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

基本信息

批准号：
8755355
负责人：
Joshua Shrout
金额：
$ 46.57万
依托单位：
UNIVERSITY OF NOTRE DAME
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8755355
关键词：
Address Affect Amino Acids Animal Model B-Lymphocytes Bacteria Bacterial Infections Behavior Biochemical Cells Characteristics Chemicals Clinical Coculture Techniques Communities Complex Detection Development Diagnostic Dimensions Escherichia coli Extracellular Matrix Gene Expression Genes Goals Growth Haemophilus influenzae Image In Situ In Vitro Individual Infection Learning Lung Maps Mass Spectrum Analysis Measurement Methodology Methods Microbe Microbial Biofilms Modeling Molecular Profiling Mucins Nature Population Production Proteins Pseudomonas Pseudomonas aeruginosa Quinolones Regulation Research Resolution Sampling Signal Transduction Signaling Molecule Spatial Distribution Surface System Therapeutic Time Variant Virulence Virulent Work design extracellular homoserine lactone imaging modality improved insight microbial microbial community pathogen pathogenic bacteria programs public health relevance quorum sensing research study rhamnolipid signal processing signature molecule spectroscopic imaging trait

项目摘要

DESCRIPTION (provided by applicant): 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 pathogen, Pseudomonas aeruginosa, 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 specifics by which multispecies groups of cells form biofilms are not yet clear. Correlated mass spectrometric and Raman imaging will be used to develop a multiplex analysis method for studying host-associated microbial communities. These methods allow the determination of individual bacterial species and their microbial products within a mixed bacterial community growing in situ on surfaces. A long-term goal is the design of detection and diagnostic strategies informed by an understanding of bacterial interactions and signature biomolecule production. Work toward this goal will begin by conducting experiments with the opportunistic pathogen Pseudomonas aeruginosa, specifically developing methods to distinguish P. aeruginosa from other bacterial species, distinguish separate P. aeruginosa strains from each other, and determine the spatial distribution of four types of P. aeruginosa molecules associated with virulence: acyl- homoserine lactones (AHL), quinolones, such as pseudomonas quinolone signal (PQS), rhamnolipids, and exopolymeric substances (EPS). Successfully implementing this research program will result in bacterial mapping methods with improved sensitivity and resolution, substantially enhancing our ability to identify specific species of bacteria in mixed samples, and to identify and map specific chemical products produced by these bacteria that are critical to host colonization, infection, and virulence.

描述（由申请人提供）：自然和临床环境中的大多数细菌随着表面附着的生物膜而生长，这些细菌群落已自组装成包含的细胞外聚合物物质（EPS）的基质。为了形成这些细菌生物膜群落并感染宿主细胞，一个被描述为Quorum Sensing（QS）的细胞间信号传导过程非常重要。对于机会性病原体，铜绿假单胞菌QS调节许多对生物膜起始，EPS产生和毒力重要的基因的表达。尽管已经了解了在体外和动物模型中调节生物膜形成的某些因素的知识，但尚不清楚形成生物膜的多物细胞组的细节。相关的质谱和拉曼成像将用于开发一种多重分析方法，用于研究宿主相关的微生物群落。这些方法允许在混合细菌群落中确定各个细菌物种及其微生物产物，在地面上生长。一个长期目标是通过了解细菌相互作用和签名生物分子产生的检测和诊断策略的设计。 Work toward this goal will begin by conducting experiments with the opportunistic pathogen Pseudomonas aeruginosa, specifically developing methods to distinguish P. aeruginosa from other bacterial species, distinguish separate P. aeruginosa strains from each other, and determine the spatial distribution of four types of P. aeruginosa molecules associated with virulence: acyl- homoserine lactones (AHL), quinolones, such as假单胞菌信号（PQS），鼠李糖脂和外聚糖物质（EPS）。成功实施这项研究计划将导致具有提高敏感性和分辨率的细菌映射方法，从而大大提高了我们在混合样品中鉴定特定细菌的能力，并识别和绘制这些细菌产生的特定化学产品，这些化学产品对于宿主定植，感染和毒力至关重要。