Multiscale coordination of planar cell polarity

平面细胞极性的多尺度协调

• 批准号: 10242655
• 负责人: Danelle N Devenport
• 金额: $ 40.91万
• 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/10242655
• 关键词: 3-Dimensional; Adhesives; Anisotropy; Architecture; Binding; Biochemical; Biological; Biological Assay; Biological Models; Biophysics; Cadherins; Cell Aggregation; Cell Communication; Cells; Characteristics; Cilia; Complex; Congenital Abnormality; Congenital Heart Defects; Cues; Cultured Cells; Defect; Development; Embryo; Embryonic Development; Epidermis; Epithelial; Epithelial Cells; Event; Feedback; Goals; Hair follicle structure; Human; Hydrocephalus; Image; Imaging Techniques; In Vitro; Intercellular Junctions; Laboratories; Length; Maps; Mechanics; Mediating; Membrane; Methods; Molecular; Monitor; Mus; Neural Tube Closure; Organ; Organism; Organoids; Pathway interactions; Pattern; Positioning Attribute; Protein Dynamics; Proteins; Regenerative Medicine; Resolution; Role; Skin; Sorting - Cell Movement; Structure; System; Tissue Engineering; Tissues; Work; cell behavior; cell motility; ciliopathy; deafness; innovation; insight; keratinocyte; molecular scale; nanoscale; planar cell polarity; reconstitution; regenerative tissue; self organization; skin organogenesis; stem cells

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.

项目总结 细胞在体外具有自组装成器官样结构的非凡能力。然而， 目前在体外衍生的器官缺乏适当的大小和更高阶的图案，这需要 菜肴中不存在的生物体级别的信息。值得注意的是，集体两极分化和 细胞在组织中的单向排列，这种现象称为平面细胞极性 (PCP)，在体外器官中缺乏，但对于正常的器官形成和功能是必不可少的。因此，要 实现组织工程学在体外产生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作用。通过开发执行以下操作的方法 体外培养、长期活体成像、生物物理扰动和器官型重建 在表皮方面，我们有了关于PCP在组织中建立的关键新发现， 细胞和分子尺度。我们发现单轴组织变形起到了对称的作用 定义表皮中PCP对齐的主轴的断开线索。我们进一步展示了 在没有全局信号情况下生长的原代角质形成细胞在局部自发形成 平面极性从头开始排列的区域。通过超分辨率成像和测绘 PCP组分的粘附性相互作用，我们确定了钙粘附素介导的顺式作用。 不对称五氯苯酚络合物聚集和分类中的相互作用。 这项工作的广泛目标是在这些以前的发现和技术的基础上 破译PCP在不同生物长度尺度上如何组织的进展。vbl.使用 哺乳动物的皮肤表皮作为一个模型系统，其特异性目标1将决定其作用距离有多远 机械提示在整个表皮上偏向和排列平面细胞的极性。特定目标2将 研究细胞皮肤细胞通过自身 组织。具体目标3将破译纳米尺度的架构和生物化学 五氯酚络合物的相互作用。使用我的实验室最近开发的技术创新 对内源性标记的PCP蛋白进行活体和超分辨率成像 和器官类型组织，这项工作将为多尺度的 PCP的协调。