Mitochondrial electron transport dysfunction: Dissecting pathomechanisms

线粒体电子传递功能障碍：剖析病理机制

基本信息

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
项目类别：
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

项目摘要

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）电子传输的失调是衰老的公认特征过程并促进细胞损伤，炎症和器官纤维化。许多慢性疾病，包括慢性肾脏疾病，与MT电子传输功能障碍有关，强调其在器官发病机理中的重要性。除了通过电子产生ATP 传输连接的氧化磷酸化，线粒体作为信号细胞器，在哪个电子传输与包括三羧酸的多个代谢途径相交酸（TCA）循环，氨基酸，脂肪酸，葡萄糖和一种碳代谢。此外，MT 电子传输产生活性氧（ROS），它们充当信号分子调节重要的细胞途径，例如缺氧诱导因子（HIF）依赖性氧气感应。尽管与许多与年龄有关和慢性疾病高度相关，但体内研究MT电子传输功能障碍有助于缺乏足够的遗传动物模型使器官发病机理混淆了。在特别是MT电子传输与TCA循环代谢与它们对衰老和慢性疾病发展的影响只不完全了解。这该探索性研究赠款的重点是发展和表征解决这些知识缺陷的新型遗传小鼠模型。我们的实验室表明，肾脏中的MT电子传输破坏抑制了TCA周期通量，氨基酸代谢和大分子的合成，影响分化和肾上皮细胞的增殖以肾段特定的方式。根据这笔赠款，我们开发肾单段特异性敲除模型，以剖析MT的机制电子转运失调会促进肾脏损伤和纤维化。具体来说，我们专注于 MT电子传输依赖性TCA循环功能障碍在肾脏发病机理中的作用。在AIM 1下，我们表征了MT电子传输的肾单段特异性遗传模型由于MT泛素醇 - 环糖型c还原酶复合酶复合酶复合体的亚基VII的灭活而造成的破坏（MT复合物III），该蛋白质QPC被称为泛素酮结合蛋白QPC。在下面 AIM 2，我们重新激活MT电子通量并恢复QPC缺陷肾脏中的TCA循环功能替代电子传输氧化酶的细胞类型特异性表达上皮细胞（AOX）。该模型将用于表征TCA循环失调的致病作用在MT复合物III中断引起的MT功能障碍的肾上皮细胞中。