MESOTHELIAL MECHANOTRANSDUCTION AS A KEY REGULATOR OF EMBRYONIC LUNG GROWTH

间皮机械传导作为胚胎肺生长的关键调节因子

• 批准号: 10000129
• 负责人: Rachel Gilbert
• 金额: $ 3.16万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000129
• 关键词: Abdomen; Affect; Animals; Birth; Cell Differentiation process; Cell Nucleus; Cells; Chest; Clinical; Confocal Microscopy; Congenital diaphragmatic hernia; Coupled; Data; Development; Devices; Disease; Embryo; Endothelium; Epicardium; Epithelial; Epithelium; FGF10 gene; Fibroblasts; Genes; Growth; Growth Factor; Heart; Impairment; In Vitro; Infant; Interruption; Invaded; Knock-out; Knockout Mice; Label; Liquid substance; Live Birth; LoxP-flanked allele; Lung; Mechanical Stimulation; Mechanics; Mediating; Membrane; Mesenchymal; Mesenchyme; Mesothelial Cell; Mesothelium; Microfluidics; Morphology; Mus; Newborn Infant; Organ; Pathway interactions; Physiological; Play; Process; Proteins; Reporter; Reporting; Respiratory Diaphragm; Role; Severities; Signal Pathway; Signal Transduction; Signaling Protein; Smooth Muscle; Stretching; Structure of parenchyma of lung; System; Techniques; Testing; Tissues; Transcription Coactivator; Tretinoin; WT1 gene; Weight; Western Blotting; Work; cell type; clinical translation; epithelial to mesenchymal transition; fibroblast growth factor 9; flexibility; improved; insight; lung lobe; mechanotransduction; morphogens; mortality; novel; pressure; prevent; progenitor; pulmonary hypoplasia; response; three dimensional cell culture; time use; transcription factor; treatment strategy

PROJECT SUMMARY/ABSTRACT Congenital diaphragmatic hernia (CDH) affects 1 in 2500 live births with mortality rates in newborns ranging from 10-40%, depending on factors such as severity. CDH involves the herniation of the abdominal organs into the chest cavity through a hole in the diaphragm and results in decreased lung growth, or pulmonary hypoplasia. The mesothelium is a specialized epithelial tissue that lines the embryonic lung and has a major influence on lung growth by secreting growth factors such as FGF9 and serving as a progenitor pool for various cells in the growing lung. However, it is unclear what regulates these two phenomena. Previous data along with our preliminary data has led to the hypothesis that these functions of the mesothelium are regulated by tissue stretch, and this stretch is dysregulated during CDH. Herein, we propose the mechanism by which this stretch is sensed by the mesothelium, along with the downstream signaling that leads to eventual FGF9 expression. Overall, the mesothelium is understudied considering its major influence on lung growth, and we are the first to propose that the mesothelium is mechanically stimulated during development. In both Aims we will use our novel microfluidic chest cavity to examine the effects of mesothelial stretch on the whole tissue level. With this device, we can control the fluid pressure internal and external of a mouse lung explant to induce controlled stretch on the lung in a physiological 3D culture system. In our first Aim, we will test our proposed pathway involving stretch mediated FGF9 signaling. We hypothesize that this stretch is sensed via the mechanosensitive molecule YAP, which in other systems is stretch responsive. YAP then induces WT1 and RA signaling to cause FGF9 expression from the tissue. In addition to using our microfluidic chest cavity studying whole tissue effects, we will also apply controlled stretch to isolated primary mesothelial cells using microfluidic stretch membranes. We will test our hypothesis by examining the effect of stretch on the various molecules in the YAP/WT1/RA/FGF9 pathway, and the effect on growth when this pathway is interrupted. In our second Aim, we hypothesize that mesothelial stretch is sensed by YAP to induce epithelial-mesenchymal transition(EMT). The mesothelium has been shown to undergo EMT as serve as a progenitor pool for various lung tissue, and in other systems, YAP is a known inducer of EMT. We will determine the effect of stretch on mesothelial EMT.

项目摘要/摘要 先天性diaphragmatic疝（CDH）影响2500名活产，新生儿死亡率的死亡率范围从 10-40％，取决于严重程度等因素。 CDH涉及腹部器官的疝气 胸腔穿过隔膜的孔，导致肺部生长降低或肺部发育不全。 间皮是一种专门的上皮组织，对胚胎肺排列，对 通过分泌生长因子（例如FGF9）和作为各种细胞中各种细胞的祖细胞库等生长因子的生长 生长肺。但是，目前尚不清楚是什么调节这两种现象。以前的数据以及我们的 初步数据导致了以下假设：间皮的这些功能受组织拉伸的调节， 在CDH期间，这种延伸失调。本文中，我们提出了感应这种拉伸的机制 通过间皮，以及最终导致FGF9表达的下游信号传导。总体而言， 考虑到其对肺部生长的主要影响，间皮细学，我们是第一个提议的人 间皮在发育过程中被机械刺激。 在这两个目标中，我们都将使用新型的微流体胸腔检查间皮伸展对 整个组织水平。使用此设备，我们可以控制小鼠肺的内部和外部流体压力 在生理3D培养系统中诱导肺部受控伸展。在我们的第一个目标中，我们将 测试我们提出的涉及拉伸介导的FGF9信号传导的途径。我们假设感觉到这种伸展 通过机械敏感的分子YAP，在其他系统中，它具有拉伸响应。 yap然后诱导WT1 RA信号传导引起组织中的FGF9表达。除了使用我们的微流体胸腔 研究整个组织效应，我们还将使用受控拉伸将受控拉伸应用于孤立的原代间皮细胞 微流体拉伸膜。我们将通过检查拉伸对各种影响的影响来检验我们的假设 YAP/WT1/RA/FGF9途径中的分子，以及该途径中断时对生长的影响。在我们的 第二个目的，我们假设通过yap感测间皮伸展以诱导上皮 - 间质 过渡（EMT）。中皮已被证明是作为各种祖细胞池的EMT 肺组织和其他系统中，YAP是EMT的已知诱导剂。我们将确定拉伸的效果 间皮EMT。