.

项目概要 我们研究的长期目标是了解控制中心粒的分子机制 生物发生以及该过程中的错误如何导致人类疾病。中心粒是结构核心 中心体，使微管成核以构建有丝分裂/减数分裂纺锤体和纤毛的细胞器。正常期间 细胞周期中，中心粒复制一次，以确保精确维持其拷贝数。的存在 多余的中心粒是人类肿瘤的一个共同特征，可以促进染色体分离 足以驱动小鼠肿瘤发展的错误。为了维持基因组的完整性，细胞不断进化 一种保护性中心粒监视途径，用于限制具有额外中心粒的细胞的增殖。目标是 我们的应用是揭示负责“感知”多余中心粒的分子机制 评估该途径的失活是否促进具有额外中心粒的细胞中的肿瘤发展。 中心粒扩增触发 PIDDosome 的激活，PIDDosome 是一种三聚蛋白复合物，充当 Caspase-2 激活平台。一旦激活，Caspase-2 就会促进 MDM2 的裂解， P53随后稳定。然而，我们对额外中心粒如何产生的理解存在差距。 感测到的信息以及如何将这些信息转发到 PIDDosome 以触发 P53 激活。为了解决这个问题 知识差距，我们开发了一种全基因组筛选方法来识别阻止疾病所需的基因 具有额外中心粒的非转化细胞的生长。我们的初步数据表明，远端附件 成熟中心粒上的形式负责在中心粒扩增后激活 PIDDosome。在 该提案的目标 1 我们将使用细胞生物学、遗传和生化方法来机械地 剖析细胞如何“感知”多余中心粒以触发 PIDDosome 激活。在目标 2 中，我们将 确定特异性失活中心粒监视途径对增殖和 体内具有额外中心粒的细胞的致癌转化。鉴于我们非常适合进行这些研究 我们在研究中心粒生物学方面的专业知识；我们开发了一种独特的小鼠模型来研究 体内中心粒扩增；以及我们与 Regot 和 Loncarek 实验室的合作关系，他们 是高分辨率活细胞成像和中心粒相关光/电磁分析方面的世界专家 超微结构。了解正常细胞如何检测中心粒扩增解决了一个基本问题 这将有助于了解非整倍体肿瘤细胞如何适应额外中心粒的强劲增殖。