Role of Neddylation in Cardiac Development

Neddylation 在心脏发育中的作用

• 批准号: 9560614
• 负责人: Rodney Eric Littlejohn
• 金额: $ 3.85万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 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/9560614
• 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; 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

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.

项目摘要 先天性心脏病仍然是最常见的先天缺陷，也是与先天缺陷有关的主要原因 世界上的死亡，至少部分源于遗传缺陷，改变了发展和成熟 心脏。尽管取得了长足的进步，但对基本理解仍然存在巨大的知识差距 控制此过程的机制。而心脏特异性转录因子的角色在心脏中的作用 已经仔细研究了开发，蛋白质修饰剂在此过程中的重要性尚未 认可。该项目的总体目标是揭示NEDD8的基本作用，NEDD8是一种新颖的泛素样 蛋白质修饰剂，心脏发育和成熟。 NEDD8以类似的方式修饰蛋白质靶标 泛素化，称为Neddylation。 Neddylation需要NEDD8特异性E1，E2和E3酶。通过 调节蛋白质靶标的功能，Neddylation参与了各种细胞过程和 病理生理事件。但是，尼迪利是否在心脏发展中起任何作用 未知。通过靶向唯一的NEDD8 E1激活酶（NAE1）的亚基，我们的初步研究发现了A 尼迪化学在维持新生儿心脏的结构完整性和功能中的关键作用。重要性 心脏中的neddylation与其在调节河马信号的调节中的新功能是一种保守的 以及控制心脏发育的关键途径。因此，该项目是为了检验中心假设 NAE1通过在YAP依赖性中维持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。 AIM 1将确定功能重要性 胚胎心脏发育中的neddylation。使用新创建的NAE1敲除鼠标线，我们将 彻底表征了NAE1缺乏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分化和缺陷（如果有）链接 增殖。此外，我们将确定在发育中的Neddylation调节下的新途径 心。在AIM 2中，我们将描述Neddylation控制CM增殖的分子机制， 专注于河马路径。我们将确定NAE1缺乏对Hippo-YAP信号的影响 Neddylation如何控制这一途径的体内和解剖分子机制。我们还将测试是否 YAP信号的重新激活挽救了NAE1失活诱导的CM增殖停滞。给定广泛的 收集的初步数据，强大而经验丰富的指导团队以及培训的高素质 计划，该提案可能会证明是成功的。这项研究的完成将为申请人综合 在心血管生物学的各个方面进行培训，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。拟议的研究将首次建立必不可少的 Neddylation在心脏发展中的作用，提供了冠心病发展的新型机制 全球与出生有关的发病率和死亡率的最重要原因。