Multiscale coordination of planar cell polarity

平面细胞极性的多尺度协调

• 批准号: 10684280
• 负责人: Danelle N Devenport
• 金额: $ 42.18万
• 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/10684280
• 关键词: 3-Dimensional; Adhesives; Anisotropy; Architecture; Binding; Biochemical; Biological; Biological Assay; Biological Models; Biophysics; Cadherins; Cell Aggregation; Cell Communication; Cell Separation; Cells; Characteristics; Cilia; Complex; Congenital Abnormality; Congenital Heart Defects; Cues; Cultured Cells; Defect; Development; Embryo; Embryonic Development; Epidermis; Epithelial Cells; Epithelium; Event; Feedback; Goals; Hair follicle structure; Human; Hydrocephalus; Image; Imaging Techniques; In Vitro; Intercellular Junctions; Laboratories; Length; Maps; Mediating; Membrane; Methods; Modeling; Molecular; Monitor; Mus; Neural Tube Closure; Organ; Organism; Organoids; Pathway interactions; Pattern; Positioning Attribute; Protein Dynamics; Proteins; Regenerative Medicine; Role; Skin; Sorting; Structure; System; Tissue Engineering; Tissues; Work; cell assembly; cell behavior; cell motility; ciliopathy; deafness; innovation; insight; keratinocyte; mechanical signal; molecular scale; nanoscale; planar cell polarity; reconstitution; regenerative tissue; self assembly; self organization; skin organogenesis; stem cells; superresolution imaging; technology development

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.

项目摘要 细胞在体外具有显着的自我组装成类型结构的能力。然而， 当前的体外衍生器官缺乏适当的尺寸和高阶图案，需要 有机体级信息不存在于菜肴中。值得注意的是，集体极化和 细胞跨组织的单向比对，这是一种称为平面细胞极性的现象 （PCP），缺乏体外器官，但对于适当的器官形成和功能至关重要。因此， 满足组织工程在体外产生功能器官的承诺，我们必须理解 细胞如何建立远程集体极化。 我们已经建立了鼠皮作为模型系统来研究多尺度 PCP在膨胀和再生组织中的协调。通过开发执行的方法 离体培养，长期实时成像，生物物理扰动和器官重建 在表皮上，我们已经对在组织中的PCP建立进行了关键的新发现， 细胞和分子尺度。我们发现单轴组织变形充当对称性 打破表皮中PCP比对的主要轴的突出提示。我们进一步显示 在没有全球提示的情况下，生长的主要角质形成细胞会自发地，局部 平面极性的对齐域。通过超分辨率成像和映射 PCP组件的粘合性相互作用，我们确定了钙粘蛋白介导的顺式的作用 在不对称PCP复合物的聚类和排序中的相互作用。 这项工作的广泛目标是建立在这些以前的发现和技术的基础上 开发如何破译PCP在不同的生物长度尺度上组织。使用 哺乳动物皮肤表皮作为模型系统，特定的目标1将确定长时间 机械提示偏置和整个表皮的平面细胞极性。具体目标2将 研究细胞皮肤细胞自发通过自我产生PCP的机制 组织。特定目标3将破译纳米级体系结构和生化 PCP复合物的相互作用。使用最近在我的实验室开发的技术创新 在两种天然 和器官组织，这项工作将为多尺度提供基本的新见解 PCP的协调。