Mechanical Forces and the Regulation of Airway Progenitor Cells

机械力和气道祖细胞的调节

• 批准号: 9788586
• 负责人: Celeste M Nelson
• 金额: $ 33.82万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9788586
• 关键词: Affect; Air; Apical; Apoptosis; Architecture; Binding; Biochemical; Biological Process; Birth; Cell Differentiation process; ChIP-seq; Chest; Chromatin; Confocal Microscopy; Cues; Defect; Development; Differentiation and Growth; Disease; Embryo; Embryonic Development; Engineering; Ensure; Epithelial; Epithelial Cells; Epithelium; Equilibrium; Fetal Lung; Gases; Gene Expression; Genes; Genetically Engineered Mouse; Human; Image; Inhalation; Ions; Liquid substance; Lung; Lung diseases; Mechanics; Microbe; Microfluidic Microchips; Microfluidics; Molecular; Morphogenesis; Mucous body substance; Mus; Neuroendocrine Cell; Neurosecretory Systems; Notch Signaling Pathway; Organ; Organogenesis; Particulate; Pathway interactions; Population; Population Control; Process; Production; Promoter Regions; Regulation; Reporter; Research Design; Resolution; Role; Signal Pathway; Signal Transduction; Specialized Epithelial Cell; Specific qualifier value; Stem cells; Surface; System; Time; Tissues; Trees; Water; Work; airway epithelium; cell type; chromatin immunoprecipitation; confocal imaging; image reconstruction; innovation; insight; lung development; mechanical force; migration; negative affect; neuroendocrine differentiation; new therapeutic target; next generation sequencing; notch protein; novel; organ growth; pathogen; physical process; pressure; prevent; progenitor; promoter; quantitative imaging; single-cell RNA sequencing; therapeutic target

PROJECT SUMMARY The branched architecture of the airways of the lungs permit the transfer of approximately six liters of air per minute between the external surroundings and the alveoli. The airway epithelial tree accomplishes gas exchange, mucus production, and pathogen clearance and blocks the entry of water, particulates, and microbes. To accomplish these diverse biological functions, the airway epithelium is comprised of several distinct cell types that differentiate from common progenitors during embryonic development, the first of which is the pulmonary neuroendocrine cell. Disrupting the differentiation of the specialized epithelial cell types negatively affects airway morphogenesis, and abnormally high numbers of pulmonary neuroendocrine cells are found in several congenital and acquired diseases of the lung. As it differentiates, the epithelium secretes ions and water across its apical surface, causing fluid to fill the lumen of the airways with a transmural pressure high enough to inflate the lungs. Defects that cause a decrease in transmural pressure are associated with both underdeveloped lungs and an increase in pulmonary neuroendocrine cells, but the specific role of pressure and the molecular signaling downstream of this mechanical cue are unknown. By combining time- lapse confocal imaging with an innovative microfluidic culture system, we found that transmural pressure controls the rate of lung development and the expression of markers of neuroendocrine cells. Using next- generation sequencing analysis, we found that low transmural pressure decreases the expression of targets of Notch, the master regulator of pulmonary neuroendocrine differentiation, and YAP, a known mechanosensor. Here, we hypothesize that transmural pressure coordinates the growth and differentiation of the different cell types within the epithelium by signaling through Notch and YAP. We will combine microfluidic devices with engineered mice, high-resolution time-lapse spinning disk confocal microscopy, and next-generation sequencing analysis to define the relative roles of pressure, Notch, and YAP in the regulation of pulmonary neuroendocrine progenitor fate decisions. In Specific Aim 1, we will use microfluidic chest cavities, engineered mice, time-lapse imaging, and single cell RNA-sequencing to define physically how transmural pressure regulates the pulmonary neuroendocrine population in the developing lung. In Specific Aim 2, we will use microfluidic chest cavities, reporter mice, and chromatin immunoprecipitation approaches to define whether and how transmural pressure regulates Notch signaling in the embryonic airway epithelium. In Specific Aim 3, we will determine whether pressure signals through YAP to affect pulmonary neuroendocrine differentiation and the Notch pathway. This work will define how mechanical signals from the microenvironment are transmitted to the first progenitor fate decision in the developing airway epithelium. We expect that our results will reveal novel insights into mechanical control of progenitor differentiation during tissue development and suggest new therapeutic targets for defects in lung development.

项目摘要 肺气道的分支建筑允许转移大约六升空气 在外部环境和肺泡之间的分钟。气道上皮树完成气体 交换，粘液产生和病原体清除率，并阻止水，微粒和 微生物。为了实现这些多样化的生物学功能，气道上皮由几个 在胚胎发育过程中与常见祖细胞区分开的不同细胞类型，其中第一个 是肺神经内分泌细胞。破坏专业上皮细胞类型的分化 负面影响气道形态发生，异常数量的肺神经内分泌细胞是 在几种先天和肺部的疾病中发现。在区分上，上皮分泌离子 和水面的水，导致流体用透射压力填充气道的腔 足够高以使肺部充气。导致透壁压力降低的缺陷与 肺部欠发育不足和肺神经内分泌细胞的增加，但 压力和该机械提示下游的分子信号传导尚不清楚。通过结合时间 - 与创新的微流体培养系统的失效共聚焦成像，我们发现透壁压力 控制肺发育速率和神经内分泌细胞的标志物的表达。使用下一步 生成测序分析，我们发现低透壁压力降低了目标的表达 Notch，肺神经内分泌分化的主要调节剂，以及已知的机械传感器YAP。 在这里，我们假设透壁压力坐在不同细胞的生长和分化 通过Notch和YAP发出信号，在上皮内的类型。我们将将微流体设备与 工程小鼠，高分辨率的延时旋转磁盘共聚焦显微镜和下一代 测序分析以定义压力，缺口和YAP在肺部调节中的相对作用 神经内分泌祖细胞命运决定。在特定的目标1中，我们将使用微流体胸腔，工程设计 小鼠，延时成像和单细胞RNA顺序，以定义透射压力如何定义 调节发育中肺中的肺神经内分泌群体。在特定目标2中，我们将使用 微流体胸腔，记者小鼠和染色质免疫沉淀方法 以及透壁压力如何调节胚胎气道上皮中的Notch信号传导。在特定的目标3中 我们将确定通过YAP的压力信号是否影响肺神经内分泌分化 和Notch路径。这项工作将定义微环境中的机械信号 传播到发展中的气道上皮的第一个祖细胞命运决定。我们期望我们的结果 将揭示对组织发育和 为肺发育缺陷的新治疗靶标提出了新的治疗靶标。