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: 10554666
• 负责人: Maria Danielle Sallee
• 金额: $ 8.4万
• 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/10554666
• 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; 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.

项目概要/摘要 细胞分裂和分化需要不同的微管排列才能发挥作用，其目标是 该提案旨在了解分化细胞如何重组其微管以进行分裂。有丝分裂 细胞使用中心体作为微管组织中心（MTOC）形成放射状微管阵列 帮助细胞分裂成两个女儿。分化细胞通常指定非中心体位点作为其 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运输很重要。然而，上皮细胞在发育过程中经常分裂，组织 维持和癌症，提出了一个障碍：它们必须暂时失去平行的微管和 重建径向微管阵列。人们对上皮细胞如何完成这种重组知之甚少， 这对于细胞成功分裂至关重要。 该提案将使用两种互补模型，即发育中的线虫肠道和原代人类 肠道“类器官”细胞，揭示上皮细胞重塑微管的机制 分裂的细胞骨架，以及在发育和疾病中破坏这种重塑的后果。这 线虫胚胎肠是一种简单的体内上皮管，易于可视化和操作，具有 进行微管重塑以分裂的细胞数量固定。此外，C. 中使用的许多蛋白质。 线虫的细胞分裂和微管组织是保守的，使其成为发现的理想环境 调节其他系统中微管组织的新基因和机制。通过结合费尔德曼 实验室最近开发的技术与经典技术相结合，所提出的实验将确定影响因素 在非中心体 MTOC 物理上保持和释放微管，以及导致 这种定位随着细胞分裂而变化。这些新发现的基因和途径将 测试了在原代人类肠道细胞中的保守作用，以及由 微管组织被破坏。 该提案解决了极化细胞如何重组为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以及指导期间的技术培训 阶段将有助于独立阶段的实验。由专家导师和合作者组成的团队将进行培训 Sallee 博士介绍了对于这项研究成功至关重要的新方法。此外，Sallee博士也将参加 在当地会议和科学会议中，参加职业规划课程，并定期与导师会面 和咨询委员会讨论她的科学进展并准备工作申请和面试。 Sallee 博士的两位导师都全力支持她成功制定研究计划，她将 与她一起创办一个独立的学术研究实验室。