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.

项目摘要/摘要 先天性心脏缺陷（CHD）是最常见的先天性畸形。但是，分子 大多数CHD基础的病因仍然知之甚少。此外，即使是手术后的冠心病也可以领导 生命后来的并发症导致心律不齐，中风和过早死亡。为了发展新颖 能够防止CHD和靶向特定心血管组织的疗法，对于Garner来说至关重要 了解指导正常心脏腔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展的保守机制 脊椎动物再生期间使用的个体心脏腔和腔室特异性机制。 已知很少有信号特定于心房开发，并具有心房的特定调节剂 再生没有被理解。该提案的具体目的是通过 同步长的非编码RNA（LNCRNA）家族限制了NR2F转录因子的表达和 NR2F蛋白水平如何影响成人发育和心房再生期间心房异质性 斑马鱼。该提案中的研究与人类健康有关，因为现在进行了许多基因组分析 表明NR2F2中的突变与CHD有关，尤其是人类的ASD。而NR2F2淘汰赛 小鼠和人类干细胞的体外研究揭示了对心房中NR2F1和NR2F2的要求 开发，NR2F蛋白直接适当心房发育的机制并非完全 理解。重要的是，目前尚不了解调节NR2F的LNCRNA依赖机制 蛋白质。我们对lncRNA的分析称为AS-OCA，表明它在体内抑制了NR2F1A的翻译 哺乳动物NR2F2的功能当量。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的未识别异质性。在AIM 1中， 我们将研究AS-OCA抑制NR2F1A翻译及其保护的具体机制 在人类诱导的多能干细胞中与NR2F相关的LNCRNA家族之间的机制。在AIM 2中，我们 将检查NR2F1A和规范Wnt信号传导的要求 先前未认识到的房屋亚群的多样性和转录特征。在AIM 3中，我们将 检查心房再生和心房内NR2F1A的需求的需求 心外膜。由于NR2F转录因子在所有脊椎动物的心房发育中起着保守的作用，所以 这些研究将极大地提高我们对调节正常情况的转录后机制的理解 心房再生期间采用的脊椎动物心房发育和独特的机制。最终，这些 研究将获得知识的基础，可用于改善能够预防和 改善CHD并有效修复受伤的心脏。