Molecular Mechanisms of Atrial Development and Regeneration

心房发育和再生的分子机制

• 批准号: 10601607
• 负责人: Joshua Waxman
• 金额: $ 60.19万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-21 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10601607
• 关键词: Ablation; Adult; Affect; Affinity Chromatography; Area; Arrhythmia; Atrial Function; Atrial Heart Septal Defects; Biological Assay; Buffers; CRISPR/Cas technology; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cells; Cessation of life; Child; Complex; Congenital Abnormality; Congenital Heart Defects; Coupled; Development; Diagnosis; Embryo; Embryonic Atrium; Embryonic Heart; Enhancers; Epicardium; Etiology; Family; Fatigue; Foundations; Gene Expression Profile; Generations; Genes; Genetic Epistasis; Genomics; Glean; Goals; Health; Heart; Heart Atrium; Heart Injuries; Heterogeneity; Human; Hypertrophy; Immunoprecipitation; In Vitro; Individual; Infant Health; Infant Mortality; Injury; Invaded; Knockout Mice; Knowledge; Life; Mammals; Mediating; Metronidazole; Molecular; Monitor; Mutation; Natural regeneration; Operative Surgical Procedures; Play; Population; Proteins; RNA; Reagent; Repression; Ribosomes; Right ventricular structure; Role; Signal Transduction; Sorting; Specific qualifier value; Stroke; Testing; Tissues; Transcript; Transgenic Organisms; Translational Repression; Translations; Tretinoin; Untranslated RNA; Venous; Ventricular; Vertebrates; WNT Signaling Pathway; Zebrafish; apoAI regulatory protein-1; cardiac regeneration; cardiogenesis; coronary vasculature; gain of function; genome editing; human stem cells; improved; in vivo; induced pluripotent stem cell; injured; injury and repair; loss of function; malformation; mutant; novel; novel therapeutic intervention; novel therapeutics; pharmacologic; posttranscriptional; premature; prevent; progenitor; promoter; repaired; scaffold; single-cell RNA sequencing; targeted treatment; transcription factor; vertebrate embryos

Project Summary/Abstract Congenital heart defects (CHDs) are the most common congenital malformations. However, the molecular etiology underlying most CHDs remain poorly understood. Furthermore, CHDs even following surgery can lead to complications later in life that result in arrhythmias, stroke, and premature death. In order to develop novel therapies able to prevent CHDs and target therapies to specific cardiovascular tissues, it is critical to garner understanding of fundamental mechanisms directing normal cardiac chamber development and regeneration. Therefore, long-term goals of our lab are to understand conserved mechanisms that direct the development of individual cardiac chambers and chamber-specific mechanisms utilized during regeneration in vertebrates. Few signals are known to be required that specifically direct atrial development, with specific regulators of atrial regeneration not being understood. The specific aims of this proposal are to elucidate the mechanisms by which a syntenic long non-coding RNA (lncRNA) family limits the expression of Nr2f transcription factors and decipher how Nr2f protein levels affect atrial heterogeneity during development and atrial regeneration in adult zebrafish. The studies in this proposal are relevant to human health as numerous genomic analyses now indicate that mutations in Nr2f2 are associated with CHDs, in particular ASDs in humans. While Nr2f2 knockout mice and in vitro studies with human stem cells have revealed requirements for both Nr2f1 and Nr2f2 in atrial development, the mechanisms by which Nr2f proteins direct proper atrial development are not completely understood. Importantly, there is currently no understanding of lncRNA-dependent mechanisms regulating Nr2f proteins. Our analysis of a lncRNA we call as-oca shows that in vivo it represses the translation of nr2f1a, the functional equivalent of mammalian Nr2f2. Moreover, we find that Nr2f1a levels regulate previously unrecognized heterogeneity of atrial cardiomyocytes in the embryonic atrium and atrial regeneration. In Aim 1, we will examine the specific mechanism that as-oca inhibits nr2f1a translation and the conservation of this mechanism among the NR2F-associated lncRNA family in human induced pluripotent stem cells. In Aim 2, we will examine the requirements of Nr2f1a and canonical Wnt signaling in generating atrial cardiomyocyte diversity and the transcriptional signature of a previously unrecognized atrial subpopulation. In Aim 3, we will examine the requirement of the epicardium in atrial regeneration and requirement of Nr2f1a within the atrial epicardium. Because Nr2f transcription factors play conserved roles in atrial development of all vertebrates, these studies will dramatically improve our understanding of post-transcriptional mechanisms regulating normal vertebrate atrial development and unique mechanisms employed during atrial regeneration. Ultimately, these studies will garner a foundation of knowledge that can be used to improve therapies capable of preventing and ameliorating CHDs and efficiently repairing injured hearts.

项目概要/摘要 先天性心脏缺陷（CHD）是最常见的先天畸形。然而，分子 大多数先心病的病因学仍然知之甚少。此外，即使在手术后，冠心病也可能导致 导致晚年并发症，导致心律失常、中风和过早死亡。为了开发新颖 能够预防冠心病的疗法和针对特定心血管组织的靶向疗法，至关重要的是 了解指导正常心室发育和再生的基本机制。 因此，我们实验室的长期目标是了解指导发育的保守机制。 脊椎动物再生过程中使用的各个心室和心室特异性机制。 已知很少有信号需要专门指导心房发育，并且具有心房的特定调节器 再生不被理解。该提案的具体目标是通过以下方式阐明机制： 同线性长非编码 RNA (lncRNA) 家族限制 Nr2f 转录因子的表达 破译 Nr2f 蛋白水平如何影响成人发育和心房再生过程中的心房异质性 斑马鱼。该提案中的研究与人类健康相关，因为现在有大量的基因组分析 表明 Nr2f2 的突变与先心病（CHD）有关，特别是人类自闭症谱系障碍（ASD）。而Nr2f2淘汰赛 小鼠和人类干细胞的体外研究揭示了心房中 Nr2f1 和 Nr2f2 的需求 Nr2f 蛋白指导心房正常发育的机制尚不完全 明白了。重要的是，目前尚不清楚 lncRNA 依赖性调节 Nr2f 的机制 蛋白质。我们对称为 as-oca 的 lncRNA 的分析表明，它在体内抑制 nr2f1a 的翻译，nr2f1a 是 哺乳动物 Nr2f2 的功能等同物。此外，我们发现 Nr2f1a 水平先前调节 胚胎心房和心房再生中心房心肌细胞未被识别的异质性。在目标 1 中， 我们将研究as-oca抑制nr2f1a翻译的具体机制及其保守性 人类诱导多能干细胞中 NR2F 相关 lncRNA 家族的机制。在目标 2 中，我们 将检查 Nr2f1a 和经典 Wnt 信号传导在生成心房心肌细胞中的要求 以前未被识别的心房亚群的多样性和转录特征。在目标 3 中，我们将 检查心房再生中心外膜的需求以及心房内 Nr2f1a 的需求 心外膜。由于 Nr2f 转录因子在所有脊椎动物的心房发育中发挥保守作用， 这些研究将极大地提高我们对调节正常的转录后机制的理解 脊椎动物心房发育和心房再生过程中采用的独特机制。最终，这些 研究将获得知识基础，可用于改进能够预防和治疗的疗法 改善冠心病并有效修复受伤的心脏。