Cellular and molecular mechanisms of atrial cardiomyocyte lineage commitment.

项目摘要 这一五年计划的目的是为以下人员提供综合和个性化的培训计划 申请人须转为心脏发展及心脏基础科学家的独立学术职位 再生。长期目标是了解心房CM谱系发育的潜在机制 以及与心房相关的先天性心脏病。申请者在单细胞基因方面有很强的背景 基因表达分析、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。这份职业发展计划 (K99阶段)将提供胚胎组织学分析、CHIP-SEQ数据分析和心脏 转基因小鼠胚胎表型分析探讨RA信号促进机制 心房CM谱系发育。申请者还将在以下地点接受大量非正式的教学培训 斯坦福大学在专业领域的专业发展和写作技能，这将是至关重要的 让申请者获得自主权，并作为一名独立调查员开始富有成效的职业生涯。在.之下 Sean Wu博士的专家指导，医学博士和集结的咨询委员会(Dr.Querterous， Ruiz-Lozano博士、Krasnow博士、Bernstein博士和Martin博士)，申请者将得到必要的指导 和资源来实现这些目标，并在R00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，维甲酸(RA)信号转导已被报道。 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 很大程度上还不清楚。申请人已经开发出一种单细胞子代分析方法来分析猪的谱系特征 心脏祖细胞，并用单细胞RNA鉴定了一系列极有希望的心房肌特异基因 测序。在这项提案中，他将使用条件基因敲除小鼠、单细胞后代试验、RA信号 报告试验、基因过表达或下调试验以及芯片序列技术分析细胞 RA信号促进心房CM谱系发育的分子机制。由此产生的结果 研究有望为心房组织相关先天性心脏病的发病机制提供洞察力。 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 世系发展。目的2(R00)：明确RA信号促进房颤的分子机制 世系发展。

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.