Role of Neddylation in Cardiac Development

Neddylation 在心脏发育中的作用

基本信息

批准号：
9766369
负责人：
Rodney Eric Littlejohn
金额：
$ 3.9万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9766369
关键词：
Affinity Binding Biological Process Biology Birth Caliber Cardiac Cardiac Myocytes Cardiac development Cardiotoxicity Cardiovascular Diseases Cardiovascular system Cell Cycle Cell physiology Cessation of life Clinical Trials Congenital Abnormality DNA Data Defect Development Embryo Embryonic Heart Enzymes Event Genetic Goals Heart Heart failure In Vitro Investigation Knock-out Knockout Mice Knowledge Link Mentors Molecular Molecular and Cellular Biology Morbidity - disease rate Mus Muscle Cells Mutation Neonatal Pathway interactions Peptide Hydrolases Perinatal Perinatal mortality demographics Phenotype Physicians Pilot Projects Play Process Proteins Regulation Role Scientist Series Signal Transduction Signaling Protein Stem cells Structure Testing Time Training Transcription Coactivator Ubiquitin Like Proteins Ubiquitination Ventricular cardiogenesis career clinically relevant congenital heart disorder enzyme deficiency experience in vivo inhibitor/antagonist insight lipid biosynthesis mortality mouse model neurodevelopment novel protein function transcription factor

项目摘要

PROJECT SUMMARY Congenital heart disease remains the most common birth defect and the major cause of birth defect-related deaths in the world, and originates at least in part from genetic defects altering the development and maturation of the heart. Despite substantial progress, there are still huge knowledge gaps in basic understanding of mechanisms controlling this process. While the roles of key cardiac-specific transcription factors in heart development have been carefully studied, the significance of protein modifiers in this process has not been recognized. The overall goal of this project is to uncover the essential role of NEDD8, a novel ubiquitin-like protein modifier, in heart development and maturation. NEDD8 modifies protein targets in a way similar to ubiquitination, termed neddylation. Neddylation requires NEDD8 specific E1, E2, and E3 enzymes. Through regulating the function of protein targets, neddylation participates in diverse cellular processes and pathophysiological events. However, whether neddylation plays any role in cardiac development is completely unknown. By targeting a subunit of the only NEDD8 E1 activating enzyme (NAE1), our pilot studies uncover a critical role of neddylation in maintaining the structural integrity and function of neonatal hearts. The importance of neddylation in the heart is connected to its novel function in regulation of Hippo-YAP signaling, a conserved and crucial pathway that controls heart development. Therefore, this project is to test the central hypothesis that NAE1 regulates embryonic cardiac development through sustaining CM proliferation in a YAP-dependent manner. Two specific aims are proposed to test this hypothesis. Aim 1 will establish the functional importance of neddylation in embryonic heart development. Using a newly created NAE1 knockout mouse line, we will thoroughly characterize the impact of NAE1 deficiency on a series of concordant and sequential cardiac morphogenic events. The cardiac phenotype will be linked to defects, if any, in CM differentiation and proliferation. Moreover, we will identify novel pathways under the regulation of neddylation in the developing heart. In Aim 2, we will delineate molecular mechanisms by which neddylation controls CM proliferation, with a focus on the Hippo-YAP pathway. We will determine the effect of NAE1 deficiency on Hippo-YAP signaling in vivo and dissect molecular mechanisms of how neddylation controls this pathway. We will also test whether reactivation of YAP signaling rescues NAE1 inactivation-induced CM proliferation arrest. Given the extensive preliminary data gathered, the strong and experienced mentoring team, as well as the high caliber of the training plan, this proposal is likely to prove successful. Completion of this study will provide the applicant comprehensive training in various aspects of cardiovascular biology, which forms the basis of setting his long-term career goal of becoming an independent physician scientist. The proposed study will, for the first time, establish the essential role of neddylation in cardiac development, offering novel mechanisms underlying the development of CHD, one of the most important causes of birth-related morbidity and mortality worldwide.

项目概要先天性心脏病仍然是最常见的出生缺陷，也是出生缺陷相关的主要原因世界上的死亡人数，至少部分源于改变发育和成熟的基因缺陷的心。尽管取得了重大进展，但在基本理解方面仍然存在巨大的知识差距控制这一过程的机制。而关键的心脏特异性转录因子在心脏中的作用的发展已经被仔细研究过，但蛋白质修饰剂在此过程中的重要性尚未得到证实。认可。该项目的总体目标是揭示 NEDD8 的重要作用，NEDD8 是一种新型的类泛素蛋白质调节剂，在心脏发育和成熟中。 NEDD8 以类似于泛素化，称为neddylation。 Neddylation 需要 NEDD8 特定的 E1、E2 和 E3 酶。通过 neddylation 调节蛋白质靶标的功能，参与多种细胞过程病理生理事件。然而，neddylation 是否在心脏发育中发挥作用尚不清楚。未知。通过针对唯一 NEDD8 E1 激活酶 (NAE1) 的亚基，我们的初步研究揭示了 neddylation 在维持新生儿心脏结构完整性和功能中的关键作用。重要性心脏中的neddylation与其调节Hippo-YAP信号传导的新功能有关，Hippo-YAP信号传导是一种保守的信号传导和控制心脏发育的关键途径。因此，这个项目是为了检验中心假设： NAE1 通过维持 YAP 依赖性的 CM 增殖来调节胚胎心脏发育方式。提出了两个具体目标来检验这一假设。目标 1 将确定以下功能的重要性胚胎心脏发育中的neddylation。使用新创建的 NAE1 敲除小鼠品系，我们将彻底描述 NAE1 缺陷对一系列一致和顺序心脏的影响形态发生事件。心脏表型将与 CM 分化和发育缺陷（如果有的话）相关。增殖。此外，我们将在发展中国家的 neddylation 调控下确定新的途径。心。在目标 2 中，我们将描述 neddylation 控制 CM 增殖的分子机制，其中重点关注 Hippo-YAP 通路。我们将确定 NAE1 缺陷对 Hippo-YAP 信号传导的影响体内并剖析 neddylation 如何控制该途径的分子机制。我们还将测试是否 YAP 信号的重新激活可挽救 NAE1 失活诱导的 CM 增殖停滞。鉴于广泛初步数据收集、强大且经验丰富的导师团队以及高水平的培训计划，这个提议很可能会成功。完成本研究将为申请人提供全面的心血管生物学各个方面的培训，这是设定长期职业目标的基础成为一名独立的医师科学家。拟议的研究将首次确定必要的 neddylation 在心脏发育中的作用，提供了 CHD 发展的新机制，其中之一是全世界出生相关发病率和死亡率的最重要原因。