Interplay between mechanical forces and retinoic acid in lung development

肺发育中机械力和视黄酸之间的相互作用

• 批准号: 10367647
• 负责人: Celeste M Nelson
• 金额: $ 55.33万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367647
• 关键词: Address; Affect; Air; Anabolism; Biochemical; Biological Models; Birth; Breathing; Cells; Chest; Congenital Abnormality; Congenital diaphragmatic hernia; Critical Care; Data; Defect; Development; Embryo; Environment; Epithelial; Equilibrium; Fetal Tissues; Gene Expression Profile; Genetic Transcription; Growth; Image; Image Analysis; Immunofluorescence Immunologic; Immunohistochemistry; In Situ Hybridization; Investigation; Knock-out; Knockout Mice; Lead; Liquid substance; Long-Term Survivors; Lung; Lung diseases; Measurement; Measures; Mechanical Stress; Mechanics; Mesenchyme; Mesothelium; Microfluidics; Molecular; Morbidity - disease rate; Morphogenesis; Mus; Neonatal Mortality; Nutritional; Pathogenesis; Pathway interactions; Pattern; Phenocopy; Publishing; Reporter; Research Design; Respiratory Diaphragm; Retinoids; Reverse Transcriptase Polymerase Chain Reaction; Rodent Model; Role; Secondary to; Signal Pathway; Signal Transduction; System; Testing; Time; Tissues; Transgenic Mice; Transgenic Organisms; Trees; Tretinoin; Work; airway epithelium; experience; fetal; fluidity; force sensor; innovation; lung development; lung imaging; mechanical force; mechanical signal; mortality; mouse model; neonate; new therapeutic target; novel; pressure; pulmonary hypoplasia; quantitative imaging; respiratory smooth muscle; response; sensor; single-cell RNA sequencing; spatiotemporal; transcriptomics

PROJECT SUMMARY Breathing after birth requires coordinated development of the airway epithelium and its surrounding mesenchyme and mesothelium. In the fetus, these tissues form in response to exogenous forces from fluid pressure within and around the developing lungs that is normally high in the lumen of the airways, thus generating a positive transpulmonary pressure. The mechanical environment of the fetal chest cavity is disrupted by conditions such as congenital diaphragmatic hernia (CDH), which reduces or reverses the pressure across the developing lungs and leads to pulmonary hypoplasia, a major cause of neonatal mortality. Although several biochemical signals, including retinoic acid (RA), have been implicated in the pathogenesis of CDH, the mechanical forces and signaling downstream of transpulmonary pressure that regulate lung development are unknown. Our preliminary and published data suggest that transpulmonary pressure itself regulates the RA-biosynthesis pathway, airway epithelial branching morphogenesis, and airway smooth muscle differentiation. Here, we propose to take advantage of our innovative microfluidic platforms, tissue- specific knockout mice, fluorescent reporter mice, and mouse models of CDH to uncover the molecular mechanisms that connect pressure, RA signaling, and morphogenesis and differentiation within the embryonic lung. We will combine these approaches with time-lapse imaging of lung explants, single-cell transcriptomic analysis, and real-time fluorescent force sensors. In Specific Aim 1, we will test the hypothesis that pressure activates the mechanosensor Yap in a tissue-specific manner to regulate the spatiotemporal pattern of expression of genes involved in the RA-biosynthesis pathway. In Specific Aim 2, we will uncover the effects of pressure and tissue-specific synthesis of RA on airway epithelial growth and morphogenesis and airway smooth muscle differentiation. In Specific Aim 3, we will measure the relative effects of pressure and RA signaling on tension, strain, and fluidity within the epithelium, mesenchyme, and mesothelium. This work will, for the first time, identify the tissue-specific mechanical forces and molecular signaling downstream of transpulmonary pressure that regulate early morphogenesis of the lung. We expect that our findings will suggest novel therapeutic targets for the treatment of defects in lung development.

项目概要 出生后的呼吸需要气道上皮及其周围的协调发育 间质和间皮。在胎儿中，这些组织是响应液体的外源力而形成的 发育中的肺部内部和周围的压力通常在气道内较高，因此 产生跨肺正压。胎儿胸腔的力学环境是 因先天性膈疝 (CDH) 等疾病而中断，从而减少或逆转 发育中的肺部承受压力，导致肺发育不全，这是新生儿死亡的主要原因。 尽管包括视黄酸（RA）在内的多种生化信号与糖尿病的发病机制有关 CDH，调节肺的跨肺压下游的机械力和信号传导 发展未知。我们的初步和已发表的数据表明跨肺压本身 调节 RA 生物合成途径、气道上皮分支形态发生和气道平滑 肌肉分化。在这里，我们建议利用我们的创新微流体平台、组织- 特异性敲除小鼠、荧光报告小鼠和 CDH 小鼠模型以揭示分子 将压力、RA 信号传导以及胚胎内的形态发生和分化联系起来的机制 肺。我们将把这些方法与肺外植体的延时成像、单细胞转录组学相结合 分析和实时荧光力传感器。在具体目标 1 中，我们将检验以下假设：压力 以组织特异性方式激活机械传感器 Yap，以调节时空模式 参与 RA 生物合成途径的基因的表达。在具体目标 2 中，我们将揭示以下效果： 压力和 RA 的组织特异性合成对气道上皮生长和形态发生以及气道的影响 平滑肌分化。在具体目标 3 中，我们将测量压力和 RA 的相对影响 上皮、间质和间皮内的张力、应变和流动性的信号传导。这项工作将， 首次确定组织特异性机械力和下游分子信号传导 调节肺早期形态发生的跨肺压。我们希望我们的发现能够 提出了治疗肺部发育缺陷的新治疗靶点。