Cyclophilin D Regulates Neonatal Cardiac Bioenergetics and Function

亲环蛋白 D 调节新生儿心脏生物能和功能

• 批准号: 9975891
• 负责人: George A Porter
• 金额: $ 50.55万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975891
• 关键词: Acetylation; Affect; Bioenergetics; Biological Assay; Biology; Biometry; Birth; Blood Pressure; Cardiac; Cardiac Myocytes; Cardiac development; Cardiomyopathies; Chaperone Protein Inhibition; Congenital Heart Defects; Coupling; Data; Development; Disease; Electron Transport; Embryo; Environment; Equilibrium; Exposure to; Future; Goals; Health; Heart; Human; Hypoxia; In Vitro; Inner mitochondrial membrane; Life; Lung diseases; Measures; Metabolism; Mitochondria; Modeling; Molecular Chaperones; Mus; Muscle Cells; Neonatal; Organelles; Output; Oxygen; Pathology; Pathway interactions; Permeability; Pharmacology; Physiological; Physiology; Play; Process; Production; Proliferating; Proteins; Publishing; Reactive Oxygen Species; Reporting; Role; Specimen; Structure; System; Techniques; Testing; base; clinically relevant; congenital heart disorder; cyclophilin D; experimental study; falls; fatty acid oxidation; genetic approach; heart function; hypoxia neonatorum; in vivo; infection risk; neonatal exposure; neonate; novel; premature; pulmonary function; targeted treatment; uptake

Birth is the most abrupt transition during life, and the neonatal heart must accommodate to this dramatic change in environment by increasing its output to the body. Exposure to higher levels of oxygen at birth likely activates intracellular pathways that allow cardiac myocytes to rapidly proliferate and then differentiate to cause the final maturation of cardiac structure and function that is required for this increased output and survival. However, major gaps in our understanding of this process remain. It is apparent that mitochondria play an important role in this process. We have found that mitochondria regulate cardiac development in the embryo and neonate and that the mitochondrial chaperone protein, cyclophilin D (CyPD), regulates changes in mitochondrial function and reactive oxygen species (ROS) production that control cardiomyocyte proliferation and differentiation. Our preliminary data have begun to define changes in this CyPD-mitochondrial-ROS-differentiation pathway that occur in the neonatal heart. In addition, these data provide novel models to dissect the mechanisms of this pathway. These findings suggest the hypothesis that increased O2 at birth initiates a rise and then fall in CyPD activity, which regulates mitochondrial function, particularly ROS production, to control neonatal myocyte proliferation and differentiation and cardiac function. The scientific premise of this proposal is supported by data discussed above, but the mechanisms involved have not been fully elucidated. Our overall goal is to use our expertise in cardiac development and mitochondrial biology to dissect the mechanisms that control this important physiologic pathway in the neonatal heart and determine if CyPD inhibition can be used to ameliorate pathology in clinically relevant models. To achieve these goals, we propose 3 Specific Aims: 1. Determine how CyPD controls the neonatal cardiac mitochondrial-ROS-differentiation pathway. 2. Determine effects of disrupting CyPD activity in the neonatal heart. 3. Determine effects of hypoxia on the neonatal CyPD- mitochondria-ROS-differentiation pathway. The proposed experiments use a novel set of pharmacologic and genetic approaches that manipulate oxygen, CyPD, inner mitochondrial membrane coupling, and ROS in the neonatal heart. Specimens will be processed using a battery of assays to measure CyPD expression, acetylation, and activity; mitochondrial structure and function, ETC activity and assembly, ROS production; myocyte proliferation and differentiation; and cardiac function. Our team has unique expertise in cardiac, developmental, and mitochondrial biology and in biostatistics and we employ novel concepts and cutting-edge techniques to study mitochondria during late cardiac development. The anticipated results will significantly change our understanding of bioenergetics in the neonatal heart and will lead to future studies that use mitochondrial targeted therapies to enhance cardiac function and cardiac myocyte differentiation in a variety of disease states in the neonatal and mature heart.

出生是一生中最突然的转变，新生儿的心脏必须适应这种戏剧性的变化 通过增加对身体的输出来改变环境。出生时可能接触较高水平的氧气 激活细胞内通路，使心肌细胞快速增殖，然后分化为 导致输出量增加所需的心脏结构和功能最终成熟 生存。然而，我们对这一过程的理解仍然存在重大差距。 显然，线粒体在此过程中发挥着重要作用。我们发现线粒体 调节胚胎和新生儿的心脏发育，线粒体伴侣蛋白， 亲环蛋白 D (CyPD)，调节线粒体功能和活性氧 (ROS) 的变化 产生控制心肌细胞增殖和分化的物质。我们的初步数据已经开始 定义新生儿心脏中发生的 CyPD-线粒体-ROS 分化途径的变化。在 此外，这些数据提供了新的模型来剖析该途径的机制。 这些发现表明出生时氧气含量增加会导致 CyPD 升高然后下降的假设 活性，调节线粒体功能，特别是 ROS 产生，以控制新生儿肌细胞 增殖和分化与心脏功能。该提案的科学前提得到以下支持： 上面讨论了数据，但所涉及的机制尚未完全阐明。我们的总体目标是使用 我们在心脏发育和线粒体生物学方面的专业知识来剖析控制这一过程的机制 新生儿心脏中重要的生理通路，并确定 CyPD 抑制是否可用于 改善临床相关模型的病理学。为了实现这些目标，我们提出 3 个具体目标： 1. 确定 CyPD 如何控制新生儿心脏线粒体 ROS 分化途径。 2. 确定 破坏新生儿心脏中 CyPD 活性的影响。 3. 确定缺氧对新生儿 CyPD-的影响 线粒体-ROS-分化途径。 拟议的实验使用了一套新颖的药理学和遗传学方法来操纵 新生儿心脏中的氧气、CyPD、线粒体内膜偶联和 ROS。标本将是 使用一系列测定法进行处理以测量 CyPD 表达、乙酰化和活性；线粒体 结构和功能、ETC活性和组装、ROS产生；心肌细胞增殖和分化； 和心脏功能。我们的团队在心脏、发育和线粒体生物学方面拥有独特的专业知识， 在生物统计学中，我们采用新颖的概念和尖端技术来研究线粒体 心脏发育。预期的结果将显着改变我们对生物能学的理解 新生儿心脏，并将导致未来使用线粒体靶向治疗来增强心脏功能的研究 新生儿和成熟心脏在各种疾病状态下的功能和心肌细胞分化。