Bacterial Sensing and Response to Chloride as a Novel Tuberculosis Drug Target

细菌对氯化物作为新型结核病药物靶点的感知和反应

• 批准号: 8966959
• 负责人: Shumin Tan
• 金额: $ 10.36万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8966959
• 关键词: Address; Advisory Committees; Affect; Attenuated; Award; Bacteria; Biological; Biological Assay; Biology; Bypass; Cells; Cessation of life; Characteristics; Chemicals; Chloride Channels; Chloride Ion; Chlorides; Chronic; Communication; Cues; Data; Dedications; Development; Development Plans; Drug Targeting; Drug resistance; Educational process of instructing; Ensure; Environment; Equilibrium; Foundations; Future; Genes; Genetic; Global Change; Goals; Growth; Housekeeping; Human; Immune; In Vitro; Infection; Institution; Ions; Iron; Knowledge; Libraries; Life Cycle Stages; Light; Link; Maintenance; Mentors; Methods; Modeling; Mus; Mutation; Mycobacterium tuberculosis; Nutrient; Pathogenesis; Phagosomes; Phase; Phenotype; Physiology; Population; Population Heterogeneity; Positioning Attribute; Proteins; Public Health; Regulation; Regulator Genes; Reporter; Research; Research Personnel; Research Training; Resources; Role; Signal Transduction; Testing; Treatment Efficacy; Tuberculosis; Universities; Vaccines; Work; attenuation; base; career; career development; cellular imaging; chemical genetics; genome-wide; global health; in vivo; innovation; insight; interest; macrophage; microbial; mutant; novel; pathogen; programs; public health relevance; research and development; research study; response; skills; small molecule; tool; treatment strategy; tuberculosis drugs; tuberculosis treatment

 DESCRIPTION (provided by applicant): Mycobacterium tuberculosis (Mtb) is an extremely successful pathogen that remains a key public health problem, infecting one-third of the human population and causing ~1.5 million deaths a year. The macrophage serves as a major host cell for Mtb, and the bacterium's ability to turn the phagosome into a replicative niche is critical for the establishment and maintenance of chronic infection. This is dependent on Mtb sensing and responding appropriately to intraphagosomal signals, and the research proposed here builds on my discovery of chloride (Cl-) as a novel environmental cue that Mtb responds to during infection. I have further identified a Cl- channel in Mtb, and observe that a Mtb mutant deficient in this channel is severely attenuated for colonization in a murine infection model. My studies raise the novel concept that the sensing and/or maintenance of an appropriate balance of an abundant ion like Cl- can profoundly impact a pathogen's ability to establish a successful infection, and suggest that this aspect of Mtb physiology represents a new target for the development of original and potentially potent tuberculosis treatment strategies synergistic with current approaches. Here, I seek to (i) elucidate the regulatory network governing Mtb's sensing and response to Cl-, using both genetic and chemical biology approaches that exploit my Cl--responsive reporter Mtb strain, and (ii) examine how environmental cues, like Cl-, can impact on Mtb's replication status, using genome-wide mutational analyses together with a novel replication reporter Mtb strain that I recently constructed. I have a longstanding dedication to research, and my career goal is to direct my own research group at an academic institution. My research will take a multi-disciplinary approach to probe host-pathogen interactions especially from the pathogen's perspective, to understand fundamental aspects of how it can colonize its specific microniche, and exploit this knowledge to examine how these critical nodes may be manipulated to shift the balance during infection in favor of the host. The mentored phase of this award will allow me to expand my methodological repertoire, and further develop my research skills and research program. Completion of genetic and chemical screens focused on elucidating Mtb's Cl- regulatory network will further provide me with tools (Mtb mutants and chemical probes that modulate Mtb's Cl- response) and results from which new projects will be developed as I transition to an independent position. During the independent phase, I will also begin to address Mtb's sensing and response to specific environmental cues in a different conceptual framework, in particular interrogating how it may be linked to regulation of bacterial replication, independent of nutrient availability. The mentored phase of this award will be carried out in the lab of Dr. David Russell at Cornell University. The Russell lab and Cornell University provide an ideal environment for my research and training, with unique resources that will enable me to carry out the proposed research, and continue to acquire skills in other facets crucial to my goal of becoming a successful independent investigator, such as lab management, teaching, and scientific communication skills. In addition, I have assembled an advisory committee with diverse expertise in Cl- channels and transporters, in chemical biology underlying signal transduction systems, and in Mtb replication and single cell imaging, to provide further support during the K99 phase and as I transition to the R00 phase. The research proposed here integrates genetic and chemical biology studies that will shed light on the interplay between host cellular environmental cues and Mtb's establishment and maintenance of infection, insights essential to understanding Mtb's life cycle, and for identifying potential targts for the development of new synergistic treatment methods. Combined with other aspects of the research and career development plan proposed, it will ensure that I have both the skill set and groundwork data needed to successfully accomplish my goals of establishing my own lab and pursuing microbial pathogenesis research using a multi-disciplinary approach.

 描述（由申请人提供）：结核分枝杆菌（Mtb）是一种非常成功的病原体，仍然是一个关键的公共卫生问题，感染三分之一的人口，每年造成约150万人死亡。巨噬细胞作为结核分枝杆菌的主要宿主细胞，细菌将吞噬体转化为复制生态位的能力是 对慢性感染的建立和维持至关重要。这取决于结核分枝杆菌对吞噬体内信号的感知和适当反应，这里提出的研究建立在我发现的氯化物（Cl-）作为结核分枝杆菌在感染过程中反应的一种新的环境线索的基础上。我进一步鉴定了Mtb中的Cl-通道，并观察到该通道缺陷的Mtb突变体在鼠感染模型中的定殖严重减弱。我的研究提出了一个新的概念，即感测和/或维持一个适当的平衡，丰富的离子，如Cl-可以深刻地影响病原体的能力，建立一个成功的感染，并表明结核分枝杆菌生理学的这方面代表了一个新的目标，为发展的原始和潜在的有效的结核病治疗策略协同与目前的方法。在这里，我试图（一）阐明管理结核分枝杆菌的传感和响应Cl-的监管网络，使用遗传和化学生物学的方法，利用我的Cl-响应报告结核分枝杆菌菌株，和（ii）研究如何环境的线索，如Cl-，可以影响结核分枝杆菌的复制状态，使用全基因组突变分析连同一个新的复制报告结核分枝杆菌菌株，我最近构建。我长期致力于研究，我的职业目标是在学术机构指导我自己的研究小组。我的研究将采取多学科的方法来探测宿主-病原体的相互作用，特别是从病原体的角度，了解它如何能够殖民其特定的microniche的基本方面，并利用这些知识来研究如何操纵这些关键节点，以改变感染期间有利于宿主的平衡。这个奖项的指导阶段将使我能够扩大我的方法论曲目，并进一步发展我的研究技能和研究计划。完成遗传和化学筛选，重点阐明结核分枝杆菌的Cl-调控网络将进一步为我提供工具（结核分枝杆菌突变体和化学探针，调节结核分枝杆菌的Cl-反应）和结果，新的项目将开发为我过渡到一个独立的位置。在独立阶段，我也将开始解决结核分枝杆菌的传感和响应特定的环境线索在不同的概念框架，特别是询问它如何可能与细菌复制的调节，独立的营养供应。该奖项的指导阶段将进行 在康奈尔大学的大卫罗素博士的实验室里。罗素实验室和康奈尔大学为我的研究和培训提供了一个理想的环境，拥有独特的资源，使我能够开展拟议的研究，并继续获得对我成为一名成功的独立调查员的目标至关重要的其他方面的技能，如实验室管理，教学和科学沟通技巧。此外，我还组建了一个咨询委员会，该委员会在Cl-通道和转运蛋白、化学生物学基础信号转导系统以及结核分枝杆菌复制和单细胞成像方面具有不同的专业知识，以便在K99阶段和我过渡到R 00阶段时提供进一步的支持。这里提出的研究整合了遗传和化学生物学研究，将揭示宿主细胞环境线索与结核分枝杆菌感染的建立和维持之间的相互作用，对理解结核分枝杆菌的生命周期至关重要的见解，并确定开发新的协同治疗方法的潜在目标。结合所提出的研究和职业发展计划的其他方面，它将确保我拥有成功实现建立自己的实验室和使用多学科方法进行微生物致病机理研究的目标所需的技能和基础数据。

项目成果

Growth of Mycobacterium tuberculosis in vivo segregates with host macrophage metabolism and ontogeny.

• DOI: 10.1084/jem.20172020
• 发表时间: 2018-04-02
• 期刊: The Journal of experimental medicine
• 作者: Huang L;Nazarova EV;Tan S;Liu Y;Russell DG
• 通讯作者: Russell DG

The Deconstructed Granuloma: A Complex High-Throughput Drug Screening Platform for the Discovery of Host-Directed Therapeutics Against Tuberculosis.

• DOI: 10.3389/fcimb.2018.00275
• 发表时间: 2018
• 期刊: Frontiers in cellular and infection microbiology
• 影响因子: 5.7
• 作者: Huang L;Kushner NL;Theriault ME;Pisu D;Tan S;McNamara CW;Petrassi HM;Russell DG;Brown AC
• 通讯作者: Brown AC