Remodeling the microtubule cytoskeleton during epithelial cell division and differentiation

上皮细胞分裂和分化过程中微管细胞骨架的重塑

• 批准号: 10115769
• 负责人: Maria Danielle Sallee
• 金额: $ 10万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115769
• 关键词: Ablation; Address; Advisory Committees; Animal Organ; Animal Sources; Apical; Award; Behavior; Biochemical Genetics; Biological; CRISPR/Cas technology; Caenorhabditis elegans; Cancer Etiology; Carcinoma; Cell Culture Techniques; Cell Cycle; Cell Cycle Regulation; Cell Cycle Stage; Cell Differentiation process; Cell Line; Cell Polarity; Cell division; Cells; Centrosome; Complex; Couples; Cytoskeleton; Daughter; Defect; Development; Disease; Embryo; Epithelial; Epithelial Cells; Excision; Genes; Genetic; Goals; Human; Hyperplasia; Image; Interphase; Interview; Intestines; Intracellular Transport; Job Application; Label; Lasers; Light; Link; Location; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mentors; Methods; Microtubule-Organizing Center; Microtubules; Mitosis; Mitotic; Modeling; Molecular; Organ; Organoids; PAR-6 protein; Pathway interactions; Pharmacology; Phase; Phenotype; Phosphotransferases; Positioning Attribute; Proteins; Radial; Regulation; Research; Rod; Role; Sampling; Signal Transduction; Site; Structure; Surface; System; Techniques; Testing; Tissues; Training; Tube; Work; apical membrane; base; c new; career; experimental study; gene discovery; genetic approach; in vivo; in vivo Model; innovation; insight; link protein; marker transgenes; meetings; polarized cell; protein degradation; recruit; success; symposium; tool

Project Summary/Abstract Dividing and differentiating cells require different arrangements of microtubules to function, and the goal of this proposal is to understand how differentiating cells reorganize their microtubules in order to divide. Mitotic cells use centrosomes as their microtubule organizing centers (MTOCs) to form radial microtubule arrays that help split the cell into two daughters. Differentiating cells often designate a non-centrosomal sites as their MTOC. In polarized epithelial cells, the apical membrane is the MTOC, and it forms parallel microtubules arrays that are important for cell polarity and intracellular transport. However, epithelial cells often divide in development, tissue maintenance, and cancer, presenting an obstacle: they must temporarily lose their parallel microtubules and reestablish radial microtubule arrays. Little is known about how epithelial cells accomplish this reorganization, which is critical for successful cell division. This proposal will use two complementary models, the developing C. elegans intestine and primary human intestinal “organoid” cells, to uncover the mechanisms that epithelial cells use to remodel their microtubule cytoskeleton for division, and the consequence of disrupting this remodeling in development and disease. The C. elegans embryonic intestine is a simple in vivo epithelial tube that is easy to visualize and manipulate, with a fixed number of cells that undergo microtubule remodeling to divide. In addition, many of the proteins used in C. elegans for cell division and microtubule organization are conserved, making it an ideal context for discovering new genes and mechanisms that regulate microtubule organization in other systems. By combining the Feldman lab’s recently developed techniques with classic ones, the proposed experiments will identify the factors that physically hold and release microtubules at the non-centrosomal MTOC, and the molecular signals that cause this localization to change concordant with cell division. These newly discovered genes and pathways will be tested for a conserved role in primary human intestinal cells, and for cancer-related defects resulting from disrupted microtubule organization. This proposal addresses the fundamental biological question of how polarized cells reorganize for cell division. The proposed experiments will cover the entire award period, and technical training during the mentored phase will facilitate experiments in the independent phase. A team of expert mentors and collaborators will train Dr. Sallee in new methods that are critical to the success of this research. In addition, Dr. Sallee will participate in local meetings and scientific conferences, attend career planning courses, and meet regularly with her mentors and advisory committee to discuss her scientific progress and to prepare for job applications and interviews. Both of Dr. Sallee’s mentors are fully committed to her success in establishing her research plan that she will take with her to start an independent academic research lab.

项目摘要/摘要 分隔和区分细胞需要不同的微管的布置才能起作用，而目标的目的是 该建议是了解如何区分细胞重组其微管以分裂。有丝分裂 细胞使用中心体作为微管组织中心（MTOC）形成径向微管阵列 帮助将牢房分成两个女儿。区分细胞通常将非中心位点表示为MTOC。 在极化上皮细胞中，顶膜是MTOC，它形成平行的微管阵列 对于细胞极性和细胞内转运很重要。但是，上皮细胞通常在发育中分裂 维护和癌症，出现障碍：他们必须暂时失去平行的微管和 重新建立径向微管阵列。关于上皮细胞如何完成这种重组的知之甚少 这对于成功的细胞分裂至关重要。 该提案将使用两个完整的型号，即秀丽隐杆线虫肠道和主要人类 肠道“器官”细胞，以发现上皮细胞用于重塑其微管的机制 用于分裂的细胞骨架，以及破坏这种重塑的发展和疾病的结果。这 秀丽隐杆线虫胚胎肠是一种简单的体内上皮管，易于可视化和操纵，并带有一个 修复了经历微管重塑以分裂的细胞数量。另外，C中使用的许多蛋白质。 秀丽隐杆线虫用于细胞分裂和微管组织是保守的，使其成为发现的理想背景 在其他系统中调节微管组织的新基因和机制。通过结合费尔德曼 实验室最近开发了经典技术，拟议的实验将确定因素 在非中心MTOC上物理持有并释放微管，以及引起的分子信号 与细胞分裂一致的本地化。这些新发现的基因和途径将是 测试了在原代人肠细胞中的配置作用，以及与癌症相关的缺陷。 微管组织中断。 该提案解决了细胞如何重组细胞的基本生物学问题 分配。拟议的实验将涵盖整个奖励期，并在此问题期间进行技术培训 阶段将促进独立阶段的实验。由专家导师和合作者组成的团队将培训 Sallee博士的新方法对这项研究的成功至关重要。此外，萨利博士将参加 在当地会议和科学会议中，参加职业计划课程，并定期与她的导师会面 和咨询委员会讨论她的科学进步并为工作申请和访谈做准备。 萨利博士的两位导师都完全致力于建立她的研究计划的成功，她将 与她一起开始一个独立的学术研究实验室。