Molecular and Cellular Mechanisms in Coronary Artery Development and Anomalies

冠状动脉发育和异常的分子和细胞机制

• 批准号: 10595393
• 负责人: BIN ZHOU
• 金额: $ 76.48万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10595393
• 关键词: Address; Affect; Biological Assay; Birth; Breeding; Cardiac; Cells; Complement; Congenital Abnormality; Coronary; Coronary Circulation; Coronary Vessel Anomalies; Coronary Vessels; Coronary artery; Data; Data Analyses; Development; Disease; Embryo; Embryonic Development; Endocardium; Endothelium; Fluorescence-Activated Cell Sorting; Genes; Genetic; Goals; Growth; Heart; Histology; Hypoxia; Immunofluorescence Immunologic; In Situ Hybridization; Individual; Intervention; Knowledge; Label; Life; Mesenchymal; Metabolic; Molecular; Mus; Myocardial; Myocardial Ischemia; Myocardium; NOTCH1 gene; Names; Pathogenesis; Pathway Analysis; Perinatal; Phenotype; Process; Proliferating; Regulation; Reporter; Research Project Grants; Signal Transduction; Sinus; TGFB2 gene; TGFBR3 gene; Testing; Transforming Growth Factor Beta 2; VEGFA gene; Vascular Endothelial Growth Factor Receptor-3; Vascular Endothelial Growth Factors; Vascularization; Ventricular; Visualization; angiogenesis; biomarker validation; clinically significant; combat; data integration; differential expression; effective intervention; experimental study; gene network; in vivo; insight; loss of function; malformation; new therapeutic target; novel therapeutics; perinatal period; progenitor; single-cell RNA sequencing; stem cells; sudden cardiac death; translational medicine

PROJECT SUMMARY Normal coronary artery formation is essential for heart growth and function. Malformed coronary arteries are a clinically significant birth defect that can cause life-threatening cardiac complications, including ventricular noncompaction, myocardial ischemia, and sudden cardiac death. Yet, developmental mechanisms that drive proper coronary artery formation are incompletely understood, which has hindered our ability to develop the heart-specific interventions for this devastating disease. The long-term goal of this project is therefore to reveal the molecular and cellular mechanisms underlying coronary artery development so that we may identify key regulatory factors for developing new targeted therapies to combat this congenital condition. We have addressed this goal during previous finding period. Our studies have shown that embryonic coronary arteries in the inner compact myocardium are formed by ventricular endocardial cells through angiogenesis regulated by the VEGF- NOTCH signaling. Furthermore, our studies have revealed that these embryonic coronary arteries undergo angiogenic expansion perinatally to add the neovessels to the growing compact myocardium. However, in contrast to the vascularization of the compact myocardium, we know little about vascularization of trabecular myocardium which remains largely avascular until birth. We have recently identified a subpopulation of coronary progenitor cells among ventricular endocardial cells which are committed to the coronary arteries in the trabecular myocardium. We named these cells as the second wave coronary progenitors (SCPs) to separate them from the first wave coronary progenitors (FCPs) for the coronary vessels at the compact myocardium. SCPs acquire angiogenic potential earlier in embryonic development through a previously unknown endocardial to mesenchymal transformation (EMT) long before they undergo angiogenesis later during perinatal periods to vascularize the trabecular myocardium. In this renewal application, we propose to characterize this new angiogenic-EMT paradigm (angioEMT) by SCPs. Our overarching hypothesis is that vascularization of trabecular myocardium by SCPs is regulated by a “two-hit” mechanism involving sequential angioEMT and hypoxia signaling. We plan to test this hypothesis in three Specific Aims. Aim 1 will characterize SCPs by distinguishing them from FCPs using a lineage-based single cell RNA-sequencing (scRNA-seq) analysis and a modified functional angioEMT assay. Aim 2 will define the angioEMT signaling in the early fate decision by SCPs using genetic loss-of-function approaches investigating the TGFb signaling. Aim 3 will decipher the angiogenic signaling in the later angiogenic activation of SCPs focusing on VEGFA-VEGFR3 and DLL4-NOTCH1 signaling. Vascularization of trabecular myocardium as well as trabecular compaction in the individual nulls will be examined by histology, immunostaining, and RNAscope in situ hybridization. The changes in the SCP lineages will be determined by scRNA-seq analysis, whereas the key factors underlying the two-hit angioEMT process will be identified through gene network analysis. By completing these aims, we expect to provide new mechanistic insights into coronary artery development that inform developmental pathogenesis of coronary artery anomalies and ventricular noncompaction.

项目摘要 正常的冠状动脉形成对于心脏的生长和功能至关重要。畸形的冠状动脉是 临床上显着的先天缺陷，可能导致威胁生命的心脏并发症，包括心室 不采用，心肌缺血和心脏猝死。然而，发展机制的发展机制 正确理解了正确的冠状动脉形成，这阻碍了我们发展的能力 这种毁灭性疾病的心脏特异性干预措施。因此，该项目的长期目标是揭示 冠状动脉发育的基础分子和细胞机制，以便我们可以识别钥匙 开发新的靶向疗法以应对这种先天性疾病的监管因素。我们已经解决了 在上一个发现期间的目标。我们的研究表明，内部的胚胎冠状动脉 紧凑的心肌是由心室心内膜细胞通过VEGF-调节的血管生成形成的 Notch信令。此外，我们的研究表明这些胚胎冠状动脉发生 血管生成的周围膨胀，将新系列添加到日益紧凑的心肌中。但是，在 与紧凑型心肌的血管形成对比，我们对小径的血管化知之甚少 心肌在很大程度上保持尸体直至出生。我们最近确定了冠状动脉的亚群 心室心内膜细胞中的祖细胞，这些细胞致力于冠状动脉 小梁心肌。我们将这些细胞命名为第二波冠状动脉祖细胞（SCP） 它们来自紧凑型心肌处的冠状动脉血管的第一波冠状动脉祖细胞（FCP）。 SCP通过先前未知的心内膜发育中获得胚胎发育中的血管生成潜力 在间充质转化（EMT）之前很早就在围产期发生血管生成 血管性小梁心肌。在此续签应用中，我们建议描述这个新的 SCPS的血管生成 - EMT范式（血管头）。我们的总体假设是小梁的血管化 SCPS的心肌受涉及顺序血管头和缺氧的“两击”机制调节 信号。我们计划以三个具体目标来检验这一假设。 AIM 1将通过区分SCP来表征SCP 它们使用基于谱系的单细胞RNA-sequest（SCRNA-SEQ）分析和修改的FCPS。 功能性血管比测定法。 AIM 2将在SCP的早期脂肪决定中定义血管头信号传导 遗传丧失功能方法研究了TGFB信号传导。 AIM 3将破译血管生成 SCP的后来血管生成激活中的信号传导，重点是VEGFA-VEGFR3和DLL4-NOTCH1信号传导。 小梁心肌的血管化以及单个无效的小梁压实将是 通过组织学，免疫染色和原位杂交检查。 SCP谱系的变化 将通过SCRNA-seq分析确定，而两击血管内过程的关键因素 将通过基因网络分析来识别。通过完成这些目标，我们希望提供新的 对冠状动脉发育的机械洞察力，为冠状动脉发育的发病机理提供了信息 动脉异常和心室不合作。