A Community Mycobacterial Systems Resource

描述（由申请人提供）：分枝杆菌疾病，主要是结核病，每年慢性感染数十亿人，造成近200万人死亡。这一全球性危机因治疗不足而加剧：卡介苗的效力值得怀疑，多重耐药和极端耐药的M。结核病的增长令人担忧。我们将应用基因组学和高通量技术开发分枝杆菌系统资源（MSR），这将大大加快整个分枝杆菌社区的研究。MSR将免费提供，并将提供所有分枝杆菌共同的生物过程的全面，综合的基因组和视觉总结，可立即应用于进一步了解分枝杆菌疾病。从MSR获得的见解将产生大量新的生物学和比较工具，并确定许多新的目标，适用于新的抗分枝杆菌治疗的发展。因此，MSR将刺激分枝杆菌研究界，并促进基本发现的转化，以对抗造成巨大全球健康负担的物种。这些目标将通过利用研究人员的综合专业知识和尖端技术来实现，以形成这个多学科，多机构的团队。 致病性分枝杆菌生长极其缓慢，使用起来非常危险，因此不适合用于高通量基因组分析。我们将通过使用一个密切相关的物种M来解决这个问题。耻垢分枝杆菌，其是快速生长的非致病性分枝杆菌，其是公认的遗传上易处理的模型分枝杆菌。MSR的焦点约为1，300 M。在M.结核复合体、M. avium，M. ulcerans和M.麻风病人这些高度保守的基因将介导基本的细胞过程，因此它们的分析将适用于所有分枝杆菌物种。 MSR将有两个组成部分：物理和电子数据库（或eSource）。物理资源将包括约1，300个高度保守的编码和非编码分枝杆菌基因的靶向缺失或敲除，以及一系列表达载体，其中编码基因与功能标签融合，以便于未来的分析。eSource将描述物理资源中创建的菌株和质粒的各种初始表型特征。这些包括蛋白质定位和细胞形态的显微镜分析，识别与抗分枝杆菌药物敏感性相关的基因，以及影响细胞表面-环境界面的基因。这些研究产生的数据将汇集并整合到在线结核病数据库（TBDB）上，并将免费提供给科学界。总之，MSR将为分枝杆菌生物学提供前所未有的见解，并将作为分枝杆菌社区研究人员未来研究的发射台。 公共卫生相关性：分枝杆菌疾病长期感染全球数十亿人，每年造成近200万人死亡，艾滋病毒和分枝杆菌的协同作用加剧了这一危机。多药耐药（MDR）和极端耐药（XDR）菌株的出现。结核全面了解分枝杆菌的生物学特性对于致病机理的表征、新型药物靶点的鉴定和疫苗的开发至关重要。该提案将应用基因组学和其他高通量技术来建立分枝杆菌系统资源，该资源将提供所有分枝杆菌共有的生物过程的全面，综合的基因组总结，这将大大加速整个分枝杆菌群落的研究。

DESCRIPTION (provided by applicant): Mycobacterial diseases, predominantly tuberculosis, chronically infect billions and kill nearly two million people annually. This global crisis is exacerbated by inadequate treatments: the BCG vaccine is of questionable efficacy, and the incidence of multi-drug and extremely-drug resistant strains of M. tuberculosis is increasing alarmingly. We will apply genomics and high throughput technologies to develop a Mycobacterial Systems Resource (MSR) that will dramatically accelerate research throughout the entire mycobacterial community. The MSR will be freely available and will provide a comprehensive, integrated genomic and visual summation of biological processes common to all mycobacteria that can be immediately applied to further our understanding of mycobacterial diseases. Insights gained from the MSR will generate a plethora of new biological and comparative tools and identify many new targets suitable for the development of novel anti- mycobacterial therapeutics. Thus, the MSR will stimulate the mycobacterial research community, and facilitate the translation of basic discoveries to combat species that cause an enormous global health burden. These goals will be achieved by taking advantage of the combined expertise and cutting-edge technologies available to the researchers assembled to form this multi-disciplinary, multi-institutional team. Pathogenic mycobacteria are extremely slow-growing and hazardous to work with, making them inappropriate for high-throughput genomic analyses. We will obviate this problem by using a closely related species, M. smegmatis, which is a fast-growing, non-pathogenic mycobacterium that is the acknowledged genetically tractable model Mycobacterium. The focus of the MSR will be ~1,300 M. smegmatis coding and non-coding genes that are highly conserved between members of the M. tuberculosis complex, M. avium, M. ulcerans and M. leprae. These highly conserved genes will mediate fundamental cellular processes and therefore their analysis will be applicable to all mycobacterial species. The MSR will have two components: physical and electronic database (or eSource). The physical resource will consist of a targeted deletion or knockdown of ~1,300 highly conserved coding and non-coding mycobacterial genes, and a collection of expression vectors with the coding genes fused to functional tags to facilitate future analyses. The eSource will describe a wide variety of initial phenotypic characterizations of the strains and plasmids created in the physical resource. These include microscopic analyses of protein localization and cellular morphology, identification of genes associated with sensitivity to anti-mycobacterial drugs, and genes influencing the cell's surface-environment interface. The data generated from these studies will be assembled and integrated on the online Tuberculosis Database (TBDB) and will be freely available to the scientific community. In summary, the MSR will provide unprecedented insight into mycobacterial biology and will serve as a launch pad for future studies by researchers spanning the mycobacterial community. PUBLIC HEALTH RELEVANCE: Mycobacterial diseases chronically infect billions of the global populace and kill nearly two million annually, a crisis that is exacerbated by the synergistic association of HIV and M. tuberculosis, and the appearance of multi-drug (MDR) and extremely-drug (XDR) resistant strains of M. tuberculosis. A comprehensive understanding of the biology of mycobacteria is critical for the characterization of pathogenesis, the identification of novel drug targets, and the development of vaccines. This proposal will apply genomics and other high throughput technologies to establish a Mycobacterial Systems Resource that will provide a comprehensive, integrated genomic summation of biological processes common to all mycobacteria, which will dramatically accelerate research throughout the entire mycobacterial community.