Spatiotemporal analysis of gene regulatory programs associated with GBS infection during pregnancy

项目总结 怀孕期间或出生后立即感染与相当大的发病率和 全球范围内的死亡率。B组链球菌(GBS)是引起绒毛膜羊膜炎(CAM)和新生儿的主要原因 败血症。胎盘是GBS和宿主免疫反应之间的初始界面。 绒毛膜羊膜炎，构成宿主-病原体相互作用的场所。目前对该问题的理解 CAM的发病机制一直受到缺乏揭示其空间和时间成分的工具的限制 胎盘感染。具体地说，我们缺乏对感染如何随着时间和跨地区展开的了解 胎盘式建筑。单细胞基因组学和空间转录组学的突破现在使 对感染过程进行公正和系统的分析。在这里，我们建议将这些方法应用于GBS 胎盘感染：检查宿主特异性反应，宿主-病原体界面，以及 GBS感染项目。在目标1中，我们将结合空间转录学和单细胞rna-seq进行研究。 在一种新的小鼠上行感染和CAM模型中GBS感染的进展。在《目标2》中我们将 同时研究原代人类感染过程中宿主和病原体的基因表达程序 胎盘巨噬细胞。在目标3中，我们将通过系统评估来测试GBS感染的概念模型 靶向缺失特定GBS基因的效果。集成这些方法将提供丰富的视图 在CAM过程中宿主与病原体的相互作用，并将导致新的干预靶点的确定。

PROJECT SUMMARY Infection during pregnancy or in the immediate postnatal period is associated with substantial morbidity and mortality worldwide. Group B Streptococcus (GBS) is a major cause of chorioamnionitis (CAM) and neonatal sepsis. The placenta acts as the initial interface between GBS and the host immune response during chorioamnionitis, constituting the locus of host-pathogen interaction. Current understanding of the pathogenesis of CAM has been limited by a lack of tools revealing the spatial and temporal components of placental infection. Specifically, we lack an understanding of how the infection unfolds over time and across the placental architecture. Breakthroughs in single-cell genomics and spatial transcriptomics now enable the unbiased and systematic analysis of the process of infection. Here we propose to apply these methods to GBS infection of the placenta: examining host-specific responses, the host-pathogen interface, and perturbations of the GBS infection program. In Aim 1 we will combine spatial transcriptomics and single-cell RNA-Seq to study the progression of GBS infection in a novel murine model of ascending infection and CAM. In Aim 2 we will simultaneously study host and pathogen gene expression programs during infection in primary human placental macrophages. In Aim 3 we will test a conceptual model of GBS infection by systematically assessing the effect of targeted deletions of specific GBS genes. Integrating these approaches will provide a rich view of host-pathogen interactions during CAM and will lead to the identification of new targets for intervention.