Exploring the mechanisms of dysfunctional mitochondrial quality control in cerebrovascular disease and the aging brain

探索脑血管疾病和大脑老化中线粒体质量控制功能失调的机制

基本信息

批准号：
10560158
负责人：
Garrett McGuire Fogo
金额：
$ 4.06万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10560158
关键词：
Affect Aging Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Architecture Bioenergetics Biogenesis Biology Blood flow Brain Cause of Death Cell Death Cells Cerebral Ischemia Cerebrovascular Disorders Characteristics Classification Complex Computational Technique Computer Models Development Disease Elements Ensure Equilibrium Event Exposure to Functional disorder Future Glucose Homeostasis Image Imaging Device Impaired cognition In Vitro Individual Injury Investigation Ischemia Knock-out Machine Learning Mammalian Cell Mediating Metabolic dysfunction Methods Mitochondria Mitochondrial Proteins Modeling Molecular Molecular Computations Morphology Mus Nature Nerve Degeneration Nervous System Trauma Neurodegenerative Disorders Neurons Nutrient Oxidative Stress Oxygen Parkinson Disease Pathologic Pathology Pattern Phase Phenotype Physiological Process Production Protein Dynamics Proteomics Quality Control Reperfusion Injury Reperfusion Therapy Reporter Reproducibility Research Respiration Risk Factors Role Swelling Synapses System Technical Expertise Technology Testing Therapeutic Therapeutic Intervention Time TimeLine Transgenic Organisms United States Work active control age related age related neurodegeneration aging brain analysis pipeline base biological adaptation to stress cerebrovascular computerized tools conditional knockout cost deprivation disability experimental study in silico in vitro Model innovation live cell imaging mitochondrial dysfunction nervous system disorder neurological pathology normal aging novel oxidation pre-doctoral preservation proteostasis real-time images simulation temporal measurement tool

项目摘要

PROJECT SUMMARY Mitochondrial dysfunction is a prominent element of many leading causes of disability, namely cerebrovascular and neurodegenerative diseases. The function and stability of mitochondria are tightly regulated by the mechanisms of mitochondrial dynamics and quality control (QC). The dynamic nature of mitochondria is maintained by the balancing forces of fission and fusion. These processes of mitochondrial dynamics operate to preserve the functional architecture of the mitochondrial network. The mechanisms of mitochondrial QC, including mitophagy, proteostasis, and biogenesis, work to regulate the components of the mitochondrial network through synthesis and degradation. These forces actively control the functionality of the mitochondrial network to ensure efficient energy production. In cerebral ischemia/reperfusion (I/R) injury, the processes of mitochondrial dynamics and QC become dysregulated, contributing to metabolic dysfunction and neurological damage. The F99 phase of this proposal aims to identify the phases of disrupted mitochondrial dynamics and QC in cerebral I/R injury, and their respective molecular mechanisms. Utilizing advanced technologies related to machine learning, computational modeling, and live cell imaging, I have created an agent-based model of mitochondrial dynamics for these investigations. This model allows for the simulation of the dynamic actions of individual mitochondrial units to culminate in the complex patterns normally observed in mammalian cells. Live cell imaging of mouse primary cortical neurons from novel transgenic reporter lines (i.e., MitoTimer, MitoQC) and conditional knockout lines will be utilized to observe the respective contributions of individual dynamics proteins to the patterns of mitochondrial morphology. Knockout neurons will be exposed to oxygen glucose deprivation (OGD), an in vitro model of I/R injury, and mitochondrial parameters (i.e., morphology, oxidation) will be imaged in real time to generate a mechanistic timeline of mitochondrial dynamics. These live cell recordings will be used to optimize and expand our agent-based model to allow for in silico experimental manipulation of mitochondrial proteins. Our expanded model will have the ability to test hypotheses regarding the basal and pathological rates of mitochondrial dynamics and quality control, as well as inform future experiments with decreased costs and increased efficiency. In the K00 phase of this proposal, I will transition from studying mitochondrial quality control in I/R to its study in neurodegeneration. Utilizing the technical skills acquired in the predoctoral phase, I will investigate age-related changes in mitochondrial proteostasis and critical long-lived mitochondrial proteins at the synapse. The K00 phase aims to determine how aging affects the turnover of synaptic mitochondrial proteins, with specific emphasis on the roles of intramitochondrial proteostasis and the integrated stress response. I intend to determine the contribution of mitochondrial proteostasis to synaptic stability and related neurodegeneration. This work has significant implications in aging and neurodegeneration research, as synaptic loss and mitochondrial dysfunction are both hallmarks of age-related neurological diseases.

项目概要线粒体功能障碍是导致残疾的许多主要原因的一个突出因素，即脑血管疾病和神经退行性疾病。线粒体的功能和稳定性受到线粒体的严格调控线粒体动力学和质量控制（QC）机制。线粒体的动态性质是由裂变和聚变的平衡力维持。线粒体动力学的这些过程的作用是保留线粒体网络的功能结构。线粒体QC的机制，包括线粒体自噬、蛋白质稳态和生物发生，致力于调节线粒体网络的组成部分通过合成和降解。这些力量主动控制线粒体网络的功能以确保高效的能源生产。在脑缺血/再灌注（I/R）损伤中，线粒体的过程动力学和质量控制失调，导致代谢功能障碍和神经损伤。这该提案的 F99 阶段旨在确定大脑中线粒体动力学和 QC 被破坏的阶段 I/R 损伤及其各自的分子机制。利用与机器相关的先进技术通过学习、计算建模和活细胞成像，我创建了一个基于代理的线粒体模型这些调查的动态。该模型可以模拟个体的动态行为线粒体单位最终形成通常在哺乳动物细胞中观察到的复杂模式。活细胞成像来自新型转基因报告系（即 MitoTimer、MitoQC）和条件条件的小鼠原代皮质神经元敲除系将用于观察各个动态蛋白对线粒体形态的模式。敲除神经元将暴露于氧糖剥夺（OGD）， I/R 损伤的体外模型和线粒体参数（即形态、氧化）将实时成像是时候生成线粒体动力学的机械时间表了。这些活细胞记录将用于优化和扩展我们基于代理的模型，以允许对线粒体进行计算机实验操作蛋白质。我们的扩展模型将能够检验有关基础率和病理率的假设线粒体动力学和质量控制，并为未来的实验提供降低成本和提高效率。在这个提案的K00阶段，我将从研究线粒体质量控制转向从事 I/R 神经退行性疾病研究。利用博士前阶段获得的技术技能，我将研究线粒体蛋白质稳态和关键长寿线粒体蛋白质与年龄相关的变化突触。 K00 阶段旨在确定衰老如何影响突触线粒体蛋白的周转，特别强调线粒体内蛋白质稳态和综合应激反应的作用。我打算确定线粒体蛋白质稳态对突触稳定性和相关神经变性的贡献。这项工作对衰老和神经退行性疾病研究具有重要意义，因为突触丢失和线粒体功能障碍是与年龄相关的神经系统疾病的两个标志。