Carbon monoxide in Mycobacterium tuberculosis pathogenesis

描述(由申请者提供)：项目摘要：Michael Shiloh博士是一位传染病内科医生，对宿主与病原体的相互作用有着明确的兴趣，该兴趣是作为纽约市三机构医学科学家培训计划的成员培养出来的。他的整个职业目标是通过在学术医学中心研究结核分枝杆菌潜伏的潜在机制，将他的临床和基础科学兴趣结合起来。构建此应用程序就是为了提供实现这一目标的框架。通过一系列特定的目标，Shiloh博士将测试关于一氧化碳(CO)在结核分枝杆菌发病机制中的作用的几个假说。具体地说，他将测试这一假设，即受感染的巨噬细胞产生的CO会改变结核分枝杆菌的代谢和基因表达模式。除了在体外和体内暴露于一氧化碳的结核分枝杆菌微阵列外，这将使用体外细胞分析来完成。为了验证结核分枝杆菌利用CO在体内生长的假设，将测试结核分枝杆菌CO摄取基因突变株在体外、巨噬细胞内和受感染小鼠体内对CO的生长能力。最后，为了验证结核分枝杆菌抵抗一氧化碳毒性效应的假设，将一个突变体库暴露在一氧化碳中，并对那些在一氧化碳存在下无法存活的突变株进行进一步的鉴定。这项拟议的工作与确定结核分枝杆菌如何能够建立和维持肺部内的潜伏感染有关。此外，这项应用旨在补充Shiloh博士以前的实验室经验，并为他提供必要的微生物发病机制方面的技术和知识背景，以发挥独立研究人员的作用。一个由医生和科学家组成的委员会将监督希洛博士走向独立的过程；他的科学发展也将通过参加加州大学旧金山分校的几个系列研讨会、课程、部门务虚会和全国会议来丰富。加州大学旧金山分校致力于学术医学事业的发展，在资助期结束时，Shiloh博士将准备开始他的职业生涯，成为一名独立的内科科学家，研究感染宿主体内生存的微生物机制。相关性：这项拟议的工作与确定结核分枝杆菌如何能够建立和维持肺部潜伏感染有关。由于世界上大约三分之一的人口存在结核分枝杆菌的潜伏感染，了解潜伏感染的机制对于开发预防、治疗和根除结核病的新方法至关重要。

DESCRIPTION (provided by applicant): Project Summary: Dr. Michael Shiloh is an infectious disease physician with a well-defined interest in host-pathogen interactions that was fostered as a member of the Tri-lnstitutional medical scientist training program in New York City. His overall career objective is to combine his clinical and basic science interests by studying the underlying mechanisms of M. tuberculosis latency in an academic medical center. This application is constructed to provide a framework to achieve this goal. Through a series of specific aims, Dr. Shiloh will test several hypotheses regarding the role of carbon monoxide (CO) in M. tuberculosis pathogenesis. Specifically, he will test the hypothesis that CO produced by infected macrophages alters the metabolism and gene expression pattern of M. tuberculosis. This will be accomplished using in vitro cellular assays in addition to microarrays from M. tuberculosis exposed to CO in vitro and in vivo. To test the hypothesis that M. tuberculosis utilizes CO for growth in vivo, mutants in the M. tuberculosis CO uptake genes will be tested for their ability to grow on CO in vitro, inside macrophages and inside infected mice. Finally, to test the hypothesis that M. tuberculosis resists the toxic effects of CO, a library of mutants will be exposed to CO and those mutants that fail to survive in the presence of CO will be characterized further. The proposed work is relevant to determining how M. tuberculosis is able to both establish and maintain a dormant infection within the lung. Additionally, this application is designed to complement Dr. Shiloh's prior laboratory experience and provide him with the requisite technical and intellectual background in microbial pathogenesis to function as an independent investigator. A committee of physicians and scientists will oversee Dr. Shiloh's progression towards independence; his scientific development will also be enriched through attendance at several seminar series at UCSF, courses, department retreats and national meetings. UCSF is committed to the development of careers in academic medicine, and at the end of the grant period, Dr. Shiloh will be prepared to embark on a career as an independent physician scientist investigating microbial mechanisms for surviving within the infected host. Relevance: The proposed work is relevant to determining how M. tuberculosis is able to both establish and maintain a latent infection within the lung. Since roughly a third of the world's population harbors a latent infection with M. tuberculosis, understanding the mechanisms involved in latency is vital to developing new ways to prevent, treat and eradicate tuberculosis.