Multiplexed Strain Phenotyping Defines Consequences of Genetic Diversity in Mycobacterium tuberculosis for Infection and Vaccination Outcomes.
Multiplexed Strain Phenotyping Defines Consequences of Genetic Diversity in Mycobacterium tuberculosis for Infection and Vaccination Outcomes.
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DOI：
10.1128/msystems.00110-22
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发表时间：
2022-06-28
期刊：
影响因子：
6.4
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中科院分区：
文献类型：
--
作者：
越来越多的证据表明，结核分枝杆菌（结核病的病原体）的遗传多样性有助于感染和公共卫生干预措施（如疫苗接种）的结果。流行病学研究表明，在M.在结核病中，属于谱系2（mL2）的亚谱系的菌株与相关的临床特征相关，包括高毒力、治疗失败和疫苗逃逸。这种亚系的全球扩张和日益增加的患病率归因于这些特征赋予的选择性优势，然而混淆的宿主和环境因素使得难以在人类研究中确定驱动这些关联的细菌决定因素。在这里，我们开发了一种分子条形码策略，以促进高通量，实验表型M。结核病临床分离株。这种方法使我们能够表征一组遗传多样性M。在小鼠模型中，在感染期间和接种疫苗后，我们发现mL2菌株在体内表现出不同的生长动力学，并且对卡介苗（BCG）接种所赋予的免疫保护具有抗性。后一项发现证实了流行病学观察结果，并表明分枝杆菌的特征有助于疫苗的效力。为了研究mL2菌株独特表型的遗传和生物学基础，我们进行了变异分析、转录研究和全基因组转座子测序。我们在一个代表性的mL2菌株中确定了多个应激和宿主反应途径中的功能性遗传变化，这些变化与调控基因的变体相关。这些适应性变化可能是该谱系独特的临床特征和流行病学成功的基础。结核病由结核分枝杆菌引起，是一种非常异质性的疾病，这一特点使临床护理和公共卫生干预变得复杂。病原体遗传多样性对这种异质性的贡献是不确定的，部分原因是实验操作M。结核病，一种生长缓慢的生物安全等级为3级的微生物。为了克服这些挑战，我们将分子条形码策略应用于一组M.结核病临床分离株。条形码的这种新应用允许高通量表征M。结核菌株在小鼠感染模型中的生长动力学和疫苗抗性。将这些结果与基因组分析相结合，我们发现了导致感染结果的细菌途径，为改进的治疗方法和疫苗提供了靶点。
There is growing evidence that genetic diversity in Mycobacterium tuberculosis, the causative agent of tuberculosis, contributes to the outcomes of infection and public health interventions, such as vaccination. Epidemiological studies suggest that among the phylogeographic lineages of M. tuberculosis, strains belonging to a sublineage of Lineage 2 (mL2) are associated with concerning clinical features, including hypervirulence, treatment failure, and vaccine escape. The global expansion and increasing prevalence of this sublineage has been attributed to the selective advantage conferred by these characteristics, yet confounding host and environmental factors make it difficult to identify the bacterial determinants driving these associations in human studies. Here, we developed a molecular barcoding strategy to facilitate high-throughput, experimental phenotyping of M. tuberculosis clinical isolates. This approach allowed us to characterize growth dynamics for a panel of genetically diverse M. tuberculosis strains during infection and after vaccination in the mouse model. We found that mL2 strains exhibit distinct growth dynamics in vivo and are resistant to the immune protection conferred by Bacillus Calmette-Guerin (BCG) vaccination. The latter finding corroborates epidemiological observations and demonstrates that mycobacterial features contribute to vaccine efficacy. To investigate the genetic and biological basis of mL2 strains’ distinctive phenotypes, we performed variant analysis, transcriptional studies, and genome-wide transposon sequencing. We identified functional genetic changes across multiple stress and host response pathways in a representative mL2 strain that are associated with variants in regulatory genes. These adaptive changes may underlie the distinct clinical characteristics and epidemiological success of this lineage. IMPORTANCE Tuberculosis, caused by the bacterium Mycobacterium tuberculosis, is a remarkably heterogeneous disease, a feature that complicates clinical care and public health interventions. The contributions of pathogen genetic diversity to this heterogeneity are uncertain, in part due to the challenges of experimentally manipulating M. tuberculosis, a slow-growing, biosafety level 3 organism. To overcome these challenges, we applied a molecular barcoding strategy to a panel of M. tuberculosis clinical isolates. This novel application of barcoding permitted the high-throughput characterization of M. tuberculosis strain growth dynamics and vaccine resistance in the mouse model of infection. Integrating these results with genomic analyses, we uncover bacterial pathways that contribute to infection outcomes, suggesting targets for improved therapeutics and vaccines.
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影响因子：
6.7
作者：
Carey AF;Rock JM;Krieger IV;Chase MR;Fernandez-Suarez M;Gagneux S;Sacchettini JC;Ioerger TR;Fortune SM
通讯作者：
Fortune SM
影响因子：
3.2
作者：
Domenech, Pilar;Zou, Jason;Reed, Michael B.
通讯作者：
Reed, Michael B.
影响因子：
3.2
作者：
Fallow, Ashley;Domenech, Pilar;Reed, Michael B.
通讯作者：
Reed, Michael B.
影响因子：
11.8
作者：
Cowley, Donna;Govender, Dhirendra;Nicol, Mark P.
通讯作者：
Nicol, Mark P.
结核分枝杆菌走出非洲的迁徙以及新石器时代与现代人类的共同扩张。
影响因子：
30.8
作者：
Comas, Inaki;Coscolla, Mireia;Luo, Tao;Borrell, Sonia;Holt, Kathryn E.;Kato-Maeda, Midori;Parkhill, Julian;Malla, Bijaya;Berg, Stefan;Thwaites, Guy;Yeboah-Manu, Dorothy;Bothamley, Graham;Mei, Jian;Wei, Lanhai;Bentley, Stephen;Harris, Simon R.;Niemann, Stefan;Diel, Roland;Aseffa, Abraham;Gao, Qian;Young, Douglas;Gagneux, Sebastien
通讯作者：
Gagneux, Sebastien