Mechanisms governing the differentiation and maintenance of atrial identity

心房特性分化和维持的机制

• 批准号: 10676430
• 负责人: Joshua Waxman
• 金额: $ 64.52万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676430
• 关键词: Adopted; Adult; Alleles; Arrhythmia; Atrial Heart Septal Defects; Binding Sites; Biological Assay; Cardiac; Cardiac Myocytes; Cause of Death; Child; Chromatin; Complication; Congenital Abnormality; Congenital Heart Defects; Data; Defect; Development; Differentiated Gene; EMSA; Electrophysiology (science); Embryo; Enhancers; Epigenetic Process; Etiology; FOXF1 gene; Failure; Foundations; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Epistasis; Genomics; Glean; Goals; Heart; Heart Atrium; Human; Luciferases; Maintenance; Molecular; Morbidity - disease rate; Mus; Mutation; Newborn Infant; Nuclear Hormone Receptors; Operative Surgical Procedures; Organ; Organogenesis; Orphan; Pacemakers; Patients; Prevalence; Reagent; Regulation; Regulatory Element; Repression; Signal Transduction; Sinoatrial Node; Sorting; Structural defect; Techniques; Testing; Tissues; Transgenic Organisms; Venous; Ventricular; Vertebrates; WNT Signaling Pathway; Work; Zebrafish; apoAI regulatory protein-1; cardiogenesis; chicken ovalbumin upstream promoter-transcription factor; gene regulatory network; genome editing; in vivo; loss of function; malformation; mortality; mutant; novel; novel strategies; novel therapeutics; overexpression; pharmacologic; prevent; single-cell RNA sequencing; transcription factor; transcriptomics; transdifferentiation; Adopted; Adult; Alleles; Arrhythmia; Atrial Heart Septal Defects; Binding Sites; Biological Assay; Cardiac; Cardiac Myocytes; Cause of Death; Child; Chromatin; Complication; Congenital Abnormality; Congenital Heart Defects; Data; Defect; Development; Differentiated Gene; EMSA; Electrophysiology (science); Embryo; Enhancers; Epigenetic Process; Etiology; FOXF1 gene; Failure; Foundations; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Epistasis; Genomics; Glean; Goals; Heart; Heart Atrium; Human; Luciferases; Maintenance; Molecular; Morbidity - disease rate; Mus; Mutation; Newborn Infant; Nuclear Hormone Receptors; Operative Surgical Procedures; Organ; Organogenesis; Orphan; Pacemakers; Patients; Prevalence; Reagent; Regulation; Regulatory Element; Repression; Signal Transduction; Sinoatrial Node; Sorting; Structural defect; Techniques; Testing; Tissues; Transgenic Organisms; Venous; Ventricular; Vertebrates; WNT Signaling Pathway; Work; Zebrafish; apoAI regulatory protein-1; cardiogenesis; chicken ovalbumin upstream promoter-transcription factor; gene regulatory network; genome editing; in vivo; loss of function; malformation; mortality; mutant; novel; novel strategies; novel therapeutics; overexpression; pharmacologic; prevent; single-cell RNA sequencing; transcription factor; transcriptomics; transdifferentiation

Project Summary/Abstract Appropriate differentiation and maintenance of cellular identity are required for normal development of all organs. In the heart, mutations in genes that are necessary to maintain cardiomyocyte identity are associated with structural congenital heart defects, which are the most common malformations found in newborns. Despite their frequency, the etiology of most congenital heart defects remains poorly understood. Furthermore, numerous structural congenital heart defects are associated with arrythmias. Although advances in surgical techniques have been successful in allowing patients to survive to adulthood, the surgeries do not repair arrythmias associated with the structural defects. Thus, it is essential to understand fundamental mechanisms directing normal vertebrate heart development, in order to inform us of the etiology of congenital heart defects and their associated arrythmias. A long-term goal of our lab is to understand the conserved molecular and genetic mechanisms that direct cardiac chamber size during early vertebrate development. Nr2f transcriptions factors have highly conserved requirements in vertebrate heart development. Furthermore, mutations in Nr2f genes in humans are associated with a spectrum of congenital heart defects, including atrial septal defects. This proposal will investigate fundamental mechanisms determining atrial chamber size through investigating Nr2f-dependent mechanisms controlling atrial cardiomyocyte differentiation and the maintenance of atrial cardiomyocyte identity. While requirements for Nr2f factors are well-established in atrial development, the mechanisms controlling Nr2f gene expression in atrial cardiomyocytes and by which Nr2f transcription factors direct atrial cardiomyocyte development remain poorly understood. Our work has shown that zebrafish Nr2f1a is the functional equivalent of Nr2f2 in atrial development. Our preliminary data has identified a conserved enhancer that that is sufficient to promote Nr2f1a expression in atrial cardiomyocytes zebrafish and that Nr2f1a has a previously unrecognized requirement concurrently maintaining atrial cardiomyocyte and inhibiting the acquisition of pacemaker cardiomyocyte identity. In Aim 1, we will interrogate the signals that regulate the conserved nr2f1a cis-regulatory enhancer that promotes atrial cardiomyocyte expression. In Aim 2, we will determine the temporal requirements of nr2f1a and the differentiation state of cardiomyocytes within the atria of nr2f1a mutants. In Aim 3, we will elucidate the Nr2f-dependent gene regulatory networks that repress pacemaker cardiomyocyte identity in venous atria. Our studies may provide a foundation of information that may inform us of the etiology of congenital heart defects and their associated arrythmias, which ultimately may lead to novel therapies that can prevent or ameliorate congenital heart defects and associated arrythmias in humans.

项目摘要/摘要 所有器官的正常发育都需要适当的细胞身份分化和维持。 在心脏中，维持心肌细胞身份所必需的基因突变与 结构性先天性心脏缺陷，这是新生儿中最常见的畸形。尽管他们 频率，大多数先天性心脏缺陷的病因仍然很少了解。此外，很多 结构性先天性心脏缺陷与雅利氏症有关。虽然手术技术的进步 已经成功地允许患者生存至成年，手术不修复雅利木马 与结构缺陷有关。因此，必须了解指导基本机制 正常的脊椎动物心脏发育，以告知我们先天性心脏缺陷及其 相关的芳香族。我们实验室的长期目标是了解保守的分子和遗传 在早期脊椎动物发育过程中导致心脏腔室大小的机制。 NR2F转录因子 在脊椎动物心脏发育中有高度保守的要求。此外，NR2F基因中的突变 人类与先天性心脏缺陷相关，包括心房间隔缺陷。这个建议 将研究通过研究NR2F依赖性的基本机制来确定心房腔室的大小 控制心房心肌细胞分化的机制和心房心肌细胞身份的维持。 虽然对NR2F因素的要求在心房发展中建立了良好的，但控制NR2F的机制 心房心肌细胞中的基因表达和NR2F转录因子直接心房心肌细胞 发展仍然很少理解。我们的工作表明斑马鱼NR2F1A是功能等效的 心房发育中的NR2F2。我们的初步数据已经确定了一个保守的增强器，足以 促进心房心肌细胞中的NR2F1A表达 要求同时维持心房心肌细胞并抑制起搏器的获取 心肌细胞身份。在AIM 1中，我们将询问调节保守的NR2F1A顺式调节的信号 增强子促进心房心肌细胞表达。在AIM 2中，我们将确定时间要求 NR2F1A和NR2F1A突变体心房内心肌细胞的分化状态。在AIM 3中，我们将 阐明依赖NR2F的基因调节网络，该网络抑制静脉的起搏器心肌细胞身份 心房。我们的研究可能会提供信息的基础，以告知我们先天性心脏的病因 缺陷及其相关的雅典，最终可能导致可以预防或 改善先天性心脏缺陷和人类相关的芳香族。