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模型动物系统 它们既易于处理，又忠实地复制了在人类中观察到的结核病状态。克服 有限的遗传和表型多样性显示在标准C57 BL/6J，在这里，我们建议利用 协作杂交（CC）小鼠的综合小组，一组遗传多样的重组近交系小鼠， 小鼠我们以前已经表明，CC包括广泛的结核病特征， 感染微环境，超过了在经典近交系中观察到的表型谱。基于 在先前的TnSeq研究中以及作为母TBRU资助的一部分（"Metabolic determinants of Mtb virtuality， 脆弱性和变异"; 1U19AI162584 - 02），我们对与结核分枝杆菌相关的结核分枝杆菌代谢基因进行了排序， 感染人类的Mtb菌株中的毒力或控制脂质变异。19个高价值结核分枝杆菌代谢基因 在常规C57 BL/6J（BL6）小鼠品系中，仅一种控制生长。但超过 当在CC小鼠品系中筛选时，所研究的一半突变体显示出体内生长表型。此外，本发明还 通过研究几种CC宿主的遗传背景，其中单个细菌基因做或不做 通过控制体内Mtb存活，我们能够开始定义控制Mtb应答的宿主因素。的 目前的申请现在将扩展这些研究，以专注于CRISPRi的靶向细菌脂质文库 突变体与CC小鼠的综合小组一起。通过感染60种不同的CC基因型， 靶向脂质CRISPRi文库，Mtb的代谢脆弱性将在 基因多样的宿主通过使用细菌突变体丰度作为表型性状，我们将进行 在CC面板上进行数量性状基因座（QTL）作图，以鉴定脂质代谢相关的宿主基因座。 Mtb感染的要求。总之，这些双重宿主和病原体的方法将确定宿主-Mtb 脂质相互作用，可以针对结核病治疗。

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.