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: 10406606
• 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/10406606
• 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; base; cofactor; cytotoxic; human disease; human pathogen; innovation; novel; overexpression; pathogen; ratiometric; sensor; spatiotemporal; tool; trafficking; 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.

项目概要 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传导。血红素是一种必需但具有潜在细胞毒性的金属辅因子， 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是安全的 从摄取或合成位点运输和贩运到位于血红素依赖或调节的蛋白质 整个细胞。然而，介导血红素动员的分子和机制 人们对贩运和信号传递知之甚少。此外，血红素贩运的时空动态是 未知。这在很大程度上是由于缺乏可用于对其相关的不稳定血红素池进行成像的工具 生物分布。 Reddi 实验室最近通过生成基因技术克服了这些技术障碍 编码比例荧光血红素传感器和表征血红素的血液蛋白质组学方法 贩运和信号动态。该提案将侧重于整合分子遗传学、生物化学、 生物物理、细胞生物学和化学生物学方法来阐明：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运输，基因编码的血红素传感器 将用于基因删除和过度表达筛选，以确定血红素分布和的新途径 贩运。为了探测血红素信号传导，将部署血液蛋白质组学方法和血红素螯合剂 确定血红素信号如何启动、传输和整合以控制新陈代谢和生理机能。到 人类病原体、血红素传感器和血液蛋白质组学方法对血红素采集的探测将是 整合以确定结核分枝杆菌摄取和利用血红素的机制 白色念珠菌。总而言之，拟议的研究将全面解决血红素的机制 与健康和疾病相关的各种背景下的贩运和信号传递。