Establishing the role of pathogenetic mechanisms in the evolution of epidemic K. pneumoniae lineages.

KP流行谱系的系统发育分析利用现有的基因组序列，我们将利用时间和地理数据对不同流行病和ST 15谱系中的RamR突变进行群体水平的基因组和系统发育分析，以确定（i）额外的突变事件（ii）具有有利突变的新兴流行病亚谱系和（iii）绘制突变事件的潜在驱动因素。在ST 15和非ST 15流行谱系中进行拉马调节子的比较基因组学。使用来自生物信息学分析的数据，我们将鉴定相关菌株，以进行已建立的基于转录组学和ChIP-seq的分析，以确定DTP精准医学项目呼叫21/22第3页（共4页）的调节级联和拉马的结合相互作用。这些数据将是至关重要的，在定义的监管影响，拉马和促进机制分析如何这些变化的主机-病原体相互作用的ST 15流行谱系。在模拟全身性感染的特定鼠模型中建立ST 15（拉马过表达）和ST 15 R（使用遗传操作修复拉马过表达）的感染动力学。使用感染的鼠模型，除了评估这些菌株在抗生素存在下的活力之外，我们还将建立与对抗ST 15和ST 15 R变体感染相关的免疫检查点。这些数据将与大卫多克雷尔教授在现有的SHIELD联盟筛选中合作使用，以建立先天免疫应答作为诊断或治疗KPN感染的工具。

Phylogenomic-Analyses of Epidemic lineages of KP. Using existing genome sequences, we will undertake a population level genomic and phylogenetic analyses of RamR mutations in different epidemic and ST15 lineages with temporal and geographic data to identify (i) additional mutational events (ii) emerging epidemic sub-lineages with advantageous mutations and (iii) map the potential drivers of mutational events. Undertake comparative genomics of the RamA regulon in ST15 and non-ST15 epidemic lineages. Using data derived from the bioinformatic analyses, we will identify the relevant strains to undertake established transcriptomic and ChIP-seq based analyses, to ascertain the DTP Precision Medicine Project Call 21/22 Page 3 of 4 regulatory cascade and binding interactions of RamA. This data will be critical in defining the regulatory impact of RamA and facilitate mechanistic analyses of how these changes host-pathogen interactions in the ST15 epidemic lineage. Establish the infection dynamics of ST15 (RamA overexpressed) and ST15R (RamA overexpression is repaired using genetic manipulation) in specific murine models which mimic systemic infection. Using the murine model of infection, in addition to assessing the viability of these strains in the presence of antibiotics, we will establish the immune checkpoints which are relevant to counter the infection of ST15 and ST15R variants. These data will be utilized in collaboration with Prof. David Dockrell in existing SHIELD consortium screens to establish innate immune responses as tools for diagnosis or treatment of KPN infection.