In vivo Transcriptional Analysis of Latent Tuberculosis

潜伏性结核病的体内转录分析

• 批准号: 7209529
• 负责人: ADEL M TALAAT
• 金额: $ 25.73万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7209529
• 关键词: Aerosols; Bacillus (bacterium); Chronic; Condition; DNA Microarray Chip; DNA Microarray format; Disease; Foundations; Future; Gene Expression; Gene Expression Profile; Genes; Genus Mycobacterium; Growth; Hand; Human; Immune; In Vitro; Inbred BALB C Mice; Infection; Label; Metabolic; Microarray Analysis; Molecular; Molecular Profiling; Mus; Mycobacterium tuberculosis; Nature; Organ; Outcome; Population; RNA; Regulator Genes; Role; Staging; Stress; Transcript; Tuberculosis; base; comparative; day; drug development; human tissue; improved; in vivo; innovative technologies; mouse model; mutant; mycobacterial; pathogen

DESCRIPTION (provided by applicant): Our understanding of the chronic stages of tuberculosis on a molecular level is incomplete and definitely needs improvement. Comparative analysis of gene expression profiled during early vs. late stages of tuberculosis will enrich our understanding of various stages of tuberculosis. The main objective of this project is to identify gene regulatory networks responsible for establishing chronic tuberculosis in mice. Based on a strong set of preliminary results, we hypothesize that the tuberculous bacilli are actively transcribing specific sets of genes to regulate their metabolic activity inside the host despite remaining at a constant level during the chronic stase of the disease. To identify key components of the mycobacterial gene regulatory network during chronic infection, we will employ the in vivo microarray analysis to: 1. Identify M.tb. genes responsible for entering into the chronic stage of tuberculosis. We will employ the mouse model of chronic tuberculosis combined with microarray analysis to identify differentially expressed genes in mice during the first 220 days of infection as well as during in vitro cultures. A reactivation model of mouse tuberculosis also will be assessed to mimic human tuberculosis during immune suppression. 2. Characterize the role of putative persistence genes in establishing chronic tuberculosis. We will follow organ colonization of M.tb. strains with defective persistence genes (generated by allelic exchange) after aerosol infection of BALB/c mice in comparison to the wild type strain of M.tb. H37Rv. Also, the transcriptional profile of mutants growing under defined in vitro stress conditions will be assessed using DNA microarrays. We believe that the approaches exploited in this proposal will generate a wealth of information related to the metabolic states of M.tb. during chronic stages of infection. The proposed studies will further improve our understanding of the nature of the host-pathogen interactions that can provide potential targets for drug development. Outcomes from this project will provide the necessary foundation for more detailed and focused studies on specific sets of genes utilizing human tissues. Transcriptional regulators that could contribute to both chronic and reactivation stages of tuberculosis will be the target for future analysis.

描述（由申请人提供）：我们对分子水平上结核病的慢性阶段的理解是不完整的，绝对需要改善。对结核病早期与晚期阶段相比的基因表达的比较分析将使我们对结核病的各个阶段的理解。该项目的主要目的是确定负责在小鼠中建立慢性结核病的基因调节网络。基于一系列强大的初步结果，我们假设结核菌正在积极转录特定的基因集，以调节其在宿主体内的代谢活性，尽管在疾病的慢性基础上保持恒定水平。为了确定慢性感染过程中分枝杆菌基因调节网络的关键组成部分，我们将使用体内微阵列分析来： 1。识别M.TB。负责进入结核病的慢性阶段的基因。我们将采用慢性结核病的小鼠模型与微阵列分析相结合，在感染的最初220天以及体外培养物中鉴定小鼠中差异表达的基因。在免疫抑制期间，还将评估小鼠结核病的重新激活模型。 2。表征推定持久基因在建立慢性结核病中的作用。我们将跟随M.TB的器官定植与M.TB的野生型菌株相比，BALB/C小鼠气溶胶感染后具有缺陷的持久性基因的菌株（由等位基因交换产生）。 H37RV。同样，将使用DNA微阵列评估在定义的体外应力条件下生长的突变体的转录曲线。 我们认为，本提案中利用的方法将产生与M.TB的代谢状态有关的大量信息。在感染的慢性阶段。拟议的研究将进一步提高我们对可以为药物开发提供潜在目标的宿主 - 病原体相互作用的性质的理解。该项目的结果将为利用人体组织的特定基因组集提供必要的基础。可能有助于结核病的慢性和重新激活阶段的转录调节剂将成为未来分析的目标。