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物质，并除去 代谢废物。这个连续网络的组织需要血管的成熟 内皮细胞（EC）根据其在心脏中的位置分为动脉和静脉命运。尽管 许多控制血管模式的指导因素已经被定义，但尚不清楚 空间信息如何控制细胞行为和身份。心外膜是单层 心脏表面的间皮细胞含有重要的心血管细胞群 祖先。我们之前报道过心外膜上皮间质转化（EMT） 冠状动脉 EC 成熟所需的。新的初步数据揭示了深刻的 EC 模式和 心外膜破裂时出现规格缺陷，最终导致不适当的 心外膜下血管生成 EC 的定位。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) 从小鼠胚胎心脏分离的心外膜来源细胞和 EC 的 RNA 测序 在关键的发育时间点。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“路标”细胞。我们提供体外和 体内证据表明存在控制 EMT 和基因表达的共同机制 编码重要的指导线索。我们还发现 EC 定位和动静脉命运 规范可以由共同的分子机制控制。根据之前的 根据已发表的初步数据，我们假设 EMT 控制表达和 协调冠状动脉 EC 模式的心外膜衍生趋化因子的定位 和AV命运在胎儿心脏的规范。当前的研究将审问这部小说 心外膜定向冠状动脉 EC 模式（定位和分支）的范式和 成熟（动静脉规格）。在这里，我们将使用转基因小鼠，延时拍摄 活体胚胎多光子成像、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 和表达的共同机制 编码 EC 引导线索的基因； 2）询问协调机制 心外膜定向 EC 模式和 AV 命运规范。我们期望这些研究将提供 对控制血管模式的机制的重要见解。这项研究还可能 增进我们对冠状动脉疾病发育起源的理解，并导致 刺激缺血心脏组织血运重建和修复的治疗策略。