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Imaging heme based mitochondrial-cell signaling networks in cell and animal models of heavy metal toxicity

重金属毒性细胞和动物模型中基于血红素的线粒体细胞信号网络成像

基本信息

批准号：
9059090
负责人：
Amit Ram Reddi
金额：
$ 21.78万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9059090
关键词：
Animal Model Animals Antioxidants Applications Grants Arsenic Binding Biological Cadmium Caenorhabditis elegans Cell Line Cell Nucleus Cell Proliferation Cell model Cell physiology Cells Code Color Cytosol Drug Metabolic Detoxication Energy Metabolism Enterocytes Environmental Hazards Environmental Pollutants Eukaryota Fluorescent Probes Future Gases Goals Health Heavy Metals Heme Hemeproteins Histocompatibility Testing Homeostasis Image Imaging Device Institutes Iron Lead Lead Poisoning Life Mammalian Cell Maryland Measures Membrane Mercury Metabolism Methods MicroRNAs Mitochondria Mitochondrial Matrix Modeling Monitor Nature Neurons Nuclear Organ Organelles Organism Oxygen Palate Pathway interactions Phase Physiological Processes Process Proteins Public Health Regulation Reporter Research Role Saccharomyces cerevisiae Signal Transduction Signaling Molecule Site Stress Technology Thermodynamics Tissues Toxic Environmental Substances Toxic effect Translations Universities Whole Organism Xenobiotics Yeasts base biological adaptation to stress biophysical properties chemical genetics chemical property circadian pacemaker cofactor cytotoxic cytotoxicity detection of nutrient fluorescence imaging genetic manipulation heme a heme biosynthesis innovation metal poisoning novel oxidant stress oxidation quantitative imaging ratiometric response sensor stressor tool trafficking transcription factor transmission process

项目摘要

DESCRIPTION (provided by applicant): As mitochondria are central hubs for energy metabolism and formation of intermediate metabolites, they must be able to transmit signals that communicate alterations in its functioning to the nucleus so that cells can remodel metabolism in response. Mitochondrial-nuclear signaling is particularly important when mitochondrial function is compromised by environmental toxicants, including heavy metals. However, the nature and dynamics of these inter-organelle signals and the mechanisms by which they are transduced are poorly understood. Given the importance of heme-regulated transcription factors that control diverse cellular processes from energy metabolism to the anti-oxidant stress response to cell proliferation, and the exquisite sensitivity of mitochondrial heme homeostasis to heavy metals, we propose that mitochondrial- derived heme signals the cellular response to heavy metal toxicity through mitochondrial-nuclear retrograde regulation. However, the chemical and genetic tools available to study the transmission of heme-based signals do not exist. In the current grant application, the focus is to generate and apply a genetically encoded ratiometric fluorescent heme sensors in cell and animal models to study the transduction of mitochondrial-derived intra- and inter-organ heme signals in response to heavy metal toxicity. During the R21 phase, the intent is develop and apply the first ratiometric heme sensors for quantitative heme imaging in yeast and mammalian cell models of heavy metal toxicity. Finally, the goal is to (a) characterize the thermodynamics of heme binding to the heme sensors and apply them site-specifically in the yeast mitochondrial matrix, inter-membrane space, cytosol, and nucleus to quantitatively image heme signals in response to lead toxicity; and (b) apply sensors in a compartment specific manner in mammalian cell models of lead toxicity. Successful completion of these aims will provide the impetus to begin the R33 phase in which studies will be expanded to develop heme sensors that can be utilized for simultaneous imaging of heme between cellular compartments, and that are oxidation state-specific, and apply them to animal models of Pb toxicity. In this phase the focus is to (a) diversify the color palate of the heme sensors for simultaneous fluorescence imaging between subcellular compartments and develop Fe3+ and Fe2+ heme specific sensors; and (b) deploy these sensors in mammalian cell lines and a C. elegans model of environmental toxicity. Altogether, these studies will result in the first sensors for quantitative imaging of labile heme relevant to its role in cell signaling and establish heme a a vital mitochondria-derived signaling molecule that initiates the adaptation to heavy metal toxicity in both cell and animal models.

描述（由申请人提供）：由于线粒体是能量代谢和中间代谢物形成的中心枢纽，因此它们必须能够将其功能变化的信号传递给细胞核，以便细胞能够相应地重塑代谢。当线粒体功能受到包括重金属在内的环境毒物损害时，线粒体核信号传导尤为重要。然而，人们对这些细胞器间信号的性质和动态及其转导机制知之甚少。鉴于血红素调节转录因子控制从能量代谢到细胞增殖的抗氧化应激反应等多种细胞过程的重要性，以及线粒体血红素稳态对重金属的敏锐敏感性，我们提出线粒体来源的血红素通过线粒体核逆行调节来发出细胞对重金属毒性反应的信号。然而，可用于研究血红素信号传递的化学和遗传工具尚不存在。在当前的拨款申请中，重点是在细胞和动物模型中生成和应用基因编码的比率荧光血红素传感器，以研究线粒体衍生的器官内和器官间血红素信号响应重金属毒性的转导。在 R21 阶段，目的是开发和应用第一个比率血红素传感器，用于重金属毒性的酵母和哺乳动物细胞模型中的定量血红素成像。最后，目标是（a）表征血红素与血红素传感器结合的热力学，并将其应用于酵母线粒体基质、膜间空间、细胞质和细胞核中，以定量成像响应铅毒性的血红素信号； (b) 在铅毒性的哺乳动物细胞模型中以隔室特定方式应用传感器。这些目标的成功完成将为R33阶段的开始提供动力，该阶段的研究将扩大到开发血红素传感器，该传感器可用于细胞区室之间血红素的同步成像，并且具有氧化态特异性，并将其应用于铅毒性的动物模型。在此阶段的重点是 (a) 使血红素传感器的色板多样化，以便在亚细胞区室之间同时进行荧光成像，并开发 Fe3+ 和 Fe2+ 血红素特异性传感器； (b) 将这些传感器部署在哺乳动物细胞系和环境毒性的秀丽隐杆线虫模型中。总而言之，这些研究将产生第一个传感器对与其在细胞信号传导中的作用相关的不稳定血红素进行定量成像，并确定血红素是一种重要的线粒体衍生信号分子，可启动细胞和动物模型中对重金属毒性的适应。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Human ribosomal G-quadruplexes regulate heme bioavailability.

人核糖体 G-四链体调节血红素生物利用度。

DOI：
10.1074/jbc.ra120.014332
发表时间：
2020
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Mestre-Fos,Santi;Ito,Chieri;Moore,CourtneyM;Reddi,AmitR;Williams,LorenDean
通讯作者：
Williams,LorenDean

Heme Gazing: Illuminating Eukaryotic Heme Trafficking, Dynamics, and Signaling with Fluorescent Heme Sensors.

DOI：
10.1021/acs.biochem.7b00007
发表时间：
2017-04-04
期刊：
Biochemistry
影响因子：
2.9
作者：
Hanna DA;Martinez-Guzman O;Reddi AR
通讯作者：
Reddi AR

Heme dynamics and trafficking factors revealed by genetically encoded fluorescent heme sensors

DOI：
10.1073/pnas.1523802113
发表时间：
2016-07-05
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Hanna, David A.;Harvey, Raven M.;Reddi, Amit R.
通讯作者：
Reddi, Amit R.

Using genetically encoded heme sensors to probe the mechanisms of heme uptake and homeostasis in Candida albicans.

使用基因编码的血红素传感器来探究白色念珠菌的血红素摄取和稳态机制。