A systems approach to decode mitochondrial metabolite transport

解码线粒体代谢物运输的系统方法

• 批准号: 10713145
• 负责人: Hongying Shen
• 金额: $ 41.88万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713145
• 关键词: Adopted; Area; Biochemistry; Biology; Brain Diseases; Cells; Cellular biology; Family; Functional disorder; Funding Mechanisms; Genes; Glutathione; Goals; Human; In Vitro; Induced pluripotent stem cell derived neurons; Laboratories; Life; Ligands; Mediating; Metabolism; Mitochondria; Molecular; Neurons; Organelles; Process; Protein Family; Proteins; Research; Resolution; Role; Structure; Synapses; System; career; flexibility; frontier; gain of function; human disease; in vitro Model; loss of function; metabolomics; mitochondrial membrane; mitochondrial metabolism; programs; uptake

Project Summary/Abstract Mitochondria house the metabolism of all building blocks of life and are central for cellular metabolism, however, how metabolites translocate across the mitochondrial membrane is poorly understood; and the impact on neuronal cellular metabolism is yet to be explored. The mitochondrial carrier SLC25 family represents the largest protein family critical for mitochondrial metabolite transport, in which approximately 20 out of the 53 human SLC25 genes remain of unknown function. Our overarching research program is to characterize mitochondrial metabolite transport by identifying the endogenous metabolite ligands for SLC25 transporters. Recently, my laboratory applied mitochondrial metabolomics to discover the role of a poorly characterized mitochondrial transporter SLC25A39 in glutathione uptake (supported by R00). Here, building upon this recent progress, we propose to expand the strategy to other SLC25 transporters using loss-of-function and gain-of-function study in cells. As a distinct and complementary approach, we will apply sequence and structure analysis towards SLC25 transporter to guide in vitro mitochondrial metabolite uptake screen with a goal to characterize ligand recognition mechanism. To study mitochondrial metabolism in neurons, we will adopt iPSC-derived neurons as an in vitro model to profile the impact of SLC25 transporters loss on mitochondrial and synaptic metabolism. Together, our study of SLC25 transporters would reveal fundamental biology in mitochondrial metabolite transport, an emerging new frontier in cellular metabolism. Our expertise in cellular metabolism, uniquely in applying state-of-art, high- resolution mass spec metabolomics to fundamental biochemistry and cell biology, is perfectly suited for the research direction. The ESI-MIRA program enables launching my research career in cellular metabolism as well as moving into a new research area in neuronal cell biology. The flexibility in the funding mechanism allows this ambitious project to systematically decode mitochondrial metabolite transport.

项目摘要/摘要 线粒体是生命的所有组成部分的新陈代谢，是细胞新陈代谢的中心， 然而，代谢物如何通过线粒体膜转运却知之甚少；而 对神经细胞新陈代谢的影响尚不清楚。线粒体载体SLC25家族 代表对线粒体代谢物运输至关重要的最大蛋白质家族，其中大约 53个人类SLC25基因中有20个功能未知。我们的主要研究计划是 通过鉴定内源性代谢物配体来表征线粒体代谢物的转运 SLC25转运蛋白。最近，我的实验室应用线粒体代谢组学发现了一种 线粒体转运蛋白SLC25A39在谷胱甘肽摄取中的特性不佳(由R00支持)。这里, 在这一最新进展的基础上，我们建议将该战略扩展到其他使用SLC25的运输商 细胞内功能丧失和功能获得的研究。作为一种独特和相辅相成的方法，我们将 应用SLC25转运蛋白的序列和结构分析来指导体外线粒体 代谢产物摄取筛选，目的是表征配基识别机制。研究线粒体 神经元的代谢，我们将采用IPSC来源的神经元作为体外模型来描述 SLC25转运体失去线粒体和突触代谢。总之，我们对SLC25的研究 转运蛋白将揭示线粒体代谢物运输的基础生物学，这是一种新兴的新技术 细胞新陈代谢的前沿。我们在细胞新陈代谢方面的专业知识，特别是在应用最先进的、高度 将质谱仪代谢组学分解到基础生物化学和细胞生物学，非常适合于 提出了研究的方向。ESI-MIRA计划使我能够在蜂窝领域开始研究生涯 正在进入神经细胞生物学的一个新的研究领域。资金的灵活性 这种机制使得这个雄心勃勃的项目能够系统地解码线粒体代谢物的运输。