Metabolic adaptions of Mycobacterium tuberculosis at diverse host-pathogen interfaces

结核分枝杆菌在不同宿主-病原体界面上的代谢适应 摘要 确定结核分枝杆菌(Mtb)代谢产物影响人类的机制 结核病的诊断和治疗将需要实施动物模型系统。 它们既易于处理，又能忠实地复制在人类身上观察到的结核病状态。要克服这个问题 有限的遗传和表型多样性在标准C57BL/6J中显示，在这里，我们建议利用 合作杂交(CC)小鼠综合小组，遗传多样性的重组近交系小组 老鼠。我们以前已经表明，CC涵盖了广泛的结核病特征和 感染微环境，超过在经典近交系中观察到的表型谱。基座 根据先前的TnSeq研究并作为母体TBRU赠款的一部分(“Mtb毒力的代谢决定因素， 脆弱性和变异“；1U19AI162584-02)，我们对与结核分枝杆菌相关的代谢基因进行了排序 控制感染人类的结核分枝杆菌菌株的毒力或控制血脂变化。19个高价值Mtb代谢基因 经鉴定，在常规C57BL/6J(BL6)小鼠品系中只有一个受控生长。然而，不止是 当在CC小鼠品系中筛选时，所研究的突变体中有一半显示出体内生长表型。此外， 通过研究CC宿主的几个遗传背景，其中单个细菌基因存在或不存在 在体内控制结核分枝杆菌的存活，我们就能够开始定义控制结核分枝杆菌应答的宿主因素。这个 目前的应用将扩展这些研究，以专注于CRISPRi的靶向细菌脂质文库 突变体与CC小鼠的综合小组相结合。通过感染60种不同的CC基因型 靶向脂质CRISPRi文库，Mtb的代谢易损性将在 遗传多样性的寄主。通过使用细菌突变丰度作为表型特征，我们将进行 跨CC面板的数量性状基因座(QTL)作图，以确定与脂质相关的宿主基因座 结核分枝杆菌感染的要求。总之，这些双重宿主和病原体方法将识别宿主-结核分枝杆菌 脂类相互作用可作为结核病治疗的靶点。

Metabolic adaptions of Mycobacterium tuberculosis at diverse host-pathogen interfaces ABSTRACT Defining the mechanisms through which Mycobacterium tuberculosis (Mtb) metabolites impact human tuberculosis (TB) disease, diagnostics and treatment will require the implementation of model animal systems that are both tractable and faithfully replicate the TB disease states observed in humans. To overcome the limited genetic and phenotypic diversity shown in standard C57BL/6J, here we propose to leverage the comprehensive panel of Collaborative Cross (CC) mice, a genetically diverse panel of recombinant inbred mice. We have previously shown that the CC encompasses a broad spectrum of TB disease traits and infection microenvironments, exceeding the phenotypic spectrum observed in classical inbred strains. Based on previous TnSeq studies and as part of the parent TBRU grant (“Metabolic determinants of Mtb virulence, vulnerability and variation”; 1U19AI162584-02), we have ranked Mtb metabolic genes that were linked to Mtb virulence or control lipid variation in Mtb strains that infect humans. Of 19 high value Mtb metabolic genes identified, only one controlled growth in the conventional C57BL/6J (BL6) mouse strain. However, more than half the mutants studied showed in vivo growth phenotypes when screened across CC mouse strains. Further, through study of several of the CC host genetic backgrounds in which individual bacterial genes do or do not control in vivo Mtb survival, we are able to start to define the host factors in control of Mtb response. The current application will now extend these studies to focus on a targeted bacterial lipid library of CRISPRi mutants in conjunction with the comprehensive panel of CC mice. By infecting 60 distinct CC genotypes with the targeted lipid CRISPRi library, the metabolic vulnerabilities of Mtb will be comprehensively defined in genetically diverse hosts. By using bacterial mutant abundance as a phenotypic trait, we will conduct quantitative trait loci (QTL) mapping across the CC panel to identify the host loci underlying the lipid requirements of Mtb infection. Altogether these dual host and pathogen approaches will identify the host-Mtb lipid interactions that can be targeted for tuberculosis treatment.