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.

项目总结/摘要 细胞的分裂和分化需要微管的不同排列才能发挥作用， 这项建议是为了了解分化中的细胞如何重组微管以进行分裂。丝分裂 细胞使用中心体作为它们的微管组织中心（MTOC）以形成放射状微管阵列， 把细胞分裂成两个子细胞分化细胞通常指定非中心体位点作为其MTOC。 在极化的上皮细胞中，顶膜是MTOC，它形成平行的微管阵列， 对细胞极性和细胞内转运很重要。然而，上皮细胞通常在发育、组织 维持，和癌症，提出了一个障碍：他们必须暂时失去他们的平行微管， 重建放射状微管阵列。关于上皮细胞是如何完成这种重组的， 这对成功的细胞分裂至关重要。 这个建议将使用两个互补的模型，发展中的C。线虫肠道和原代人 肠道“类器官”细胞，以揭示上皮细胞用于重塑其微管的机制 细胞骨架的分裂，以及破坏这种重建的发展和疾病的后果。的 C.线虫胚胎肠是一种简单的体内上皮管，易于观察和操作， 固定数量的细胞进行微管重塑分裂。此外，C. 细胞分裂和微管组织是保守的，使其成为发现的理想环境。 在其他系统中调节微管组织的新基因和机制。通过将费尔德曼 实验室最近开发的技术与经典的，拟议的实验将确定的因素， 在非中心体MTOC处物理地保持和释放微管，以及引起MTOC的分子信号。 这种定位随着细胞分裂而改变。这些新发现的基因和途径将被 测试了在原代人类肠细胞中的保守作用，以及由以下原因引起的癌症相关缺陷： 破坏微管组织。 这一提议解决了极化细胞如何重组为细胞的基本生物学问题。 师.拟议的实验将涵盖整个授标期间，以及受指导期间的技术培训。 第一阶段将促进独立阶段的实验。一个由专家导师和合作者组成的团队将培训 博士Sallee在新的方法，这是至关重要的这项研究的成功。此外，Sallee博士将参加 参加当地会议和科学会议，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，并为工作申请和面试做准备。 Sallee博士的两位导师都完全致力于她成功地建立她的研究计划，她将 带着她开始了一个独立的学术研究实验室。