Cilia in Heart Development and Disease

纤毛在心脏发育和疾病中的作用

• 批准号: 10353389
• 负责人: MARTINA BRUECKNER
• 金额: $ 75.03万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-15 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10353389
• 关键词: Address; Adult; Affect; Biological; Candidate Disease Gene; Cardiac; Cardiac development; Cells; Cilia; Clinical; Clinical Research; Congenital Abnormality; Contracts; Data; Development; Embryo; Environment; Epigenetic Process; Etiology; Functional disorder; Genes; Genetic; Genetic Transcription; Genomics; Goals; Heart; Heart Diseases; Heart Valves; Histone H2B; Human; Infant; Lead; Left; Link; Molecular; Monoubiquitination; Morphogenesis; Mus; Mutation; Organ; Outcome; Patients; Pediatric Cardiac Genomics Consortium; Public Health; Research Personnel; Role; Signal Transduction; Structure; Testing; Translating; Ventricular; Work; Zebrafish; cardiogenesis; chromatin remodeling; cohort; comorbidity; congenital heart disorder; experimental study; extracellular; genomic data; improved; induced pluripotent stem cell derived cardiomyocytes; live cell imaging; mechanical force; mouse development; optogenetics; personalized medicine; single-cell RNA sequencing; zebrafish development

Congenital Heart Disease (CHD) is the most common birth defect affecting 1% of all live born infants. While ~90% of patients with CHD survive into adulthood, there are many comorbidities that make CHD an increasingly significant public health problem. Genomic analyses of large cohorts of CHD patients have identified a significant genetic contribution to CHD, but the link between etiology and clinical outcome remains an important question. When I began my search for the cause of CHD, I identified the cilium as being central to left-right (LR) axis and cardiac development, and most recently, as part of the Pediatric Cardiac Genomics Consortium (PCGC), identified significant contributions from mutations affecting cilia and chromatin remodeling genes to human CHD. However, the question of how cilia dysfunction precisely influences CHD remains unanswered. I have assembled a group of co-investigators with expertise in mouse and zebrafish development, live-cell imaging, optogenetics and genomics to take a multi-pronged approach to understanding the central role of the cilium in heart development with the long-term goal of leveraging this data with ongoing genomic and clinical studies to improve clinical outcomes of CHD. First, we will resolve the long-standing question of how cilia instruct cardiac LR asymmetry. We will use single-cell RNAseq to define the cellular composition of the left-right organizer (LRO), a transient ciliated organ that is essential for instructing cardiac asymmetry. We will then establish the molecular mechanism linking cilia signaling at the LRO to cardiac LR development in mouse and zebrafish embryos. Together these experiments will uncover the mechanism by which an embryo determines LR asymmetry, and provide gene sets that will inform the search for human CHD candidate genes. Second, we will investigate the role of cilia in cardiac valve formation. We have found dynamic, flow-sensitive cilia in the presumptive atrio-ventricular valve region of the zebrafish heart, and will test the hypothesis that valve specification is driven by the mechanical forces occurring at interfaces between differentially contracting chambers, and that valve cilia are the mechanotransducers leading to changes in transcription of klf2/klf4 and downstream valve morphogenesis. Third, we will unravel the mechanism by which epigenetic factors influence cardiac development. The important role of chromatin remodeling genes in human CHD has raised the question whether any of these transcriptionally regulate cilia in heart development. We have already found that histone H2B monoubiquitination (H2BUb1) transcriptionally regulates cilia function at the LRO. We are now testing how H2BUb1 affects cardiac development in mouse embryos and human iPSC- derived cardiomyocytes through cilia-dependent and/or cilia-independent mechanism(s). Our long-term goal of translating the basic biologic and genomic data to clinical impact will be addressed by returning mechanistic and genetic data from our cilia work to the PCGC CHD genomics project so that the resulting discoveries lead to personalized medicine for patients with CHD. !

先天性心脏病（CHD）是影响所有活出生婴儿1％的最常见的先天缺陷。尽管 〜90％的冠心病患者生存到成年，有许多合并症使CHD成为CHD 越来越重大的公共卫生问题。大量冠心病患者的基因组分析具有 确定了对CHD的重要遗传贡献，但病因和临床结果之间的联系仍然存在 一个重要的问题。当我开始寻找CHD原因时，我确定Cilium是 左右（LR）轴和心脏发育，最近作为儿科心脏基因组学的一部分 财团（PCGC）确定了影响纤毛和染色质重塑的突变的重要贡献 人类冠心的基因。但是，纤毛功能障碍如何精确影响冠心病的问题仍然存在 未得到答复。我已经组建了一组共同​​投资者，在鼠标和斑马鱼方面具有专业知识 开发，现场成像，光遗传学和基因组学以采用多方面的方法来理解 纤毛在心脏发育中的核心作用，其长期目标是利用这些数据 基因组和临床研究以改善冠心病的临床结果。首先，我们将解决长期的 纤毛如何指导心脏LR不对称性的问题。我们将使用单细胞RNASEQ定义细胞 左右组织者（LRO）的组成，这是一种瞬态纤毛器官，对于指导心脏至关重要 不对称。然后，我们将建立将LRO纤毛信号传导与心脏LR联系起来的分子机制 小鼠和斑马鱼胚胎的发育。这些实验将通过 哪个胚胎决定LR不对称性，并提供基因集，以告知人们对人类冠心的搜索 候选基因。其次，我们将研究纤毛在心脏瓣膜形成中的作用。我们找到了 斑马鱼心的推定心室室瓣区域中的动态，流动敏感的纤毛，并将测试 阀规范是由在界面之间发生的机械力驱动的假设 差分收缩的室，而阀纤毛是机械转换器，导致变化 KLF2/KLF4和下游瓣膜形态发生的转录。第三，我们将揭开该机制 表观遗传因素会影响心脏发展。染色质重塑基因在人类中的重要作用 CHD提出了一个问题，这些转录是否在心脏发育中调节纤毛。我们 已经发现组蛋白H2B单泛素化（H2BUB1）在转录中调节CILIA功能 LRO。我们现在正在测试H2BUB1如何影响小鼠胚胎和人IPSC-的心脏发展 通过纤毛依赖性和/或与纤毛独立的机制（S）衍生的心肌细胞。我们的长期目标 将基本的生物学和基因组数据转换为临床影响，将通过返回机理来解决 以及从纤毛工作到PCGC CHD基因组学项目的遗传数据，以使结果发现率很高 给冠心病患者的个性化医学。 呢