Illuminating Heme Trafficking and Signaling Pathways in Health and Disease

阐明健康和疾病中的血红素贩运和信号通路

• 批准号: 10614044
• 负责人: Amit Ram Reddi
• 金额: $ 37.47万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614044
• 关键词: Address; Biochemical; Biodistribution; Biological; Biology; Biophysics; Candida albicans; Cell Line; Cells; Chelating Agents; Chemicals; Disease; Eukaryotic Cell; Gene Deletion; Genetic; Health; Heme; Image; Iron; Mammalian Cell; Mediating; Metabolism; Modeling; Molecular Genetics; Mycobacterium tuberculosis; Nutritional; Pathogenesis; Pathway interactions; Physiology; Process; Proteins; Research; Saccharomyces cerevisiae; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Source; Technology; cofactor; cytotoxic; human disease; human pathogen; innovation; novel; overexpression; pathogen; ratiometric; sensor; spatiotemporal; tool; trafficking; transmission process; uptake

PROJECT SUMMARY The long-term objectives of this proposal are to elucidate the molecules and mechanisms that mediate heme trafficking and signaling in health and disease. Heme is an essential but potentially cytotoxic metallocofactor, signaling molecule, and nutritional source of iron. All heme-dependent processes require that heme is safely transported and trafficked from sites of uptake or synthesis to heme dependent or regulated proteins located throughout the cell. However, the molecules and mechanisms that mediate the mobilization of heme for trafficking and signaling are poorly understood. Further, the spatio-temporal dynamics of heme trafficking are unknown. This is in large part due to the lack of tools available to image labile heme pools relevant to it biodistribution. The Reddi lab has recently overcome these technological barriers by generating genetically encoded ratiometric fluorescent heme sensors and hemoproteomics approaches to characterize heme trafficking and signaling dynamics. This proposal will focus on integrating molecular genetic, biochemical, biophysical, cell biological, and chemical biology approaches towards elucidating: a. heme transport and trafficking; b. heme-based signal transduction; and c. heme acquisition across a variety of model eukaryotic cell lines, e.g. Saccharomyces cerevisiae and mammalian cell lines, and pathogens, e.g. Mycobacterium tuberculosis and Candida albicans. To probe intracellular heme trafficking, genetically encoded heme sensors will be used in gene deletion and over-expression screens to identify novel pathways for heme distribution and trafficking. To probe heme signaling, hemoproteomics approaches and heme chelators will be deployed to identify how heme signals are initiated, transmitted, and integrated to control metabolism and physiology. To probe heme acquisition by human pathogens, heme sensors and hemoproteomics approaches will be integrated to identify the mechanisms of heme uptake and utilization by Mycobacterium tuberculosis and Candida albicans. Altogether, the proposed research will comprehensively address the mechanisms of heme trafficking and signaling in various contexts relevant to health and disease.

项目摘要 该提案的长期目标是阐明介导血红素的分子和机制 健康和疾病中的贩运和信号。血红素是必不可少的但潜在的细胞毒性金属效应器， 信号分子和铁的营养来源。所有依赖血红素的过程都要求血红素安全 从摄取或合成部位运输并运输到依赖或调节的蛋白质的部位 在整个细胞中。但是，介导血红素动员的分子和机制 贩运和信号传导知之甚少。此外，血红素贩运的时空动力学是 未知。这在很大程度上是由于缺乏可用于图像不稳定血红素池的工具 生物分布。 Reddi实验室最近通过遗传来克服这些技术障碍 编码的比率计量荧光血红素传感器和血蛋白组学方法来表征血红素 贩运和信号动态。该建议将着重于整合分子遗传，生化， 生物物理，细胞生物学和化学生物学方法阐明：血红素运输和 贩运； b。基于血红素的信号转导；和c。跨多种真核模型的血红素获取 细胞系，例如酿酒酵母和哺乳动物细胞系以及病原体，例如分枝杆菌 结核病和白色念珠菌。为了探测细胞内血红素的运输，遗传编码的血红素传感器 将用于基因缺失和过表达筛选，以识别血红素分布和 贩运。为了探测血红素的信号传导，将采用血红素学方法和血红素螯合剂部署到 确定如何启动，传播和集成血红素信号以控制代谢和生理学。到 人类病原体，血红素传感器和血红素学方法的探测血红素将是 整合以识别结核分枝杆菌的血红素摄取和利用机制 白色念珠菌。总共提出的研究将全面解决血红素的机制 在与健康和疾病有关的各种情况下的贩运和信号。