Cilia in Heart Development and Disease

纤毛在心脏发育和疾病中的作用

• 批准号: 10577745
• 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/10577745
• 关键词: Address; Adult; Affect; Biological; Candidate Disease Gene; Cardiac; 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; Specific qualifier value; Structure; Testing; Translating; 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 drive; 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）是最常见的出生缺陷，影响所有活产婴儿的1%。而 约90%的CHD患者可存活至成年，有许多合并症使CHD和 日益严重的公共卫生问题。对大量CHD患者进行基因组分析， 确定了一个显着的遗传因素，冠心病，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冠心病的病因时，我发现纤毛是冠心病的核心。 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 联合会（PCGC），确定了影响纤毛和染色质重塑的突变的重要贡献 人类CHD的基因然而，纤毛功能障碍如何确切地影响冠心病的问题仍然存在， 无人回应我召集了一群在老鼠和斑马鱼方面有专长的合作研究者 发展，活细胞成像，光遗传学和基因组学采取多管齐下的方法来了解 纤毛在心脏发育中的核心作用，长期目标是利用这些数据， 基因组和临床研究，以改善CHD的临床结局。首先，我们将解决长期存在的 纤毛如何指导心脏LR不对称的问题。我们将使用单细胞RNAseq来定义细胞 左-右组织器（LRO）的组成，LRO是一种短暂的纤毛器官，对指导心脏运动至关重要。 不对称。然后我们将建立连接LRO纤毛信号与心脏LR的分子机制 小鼠和斑马鱼胚胎的发育。这些实验将通过以下方式揭示其机制： 胚胎决定LR不对称性，并提供基因集，将告知人类CHD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。我们发现 动态，流动敏感的纤毛在假定房室瓣区域的斑马鱼心脏，并将测试 假设瓣膜规格由发生在 差异收缩室，和阀纤毛是机械传感器导致的变化， klf 2/klf 4的转录和下游瓣膜形态发生。第三，我们将揭示 表观遗传因素影响心脏发育。染色质重塑基因在人类细胞中的重要作用 CHD提出了一个问题，即这些基因中是否有任何一个在心脏发育中转录调节纤毛。我们 已经发现组蛋白H2 B单泛素化（H2 BUb 1）在转录水平调节纤毛功能， LRO。我们现在正在测试H2 BUb 1如何影响小鼠胚胎和人类iPSC的心脏发育。 通过纤毛依赖性和/或纤毛非依赖性机制衍生的心肌细胞。我们的长期目标是 将基本生物学和基因组数据转化为临床影响的问题将通过返回机械 和遗传数据从我们的纤毛工作到PCGC CHD基因组学项目， 为冠心病患者提供个性化治疗。 !