Mechanisms of epicardium-directed coronary vessel patterning

心外膜定向冠状血管模式的机制

• 批准号: 10686088
• 负责人: Eric M Small
• 金额: $ 54.08万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10686088
• 关键词: ANGPTL2 gene; Acceleration; Address; Bioinformatics; Blood; Blood Vessels; Cardiac Myocytes; Cardiovascular system; Cause of Death; Cell Line; Cell Maturation; Cell Separation; Cell Surface Receptors; Cells; Cellular biology; Coronary; Coronary Arteriosclerosis; Coronary Vessels; Cues; Data; Defect; Development; Embryo; Endothelial Cells; Epicardium; Fetal Heart; Fibroblasts; Fluorescent in Situ Hybridization; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Heart; Image; In Vitro; Ischemia; Ligands; Link; Location; Mesenchymal; Mesothelial Cell; Mesothelium; Metabolic; Molecular; Molecular Biology; Morphology; Mus; Myocardial Infarction; Myocardial Ischemia; Myocardium; Nutrient; Oxygen; Pattern; Play; Population; Proliferating; Publishing; Regulation; Reporting; Risk Factors; Role; Signal Transduction; Specific qualifier value; Stretching; Surface; Therapeutic; Tissues; Transcriptional Regulation; Vascular Endothelial Cell; Vascularization; Venous; blood vessel development; cell behavior; chemokine; directed differentiation; endothelial stem cell; epithelial to mesenchymal transition; experimental study; fetal; improved; in vivo; insight; interest; multiphoton imaging; myocardin; novel; novel therapeutic intervention; progenitor; programs; repaired; single-cell RNA sequencing; stem cells; transcription factor; transcriptomics; wasting

The coronary blood vasculature provides the heart with oxygen and nutrients, and removes metabolic waste. Organization of this contiguous network requires the maturation of vascular endothelial cells (EC) into arterial and venous fates based upon their location in the heart. While many of the guidance factors that control vascular patterning have been defined, it is not clear how spatial information controls cell behavior and identity. The epicardium is a single layer of mesothelial cells on the surface of the heart that harbors an important population of cardiovascular progenitors. We previously reported that epicardial epithelial-to-mesenchymal transition (EMT) is required for coronary EC maturation. New preliminary data reveals profound EC patterning and specification defects upon disruption of the epicardium, culminating with the inappropriate localization of angiogenic ECs in the sub-epicardium. To define the cellular and molecular mechanisms linking epicardial EMT to EC patterning and maturation we performed single cell (sc) RNA-sequencing of epicardium-derived cells and ECs isolated from the embryonic mouse heart at key developmental timepoints. This study defined epicardium-derived “shepherding” and “guidepost” cells that express unique angiogenic chemokine signatures. We provide in vitro and in vivo evidence that suggest a common mechanism controls EMT and the expression of genes that encode important guidance cues. We also find that EC localization and arteriovenous fate specification may be controlled by a common molecular mechanism. Based on previously published and preliminary data, we hypothesize that EMT controls the expression and localization of epicardium-derived chemokines that coordinate coronary EC patterning and AV fate specification in the fetal heart. The current study will interrogate this novel paradigm of epicardium-directed coronary EC patterning (localization and branching) and maturation (arteriovenous specification). Here, we will use genetically modified mice, time-lapse live embryo multi-photon imaging, scRNA-seq and spatial transcriptomics, and cell and molecular biology approaches to: 1) Define a common mechanism regulating EMT and the expression of genes that encode EC guidance cues; and 2) Interrogate the mechanisms coordinating epicardium-directed EC patterning and AV fate specification. We expect these studies will provide important insights into the mechanisms that control vascular patterning. This study may also advance our understanding of the developmental origins of coronary artery disease, and lead to therapeutic strategies that stimulate revascularization and repair of ischemic heart tissue.

冠状血管为心脏提供氧气和营养，并去除 代谢废物。这个连续网络的组织需要血管成熟 内皮细胞（EC）根据其在心脏中的位置成动脉和静脉命运。尽管 控制血管模式的许多指导因素已定义，尚不清楚 空间信息如何控制细胞行为和身份。心外膜是一层 心脏表面上的间皮细胞具有重要的心血管群 祖先。我们以前报道过上皮上皮到间质转变（EMT）是 冠状动脉成熟所必需的。新的初步数据揭示了深刻的EC模式和 外膜破坏后的规格缺陷，与不适当 血管生成EC的定位在亚膜下。定义细胞和分子 将心外膜EMT与EC模式和成熟联系起来的机制我们进行了单细胞（SC） 从胚胎小鼠心脏分离的心外膜衍生细胞和EC的RNA测序 在关键的开发时间点。这项研究定义了心外膜衍生的“牧羊”和 表达独特的血管生成趋化因子特征的“指南”细胞。我们提供体外和 表明一种共同机制控制EMT的体内证据和基因的表达 该编码重要的指导提示。我们还发现EC的本地化和动静脉命运 规范可以通过常见的分子机制控制。基于以前的 发布和初步数据，我们假设EMT控制着表达式和 表达冠状动脉EC模式的心外膜衍生的趋化因子的定位 和胎儿心脏中的AV命运规格。当前的研究将审问这本小说 表达冠状动脉EC图案（本地化和分支）和 成熟（动静脉规范）。在这里，我们将使用一般修改的小鼠，延时 活胚多光子成像，SCRNA-seq和空间转录组学以及细胞和分子 生物学方法：1）定义调节EMT的常见机制和表达 编码EC指导提示的基因； 2）审问协调的机制 心外膜定向的EC模式和AV命运规格。我们希望这些研究将提供 对控制血管图案的机制的重要见解。这项研究也可能 促进我们对冠状动脉疾病的发育起源的理解，并导致 刺激缺血性心脏组织血运重建和修复的治疗策略。