Dispersion and the Biofilm Matrix of Pseudomonas aeruginosa

铜绿假单胞菌的分散和生物膜基质

• 批准号: 10376850
• 负责人: MATTHEW R. PARSEK
• 金额: $ 55.47万
• 依托单位: STATE UNIVERSITY OF NY,BINGHAMTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-25 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10376850
• 关键词: Acute; Adopted; Animal Model; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Automobile Driving; Bacteremia; Bacteria; Bacterial Adhesins; Cells; Clinic; Clinical; Communities; Complex; Cues; Deoxyribonucleases; Development; Enzymes; Event; Excision; Extracellular Matrix; Future; Gene Expression Profile; Genes; Glycoside Hydrolases; Goals; Human; Immune; Immune system; In Vitro; Individual; Infection; Knowledge; Laboratories; Lead; Microbial Biofilms; Mus; Names; Nature; Nosocomial Infections; Operative Surgical Procedures; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Play; Polymers; Predisposition; Process; Proteins; Pseudomonas aeruginosa; Psychological reinforcement; Regulon; Research; Role; Signal Transduction; Societies; Swimming; Testing; Translating; Virulence; World Health Organization; Wound models; antimicrobial; base; burn wound; cell motility; chronic infection; gene function; global health; human pathogen; in vivo; microorganism; novel; transcriptome sequencing

PROJECT SUMMARY Antimicrobial resistant, chronic infections are one of the greatest challenges facing our society today. This problem is so significant that the World Health Organization named antibiotic resistance as one of the top ten threats to global health in 2019. More than 80% of these infections have been attributed to the remarkable capabilities of bacteria to form biofilms, communities of microorganisms encased in a protective matrix of extracellular polymeric substance, that are irresponsive to antimicrobials. Pseudomonas aeruginosa is the 2nd leading cause of these infections, and routinely the most common cause of fatalities in patients with nosocomial infections. Dispersion is considered a promising avenue open for biofilm control, by assisting in resolving biofilm- associated infections. This is because dispersion is an active, regulated process by which bacteria escape biofilms as free-living cells that are once again susceptible to antimicrobials, with dispersion thus capable of enhancing the efficacy of antibiotics. However, while dispersion cue sensing by P. aeruginosa and how these cues are translated into the modulation of the c-di-GMP pool has been elucidated in great detail, little is known about downstream events enabling P. aeruginosa to egress from the biofilm, the contribution of c-di-GMP, or factors driving the virulence switch by dispersed cells. Using RNA-seq, we have identified genes that are unique to dispersed cells (termed “dispersion regulon”) and contribute to the phenotype ascribed to dispersed cells including drug susceptibility, virulence, and matrix degradation. We confirmed factors involved in matrix degradation and the reinforcement of the biofilm matrix to contribute to dispersion. However, no obvious matrix degrading enzymes have been thus far identified which target the matrix EPS Psl, suggesting differences in matrix degradation and dispersion by different classes of matrix-producing P. aeruginosa strains. Moreover, our findings suggest that the modulation of c-di-GMP is not sufficient to induce dispersion and only explains some of the phenotypes of dispersed cells. Based on these premises, we hypothesize that dispersion and dispersion- related phenotypes require c-di-GMP-(in)dependent pathways including genes and gene functions involved in matrix degradation that likely differ amongst the classes of matrix utilization. The goal of this project is to elucidate the signaling and regulatory events leading to dispersion including matrix-degradation, and dispersed cells adopting an acute virulence phenotype. We will therefore elucidate in Aim 1 pathways leading to dispersion and dispersion-related phenotypes in vitro and in vivo, and spatio-temporally assess the fate of matrix components during dispersion in Aim 2. Aim 3 aims at determining whether the dispersion and factors required for dispersion are similar or are different in non-mucoid clinical isolates representing 4 different matrix classes. We anticipate our findings to not only contribute to understanding infection but also assist in guiding the development of dispersion towards an adjunctive therapy for biofilm control.

项目概要 抗生素耐药性慢性感染是当今社会面临的最大挑战之一。这 问题如此严重，以至于世界卫生组织将抗生素耐药性列为十大问题之一 2019 年全球健康面临的威胁。其中超过 80% 的感染归因于显着的 细菌形成生物膜的能力，生物膜是包裹在保护性基质中的微生物群落 细胞外聚合物，对抗菌药物无反应。铜绿假单胞菌是第二位 这些感染的主要原因，通常也是院内感染患者死亡的最常见原因 感染。分散被认为是生物膜控制的一个有前途的途径，通过协助解决生物膜- 相关感染。这是因为分散是细菌逃逸的一个主动的、受调节的过程 生物膜作为自由生活的细胞，再次对抗菌剂敏感，因此能够分散 增强抗生素的功效。然而，虽然铜绿假单胞菌的分散线索传感以及这些如何 线索被转化为 c-di-GMP 库的调节已被详细阐明，但知之甚少 关于使铜绿假单胞菌从生物膜中逸出的下游事件、c-di-GMP 的贡献，或 驱动分散细胞毒力转换的因素。使用 RNA-seq，我们鉴定出了独特的基因 分散细胞（称为“分散调节子”）并有助于分散细胞的表型 包括药物敏感性、毒力和基质降解。我们确认了矩阵中涉及的因素 生物膜基质的降解和强化有助于分散。但没有明显的矩阵 迄今为止，已鉴定出针对基质 EPS Psl 的降解酶，这表明 不同类别的产生基质的铜绿假单胞菌菌株的基质降解和分散。此外，我们的 研究结果表明，c-di-GMP 的调节不足以诱导分散，只能解释一些 分散细胞的表型。基于这些前提，我们假设色散和色散- 相关表型需要c-di-GMP（in）依赖性途径，包括参与的基因和基因功能 基质退化可能因基质利用类别而异。该项目的目标是 阐明导致分散的信号传导和调控事件，包括基质降解和分散 采用急性毒力表型的细胞。因此，我们将在目标 1 中阐明导致分散的途径 和体外和体内分散相关的表型，并时空评估基质的命运 目标 2 中分散过程中的成分。目标 3 旨在确定是否需要分散和因子 代表 4 个不同基质类别的非粘液临床分离株的分散度相似或不同。 我们预计我们的发现不仅有助于了解感染，而且有助于指导 开发分散体以用于生物膜控制的辅助疗法。