Single Cell RNA-seq to Identify Endocardial Ontogenic Factors for the Heart

单细胞 RNA-seq 鉴定心脏的心内膜个体发育因子

基本信息

批准号：
9769109
负责人：
BIN ZHOU
金额：
$ 20.88万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9769109
关键词：
Address Anatomy Arteries Automobile Driving Basic Science Bioinformatics Biological Assay Biology Blood Circulation Candidate Disease Gene Cardiac Cardiac development Cardiovascular system Cell Separation Cell Therapy Cells Clinical Coronary Coronary Vessel Anomalies Coronary artery Cues Defect Development Endocardium Endothelial Cells Equilibrium Etiology Expression Profiling Future Genes Genetic Genetic Models Goals Heart Heart Diseases Heart Valve Diseases Heart Valves Heterogeneity In Situ In Vitro Individual Intrinsic factor Knowledge Label Lead Light Location Lung Maintenance Mediating Medical Mesenchymal Mesenchyme Messenger RNA Methodology Methods Modeling Molecular Molecular Analysis Molecular Genetics Morbidity - disease rate Morphology Mus Natural regeneration Outcome Pattern Population Population Heterogeneity Preparation Process RNA Regulation Research Research Project Grants Role Specific qualifier value Stem cells Structure Systems Biology Technology Testing To specify Vascular blood supply Ventricular angiogenesis base cDNA Library cardiogenesis cell fate specification congenital heart disorder differential expression experimental study fetal genetic signature heart cell heart function in vivo mortality mouse model novel novel therapeutic intervention prevent programs repaired single-cell RNA sequencing transcriptome transcriptome sequencing transcriptomics

项目摘要

Abstract: The goal of this research project is to identify the signature genes that define the ontogenic niche for cardiac endocardial cells to generate the critical heart structures, valves and coronary arteries, using mouse genetics and single cell transcriptomics. Our previous studies showed that cardiac endocardial cells are the progenitor cells for heart valves and coronary arteries, arising from different cardiac regions and at different stages through distinct mechanisms. Endocardial cells at the cardiac outflow tract and atrioventricular canal give rise to heart valves, whereas those at the ventricle generate coronary arteries. These observations document that the endocardial cells are a heterogeneous population with distinct developmental fates and functions. However, the intrinsic factors temporally regulating the endocardial ontogenic niche to distinguish some cells from the others and to specify cell fate and function are currently unknown, due at least to the lack of experiment models and methods to isolate and interrogate individual endocardial cells. This research program will fill this critically missing knowledge gap using a synergistic approach of mouse genetic models, developmental heart anatomy, advanced single cell transcriptome technology, and systems biology to study the expression profiles of individual cells in order to understand cell heterogeneity and uncover subpopulations. The study will lead to the identification of potential regulators for cell fate determination and for initiating coronary angiogenesis, which can be further validated and functionally confirmed. We have generated genetic mouse models that allow us to isolate ventricular versus valve endocardial cells at different ontogenic stages. In this research program we will use single cell sorting and RNA-seq on these models to identify the genetic factors differentially expressed by subpopulations of endocardial cells at valve- or coronary-forming regions at different stages (Aim 1). We will then use integrated morphologic, cellular, and molecular analysis to reveal their potential functions in heart valve formation and coronary artery development (Aim 2). The new information will shed light on endocardial biology in heart development and disease.

摘要：本研究项目的目标是确定定义个体发育生态位的特征基因。使用小鼠的心脏心内膜细胞生成关键的心脏结构、瓣膜和冠状动脉遗传学和单细胞转录组学。我们之前的研究表明，心脏的心内膜细胞是心脏瓣膜和冠状动脉的祖细胞，来自不同的心脏区域和不同的位置通过不同的机制分阶段进行。心脏流出道和房室管的心内膜细胞产生心脏瓣膜，而心室的瓣膜产生冠状动脉。这些观察证明心内膜细胞是具有不同发育命运的异质群体，并且功能。然而，内在因素暂时调节心内膜个体生态位来区分一些细胞与其他细胞的区别以及具体细胞命运和功能目前尚不清楚，至少是由于缺乏分离和询问单个心内膜细胞的实验模型和方法。本研究计划将利用小鼠遗传模型的协同方法填补这一严重缺失的知识空白，发育心脏解剖学、先进的单细胞转录组技术和系统生物学来研究单个细胞的表达谱，以了解细胞异质性并揭示亚群。这研究将导致确定细胞命运决定和启动冠状动脉的潜在调节剂血管生成，可以进一步验证和功能确认。我们已经培育出基因小鼠模型使我们能够在不同个体发生阶段分离心室与瓣膜心内膜细胞。在这个研究计划，我们将在这些模型上使用单细胞分选和 RNA-seq 来识别遗传因素瓣膜或冠状动脉形成区域的心内膜细胞亚群在不同时间有差异表达阶段（目标 1）。然后，我们将使用综合形态学、细胞和分子分析来揭示它们的心脏瓣膜形成和冠状动脉发育的潜在功能（目标 2）。新信息将阐明心脏发育和疾病中的心内膜生物学。