Deciphering the roles of Nfatc1 in developmental coronary angiogenesis

解读 Nfatc1 在发育性冠状动脉血管生成中的作用

基本信息

批准号：
9276779
负责人：
BIN ZHOU
金额：
$ 41.75万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9276779
关键词：
Alleles Animal Model Basic Science Bioinformatics Biological Assay Blood Vessels Cardiac Cardiovascular system Cell Shape Cell Therapy Cells Cessation of life ChIP-seq Coronary Coronary Arteriosclerosis Coronary Circulation Coronary Vessel Anomalies Coronary artery Coronary heart disease Defect Development Developmental Biology Distal Embryo Embryonic Development Endocardium Endothelial Cells Etiology Future Gene Expression Genes Genetic Goals Heart Heart failure Hematopoietic stem cells Immunofluorescence Immunologic In Situ In Vitro Invaded Knock-out Light Molecular Molecular Biology Mus Muscle Cells Mutagenesis Mutant Strains Mice Myocardium Natural regeneration Organ Outcome Pathogenesis Pathway interactions Process Proliferating Protein Isoforms Recruitment Activity Regulation Regulator Genes Research Research Project Grants Rest Role Signal Transduction Stem cells Sudden Death Systems Biology Testing Therapeutic Time Tissues Transcription Repressor/Corepressor Transcriptional Regulation Translating Valsalva sinus Vascular Endothelial Cell Vascular Endothelial Growth Factors Vascular Smooth Muscle Ventricular Ventricular Function Vertebrates angiogenesis base cardiac angiogenesis clinical practice design genetic signature in vivo mouse model mutant notch protein novel novel therapeutics nuclear factors of activated T-cells progenitor programs transcription factor transcriptome sequencing

项目摘要

Project Summary The goal of this research project is to understand the molecular aspects of Nfatc1 in developmental coronary angiogenesis (DCA). We attempt to achieve the goal by using a synergistic approach of mouse genetics, developmental and molecular biology, and systems biology. DCA by the ventricular endocardial cells (VECs) gives rise to coronary arteries; therefore, successful completion of the proposed research will have significant translational implications by revealing the factors underlying DCA that can be relevant to the etiology of congenital coronary artery defects, one of major causes of sudden death. Better understanding of DCA will also provide the developmental basis that is critically needed for cell-based therapies for coronary heart disease, the leading cause of heart failure and death. DCA is a cardiac-specific process different from the vascular angiogenesis in other organs of body. Factors underlying this process are understudied and remain largely unknown due to the lack of proper animal models and experimental approaches. This project aims to fill the gap by studying newly generated mouse models of defective DCA and coronary arteries, focusing on an endocardial specific transcription factor, nuclear factor of activated T-cells cytoplasmic 1 (Nfatc1) in the regulation of a VECs specific gene regulatory network (GRN) of DCA. Preliminary studies revealed that Nfatc1 is required for maintaining the expression of genes for the hematopoietic stem cells and multi-potent cardiovascular progenitor cells and limiting the expression of genes involved in endothelial differentiation and/or specification, and knockout of Nfatc1 leads to abnormal DCA. Based on these results, we hypothesize that Nfatc1 functions as a `molecular checkpoint' for timing DCA through its transcriptional regulation of a tissue specific GRN for the differentiation of progenitor VECs into coronary arterial endothelial cells. Three Aims are designed to test this hypothesis. Aim 1 will characterize the angiogenic VEC in the developing heart by their changes in the expression of angiogenic genes and cell shape, and functional angiogenesis assays. Aim 2 is to ascertain Nfatc1 functions in DCA and determine whether it interacts with the Vegf-Notch pathway that is critical for angiogenesis using mouse genetics, and in vitro functional assays. Aim 3 will identify the Nfatc1-dependent GRN and hub genes VECs using RNA-seq/ChIP-seq and bioinformatics analysis, and confirm their roles in DCA in vivo by expression and in vitro by angiogenesis assays.

项目概要该研究项目的目标是了解 Nfatc1 在发育性冠状动脉中的分子方面血管生成（DCA）。我们尝试通过使用小鼠遗传学的协同方法来实现这一目标，发育和分子生物学以及系统生物学。心室心内膜细胞 (VEC) 的 DCA 产生冠状动脉；因此，成功完成拟议的研究将具有重大意义通过揭示与 DCA 病因学相关的潜在因素来实现转化意义先天性冠状动脉缺陷，是猝死的主要原因之一。更好地理解 DCA 将还为冠心病细胞疗法提供了急需的发展基础疾病，心力衰竭和死亡的主要原因。 DCA 是一种心脏特异性过程，不同于身体其他器官的血管生成。这一过程背后的因素尚未得到充分研究，并且仍然存在由于缺乏适当的动物模型和实验方法，很大程度上未知。该项目旨在填补通过研究新生成的 DCA 和冠状动脉缺陷小鼠模型，重点关注心内膜特异性转录因子，激活 T 细胞胞质核因子 1 (Nfatc1) DCA 的 VEC 特异性基因调控网络 (GRN) 的调控。初步研究表明 Nfatc1 是维持造血干细胞和多能细胞基因表达所必需的心血管祖细胞并限制内皮分化相关基因的表达和/或规范，以及 Nfatc1 的敲除会导致 DCA 异常。根据这些结果，我们假设 Nfatc1 作为“分子检查点”，通过转录调节 DCA 的时间组织特异性 GRN，用于将祖 VEC 分化为冠状动脉内皮细胞。三目的旨在检验这一假设。目标 1 将表征发育中心脏中的血管生成 VEC 通过血管生成基因表达和细胞形状的变化以及功能性血管生成测定。目标 2 是确定 Nfatc1 在 DCA 中的功能并确定它是否与 Vegf-Notch 通路相互作用这对于使用小鼠遗传学和体外功能测定的血管生成至关重要。目标 3 将确定使用 RNA-seq/ChIP-seq 和生物信息学分析依赖 Nfatc1 的 GRN 和 hub 基因 VEC，以及通过体内表达和体外血管生成测定证实它们在 DCA 中的作用。