Metabolic Discrimination of Unknown Bacterial Pathogens

未知细菌病原体的代谢区分

基本信息

批准号：
7178451
负责人：
JOHN PETER WIKSWO
金额：
$ 103.69万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-01 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7178451
关键词：
2,4-Dinitrophenol Achievement Action Potentials Address Affect Algorithms Anthrax disease Antibodies Antidotes Bacterial Toxins Benign Binding Biological Biological Assay Biosensor Botulinum Toxins Carbon Dioxide Cardiac Cell Line Cell physiology Cells Characteristics Chemicals Citric Acid Cycle Class Clinical Clostridium botulinum Collaborations Complex Complex Mixtures Computer software Computers Condition Coupled Cyanides DNA Data Detection Development Devices Diagnosis Diagnostic Diagnostic Procedure Discrimination Electron Transport Complex III End Point Engineering Enterotoxins Enzymes Escherichia Event Exposure to Extracellular Space Fibroblasts Fluorescence Fluorides Functional disorder Generations Genetic Glucose Glycolysis Homeostasis Hydrazine Hydrazines Infection Infectious Agent Iodoacetates Ion Channel Kidney Knowledge Laboratories Laboratory Study Lactate Dehydrogenase Lead Learning Libraries Life Liver Mammalian Cell Mannoheptulose Measurement Measures Membrane Metabolic Methodology Methods Microbial Genetics Microfabrication Microprocessor Military Personnel Mitochondria Modeling Modification Molecular Monitor Nerve Noise Nucleic Acids Numbers Organism Output Oxidation-Reduction Pathway interactions Pattern Performance Pharmaceutical Preparations Phosphorylation Physiological Preclinical Drug Evaluation Principal Component Analysis Probability Process Production Prophylactic treatment Proteins Protocols documentation Pseudomonas aeruginosa Public Health Pyruvate Pyruvates Range Rate Reaction Research Research Personnel Resistance Resolution Ricin Robotics Rotenone Sampling Screening procedure Security Sensitivity and Specificity Signal Pathway Signal Transduction Silicon Site Standards of Weights and Measures Staphylococcal Enterotoxin B Staphylococcus aureus Succinate Dehydrogenase Sum System Technology Testing Therapeutic Time Tissues Today Toxic effect Toxicology Toxin Trees Tweens Uncoupling Agents Universities Virulence anthrax toxin antimycin base biodefense cell type complex IV computerized data processing cost design desire enolase expectation extracellular glucose uptake hexokinase instrument macrophage microbial microorganism miniaturize nano pathogen pathogenic bacteria programs prophylactic receptor response sensor sound tool trait vector

项目摘要

DESCRIPTION (provided by applicant): A collaboration between Vanderbilt University and the U.S. Army Edgewood Chemical and Biological Center (ECBC) at the Aberdeen Proving Grounds will develop a wide-spectrum, activity-detection technology that employs multiphasic sensing, in order to provide a biofunctional signature of a CBW agent, unknown drug, or other threat. The signatures will be used with advanced algorithms to discriminate between different agents acting on a set of target cell lines. The proposed approach is extraordinarily versatile and general, because we are measuring the biological impact of the toxins, rather than simply their presence. This will address a critically important and as-yet-unmet need for diagnostic tools capable of identifying the mechanism of action of unknown or reengineered threat agents that defy accurate detection by existing, agent-specific sensors. The diagnostic tools will be created to establish signatures of key changes in metabolic and signaling pathways that occur in cell lines responsive to Anthrax, Ricin, Staphylococcal Enterotoxin B (SEB), and Clostridium Botulinum toxins. The interaction of the toxins with cells leads to a multitude of metabolic and signaling events, as toxins disrupt normal cellular functions in specific and non-specific ways that are not yet fully understood. To provide a means to characterize the effects of unknown toxins, the proposed diagnostic system will monitor key parameters of specific metabolic and signaling pathways over a spectrum of time- and volume-scales. Newly developed well-plate protocols for end-point metabolic rates will be coupled with commercial fluorescence assays for signaling events. Additionally, the same metabolic and signaling events will be captured as dynamic biosignatures using a modified Cytosensor that simultaneously monitors multiple analytes on the time scale of minutes. This approach will be scaled down to achieve dynamic resolution on the order of seconds in a microfabricated NanoPhysiometer. This approach should find wide application in the discovery of new drugs and unexpected and/or undesired physiological activity; environmental and industrial toxicology; metabonomics and signaling; and screening of countermeasures, therapies, and prophalaxis for pathogenic bacteria and toxins.

描述（由申请人提供）：范德比尔特大学和位于阿伯丁试验场的美国陆军埃奇伍德化学和生物中心 (ECBC) 之间的合作将开发一种采用多相传感的广谱活动检测技术，以便提供生化武器制剂、未知药物或其他威胁的生物功能特征。这些特征将与先进的算法一起使用，以区分作用于一组目标细胞系的不同药物。所提出的方法非常通用且通用，因为我们正在测量毒素的生物影响，而不仅仅是它们的存在。这将解决对能够识别未知或重新设计的威胁代理的作用机制的诊断工具的极其重要且尚未满足的需求，这些威胁代理无法通过现有的特定于代理的传感器进行准确检测。将创建诊断工具来建立对炭疽、蓖麻毒素、葡萄球菌肠毒素 B (SEB) 和肉毒杆菌毒素有反应的细胞系中发生的代谢和信号传导途径关键变化的特征。毒素与细胞的相互作用导致大量代谢和信号传导事件，因为毒素以尚未完全了解的特定和非特定方式破坏正常细胞功能。为了提供一种表征未知毒素影响的方法，所提出的诊断系统将在一系列时间和体积尺度上监测特定代谢和信号传导途径的关键参数。新开发的终点代谢率孔板方案将与信号事件的商业荧光测定相结合。此外，使用改进的细胞传感器将捕获相同的代谢和信号事件作为动态生物特征，该传感器在分钟的时间尺度上同时监测多种分析物。这种方法将按比例缩小，以在微制造的纳米生理计中实现秒级的动态分辨率。这种方法应该在新药和意外和/或不需要的生理活性的发现中得到广泛应用；环境和工业毒理学；代谢组学和信号传导；病原菌和毒素的对策、治疗和预防的筛选。