Investigating the Establishment and Propagation of Planar Cell Polarity

研究平面细胞极性的建立和传播

• 批准号: 10424545
• 负责人: Jelena Popovic Basta
• 金额: $ 4.68万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10424545
• 关键词: Address; Automobile Driving; Binding; Biological; Biological Models; Biological Phenomena; Biology; Cadherins; Cell Polarity; Cells; Cilia; Communication; Complex; Coupling; Cues; Defect; Development; Disease; Drosophila genus; Endocytosis; Epithelial; Evolution; Feedback; Genes; Goals; Hair; Hair follicle structure; Homeostasis; Image; In Vitro; Individual; Integral Membrane Protein; Intercellular Junctions; Mediating; Membrane; Microscopy; Molecular; Mus; Neural Tube Defects; Neural tube; Pathway interactions; Pattern; Physiology; Process; Proteins; Resolution; Role; Side; Skin; Structure; System; Testing; Time; Tissues; Wing; Work; defined contribution; genetic manipulation; in silico; innovation; insight; keratinocyte; planar cell polarity; reconstitution; recruit; repaired; response; tool

Project Summary Planar cell polarity (PCP) is a highly conserved pathway that regulates the coordination of polarity in cells along a single tissue plane, and is fundamental in development and tissue function. A defining feature of PCP is the asymmetric localization of its core components, which transition from uniform to asymmetric membrane distribution over time. Frizzled (Fz) localizes with Dishevelled (Dvl) on one side of the cell, while Vangl and Prickle (Pk) localize to the other. It is thought that transmembrane components Vangl and Fz recruit opposing complexes between cells while the cytoplasmic components, Dvl and Pk, mediate feedback interactions that amplify asymmetry within cells. Thus, both intercellular and intracellular core PCP protein feedback interactions are central to how we think PCP is established, however, the specific feedback interactions underlying PCP establishment, and whether they primarily occur within cells or between cells, are not well understood. The overall goal of this work is to determine how the core PCP proteins self-organize via feedback interactions to coordinate the establishment and propagation of polarity. What is needed to investigate these questions is a reductionist system that can uncouple intracellular feedback interactions from those that occur between cells, and isolate these interactions from the influence of tissue level polarizing cues. Our lab has established the murine skin as a powerful model system to investigate conserved mechanisms of PCP establishment. We have shown that primary keratinocytes isolated and cultured in vitro display spontaneous and locally aligned planar polarity in the absence of any global cue. I propose to further simplify this system using micropatterning and cell- mimicking interfaces to reconstitute the establishment and propagation polarity from its minimal components. Using a combination of live imaging, super-resolution microscopy, and genetic manipulation, I will determine the molecular mechanisms underlying the PCP protein interactions that drive polarity establishment and propagation. This study will provide new and innovative tools that will push the PCP field forward. Importantly, completion of the proposed aims will address an essential, but poorly understood question, by providing insight into the molecular mechanisms driving PCP establishment and revealing fundamental features of epithelial biology and tissue organization.

项目概要 平面细胞极性（PCP）是一种高度保守的途径，可调节细胞内极性的协调 单一组织平面，对于发育和组织功能至关重要。 PCP 的一个决定性特征是 其核心成分的不对称定位，从均匀膜过渡到不对称膜 随时间的分布。 Frizzled (Fz) 与 Disheveled (Dvl) 定位在细胞的一侧，而 Vangl 和 刺痛 (Pk) 定位到另一个。人们认为跨膜成分 Vangl 和 Fz 招募相反的 细胞之间的复合物，而细胞质成分 Dvl 和 Pk 介导反馈相互作用 放大细胞内的不对称性。因此，细胞间和细胞内核心 PCP 蛋白反馈相互作用 是我们认为 PCP 建立方式的核心，然而，PCP 背后的具体反馈相互作用 其建立以及它们主要发生在细胞内还是细胞之间尚不清楚。这 这项工作的总体目标是确定核心 PCP 蛋白如何通过反馈相互作用进行自组织 协调极性的建立和传播。调查这些问题所需要的是 还原论系统可以将细胞内反馈相互作用与细胞之间发生的相互作用分开， 并将这些相互作用与组织水平极化线索的影响隔离开来。我们的实验室建立了 小鼠皮肤作为研究 PCP 建立的保守机制的强大模型系统。我们有 表明体外分离和培养的原代角质形成细胞表现出自发的和局部排列的平面 在没有任何全局提示的情况下的极性。我建议使用微图案和单元格进一步简化该系统 模仿接口从其最小组件重建建立和传播极性。 结合实时成像、超分辨率显微镜和基因操作，我将确定 PCP 蛋白相互作用的分子机制驱动极性建立和 传播。这项研究将提供新的创新工具，推动 PCP 领域向前发展。重要的是， 完成拟议的目标将通过提供洞察力来解决一个重要但人们知之甚少的问题 深入研究驱动 PCP 建立的分子机制并揭示上皮细胞的基本特征 生物学和组织组织。