Characterization of Biofilms by Correlated Mass Spectrometric and Raman Imaging

通过相关质谱和拉曼成像表征生物膜

• 批准号: 10468036
• 负责人: Joshua Shrout
• 金额: $ 50.35万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468036
• 关键词: 3-Dimensional; Address; Animal Model; Bacteria; Bacterial Infections; Behavior; Biochemical; Biological; Biological Assay; Cataloging; Cells; Characteristics; Chemicals; Chemistry; Clinical; Coculture Techniques; Communities; Complex; Deposition; Detection; Environment; Exhibits; Extracellular Matrix; Gene Expression; Genes; Goals; Grant; Image; Imaging Device; In Situ; In Vitro; Individual; Infection; Lung; Maps; Mediating; Metagenomics; Methodology; Methods; Microbe; Microbial Biofilms; Modeling; Mus; Nature; Nutrient; Pattern; Peptides; Phenazines; Polymers; Population; Positioning Attribute; Process; Production; Pseudomonas aeruginosa; Quinolones; Raman Spectrum Analysis; Regulation; Research; Resolution; Sampling; Signal Transduction; Spatial Distribution; Staphylococcus aureus; Stimulus; Surface; System; Technology; Therapeutic; Time; Tissues; Virulence; Virulence Factors; Virulent; Work; bacterial community; base; cell community; density; design; diagnostic strategy; experimental study; extracellular; gene product; homoserine lactone; host colonization; host-associated microbial communities; imaging approach; imaging modality; improved; insight; interest; interspecies communication; mass spectrometric imaging; microbial; microbial colonization; microbial community; multimodality; opportunistic pathogen; pathogen; pathogenic bacteria; quinoline; quorum sensing; response; rhamnolipid; social; spatiotemporal; trait

PROJECT SUMMARY Most bacteria in natural and clinical settings grow as surface-attached biofilms, which are bacterial communities that have self-assembled into an encased matrix of extracellular polymeric substances (EPS). To form these bacterial biofilm communities and infect host cells, an intercellular signaling process described as quorum sensing (QS) is very important. For the opportunistic pathogens Pseudomonas aeruginosa and Staphylococcus aureus, QS regulates the expression of many genes important to biofilm initiation, EPS production, and virulence. While much has been learned about select factors that regulate biofilm formation in vitro and in animal models, the specific mechanisms by which multispecies groups of cells form biofilms are not understood. Furthermore, the therapeutic capabilities needed to treat the increasingly virulent infections of the coming decades will require deeper insight into multi-species interactions and their regulation of multiple gene expression profiles as represented over extended spatial and temporal scales during host cell infection. The correlated mass spectrometric and Raman chemical imaging approaches that have been developed by our combined research group circumvent the limitations imposed by previous technology which allowed examination of either a small number of cells or entire cell populations that had been removed from the conditions of interest. In contrast our correlated chemical imaging methods allow the determination and spatial mapping of individual bacterial species and their microbial products within a mixed bacterial community growing in situ on surfaces. One of our long-term goals is the design of detection and diagnostic strategies informed by an understanding of bacterial interactions and signature biomolecule production. We will continue our work toward this goal by conducting mono-culture and co-culture experiments using Pseudomonas aeruginosa and Staphylococcus aureus as model pathogens. Correlated chemical imaging will be used to define the onset and range of intercellular quorum sensing signaling in space and time. The biosecretome and interactions of specific species in co-culture biofilms will be characterized for both intra- and inter-species interactions. Also, an ex vivo lung assay will be created to study the spatial biofilm secretome. This research will exploit the ability to spatially map specific chemical products produced by these and other pathogenic bacteria thereby yielding deep and therapeutically informative insights into host colonization, infection, and virulence.

项目概要 自然和临床环境中的大多数细菌以表面附着的生物膜的形式生长，这些生物膜是细菌 自组装成胞外聚合物 (EPS) 封装基质的群落。到 形成这些细菌生物膜群落并感染宿主细胞，细胞间信号传导过程描述为 群体感应（QS）非常重要。对于机会致病菌铜绿假单胞菌和 金黄色葡萄球菌，QS 调节许多对生物膜形成很重要的基因的表达，EPS 产量和毒力。虽然人们对调节生物膜形成的选择因素已经了解很多 在体外和动物模型中，多物种细胞形成生物膜的具体机制尚不明确。 明白了。此外，治疗日益致命的感染所需的治疗能力 未来几十年将需要更深入地了解多物种相互作用及其对多基因的调控 宿主细胞感染期间在扩展的空间和时间尺度上表示的表达谱。 已开发的相关质谱和拉曼化学成像方法 我们的联合研究小组规避了先前技术所施加的限制 检查从细胞中取出的少量细胞或整个细胞群 利益条件。相比之下，我们的相关化学成像方法允许确定和空间 混合细菌群落中单个细菌物种及其微生物产物的绘图 在表面原位生长。 我们的长期目标之一是设计基于以下信息的检测和诊断策略： 了解细菌相互作用和特征生物分子的产生。我们将继续努力 通过使用铜绿假单胞菌进行单一培养和共培养实验来实现这一目标 金黄色葡萄球菌作为模型病原体。相关的化学成像将用于定义发病和 细胞间群体感应信号在空间和时间上的范围。生物分泌组及其相互作用 共培养生物膜中的特定物种将针对物种内和物种间相互作用进行表征。还， 将创建离体肺测定来研究空间生物膜分泌组。这项研究将利用 能够在空间上绘制这些和其他病原菌产生的特定化学产品的能力 对宿主定植、感染和毒力产生深入且具有治疗信息的见解。