Mechanical Clocks During Fetal Development

项目概要 发展完全取决于时机。从受精卵到新生儿，胚胎发育是一个过程 高度协调的刻板事件序列。流程每个阶段的顺序和时间安排， 包括组织形态发生和祖细胞分化的时间，对于构建 器官成熟，适合出生。时机缺陷与先天性出生缺陷以及出生缺陷有关。 成人慢性疾病，包括哮喘、肺气肿、肾衰竭和 II 型糖尿病。什么控制 发育的节奏——胚胎的中央节拍器——是生物学中最大的谜团之一。 迄今为止，只发现了少数分子计时器，包括昼夜节律和分段计时器 时钟，两者都作为转录振荡器运行，由周期性激活的 细胞外生化刺激。然而，目前尚不清楚生物化学信号是如何传递的。 扩散可以耦合体内相距较远的器官的发育速率 胚胎。我们最近出乎意料地发现，哺乳动物胚胎的形态发生率 肺受到来自管腔流体压力的机械力的牵引，该压力控制着肺的频率 整个器官的同步上皮分支和平滑肌收缩。这些发现表明 胎儿体内存在“机械钟”。因为流体压力是瞬时传递的 在遥远的组织之间，机械时钟可以同步各个器官的发育速度， 允许出生前协调成熟。在这里，我们建议研究肺、肾、 和胰腺发育，所有器官都通过胚胎内部和周围的液体连接并形成 通过分支形态发生。我们将定义压力的大小如何控制扩散速度， 使用微流体方法进行分化和形态发生。我们还将识别振荡 由压力引起的信号通路并研究流体力如何在之间传递 远距离器官，以确保它们的发育速度相匹配。我们将结合芯片上的器官模型， 组织特异性报告动物、转基因敲除小鼠、单细胞转录组学和蛋白质组学，以及 定量延时成像分析，并通过对人类患者样本的研究来补充这些分析 夹带缺陷的小鼠模型。这项工作将揭示胎儿的共享机械环境如何 器官允许它们在出生时协调生长和成熟，这对于设计新的产品至关重要 治疗与先天性缺陷和发育性早产相关的疾病的方法，以及 成人的慢性疾病。

PROJECT SUMMARY Development is all about timing. From fertilized egg to newborn infant, embryonic development proceeds as a highly coordinated sequence of stereotyped events. The order and timing of each stage of the process, including the timing of tissue morphogenesis and differentiation of progenitor cells, are essential for building mature organs in time for birth. Defects in timing are associated with both congenital birth defects as well as chronic diseases in the adult, including asthma, emphysema, renal failure, and type II diabetes. What controls the tempo of development – the central metronome of the embryo – is one of the great mysteries of biology. Only a handful of molecular timers have been discovered to-date, including the circadian and segmentation clocks, both of which operate as transcriptional oscillators that are entrained by periodic activation of extracellular biochemical stimuli. However, it is unclear how biochemical signals that are transmitted by diffusion can couple the rates of development of organs that are separated by large distances within the embryo. We recently discovered unexpectedly that the rate of morphogenesis of the embryonic mammalian lung is entrained by mechanical forces from luminal fluid pressure, which controls the frequency of synchronized epithelial branching and smooth muscle contraction across the organ. These findings suggest the presence of a “mechanical clock” in the fetus. Because fluid pressure is transmitted instantaneously between distant tissues, a mechanical clock could synchronize the rates of development across organs, permitting coordinated maturation before birth. Here, we propose to investigate the coupling of lung, kidney, and pancreatic development, organs that are all connected by fluid within and around the embryo and that form via branching morphogenesis. We will define how the magnitude of pressure controls the rates of proliferation, differentiation, and morphogenesis using microfluidics approaches. We will also identify the oscillatory signaling pathways that are induced by pressure and investigate how fluid forces are transmitted between distant organs to ensure that their rates of development are coupled. We will combine organ-on-a-chip models, tissue-specific reporter animals, transgenic knockout mice, single-cell transcriptomics and proteomics, and quantitative time-lapse imaging analysis, and complement these with studies of human patient samples and mouse models of entrainment defects. This work will uncover how the shared mechanical environment of fetal organs permits them to grow and mature coordinately in time for birth, which is essential for designing new approaches to treat disorders associated with congenital defects and developmental prematurity, as well as chronic diseases in the adult.