Cellular and molecular mechanisms of atrial cardiomyocyte lineage commitment.

心房心肌细胞谱系定型的细胞和分子机制。

• 批准号: 9314101
• 负责人: Guang Li
• 金额: $ 12.76万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9314101
• 关键词: Address; Advisory Committees; Area; Atrial Heart Septal Defects; Award; Binding; Biological Assay; COUP transcription factor I; Cardiac; Cardiac Myocytes; Cardiac development; Cardiovascular system; Cell physiology; Cells; Cellular biology; Complex; Critical Pathways; Data Analyses; Development; Development Plans; Diagnosis; Disease; Dissection; Doctor of Philosophy; Down-Regulation; Ebstein' s Anomaly; Embryo; Embryonic Development; FGF12 gene; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genomics; Goals; Heart; Heart Atrium; Heart Diseases; Histology; In Situ Hybridization; In Vitro; Knock-out; Knockout Mice; Knowledge; Live Birth; Mentorship; Mesoderm; Mission; Molecular; Mouse Strains; Mus; National Heart, Lung, and Blood Institute; Nuclear Receptors; Pathogenesis; Pathway interactions; Pattern; Phase; Phenotype; Positioning Attribute; Prevention; Process; Reporter; Reporting; Research; Research Personnel; Resources; Retinoic Acid Response Element; Scientist; Signal Pathway; Signal Transduction; Stem cells; Technology; Tissues; Training; Training Programs; Transgenic Mice; Tretinoin; Tube; United States National Institutes of Health; Universities; VSNL1 gene; Ventricular; Work; Writing; accurate diagnosis; apoAI regulatory protein-1; cardiac regeneration; career; career development; cell type; cellular targeting; chromatin immunoprecipitation; congenital heart disorder; differential expression; experimental study; gene synthesis; genome-wide; human disease; in vivo; inhibitor/antagonist; insight; knock-down; overexpression; progenitor; response; single cell analysis; skills; stillbirth; tissue/cell culture; transcriptome sequencing

Project Summary The purpose of this five-year proposal is to provide an integrative and personalized training program for the applicant to transition into an independent academic position as a basic scientist in cardiac development and regeneration. The long-term goal is to understand the underlying mechanisms of atrial CM lineage development and atria-related congenitial heart diseases. The applicant has a strong background in single-cell gene expression analysis, tissue cell culture, and mouse embryonic development with facility at the bench in all aspects of single-cell analysis, tissue cell culture, and mouse embryos dissection. This career development plan (K99 phase) will provide additional training in embryonic histology analysis, ChIP-seq data analysis, and cardiac phenotypic analysis of transgenic mouse embryos to investigate the mechanisms of RA signaling in promoting atrial CM lineage development. The applicant will also receive a wealth of informal and didactic training at Stanford University in specialized areas such as professional development and writing skills, which will be critical for the applicant to gain autonomy and launch a productive career as an independent investigator. Under the expert mentorship of Dr. Sean Wu, MD PhD along with the assembled advisory committee (Dr. Quertermous, Dr. Ruiz-Lozano, Dr. Krasnow, Dr. Bernstein, and Dr. Martin), the applicant will receive the necessary guidance and resources to accomplish these goals and efficiently transition to independence during the R00 phase. Atrial CM lineage development is a basic cellular process, and retinoic acid (RA) signaling has been reported to be a critical pathway in promoting this process, but the underlying cellular and molecular mechanisms are still largely unclear. The applicant has developed a single cell progeny assay to analyze the lineage specification of cardiac progenitor cells, and identified a list of highly promising atrial CM specific genes with single cell RNA sequencing. In this proposal, he will use conditional knockout mice, single-cell progeny assay, RA signaling report assay, gene overexpression or downregulation assay, and ChIP-seq technology to analyze the cellular and molecular mechanisms of RA signaling in promoting atrial CM lineage development. Results from this research are expected to provide insights into the mechanisms of atrial tissue-related congenital heart diseases. His specific aims are: Aim 1 (K99): To define the cellular mechanisms of RA signaling in promoting atrial CM lineage development. Aim 2 (R00): To define the molecular mechanisms of RA signaling in promoting atrial CM lineage development.

项目摘要 这一五年计划的目的是提供一个综合和个性化的培训计划， 申请人过渡到一个独立的学术地位，作为心脏发育的基础科学家， 再生长期的目标是了解心房CM谱系发展的潜在机制 和心房相关的先天性心脏病申请人在单细胞基因方面有很强的背景 表达分析、组织细胞培养和小鼠胚胎发育， 单细胞分析、组织细胞培养和小鼠胚胎解剖等方面。这个职业发展计划 (K99阶段）将提供胚胎组织学分析，ChIP-seq数据分析和心脏 转基因小鼠胚胎的表型分析，以研究RA信号传导促进 心房CM谱系发育。申请人还将接受丰富的非正式和教学培训， 斯坦福大学在专业领域的专业发展和写作技巧，这将是至关重要的 申请人获得自主权，并作为独立调查员开展富有成效的职业生涯。下 Sean Wu博士的专家指导，MD博士沿着与聚集的咨询委员会（Quertermous博士， 博士Ruiz-Lozano、Krasnow博士、伯恩斯坦博士和马丁博士），申请人将得到必要的指导 和资源来实现这些目标，并在R 00阶段有效地过渡到独立。 心房CM谱系发育是一个基本的细胞过程，据报道视黄酸（RA）信号转导可促进心房CM谱系发育。 是促进这一过程的关键途径，但潜在的细胞和分子机制仍然是 基本上不清楚。申请人已经开发了单细胞后代测定法来分析小鼠的谱系特化。 心脏祖细胞，并确定了一系列非常有前途的心房CM特异性基因与单细胞RNA 测序在这个提案中，他将使用条件性基因敲除小鼠、单细胞子代测定、RA信号传导 报告分析、基因过表达或下调分析以及ChIP-seq技术来分析细胞内的 以及RA信号促进心房CM谱系发育的分子机制。结果从这个 预计研究将深入了解心房组织相关先天性心脏病的机制。 他的具体目标是：目标1（K99）：确定RA信号促进心房CM的细胞机制 谱系发育目的2（R 00）：明确RA信号促进心房CM的分子机制 谱系发育