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 该五年提案的目的是为 申请人在心脏发展中的基础科学家过渡到独立的学术地位， 再生。长期目标是了解房屋CM谱系发展的基本机制 和与心房有关的先天性心脏病。申请人在单细胞基因上具有强大的背景 表达分析，组织细胞培养和小鼠胚胎发育，在所有设施中都在板凳上 单细胞分析，组织细胞培养和小鼠胚胎解剖的各个方面。这个职业发展计划 （K99阶段）将在胚胎组织学分析，CHIP-seq数据分析和心脏方面提供其他培训 转基因小鼠胚胎的表型分析，以研究促进RA信号的机制 心房CM谱系发展。申请人还将在 斯坦福大学的专业领域，例如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从事独立调查员的富有成效的职业。在 MD博士Sean Wu博士的专家指导以及集会咨询委员会（Querternous博士， Ruiz-Lozano博士，Krasnow博士，Bernstein博士和Martin博士），申请人将获得必要的指导 以及实现这些目标并在R00阶段有效过渡到独立性的资源。 心房CM谱系的发展是一种基本的细胞过程，据报道，视黄酸（RA）信号传导为 成为促进这一过程的关键途径，但是潜在的细胞和分子机制仍然是 在很大程度上不清楚。申请人开发了单个细胞后代测定法，以分析谱系规范 心脏祖细胞，并鉴定出具有单细胞RNA的高度有希望的心房特异性基因的列表 测序。在此提案中，他将使用有条件的基因敲除小鼠，单细胞后代测定，RA信号传导 报告测定，基因过表达或下调测定以及CHIP-SEQ技术以分析细胞 RA信号传导的分子机制在促进心房CM谱系发展中。结果 预计研究将提供有关与房组织相关的先天性心脏病的机制的见解。 他的具体目的是：AIM 1（K99）：定义促进心房CM中RA信号的细胞机制 血统的发展。 AIM 2（R00）：定义促进心房CM中RA信号的分子机制 血统的发展。