Probing Polymicrobial Synergy Using High Throughput Genomics

多微生物感染天生比单一物种感染更复杂，这种复杂性已经 是更充分地了解致病机制和发展的重大障碍 采取有效措施控制或预防该病。牙周病是典型的多菌类疾病， 是人类最常见的感染之一。尽管某些生物，如牙龈卟啉单胞菌， 被认为是关键病原体，但正是多菌群落发起和推动了这种疾病。活体内 影像研究表明，口腔微生物群落高度结构化；然而，人们对此知之甚少 关于控制社区内空间方向的机制或空间结构在 致病性。我们正在进行的研究已经确定并量化了协同中的关键空间参数 牙周病原体Aggregatibacter放线菌伴生菌与S.gordonii的群落在这 建议，我们将表征控制空间参数的分子基础和生物学相关性 牙龈假单胞菌协会及其社区伙伴。在第一个目标中，我们将重点关注 牙龈假单胞菌和戈登葡萄球菌，这是一个分子上很好描述的过程，导致了致病性的提高 双重物种群落。我们将确定定义双物种群落的空间参数 以及它们对致病性的影响。此外，通过使用特定的突变体，我们将评估 已知的群落毒力因子和空间定向的沟通机制。在第二个目标中， 我们将使用TN-SEQ和RNA-SEQ基因组方法来全面评估适应度决定因素和 控制牙龈假单胞菌和戈登假单胞菌空间模式的转录组模式。我们将对潜力进行优先排序 代谢协同的贡献者，以进一步研究空间邻近和种间关系 沟通。在第三个目标中，我们将探索PG在感染过程中对健康至关重要的遗传途径 一组不同的共同感染细菌。这些重点和全面的方法的应用 将允许我们在以下背景下将关键空间参数与分子种间相互作用相结合 社区致病潜力。该项目的成功完成将提供基本信息 关于空间定义的协同致病力的发展和调节 多微生物群落，最终可能被转化为针对 以社区为基础的发病机制，是牙周病的基础。

Polymicrobial infections are inherently more complex than mono-species infections, and this complexity has been a significant barrier to both a fuller appreciation of pathogenic mechanisms, and to the development of effective measures to control or prevent the disease. Periodontal diseases typify polymicrobial diseases, and are among the most common infections of humans. Although certain organisms, such as Porphyromonas gingivalis, are considered key pathogens, it is the polymicrobial community that initiates and drives the disease. In vivo imaging studies have shown that oral microbial communities are highly structured; however, little is known regarding the mechanisms that control spatial orientation within communities or the role of spatial structure in pathogenicity. Our ongoing studies have identified and quantified key spatial parameters in synergistic communities of the periodontal pathogen Aggregatibacter actinomycetemcomitans with S. gordonii. In this proposal, we will characterize the molecular basis and biological relevance of spatial parameters controlling the association of P. gingivalis with its community partners. In the first Aim, we will focus on the interaction between P. gingivalis and S. gordonii, a molecularly well-characterized process resulting in elevated pathogenicity of the dual species communities. We will determine the spatial parameters that define the dual species communities and their impact on pathogenicity. Further, through the use of specific mutants, we will assess the impact of known community virulence factors and communication mechanisms on spatial orientation. In the second Aim, we will use Tn-seq and RNA-seq genomic approaches to holistically assess the fitness determinants and transcriptome patterns which control spatial patterning in P. gingivalis and S. gordonii. We will prioritize potential contributors to metabolic synergy for further study of the relationship between spatial proximity and interspecies communication. In the third aim, we will explore Pg genetic pathways important for fitness during infection with a diverse set of co-infecting bacteria. The application of these focused and comprehensively based approaches will allow us to integrate key spatial parameters with molecular interspecies interactions in the context of community pathogenic potential. Successful completion of this project will provide fundamental information regarding the development and regulation of synergistic pathogenicity displayed by spatially defined polymicrobial communities which could ultimately be translated into therapeutic strategies designed to target the community-based pathogenesis that underlies periodontal disease.