MRC AMED - Systematic analysis of persistence mechanisms by high-throughput bar-seq and single cell analyses.

抗菌素耐药性（AMR）是一个重大的全球卫生问题，需要通过对耐药性机制进行新颖的合作研究来解决。持久性，发生在细菌种群中一小部分基因相同的亚部分暴露于抗生素后，由于未知的原因，没有被杀死。它是一种表型特征（即非遗传可观察特征），而不是遗传变异，由于它发生在种群的一小部分并且不稳定，因此很难研究。然而，坚持是抗生素不能治愈许多病人疾病的原因。它也可能是AMR的前身，这是一个紧迫的全球健康问题。英国团队在分析单细胞水平的持久性细胞方面具有专业知识，这是观察这种罕见亚群的必要条件。日本团队在构建细菌单基因缺失文库方面具有互补的专业知识；ASKA条码删除集合。ASKA文库中的每个突变株都有一个人工的独特序列，可以通过一种称为深度测序的技术来识别。研究小组已经确定了13种表现出更强持久性的突变体。所涉及的机制尚不清楚，通常在亚种群水平上表现出来，例如，生长缓慢的百分比增加或形态异常。该项目将汇集英国和日本团队的互补专业知识。英国研究小组将利用微流体技术应用单细胞成像技术，在单细胞和亚群体水平上对细菌突变体进行计算机跟踪，以阐明不同形态和生长速度在持久性中的作用。这一知识可能导致开发针对持久性细胞的新药，从而导致更高的治愈率、更短的治疗方案和减少抗菌素耐药性。因此，该项目将有助于应对抗生素耐药性对全球健康的威胁。

Antimicrobial resistance (AMR) is a major global health issue which needs to be addressed by novel, collaborative research into the mechanisms of resistance. Persistence, occurs where a small, genetically-identical sub-fraction of a bacterial population that is exposed to antibiotic is, for unknown reasons, not killed. It is a feature of phenotypic (meaning non-genetic observable characteristic), rather than genetic variation and is very difficult to study because it occurs in a small sub-fraction of the population and is unstable. Yet persistence is the reason why antibiotics fail to cure many patients of disease. It may also be the progenitor of AMR, which is an urgent worldwide health issue. The UK team have expertise in analysing persister cells at the single-cell level, a requisite when observing this rare sub-population. The Japan team have complementary expertise in construction of a bacterial single-gene deletion library; the ASKA barcode deletion collection. Each mutant strain in the ASKA library has an artificial unique sequence that can be identified, via a technique called deep sequencing. The team have identified 13 mutants that exhibit increased persistence. The mechanisms involved are unknown and are often exhibited at the sub-population level, eg increased % of slow growers or aberrant morphology. This project will bring together the complementary expertise of the UK and Japan teams. The UK team will apply single cell imaging using microfluidics to perform computerized tracking of the bacterial mutants at the single cell and sub-population level to elucidate the role of differential morphology and growth rate in persistence. This knowledge could lead to development of novel drugs that target persister cell and thereby lead to higher cure rates, shorter treatment regimes and reduced AMR. The project will thereby contribute to the AMR global health threat.