Neddylation and cardiac protein quality control

Neddylation 和心脏蛋白质量控制

• 批准号: 10064097
• 负责人: Huabo Su
• 金额: $ 45.1万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064097
• 关键词: Address; Adult; Adverse event; Animal Model; Animals; Attenuated; Biochemical; Bioenergetics; Biological; Biological Assay; Cancer Patient; Cardiac; Cardiac Myocytes; Cardiac development; Cardiac health; Cardiomyopathies; Cardiotoxicity; Cell Death; Cell physiology; Cells; Cessation of life; Clinical Trials; Congestive Heart Failure; Cullin Proteins; Data; Development; Dilated Cardiomyopathy; Disease; Enzymes; Equilibrium; Functional disorder; Funding; Goals; Grant; Guanosine Triphosphate Phosphohydrolases; Health; Heart; Heart Diseases; Heart failure; Homeostasis; Human; ISG15 gene; Impairment; Investigation; Laboratories; Maintenance; Mediating; Membrane Potentials; Mitochondria; Modification; Molecular; Morphology; Mus; Myocardial dysfunction; Neonatal; Non-compaction cardiomyopathy; Oncology; Organ; Pathologic; Pathology; Pathway interactions; Patients; Performance; Perinatal; Perinatal mortality demographics; Personal Satisfaction; Pharmacology; Phenotype; Physiology; Play; Post-Translational Protein Processing; Proteins; Proteome; Publishing; Quality Control; Regenerative capacity; Regulation; Research; Respiration; Rest; Role; Stress; Structure; Testing; Time; Tissues; Ubiquitin; Ubiquitin Like Proteins; Ubiquitination; Ventricular; cancer therapy; combat; conditional knockout; heart function; inhibitor/antagonist; innovation; insight; live cell imaging; mitochondrial dysfunction; mitochondrial fitness; mitochondrial membrane; novel; organ growth; parkin gene/protein; pre-clinical; premature; pressure; proteostasis; restoration; ubiquitin ligase

PROJECT SUMMARY Disruption of protein homeostasis causes cardiomyocyte (CM) dysfunction and death, and is increasingly recognized to play a causal role in human cardiomyopathies and heart failure. Ubiquitin (Ub) and Ub-like proteins modify diverse protein substrates and expand the functional diversity of the proteome. An extensive body of work has suggested the exciting potential of targeting these protein modifiers to combat cardiac disease. However, the functional role of NEDD8, a novel Ub-like protein, remains poorly understood in the heart. Neddylation covalently attaches NEDD8 to target proteins via NEDD8-specific E1-E2-E3 enzymes. Over the previous funding cycle, we discovered that neddylation is dysregulated in the failing hearts of patients and animal models of cardiac diseases. We further demonstrated that a balance between neddylation and deneddylation is essential to cardiac homeostasis, as evidenced by the fact that perturbation of deneddylation precipitated the heart to cardiomyopathy and heart failure, whereas blockade of neddylation in the developing heart caused ventricular noncompaction and perinatal lethality. Although it is becoming apparent that neddylation is essential for organ development, its functional importance in fully differentiated, postmitotic tissues is not clear. This grant addresses a novel linkage between neddylation and mitochondrial integrity in adult CMs. Pharmacological inhibition of neddylation promoted mitochondrial fusion, disrupted mitochondrial membrane potential and impaired mitochondrial respiration in neonatal CMs. Deletion of NAE1 (a regulatory subunit of the NEDD8 E1 enzyme) in adult CMs also resulted in mitochondrial elongation and impaired mitochondrial quality. Mechanistically, neddylation modifies various cellular proteins to regulate mitochondrial dynamics and mitophagy. Conditional knockout (KO) of NAE1 in adult mouse hearts resulted in dilated cardiomyopathy, heart failure, and ultimately premature death. In clinical trials, administration of the neddylation inhibitor MLN4924 to cancer patients elicited severe cardiotoxicity. These compelling published and preliminary data form the basis of our central hypothesis that neddylation is required for normal heart function through regulation of mitochondrial dynamics and mitophagy. Using the newly generated inducible NAE1KO mice and MLN4924, Aim 1 will establish the significance of neddylation in the adult healthy and failing heart and determine the impact of neddylation on mitochondrial function. Aim 2 will dissect the molecular basis by which neddylation controls mitochondrial dynamics and will test if normalization of mitochondrial morphology benefits the NAE1KO heart. Aim 3 will elucidate novel Parkin-independent mechanisms by which neddylation regulates mitophagy and test whether restoration of neddylation attenuates cardiac dysfunction in NAE1KO mice. This study will be the first to establish an unappreciated role for neddylation in regulating mitochondrial fitness and function in adult CMs. Moreover, it will provide novel insight into the potential cardiotoxicity of neddylation inhibitors that are currently being applied in cancer therapy, thus advancing the field of cardio-oncology.

项目总结 蛋白质稳态的破坏会导致心肌细胞(CM)功能障碍和死亡，而且这种情况越来越多 被认为在人类心肌病和心力衰竭中起因果作用。泛素(Ub)及其类似蛋白 修饰不同的蛋白质底物，扩大蛋白质组的功能多样性。内容广泛的作品 已经表明了以这些蛋白质修饰物为靶点来对抗心脏病的令人兴奋的潜力。然而， NEDD8是一种新的Ub样蛋白，其功能在心脏中仍知之甚少。去瘤作用 通过NEDD8特异性的E1-E2-E3酶将NEDD8共价连接到靶蛋白上。比之前的资金 周期中，我们发现心脏衰竭患者和动物模型的心脏代谢失调。 心脏病。我们进一步证明，平衡平衡是必要的。 对心脏动态平衡的影响，事实证明，去动力的扰动促使心脏 心肌病和心力衰竭，而在发育中的心脏阻断肾病引起的室性 致密化不全和围产儿致死性。尽管越来越明显的是，新陈代谢对器官是必不可少的 在发育过程中，其在完全分化、有丝分裂后组织中的功能重要性尚不清楚。这笔赠款解决了 成年CMS患者核变性与线粒体完整性之间的新关联。药理抑制作用 糖尿病促进线粒体融合，破坏线粒体膜电位，损害线粒体 新生儿CMS的线粒体呼吸。NEDD8 E1酶的调节亚基NAE1的缺失 成年CMS还导致线粒体延长和线粒体质量受损。从机械上讲， 核酸化修饰各种细胞蛋白质以调节线粒体动力学和有丝分裂。有条件的 成年小鼠心脏中NAE1基因敲除(KO)导致扩张型心肌病、心力衰竭和最终 过早死亡。在临床试验中，给癌症患者服用代谢抑制剂MLN4924引发了 严重的心脏毒性。这些令人信服的公布和初步数据构成了我们中心假设的基础 正常的心脏功能需要通过调节线粒体动力学和 有丝分裂。使用新产生的可诱导NAE1KO小鼠和MLN4924，Aim 1将建立 糖尿病在健康成人和心力衰竭患者中的意义及糖尿病对心力衰竭的影响 线粒体功能。目标2将剖析代谢控制线粒体的分子基础 并将测试线粒体形态的正常化是否有利于NAE1KO心脏。目标3将 阐明新的Parkin非依赖的机制，通过它调节有丝分裂和测试 糖尿病的恢复可减轻NAE1KO小鼠的心功能障碍。这项研究将是第一个建立 肾病在成人CMS中调节线粒体适合性和功能中的未被认识的作用。此外，它还 将为目前正在应用的代谢抑制剂的潜在心脏毒性提供新的见解 在癌症治疗方面，从而推动了心脏肿瘤学领域的发展。