Elemental imaging of M. tuberculosis during infection.

感染期间结核分枝杆菌的元素成像。

• 批准号: 7740406
• 负责人: ADEL M TALAAT
• 金额: $ 20.61万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-19 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7740406
• 关键词: Address; Affect; Animals; Antibiotics; Attention; Bacillus (bacterium); Biochemical; Biological; Biology; Chemotherapy-Oncologic Procedure; Chronic; Copper; DNA Microarray Chip; Data; Disease; Divalent Cations; Drug Metabolic Detoxication; Electron Microscopy; Elements; Environment; Enzymes; Exploratory/Developmental Grant; Foundations; Funding; Gene Expression Profile; Genes; Goals; Homeostasis; Human; Image; Immune; Immunocompromised Host; Inbred BALB C Mice; Indium; Individual; Infection; Investigation; Ions; Iron; Knock-out; Life; Link; Lung; Magnesium; Maps; Metalloproteins; Metals; Mission; Modeling; Molecular; Molecular Biology; Mus; Mycobacterium Infections; Mycobacterium tuberculosis; Nitrogen; Operon; Outcome; Oxidoreductase; Pathogenesis; Phagosomes; Pharmaceutical Preparations; Physiological; Play; Population; Proteins; Regulation; Resistance; Resistance development; Resolution; Risk; Role; Sampling; Scanning Transmission Electron Microscopy Procedures; Staging; Superoxide Dismutase; System; Techniques; Technology; Tissues; Transcript; Tuberculosis; United States National Institutes of Health; Vaccines; base; design; lipoarabinomannan; macrophage; mutant; mycobacterial; new technology; novel; pathogen; public health relevance; response; trafficking; trend

DESCRIPTION (provided by applicant): More than 1.8 million people die annually from infection with Mycobacterium tuberculosis (M.tb), the causative agent of tuberculosis. Current vaccines or chemotherapy regimens are not able to reverse this trend. Our long-term goal is to better understand the molecular biology of M.tb and to discover new mechanisms of survival employed by this successful pathogen. An important aspect of the host-pathogen interactions is the regulation of the acquisition of metal elements in the microenvironment where tuberculous bacilli live. The ultimate goal in this project is to identify the role of metal-responsive genes to the overall basic biology of M.tb and to examine their impact on mycobacterial phagosome maturation. Preliminary data gathered so far laid the foundation for strong hypotheses related to Cu trafficking systems. Additionally, it directed our attention to other metal ions that could be equally or even more important than Cu and Fe. Fortunately, we embarked on a new technology for Scanning Transmission Electron Microscopy (STEM) for elemental analysis of biological tissues on the ionic level that could analyze multiple ions (e.g. 10 elements) of the same sample. We decided to utilize the R21 mechanism of funding to explore the advantages of this technology and to apply it towards the better understanding of the molecular pathogenesis of M.tb. We believe that our studies will open the door for more detailed investigations of the role of specific metals in M.tb survival strategies. In the first aim of this project we will utilize the power of analytical electron microscopy to profile the elemental map in lungs of mice representing 2 different models of murine tuberculosis (BALB/c and SCID). In the second aim, we will profile the mycobacterial responses to key ions significantly changed their levels during progression from early to chronic tuberculosis. Finally, in the third aim, we will examine mycobacterial phagosome maturation following infection with bacilli defective in metal ion acquisition systems. Overall, we will take full advantage of the high resolution, micro- analytical system (STEM) and DNA microarrays to dissect the contribution of metalloproteins to metal homeostasis and to the intracellular environment of M. tb. We believe that the outcomes of this project will delineate novel mechanisms of pathogenesis and could help in designing better drugs/vaccines against tuberculosis. PUBLIC HEALTH RELEVANCE: This project is very relevant to the mission of the NIH. Tuberculosis causes a tremendous risk to healthy populations in the USA as well as immunocompromised individuals.

描述（由申请人提供）：每年有180万人死于结核分枝杆菌（M.TB），这是结核病的病因。当前的疫苗或化疗方案无法扭转这一趋势。我们的长期目标是更好地了解M.TB的分子生物学，并发现该成功病原体采用的生存机制。宿主 - 病原体相互作用的一个重要方面是调节结核菌生存的微环境中金属元素的获取。该项目的最终目标是确定金属响应基因对M.TB的总体基本生物学的作用，并检查其对分枝杆菌吞噬体成熟的影响。迄今为止收集的初步数据为与CU贩运系统有关的强有力假设奠定了基础。此外，它将我们的注意力转移到其他可能比Cu和Fe更重要的金属离子上。幸运的是，我们启动了一种用于扫描透射电子显微镜（STEM）的新技术，用于对离子水平上生物组织的元素分析，该技术可以分析同一样品的多个离子（例如10个元素）。我们决定利用R21的资金机制来探索这项技术的优势，并将其应用于对M.TB的分子发病机理的更好理解。我们认为，我们的研究将为特定金属在M.TB生存策略中的作用进行更详细的研究打开大门。在该项目的第一个目的中，我们将利用分析电子显微镜的力量来介绍代表2种不同鼠结核模型的小鼠肺图（BALB/C和SCID）。在第二个目标中，我们将在从早期到慢性结核病中的进展过程中介绍对关键离子的分枝杆菌反应显着改变其水平。最后，在第三个目标中，我们将检查金属离子采集系统中有缺陷的杆菌后的分枝杆菌吞噬体成熟。总体而言，我们将充分利用高分辨率，微分析系统（STEM）和DNA微阵列，以剖析金属蛋白对金属稳态和M. TB的细胞内环境的贡献。我们认为，该项目的结果将描述发病机理的新型机制，并有助于设计更好的药物/疫苗针对结核病。公共卫生相关性：该项目与NIH的任务非常相关。结核病会给美国健康人群以及免疫功能低下的个体带来巨大的风险。