Multiscale coordination of planar cell polarity

平面细胞极性的多尺度协调

• 批准号: 10509600
• 负责人: Danelle N Devenport
• 金额: $ 3.53万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10509600
• 关键词: 3-Dimensional; Adhesives; Architecture; Biochemical; Biological; Biological Models; Biophysics; Cadherins; Cells; Complex; Cues; Defect; Development; Epidermis; Goals; Image; In Vitro; Laboratories; Length; Mediating; Methods; Mus; Organ; Organism; Pathway interactions; Pattern; Proteins; Regenerative Medicine; Resolution; Role; Skin; Sorting - Cell Movement; Structure; Tissue Engineering; Tissues; Work; innovation; insight; keratinocyte; mechanical signal; molecular scale; nanoscale; planar cell polarity; reconstitution; regenerative tissue; self organization

PROJECT SUMMARY_________________________________________________________ Cells have a remarkable capacity to self-assemble into organ-like structures in vitro. However, current in vitro-derived organs lack proper size and higher-order patterning, features that require organism-level information not present in a dish. Notably, the collective polarization and unidirectional alignment of cells across a tissue, a phenomenon known as planar cell polarity (PCP), is lacking in vitro organs, yet is essential for proper organ formation and function. Thus, to fulfill the promise of tissue engineering to generate functional organs in vitro we must understand how cells establish long-range collective polarization. We have established the murine skin as a model system to investigate the multiscale coordination of PCP in an expansive and regenerative tissue. By developing methods to perform ex vivo culturing, long-term live imaging, biophysical perturbations, and organotypic reconstitution of the epidermis, we have made key new discoveries about PCP establishment at the tissue, cellular and molecular scales. We discovered that uniaxial tissue deformation acts as a symmetry breaking cue that defines the major axis of PCP alignment in the epidermis. We further showed that that primary keratinocytes grown in the absence of global cues establish spontaneous, locally aligned domains of planar polarity de novo. Through super-resolution imaging and mapping the adhesive interactions of PCP components, we identified a role for cadherin-mediated cis- interactions in the clustering and sorting of asymmetric PCP complexes. The broad goal of this work is to build on these previous discoveries and technological developments to decipher how PCP is organized across different biological length scales. Using the mammalian skin epidermis as a model system, Specific Aim 1 will determine how long-range mechanical cues bias and align planar cell polarity across the epidermis. Specific Aim 2 will investigate the mechanisms by which cells skin cells spontaneously generate PCP through self- organization. Specific Aim 3 will decipher the nanoscale architecture and biochemical interactions of PCP complexes. Using technical innovations recently developed in my laboratory to perform live and super-resolution imaging of endogenously-tagged PCP proteins in both native and organotypic tissues, this work will provide fundamental new insights into the multiscale coordination of PCP.

项目概要_________________________________________________________ 细胞具有在体外自组装成器官样结构的非凡能力。然而， 目前的体外衍生器官缺乏适当的尺寸和高阶模式，这些特征需要 培养皿中不存在有机体水平的信息。值得注意的是，集体两极分化和 组织中细胞的单向排列，这种现象称为平面细胞极性 （PCP）缺乏体外器官，但对于正常的器官形成和功能至关重要。因此，为了 履行组织工程在体外生成功能器官的承诺，我们必须了解 细胞如何建立远程集体极化。 我们建立了小鼠皮肤作为模型系统来研究多尺度 PCP 在扩张和再生组织中的协调。通过开发方法来执行 离体培养、长期实时成像、生物物理扰动和器官重建 在表皮中，我们对 PCP 在组织中的建立有了重要的新发现， 细胞和分子尺度。我们发现单轴组织变形表现为对称性 定义表皮中 PCP 排列主轴的断裂线索。我们进一步展示了 在缺乏整体线索的情况下生长的初级角质形成细胞建立了自发的、局部的 平面极性从头对齐的域。通过超分辨率成像和测绘 PCP 成分的粘附相互作用，我们确定了钙粘蛋白介导的顺式- 不对称PCP复合物聚类和排序中的相互作用。 这项工作的总体目标是建立在这些先前的发现和技术基础上 破译 PCP 如何在不同的生物长度尺度上组织的进展。使用 以哺乳动物皮肤表皮作为模型系统，具体目标 1 将确定远距离 机械线索偏置并对齐表皮上的平面细胞极性。具体目标 2 将 研究细胞皮肤细胞通过自身作用自发产生PCP的机制 组织。具体目标 3 将破译纳米级结构和生化 PCP复合物的相互作用。使用我实验室最近开发的技术创新 对天然中的内源标记 PCP 蛋白进行实时和超分辨率成像 和器官型组织，这项工作将为多尺度提供基本的新见解 PCP 的协调。