Metabolic Discrimination of Unknown Bacterial Pathogens

未知细菌病原体的代谢区分

• 批准号: 6818160
• 负责人: JOHN PETER WIKSWO
• 金额: $ 107.72万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 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/6818160
• 关键词: Clostridium botulinum; Escherichia coli; Pseudomonas aeruginosa; Staphylococcus aureus; acid base balance; bacterial toxins; biohazard detection; biological signal transduction; biosensor device; bioterrorism /chemical warfare; calcium flux; cell line; cooperative study; glucose metabolism; lactates; membrane potentials; oxidation reduction reaction; oxygen consumption; technology /technique development; toxin metabolism; Clostridium botulinum; Escherichia coli; Pseudomonas aeruginosa; Staphylococcus aureus; acid base balance; bacterial toxins; biohazard detection; biological signal transduction; biosensor device; bioterrorism /chemical warfare; calcium flux; cell line; cooperative study; glucose metabolism; lactates; membrane potentials; oxidation reduction reaction; oxygen consumption; technology /technique development; toxin metabolism

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）：范德比尔特大学与阿伯丁证明基础的美国陆军边缘化学与生物中心（ECBC）之间的合作将开发一种采用多相传感的广泛，活动检测技术，以提供多相传感，以便提供CBW代理商的生物官能签名，不知名的药物，不知名的药物，或其他威胁或其他威胁。签名将与高级算法一起使用，以区分作用在一组目标细胞系上的不同试剂。所提出的方法非常具有广泛的用途和一般性，因为我们正在测量毒素的生物学影响，而不仅仅是它们的存在。这将解决一种至关重要的，尚未达到的诊断工具的需求，能够识别未知或重新设计的威胁代理的作用机理，这些作用机理无视现有的特定于特定于代理特定的传感器的准确检测。将创建诊断工具，以建立响应于炭疽，ricin，葡萄球菌肠毒素B（SEB）和肉毒杆菌梭菌毒素的细胞系中代谢和信号通路的关键变化的特征。毒素与细胞的相互作用导致多种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异性方式破坏了正常的细胞功能。为了提供一种表征未知毒素影响的手段，所提出的诊断系统将在一系列时间和体积尺度上监视特定代谢和信号通路的关键参数。 新开发的用于终点代谢率的修复板协议将与用于信号事件的商业荧光测定法相结合。此外，使用改良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签名，该细胞传感器同时在分钟的时间范围内同时监视多个分析物。 该方法将缩小以在微生物纳米生理计中的秒数上实现动态分辨率。这种方法应在发现新药以及意外和/或不希望的生理活动中发现广泛的应用；环境和工业毒理学；代表学和信号传导；以及对病原细菌和毒素的对策，疗法和预言的筛查。