Mechanisms for acquisition and transmission of successful antibiotic resistant pneumococcal clones pre- and post-vaccination

肺炎链球菌的抗菌素耐药性通过数量有限的克隆在全球传播。PCV疫苗接种降低了疫苗型毒株的抗菌素耐药性。抗菌素耐药性现在是通过非pcv类型的扩展而出现的。该项目侧重于AMR克隆的遗传/功能特性，目标是瞄准它们在携带者群体中的成功和传播。目标是：1)对引入PCV后出现的AMR进行基于全基因组的分析，并与PCV前进行比较。序列数据将与宿主因素相关，包括临床患者信息。将应用系统发育和通用机器学习方法，并建立数据库，以识别将AMR的成功与传播、定植和引起侵袭性疾病的能力联系起来的微生物特征。2)将使用一套动物模型来研究AMR克隆的传播、定植和疾病能力。通过改变宿主免疫防御、甲型流感病毒致敏、长期暴露于抗生素以及影响环境中的物理或化学参数，将不同的内源性和环境线索应用于这些研究。3)将使用适当的突变体确定影响传播/定植/侵袭性疾病的驱动因素，并使用无菌小鼠监测宿主微生物组的影响。将在存在和不存在呼吸微生物组的情况下研究接合转移和缺乏CRISPR/Cas9干扰5)克隆消除将在小鼠模型中尝试使用靶向AMR克隆的抗原，并通过生成在运输过程中干扰AMR克隆的CRISPR/Cas9递送系统。

AMR in Streptococcus pneumoniae is spread globally by a limited number of clones. PCV vaccination has decreased AMR among vaccine-type strains. AMR now emerges by expansion of non-PCV types. The project focuses on genetic/functional properties of AMR clones with the goal to target their success and transmission in the carrier population. The goals are to: 1) Perform whole genome based analyses on emerging AMR after PCV introduction, comparing with pre-PCV. Sequence data will be correlated to host factors including clinical patient information. Phylogenetic and general machine learning methods will be applied and a data base will be created to identify microbial traits that link success of AMR to transmission, colonization and ability to cause invasive disease 2) A set of animal models will be used to study transmission, colonization and disease capability of AMR clones. Different endogenous and environmental cues will be applied to these studies by altering host immune defense, by sensitizing with influenza A virus, by exposure to long term antibiotics, and by affecting physical or chemical parameters in the environment 3) Drivers affecting transmission/colonization/invasive disease will be identified using appropriate mutants, and monitoring the influence of the host microbiome using germ free mice 4) Resistance transfer and role of competence pili, conjugative transfer and lack of CRISPR/Cas9 interference will be studied in the presence and absence of a respiratory microbiome 5) Clonal elimination will be attempted in mice models using antigens targeting AMR clones and by generating a CRISPR/Cas9 delivery system for interference of AMR clones during carriage.