Dynamic regulation of embryonic endothelial cell migration in response to hemodynamic force

胚胎内皮细胞迁移响应血流动力学的动态调节

• 批准号: 10406161
• 负责人: Mary E Dickinson
• 金额: $ 49.6万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10406161
• 关键词: Acute; Affect; Arteries; Arteriovenous malformation; Automobile Driving; Biosensor; Blood Circulation; Blood Vessels; Blood capillaries; Blood flow; Caliber; Cardiac; Cardiovascular system; Cells; Cessation of life; Complement; Data; Defect; Development; Embryo; Endothelial Cells; Endothelium; Ensure; FOXO1A gene; Future; Gene Expression; Genes; Genetic Transcription; Growth; Image; Investigation; Lead; Methods; Microscopy; Modeling; Morphology; Mus; Mutation; Notch Signaling Pathway; Pathway interactions; Pharmacology; Phosphotransferases; Population; Process; Regulation; Reporter; Research Personnel; Signal Transduction; Testing; Three-Dimensional Imaging; Time; Tissues; Trees; Uncertainty; Work; Yolk Sac; angiogenesis; cell motility; conditional knockout; directional cell; experimental study; fetal; gain of function; hemodynamics; human disease; mechanical signal; mechanical stimulus; migration; mouse model; new technology; notch protein; novel; optogenetics; polarized cell; response; sensor; shear stress; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY To accommodate the demands of the constantly expanding embryo, vessels of the fetal circulatory system must undergo extensive morphological remodeling while simultaneously supporting pre-existing tissues. Mechanical stimuli, generated by the hemodynamic force, are critical regulators of this process, driving adaptive responses to blood flow such as vessel fusion, widening, or regression. Herein, we propose experiments to determine how early vessels remodel in response to changes in blood flow. Our hypothesis is that differential activation of a Flk1/ERK/Dll4/Notch signaling axis creates a subset of ECs competent to respond to blood flow, resulting in the directed migration and expansion of the vitelline artery. We have assembled an outstanding team of investigators, as well as novel technologies, mouse models and methods to test this hypothesis in three specific aims: 1. Use loss- and gain-of-function models of Flk1/ERK/Dll4/Notch signaling to define differences in endothelial cell (EC) migration during vessel remodeling. 2. Use a real-time ERK biosensor to determine whether ECs that migrate in response to changes in blood flow upregulate ERK activity and if altering ERK signaling, or modulating blood flow, affects cell migration. 3. Employ single cell RNA-sequencing to determine if a specific population of cells exists in the VA with high arterial gene expression and elevated levels of polarization and pro-migratory genes. The transcriptional data will complement the dynamic, 3D imaging data, allowing us to reveal the network of factors required in specialized ECs that would otherwise be lost through studies at a population level (e.g. via bulk RNA-seq) and will no doubt prompt novel future investigation into hemodynamic force signaling and EC migration in the embryo.

项目摘要 为了适应不断扩展的胚胎的需求，胎儿循环系统的船只必须 经过广泛的形态重塑，同时支撑先前存在的组织。机械的 血液动力学产生的刺激是该过程的关键调节剂，推动了适应性反应 到血流，例如血管融合，扩大或消退。 本文中，我们提出了实验，以确定早期血管对血液变化的反应 流动。我们的假设是flk1/erk/dll4/Notch信号轴的差分激活会产生 EC的子集有能力应对血流的响应，导致了定向迁移和扩张 卵黄动脉。我们组建了一个杰出的调查员团队以及新颖的技术， 鼠标模型和方法以三个特定目的检验该假设：1。使用功能损失和功能获得 flk1/erk/dll4/Notch信号的模型，以定义血管中内皮细胞（EC）迁移的差异 重塑。 2。使用实时ERK生物传感器来确定是否响应变化而迁移的EC迁移 血流上调ERK活性，如果改变ERK信号传导或调节血流会影响细胞迁移。 3。采用单细胞RNA测序来确定特定的细胞群是否存在于VA中 动脉基因表达以及极化和促迁移基因的水平升高。转录数据 将补充动态3D成像数据，使我们能够揭示 专门的EC会通过人群水平的研究（例如通过大量RNA-Seq）和 毫无疑问，将促使未来对胚胎中血液动力学信号传导和EC迁移的新调查。