Pathogen Synergy Through Cross-Species Induction of Outer Membrane Vesicle Biogenesis

通过跨物种诱导外膜囊泡生物发生的病原体协同作用

• 批准号: 10204938
• 负责人: Jeffrey Schertzer
• 金额: $ 15.02万
• 依托单位: STATE UNIVERSITY OF NY,BINGHAMTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-29 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204938
• 关键词: ANXA5 gene; Antibiotics; Apoptosis; Area; Bacteria; Behavior; Biogenesis; Cells; Coculture Techniques; Communicable Diseases; Communication; Communities; Complex; Data; Disease; Disease model; Enzymes; Epidemiology; Epithelial Cells; Escherichia coli; Foundations; Future; Gammaproteobacteria; Human; Immune response; Immune system; In Vitro; Individual; Induction of Apoptosis; Infection; Label; Lead; Life; Lipids; Lung; Mass Fragmentography; Mediator of activation protein; Membrane; Methods; Modeling; Organism; Pathogenesis; Pathogenicity; Patient-Focused Outcomes; Patients; Phenotype; Play; Population; Problem Solving; Production; Pseudomonas aeruginosa; Reporting; Role; Signal Transduction; Site; Testing; Time; Toxin; Trypan Blue; Vesicle; Virulence; Virulence Factors; Virulent; Work; antibiotic tolerance; bacterial community; cell type; co-infection; cytotoxic; cytotoxicity; experimental study; follow-up; genetic element; human pathogen; in vivo; insight; macrophage; microbiota; novel; pathogen; pathogenic bacteria; small molecule; synergism

Abstract Bacteria have been cooperating and competing for all of evolutionary time. Only recently, however, have we seriously begun to take this into account in the context of infectious disease mechanisms. The expanding ability to study bacteria in complex consortia (i.e. how they normally live) has sparked a renewed appreciation for the complexity of real bacterial populations and an examination of how they interact. Co-infection studies show that pathogen communities are more virulent, and this aligns with epidemiological reports that connect multispecies infections to worse patient outcomes. A common hypothesis is that this pathogen synergy comes about because of competition and communication between multiple species at an infection site. We hypothesize that Outer Membrane Vesicles (OMVs) serve as a major mediator of these interactions because they are known to facilitate both competition and communication between bacteria (in addition to direct virulence against host cells). We proposed a model in P. aeruginosa where OMV biogenesis is driven by the secretion and insertion of a self- produced small molecule into the outer leaflet of the outer membrane. We recently showed that this molecule could induce related species to overproduce OMVs when given at low concentration, and that those recipient species produced their own OMV cross-inducing factors. This raised the possibility that cross-species induction of OMV biogenesis could serve as a mechanism for multispecies communities to synergize for increased virulence. Guided by strong preliminary data, we will pursue two specific aims to characterize how multiple pathogens at an infection site might interact through cross-species induction of OMV biogenesis and what effects this would have on host cells: (1) We will begin by testing for small molecule-induced OMV formation using the known P. aeruginosa OMV-inducing compound PQS against a broad panel of important human pathogens likely to encounter each other at an infection site. Our established methods have already identified several such interactions among the γ-proteobacteria. We will then test whether actually growing species together (in contrast to using monocultures with exogenous compounds as above) will result in increased OMV production for the community. Preliminary results show that this is true for our pilot pairing of P. aeruginosa + E. coli. (2) OMVs from mono- vs. co-culture will be tested for their cytotoxic potential against macrophage and lung epithelial cells as well as their ability to degrade or sequester antibiotics (another disease-related function of OMVs). Preliminary results with P. aeruginosa + E. coli co-culture OMVs show that cytotoxicity and induction of apoptosis are both increased over monoculture OMVs, demonstrating feasibility of the approach and providing support for the hypothesis that cross-species OMV induction may contribute to pathogen synergy. Understanding pathogen interactions at infection sites is critical to understanding why multispecies infections lead to more severe disease. Given the ubiquity of OMV production among Gram-negative organisms, the insights gained from this study promise to broadly impact our understanding of multi-species pathogenesis.

抽象的 细菌一直在整个进化时期合作和竞争。 认真地考虑了传染病机制的背景 研究复杂的细菌（即，他们的正常生活如何）引发了对它们的重新欣赏 实际的双脚群体的复杂性和对它们相互作用的研究表明。 病原体群落更具敏感性，这与连接多种产品的流行病学报告保持一致 感染对患者预后更糟的感染。 在感染部位的竞争和沟通。 膜囊泡（OMV）是相互作用的主要中介者，因为已知它们可以促进 竞争和巴克氏菌之间的交流（除了直接针对宿主细胞的病毒性）。 我们在铜绿假单胞菌中支撑了一个模型，其中OMV生物发生是由自我的分泌和插入驱动的 产生的小分子进入外膜的外部小叶。 在低浓度下给出时，可能会诱导相关物种过度生产OMV，而那些受体 物种产生了自己的OMV交叉诱导因素。 OMV生物发生可以作为多种植者协同作用以增加的机制 在强大的初步数据的指导下，我们将追求两个特定的目标 感染部位的病原体可能通过跨物种诱导OMV生物发生以及什么影响相互作用 这将在宿主细胞上具有：（1）我们将首先测试小分子诱导的OMV形成 已知的铜绿假单胞菌OMV诱导化合物PQs针对一组重要的人类病原体可能 在感染部位相互遇到。 γ-蛋白菌之间的相互作用。 使用具有外源化合物的单一培养物将增加OMV的产生。 社区 从单与培养物中，将对其针对巨噬细胞和肺部占地细胞的细胞毒性潜力进行测试 以及降解或隔离抗生素（OMV的另一个功能）的Arity 铜绿 + e的结果。 对单一文化OMV的增加，证明了该方法的可行性并为其提供支持 跨物种OMV指示可能有助于病原体协同作用的假设。 了解感染部位的病原体相互作用对于理解为什么多物种感染至关重要 导致更严重的疾病。 洞察力从跌至巨大影响我们对多物种发病机理的理解的陷入障碍中获得的见解。