Evaluation of Mitochondrial Cardiolipin Modification in Neonatal Hypoxia/Ischemia Encephalopathy

线粒体心磷脂修饰在新生儿缺氧/缺血性脑病中的评价

• 批准号: 10679306
• 负责人: Katlynn Joy Emaus
• 金额: $ 4.08万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679306
• 关键词: Anabolism; Animal Model; Antioxidants; Apoptosis; Asphyxia Neonatorum; Automobile Driving; Biological Assay; Blood flow; Brain; Brain Hypoxia-Ischemia; Brain Injuries; Brain region; Cardiolipins; Cell Death; Cell Survival; Charge; Clinical Research; Cre-LoxP; Critical Thinking; Development; Disease model; Environment; Enzymes; Evaluation; Exposure to; Fostering; Future; Gases; Genes; Genetic; Glucose; Homeostasis; Hypoxia; Hypoxic Brain Damage; In Vitro; Infant; Injury; Ischemia; Knock-out; Lipids; Lipolysis; LoxP-flanked allele; Machine Learning; Mass Spectrum Analysis; Mentorship; Methodology; Methods; Mitochondria; Modeling; Modification; Molecular; Molecular Analysis; Morphology; Mus; Neonatal; Neurons; Outcome; Oxidative Stress; Oxygen; Perinatal; Periodicity; Phospholipase; Phospholipids; Play; Production; Protein Isoforms; Quality Control; Reactive Oxygen Species; Regulation; Reperfusion Injury; Reperfusion Therapy; Reporter; Reproducibility; Research; Research Proposals; Role; Sampling; Scientist; Specificity; Spectrometry; Structure; System; Techniques; Technology; Therapeutic; Tissues; Training; Transgenic Mice; Transgenic Organisms; Translational Research; cardiolipin synthase; clinically relevant; conditional knockout; cytochrome c; defined contribution; deprivation; experimental study; gain of function; hypoxia neonatorum; in vivo; interest; ion mobility; lipidomics; loss of function; machine learning pipeline; mitochondrial dysfunction; monolysocardiolipin; mortality; mouse model; neonatal brain; neonatal hypoxic-ischemic brain injury; neuron loss; novel; overexpression; oxidative damage; particle; peroxidation; phospholipase inhibitor; porcine model; prevent; pup; therapeutic target

Project Summary Neonatal hypoxia/ischemia can result in severe damage to the infant brain. Reestablishing blood flow and oxygen delivery (reperfusion) is crucial for survival. However, reperfusion induces an accumulation of reactive oxygen species (ROS) produced by the mitochondria, culminating in oxidative stress and irreversible tissue damage. Current studies suggest that cardiolipin (CL) and its remodeling via oxidative injury to monolysocardiolipin (MLCL) directly participate in activation of mitophagy and lipidic pore formation to regulate cytochrome c release and facilitate induction of programmed cell death. The objectives of this research proposal are to investigate of the roles of CL and CL modification as molecular mechanisms in hypoxic brain injury. Aim 1 will focus on establishing the clinical relevance of CL modification and investigate the role of CL modification in brain injury following HIE. Utilizing the cutting-edge technology of cyclic ion mobility spectrometry mass spectrometry (cIMS-MS) CL and its isoforms will be quantitatively analyzed following hypoxia/ischemia in a neonatal piglet model of HIE. Unlike traditional mass spectrometry, cIMS-MS separates molecules based on structure as well as mass to charge ratio (m/z) allowing detailed analysis of CL species and to directly investigate its remodeling in I/R injury. Several novel transgenic mouse-lines will also be used to manipulate CL biosynthesis and remodeling in vivo. Mouse pups will be exposed to hypoxia/ischemia, then brains will be analyzed for injury to evaluate the contribution of CL modification on outcomes following HIE. Aim 2 will further explore the mechanisms regulated by CL modification following hypoxia/ischemia, including disruption to the mitochondrial network through mitophagy and mitochondrial dynamics. Mitophagic flux will be characterized in primary culture from mice possessing the mitochondrial quality control (mitoQC) reporter and disruptions to mitochondrial morphology will be assessed with our cutting-edge machine learning based quantification methodology. Finally, the mitochondrial specific antioxidant mitoTEMPO and phospholipase inhibitors will be used in combination with our genetic mouse models to define the contribution of oxidative modification and lipolysis to CL modification following hypoxia/ischemia. Together, these experiments will establish the mechanism of post-hypoxic CL modification, interrogated the casual role of CL and MLCL in the progression of brain injury , and define potential therapeutic targets. With exceptional scientific mentorship and rigorous academic study, this research will develop a deeper understanding of the role lipidomics play in HIE. This project will allow extensive training and allow me to gain expertise in a diverse range of animal models and molecular analysis of mechanisms of brain damage. The proposed project creates a unique environment to foster the development and critical thinking of a young scientist.

项目摘要 新生儿缺氧/缺血可能会对婴儿大脑造成严重损害。重新建立血液和 氧递送（再灌注）对于存活至关重要。但是，再灌注会诱导反应性的积累 线粒体产生的氧（ROS），最终氧化应激和不可逆组织 损害。目前的研究表明，心磷脂（CL）及其通过氧化损伤进行重塑 单一心动脂（MLCL）直接参与线粒体和脂质孔的激活以调节 细胞色素C释放并促进诱导程序性细胞死亡。该研究建议的目标 将CL和CL修饰作为分子机制在低氧脑损伤中的作用。目的 1将着重于建立CL修饰的临床相关性并研究CL修饰的作用 Hie之后的脑损伤。利用循环离子迁移率光谱量的尖端技术 光谱（CIMS-MS）CL及其同工型将在A中的缺氧/缺血后进行定量分析 Hie的新生儿小猪模型。与传统的质谱法不同，CIMS-MS基于 结构以及质量与电荷比（m/z），允许对Cl物种进行详细分析，并直接研究 它在I/R伤害中进行了改造。几种新型的转基因小鼠线也将用于操纵CL生物合成 并在体内重塑。小鼠幼崽将暴露于缺氧/缺血，然后将分析损伤的大脑 评估CL修饰对HIE后结果的贡献。 AIM 2将进一步探索 缺氧/缺血后通过CL修饰调节的机制，包括对线粒体的破坏 通过线粒体和线粒体动力学网络。线粒体通量将在原发性培养物中表征 来自具有线粒体质量控制（MITOQC）记者的小鼠，并破坏了线粒体 我们的基于机器学习的量化方法将评估形态。最后， 线粒体特异性抗氧化剂介摄氏菌和磷脂酶抑制剂将与 我们的遗传小鼠模型定义了氧化修饰和脂解对CL修饰的贡献 遵循低氧/缺血。这些实验将共同建立催眠后CL的机制 修改，询问CL和MLCL在脑损伤进展中的随意作用，并定义潜力 治疗靶标。通过出色的科学指导和严格的学术研究，这项研究将 对Hie中的脂质组学作用有更深入的了解。该项目将允许广泛的培训和 允许我在各种动物模型和大脑机制的分子分析中获得专业知识 损害。拟议的项目创造了一个独特的环境，以促进对 年轻科学家。