Investigating the physical mechanisms that drive multicellular lumen morphogenesis

研究驱动多细胞管腔形态发生的物理机制

• 批准号: 9393715
• 负责人: Claudia Gabriela Vasquez
• 金额: $ 5.71万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9393715
• 关键词: Address; Architecture; Biochemical; Biological; Biological Models; Biophysical Process; Canis familiaris; Cell-Cell Adhesion; Cells; Collaborations; Complement; Congenital Abnormality; Cyst; Cystic kidney; Data; Elasticity; Embryo; Embryonic Development; Employee Strikes; Epithelial; Epithelial Cells; Ethics; Generations; Genetic; Growth; Human; Human Development; Kidney; Kinetics; Knowledge; Laboratories; Liquid substance; MDCK cell; Malignant Neoplasms; Measurement; Modeling; Morphogenesis; Morphology; Mus; Organ; Organogenesis; Outcome; Physical shape; Physics; Physiological; Plant Roots; Positioning Attribute; Shapes; Solid; Specificity; Structure; System; Tissues; Work; biophysical properties; career; comparative; expectation; experimental study; human disease; human embryonic stem cell; insight; kidney epithelial cell; mouse development; movie; physical model; physical process; pressure; temporal measurement; therapy development; three dimensional cell culture; tool

PROJECT SUMMARY How tissues assume their correct shape and topology is a central biological question with immediate relevance to both embryonic development and human disease. This project will elucidate the fundamental physical mechanisms by which solid cell masses form a central cavity, termed a lumen. Lumen formation occurs repeatedly during both early embryonic development and organogenesis, and defines both a functional “inside” and “outside” as well as the physical shape of the associated tissue. Previous work has established many of the genetic and biochemical requirements for epithelial polarity and de novo lumen generation. However, lumen formation is an intrinsically physical process – the transition from a solid cell mass to a hollow, spherical shell. Due to previous technical limitations, surprisingly little is known about the biophysical mechanisms that drive lumen formation. To address this knowledge gap, this project will combine quantitative biophysical measurements with classical cell biological approaches to determine the fundamental physical mechanisms that drive lumen formation and expansion in Madin Darby Canine Kidney (MDCK) epithelial cells, a standard model in the field. Preliminary data indicate that, contrary to expectation, cells can form multiple initial openings, suggesting that lumen formation occurs over two stages: initiation, defined as the formation of at least one small opening, followed by establishment, the growth and stabilization of a single central lumen. Recent work has revealed that human embryonic stem cell (hESCs) under proper 3D culture conditions form hollow spheres with striking similarities to the proamniotic cavity, a lumen that is essential for the initiation of the body plan. In the second part of this project, I will use the experimental approach refined using the MDCK model system to determine the physical mechanism of lumen formation employed by hESCs. This work will determine if the physical processes that govern lumen formation in these two systems, which reflect distinct biological origins and functions, are fundamentally similar or different. In addition, these latter experiments provide insight into the physical mechanisms that drive early human embryogenesis, which is not otherwise accessible due to unavoidable technical and ethical barriers.

项目概要 组织如何呈现正确的形状和拓扑结构是一个具有直接相关性的核心生物学问题 胚胎发育和人类疾病。该项目将阐明基本的物理 固体细胞团形成中央空腔（称为内腔）的机制。发生管腔形成 在早期胚胎发育和器官发生过程中反复出现，并定义了功能性“内部” 和“外部”以及相关组织的物理形状。以前的工作已经建立了许多 上皮极性和从头产生管腔的遗传和生化要求。然而， 管腔形成本质上是一个物理过程——从固体细胞团到空心球形的转变 壳。由于之前的技术限制，令人惊讶的是，人们对这种现象的生物物理机制知之甚少。 驱动管腔形成。为了解决这一知识差距，该项目将结合定量生物物理 使用经典细胞生物学方法进行测量以确定基本物理机制 驱动 Madin Darby 犬肾 (MDCK) 上皮细胞管腔形成和扩张，这是一种标准 领域的典范。初步数据表明，与预期相反，细胞可以形成多个初始 开口，表明管腔形成发生在两个阶段：起始，定义为 at 的形成 至少一个小开口，然后是单个中央管腔的建立、生长和稳定。 最近的工作表明，人胚胎干细胞 (hESC) 在适当的 3D 培养条件下形成 空心球体与前羊膜腔具有惊人的相似性，前羊膜腔是启动羊膜腔所必需的内腔 身体计划。在该项目的第二部分中，我将使用使用 MDCK 改进的实验方法 模型系统来确定 hESC 所采用的管腔形成的物理机制。这项工作将 确定这两个系统中控制管腔形成的物理过程是否反映了不同的 生物起源和功能从根本上相似或不同。此外，后面这些实验 提供对驱动早期人类胚胎发生的物理机制的深入了解，否则这是不可能的 由于不可避免的技术和道德障碍而无法访问。