Mitochondrial electron transport dysfunction: Dissecting pathomechanisms

线粒体电子传递功能障碍：剖析病理机制

• 批准号: 10679988
• 负责人: Volker Hans Haase
• 金额: $ 26.25万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679988
• 关键词: Address; Adult; Aging; Amino Acids; Animal Genetics; Animal Model; Apoptosis; Applications Grants; Binding Proteins; Carbon; Cell Separation; Cells; Chronic; Chronic Disease; Chronic Kidney Failure; Ciona intestinalis; Citric Acid Cycle; Coenzyme Q10; Complex; Cre lox recombination system; Cytosol; Data; Degenerative Disorder; Development; Diabetic Nephropathy; Disease; Electron Transport; Electron Transport Complex III; Electrons; Epigenetic Process; Epithelial Cells; Epithelium; Fatty Acids; Fibrosis; Functional disorder; Future; Gene Expression Regulation; Generations; Genetic; Genetic Models; Glucose; Grant; Hypoxia Inducible Factor; In Vitro; Inflammation; Injury to Kidney; Kidney; Kidney Diseases; Knock-out; Knockout Mice; Knowledge; Laboratories; Limb structure; Link; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Modeling; Mus; Nephrons; Organ; Organelles; Oxidases; Oxidative Phosphorylation; Oxygen; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Physiology; Play; Prevention; Process; Production; Proliferating; Public Health; Reactive Oxygen Species; Research Project Grants; Research Proposals; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Superoxides; Tetracyclines; Therapeutic; Thick; Three-Dimensional Imaging; Tissue imaging; Tricarboxylic Acids; Urochordata; age related; alternative oxidase; amino acid metabolism; cell injury; cell type; cytochrome c; electron energy; feature detection; genetic approach; in vivo; insight; kidney fibrosis; macromolecule; mass spectrometric imaging; mitochondrial dysfunction; mouse model; novel; public health relevance; renal epithelium; superresolution microscopy; ubiquinol; ubiquinone-binding proteins

Dysregulation of mitochondrial (mt) electron transport is a well-recognized feature of the aging process and promotes cellular injury, inflammation, and organ fibrosis. Many chronic diseases, including chronic kidney diseases, are associated with mt electron transport dysfunction, underscoring its importance in organ pathogenesis. In addition to ATP production via electron transport-linked oxidative phosphorylation, mitochondria operate as signaling organelles, in which electron transport intersects with multiple metabolic pathways including the tricarboxylic acid (TCA) cycle, amino acid, fatty acid, glucose and one carbon metabolism. Furthermore, mt electron transport generates reactive oxygen species (ROS), which act as signaling molecules that regulate important cellular pathways, such as hypoxia-inducible factor (HIF)-dependent oxygen sensing. Although highly relevant to many age-related and chronic diseases, in vivo studies investigating the mechanisms by which mt electron transport dysfunction contributes to organ pathogenesis have been confounded by the lack of adequate genetic animal models. In particular, the interconnections between mt electron transport and TCA cycle metabolism and their impact on aging and chronic disease development are only incompletely understood. The focus of this exploratory research grant application is on the development and characterization of novel genetic mouse models that address these knowledge deficits. Our laboratory has shown that mt electron transport disruption in kidney suppresses TCA cycle flux, amino acid metabolism and synthesis of macromolecules, impacting on differentiation and proliferation of renal epithelial cells in a nephron-segment specific manner. Under this grant we develop nephron segment-specific knock-out models to dissect the mechanisms by which mt electron transport dysregulation promotes kidney injury and fibrosis. Specifically, we focus on the role of mt electron transport-dependent TCA cycle dysfunction in kidney pathogenesis. Under aim 1, we characterize nephron segment-specific genetic models of mt electron transport disruption due to inactivation of subunit VII of the mt ubiquinol-cytochrome c reductase complex (mt complex III), which is known as ubiquinone-binding protein Q-binding protein QPC. Under aim 2, we reactivate mt electron flux and restore TCA cycle function in Qpc-deficient renal epithelial cells by cell type-specific expression of an alternative electron-transporting oxidase (AOX). This model will be used to characterize the pathogenic role of TCA cycle dysregulation in renal epithelial cells with mt dysfunction due to mt complex III disruption.

线粒体（mt）电子传递的失调是一个公认的衰老特征 处理并促进细胞损伤、炎症和器官纤维化。许多慢性疾病， 包括慢性肾病，与MT电子传递功能障碍有关， 强调了其在器官发病机制中的重要性。除了通过电子产生ATP外， 在与转运相关的氧化磷酸化中，线粒体作为信号细胞器发挥作用， 该电子传递与多种代谢途径相交，包括三羧酸 三羧酸（TCA）循环、氨基酸、脂肪酸、葡萄糖和一碳代谢。此外，MT 电子传递产生活性氧（ROS），其充当信号分子 调节重要的细胞通路，如缺氧诱导因子（HIF）依赖的 氧传感尽管与许多与年龄相关的慢性疾病高度相关，但在体内 研究MT电子传递功能障碍有助于 器官发病机制由于缺乏适当的遗传动物模型而受到混淆。在 特别是mt电子传递和TCA循环代谢之间的相互联系， 它们对衰老和慢性病发展的影响还不完全清楚。的 这项探索性研究资助申请的重点是开发和表征 新的遗传小鼠模型来解决这些知识缺陷。 我们的实验室已经表明，在肾脏mt电子传递中断抑制TCA循环 通量、氨基酸代谢和大分子合成，影响分化和 肾上皮细胞以肾单位段特异性方式增殖。在这一资助下，我们 发展肾单位片段特异性敲除模型，以剖析mt 电子传递失调促进肾损伤和纤维化。具体来说，我们专注于 线粒体电子转运依赖性TCA循环功能障碍在肾脏发病机制中的作用。 在aim 1下，我们描述了mt电子传递的肾单位节段特异性遗传模型 由于mt泛喹啉-细胞色素c还原酶复合物的亚基VII失活引起的破坏 (mt复合物III），被称为泛醌结合蛋白Q结合蛋白QPC。下 目的2，我们重新激活mt电子流，恢复TCA循环功能，在Qpc缺陷的肾， 上皮细胞通过细胞类型特异性表达替代电子转运氧化酶 （AOX）.该模型将用于描述TCA循环失调的致病作用 在由于mt复合物III破坏而具有mt功能障碍的肾上皮细胞中。