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.

项目总结