A High-Throughput Screen for Modulators of Quorum Sensing in Vibrio cholerae

描述（由申请人提供）：本研究的长期目标是探索细菌用于细胞间通讯的分子机制（即群体感应）。直到最近，细菌的交流能力被认为是一种异常现象，只发生在少数海洋弧菌物种中。现在很清楚，细胞间的交流是细菌世界的常态，理解这个过程是所有微生物学的基础，包括工业和临床微生物学。在这里，我们提出了一个综合的化学生物学研究群体感应电路的人类病原体霍乱弧菌，疾病的病原体。霍乱弧菌群体感应电路依赖于一种被称为自诱导体的小分子信号的产生和检测。我们建议与MLPCN合作，通过执行既定的高通量筛选来识别在群体感应级联中调制三个不同步骤的小分子。后续的二次筛选将确定这些先导化合物的作用模式，并将其分类为：群体感应受体激动剂；2. LuxO抑制剂;3. Hfq抑制剂。通过这项工作鉴定的分子将为探索霍乱弧菌和其他微生物群体感应调节网络的分子机制提供新的工具。此外，这些化合物将被用作开发旨在干扰群体感应的广谱抗菌药物的先导物。这种疗法可能对改善人类健康产生巨大影响。

DESCRIPTION (provided by applicant): The long-term goal of this research is to explore the molecular mechanisms that bacteria use for cell-cell communication (i.e., quorum sensing). Until recently, the ability of bacteria to communicate was considered an anomaly that occurred only in a few marine Vibrio species. It is now clear that cell-cell communication is the norm in the bacterial world and that understanding this process is fundamental to all of microbiology, including industrial and clinical microbiology. Here we propose an integrative chemical biological study of the quorum-sensing circuit of the human pathogen Vibrio cholerae, the causative agent of the disease cholera. The V. cholerae quorum-sensing circuit depends on the production and detection of a small molecule signals called autoinducers. We propose to collaborate with the MLPCN to identify small molecules that modulate three distinct steps in the quorum-sensing cascade by performing an established high- throughput screen. Follow up secondary screens will define the modes-of-action of these lead compounds and categorize them as: 1. Quorum-sensing receptor agonists; 2. LuxO inhibitors; 3. Hfq inhibitors. Molecules identified through this work will provide new tools to probe the molecular mechanisms governing the quorum-sensing regulatory networks of V. cholerae and other microorganisms. Additionally, these compounds will be used as leads for the development of broad-spectrum antibacterial drugs designed to interfere with quorum sensing. Such therapies could have enormous ramifications for improving human health. PUBLIC HEALTH RELEVANCE: V. cholerae is a globally important pathogen that causes several million cases of disease per year. Our studies have shown that, in order to be pathogenic, V. cholerae must possess an intact quorum-sensing system because cell-cell communication controls the timing underlying production and release of virulence factors and the formation of biofilms. The investigations proposed here will facilitate the development of synthetic strategies for controlling quorum sensing in this important pathogenic microbe and possibly other pathogenic bacteria that use quorum sensing to control virulence.