Embryonic origins of endothelial heterogeneity

内皮异质性的胚胎起源

• 批准号: 10328511
• 负责人: NATHAN D LAWSON
• 金额: $ 100.5万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10328511
• 关键词: Anatomy; Biological Models; Blood Vessels; Caliber; Cardiovascular system; Cell Differentiation process; Cell Line; Characteristics; Coupled; Development; Developmental Biology; Embryo; Embryonic Development; Endothelial Cells; Endothelium; Genes; Genetic Enhancer Element; Genetic Transcription; Growth; Heterogeneity; Knock-out; Link; Location; Modeling; Molecular; Molecular Profiling; Morphogenesis; Pattern; Phenotype; Physiological; Process; Regulatory Pathway; Severities; Signal Pathway; Signal Transduction; Site; Techniques; Time; Vascular Diseases; Venous; Zebrafish; blood vessel development; capillary bed; cell type; genome editing; insight; molecular phenotype; progenitor; single-cell RNA sequencing; transcriptome

PROJECT SUMMARY Endothelial heterogeneity is a defining characteristic of the mature circulatory system. Endothelial cells that line capillary beds or other small caliber vessels have distinct phenotypes and molecular signatures that relate to their various functions in different anatomical locations. In larger vessels, there are clear molecular and phenotypic differences between arterial and venous endothelial cells. In all of these cases, endothelial differentiation is essential for normal physiological function of the circulatory system. Importantly, endothelial heterogeneity can have a major influence on the site and severity of vascular disease. Thus, a better understanding of how endothelial cell types are determined is highly relevant. For the past 15 years, we have used the zebrafish as a model system to investigate basic mechanisms of vascular morphogenesis and patterning during embryonic development. Our efforts have revealed new insights into how blood vessels are formed and underscore the importance of endothelial differentiation in this process. Importantly, we have found that endothelial cell differentiation is a primary step that is essential for blood vessel formation and assembly. However, the developmental origins of endothelial identities and the signaling pathways that drive differentiation are largely unknown. In the studies proposed here, we will apply a number of traditional developmental biology approaches coupled with cutting-edge molecular techniques to define the hierarchy of endothelial ontogeny during embryonic development. Through Cre/lox lineage tracing we will identify where and when endothelial cell types are established. In parallel, we will apply single cell RNA sequencing on endothelial progenitors at multiple developmental stages to identify transcriptome signatures that define endothelial subtypes. At the same time, efforts to identify enhancer elements flanking subtype-specific genes will contribute to our knowledge of transcriptional regulatory pathways and upstream signals that drive differentiation. Finally, we will continue to investigate the link between endothelial differentiation and vascular morphogenesis through functional interrogation of subtype specific genes using knockout zebrafish models generated through genome editing. Together, our efforts will define the developmental endothelial hierarchy and allow us to identify essential signaling pathways responsible for endothelial heterogeneity.

项目概要 内皮异质性是成熟循环系统的一个决定性特征。内皮细胞 毛细血管床或其他小口径血管内衬的细胞具有不同的表型和分子 与不同解剖位置的各种功能相关的特征。在较大的船只中， 动脉内皮细胞和静脉内皮细胞之间存在明显的分子和表型差异。 在所有这些情况下，内皮分化对于正常的生理功能至关重要。 循环系统。重要的是，内皮异质性会对位点和 血管疾病的严重程度。因此，更好地了解内皮细胞类型如何 确定是高度相关的。在过去的 15 年里，我们一直使用斑马鱼作为模型系统 研究胚胎时期血管形态发生和模式形成的基本机制 发展。我们的努力揭示了关于血管如何形成和如何形成的新见解 强调内皮分化在此过程中的重要性。重要的是，我们发现 内皮细胞分化是血管形成所必需的第一步 集会。然而，内皮特性和信号通路的发育起源 推动差异化的因素在很大程度上是未知的。在这里提出的研究中，我们将应用一些 传统的发育生物学方法与尖端分子技术相结合 定义胚胎发育过程中内皮个体发育的层次结构。通过 Cre/lox 谱系 通过追踪，我们将确定内皮细胞类型何时何地建立。与此同时，我们将 对多个发育阶段的内皮祖细胞应用单细胞 RNA 测序 识别定义内皮亚型的转录组特征。同时，努力查明 亚型特异性基因侧翼的增强子元件将有助于我们对转录的认识 驱动分化的调控途径和上游信号。最后，我们将继续 通过功能研究研究内皮分化和血管形态发生之间的联系 使用通过基因组生成的敲除斑马鱼模型来询问亚型特定基因 编辑。我们的共同努力将定义发育内皮层次结构，并使我们能够 确定负责内皮异质性的重要信号通路。