Defining the developmental mechanisms of pericardium formation

定义心包形成的发育机制

• 批准号: 10605059
• 负责人: Hannah Rose Moran
• 金额: $ 3.61万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605059
• 关键词: AXIN2 gene; Binding; Binding Sites; Biology; CRISPR/Cas technology; Cardiac; Cardiovascular system; Cell Lineage; Cells; Color; Complex; Data; Development; Embryo; Embryonic Development; Encapsulated; Epicardium; Feedback; Friction; Gene Proteins; Genes; Genetic; Genetic Markers; Genetic Transcription; Goals; Heart; Heart Injuries; Homeostasis; Human; Image; Infection; Injury; Investigation; Label; Lateral; Link; Maps; Mesoderm; Mesothelial Cell; Mesothelium; Microscopy; Mining; Molecular; Morphogenesis; Mutagenesis; Mutate; Natural regeneration; Nucleic Acid Regulatory Sequences; Parietal; Pathway interactions; Pattern; Pericardial body location; Phenotype; Population; Process; Property; Regulation; Regulator Genes; Regulatory Element; Reporter; Research; Research Proposals; Role; Signal Transduction; Source; Structure; Testing; Time; Tissues; Transgenic Organisms; Tube; Visceral; Visualization; WNT Signaling Pathway; Work; Zebrafish; beta catenin; body system; cardiogenesis; cell type; congenital anomaly; congenital heart disorder; genomic locus; in vivo; inhibitor; insight; knock-down; migration; mutant; overexpression; pericardial sac; predictive modeling; progenitor; single-cell RNA sequencing; spatiotemporal; tool; transcription factor

PROJECT SUMMARY The pericardium is a mesothelial sac that encapsulates the heart to support its development and maintain cardiac homeostasis over time. Throughout the process of heart looping, the pericardium serves as a cell source to form various cardiac structures, making it a crucial player in heart morphogenesis. However, little is known about its developmental origins and how it acquires its unique properties in cardiac development, homeostasis, and regeneration post-injury. While early mesothelial origins overall have only been recently described in detail using the zebrafish, the pericardium has been particularly difficult to study due to its dynamic development and a lack of genetic markers. Our lab has mapped mesothelial progenitors to the lateral plate mesoderm expressing the transcription factor Hand2, and further linked Hand2 function to mesothelium and pericardium formation. These conceptual and technical advances in mesothelial biology enable us to investigate how pericardial and cardiac structures come together to form a single functional organ system during development. My expansion of this data suggests that pericardial and cardiac progenitors are distinct progenitor populations that later come together to fuse together during embryogenesis. The objective of this proposal is to uncover the developmental mechanisms governing progenitor patterning into the heart and pericardium and identify the role of canonical Wnt signaling that controls the pericardial emergence. In Aim 1, I will use a combination of transgenic zebrafish lines that label the developing heart and pericardium, in vivo time-lapse imaging, stable genetic knockdowns of cardiac components, and multi-color lineage tracing reporters to examine the cellular origins and dynamics of the developing pericardium. In Aim 2, I will investigate the interplay between Hand2 and canonical Wnt signaling by documenting the expression dynamics of Wnt targets in Hand2 mutant zebrafish, overexpressing Wnt components, and using CRISPR-Cas9 mutagenesis to mutate Hand2 binding sites. Together, this project will be the first to fully characterize how pericardial development proceeds and incorporates the heart and the interplay between Hand2 and canonical Wnt signaling in controlling pericardium development.

项目概要 心包是一种间皮囊，包裹心脏以支持其发育和维持 随着时间的推移心脏稳态。在整个心脏循环过程中，心包作为细胞来源 形成各种心脏结构，使其成为心脏形态发生的关键参与者。然而，鲜为人知 关于其发育起源以及它如何在心脏发育、体内平衡、 以及损伤后的再生。虽然早期间皮起源总体上直到最近才被详细描述 使用斑马鱼来研究心包特别困难，因为它的动态发育和 缺乏遗传标记。我们的实验室已将间皮祖细胞映射到侧板中胚层表达 转录因子 Hand2，并进一步将 Hand2 功能与间皮和心包形成联系起来。 间皮生物学的这些概念和技术进步使我们能够研究心包和 心脏结构在发育过程中聚集在一起形成单一的功能器官系统。我的扩展 该数据表明心包和心脏祖细胞是后来出现的不同祖细胞群 在胚胎发生过程中融合在一起。该提案的目的是揭示发展的 控制心脏和心包的祖细胞模式的机制，并确定经典的作用 Wnt 信号控制心包的出现。在目标 1 中，我将使用转基因斑马鱼的组合 标记发育中的心脏和心包的线、体内延时成像、稳定的基因敲低 心脏成分和多色谱系追踪记者来检查细胞起源和动态 正在发育的心包。在目标 2 中，我将研究 Hand2 和规范 Wnt 信号之间的相互作用 通过记录 Hand2 突变斑马鱼中 Wnt 靶标的表达动态，过度表达 Wnt 组件，并使用 CRISPR-Cas9 诱变来突变 Hand2 结合位点。总之，该项目将 第一个全面描述心包发育如何进行并结合心脏和相互作用的人 Hand2 和经典 Wnt 信号在控制心包发育中的关系。