Molecular Dissection of Mitochondria-Organelle Interactions

线粒体-细胞器相互作用的分子解剖

• 批准号: 10000604
• 负责人: Yasemin S Sancak
• 金额: $ 233.25万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10000604
• 关键词: Aging; Biochemical; Calcium Signaling; Cell Death; Cell Line; Cell membrane; Cell model; Cells; Cellular Structures; DNA biosynthesis; Development; Disease; Dissection; Goals; Immunity; Inherited; Investigation; Location; Malignant Neoplasms; Mediator of activation protein; Metabolic Diseases; Metabolism; Methods; Mitochondria; Mitochondrial DNA; Mitochondrial Diseases; Molecular; Neurodegenerative Disorders; Non-Insulin-Dependent Diabetes Mellitus; Organelles; Parkinson Disease; Pathogenesis; Physiological; Play; Post-Translational Protein Processing; Proteome; Regulation; Role; Signal Transduction; Site; base; experimental study; innovation; mitochondrial dysfunction; novel; proteostasis; targeted treatment

PROJECT SUMMARY/ ABSTRACT Mitochondria are multi-functional organelles that play central roles in metabolism, protein homeostasis, calcium signaling, immunity, and cell death. Perturbation of these mitochondrial functions is an underlying feature of several common diseases and physiological conditions such as cancer, neurodegenerative diseases and aging. Interaction of mitochondria with other organelles plays a pivotal role in regulation of mitochondrial functions. However these mitochondria-organelle interactions remain poorly understood, partly due to lack of methods that enable isolation and characterization of mitochondria that associate with different organelles. This also hinders investigation of mitochondria-organelle interactions in diseases and their contribution to disease pathogenesis. The goal of this proposal is to overcome this technical challenge in order to investigate the mechanisms of mitochondria regulation through organelle interactions in cell lines and cellular models of mitochondrial diseases. Here, I propose the development of an innovative biochemical workflow that will allow differential tagging and rapid isolation of mitochondria based on their cellular location and organellar interactions. Using this method, we will first characterize the proteome and metabolite composition of mitochondria that are in close proximity to four organelles and the plasma membrane. Next, we will investigate whether specific metabolites and protein modifications regulate mitochondrial functions at organelle contact sites. We will also determine how mitochondria- organelle interactions are altered in cellular models of mitochondrial disease and whether these interactions can be targeted for therapeutic gain. These experiments will be the first comprehensive biochemical analysis of mitochondria-organelle interactions in healthy cells and cells with mitochondrial dysfunction. Our results will lead to identification of signals that originate from different cellular structures to regulate mitochondrial functions and will elucidate their mechanisms. In addition, investigation of mitochondria-organelle interactions in cells with mitochondrial dysfunction has the potential to transform our understanding of mechanisms of mitochondrial diseases, which will lead to novel treatment approaches. The methods developed can easily be applied to investigate other inter-organellar interactions to reveal their mediators and mechanisms at unprecedented biochemical detail.

项目概要/摘要 线粒体是多功能细胞器，在新陈代谢、蛋白质稳态、钙 信号传导、免疫和细胞死亡。这些线粒体功能的扰动是 一些常见的疾病和生理状况，例如癌症、神经退行性疾病和 老化。线粒体与其他细胞器的相互作用在线粒体的调节中起着关键作用 功能。然而，这些线粒体-细胞器相互作用仍然知之甚少，部分原因是缺乏 能够分离和表征与不同细胞器相关的线粒体的方法。 这也阻碍了对疾病中线粒体-细胞器相互作用及其对疾病的贡献的研究 疾病发病机制。该提案的目标是克服这一技术挑战，以便调查 通过细胞系和细胞模型中细胞器相互作用调节线粒体的机制 线粒体疾病。在这里，我建议开发一种创新的生化工作流程，该工作流程将允许 根据线粒体的细胞位置和细胞器对线粒体进行差异标记和快速分离 互动。使用这种方法，我们将首先表征蛋白质组和代谢物组成 线粒体靠近四个细胞器和质膜。接下来我们将进行调查 特定代谢物和蛋白质修饰是否调节细胞器接触时的线粒体功能 网站。我们还将确定线粒体-细胞器相互作用在细胞模型中如何改变 线粒体疾病以及这些相互作用是否可以作为治疗收益的目标。这些实验 这将是对健康细胞中线粒体-细胞器相互作用的第一个全面的生化分析 线粒体功能障碍的细胞。我们的结果将导致识别来自不同来源的信号 调节线粒体功能的细胞结构并将阐明其机制。此外， 对线粒体功能障碍细胞中线粒体与细胞器相互作用的研究有可能 改变我们对线粒体疾病机制的理解，这将带来新的治疗方法 接近。开发的方法可以很容易地应用于研究其他细胞器间相互作用 以前所未有的生化细节揭示其介质和机制。