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.

项目摘要 抗菌素，慢性感染是当今我们社会面临的最大挑战之一。这 问题是如此重要，以至于世界卫生组织将抗生素耐药性称为前十名 2019年对全球健康的威胁。这些感染中有80％以上归因于非凡的感染 细菌形成生物膜的能力，被包裹在受保护基质中的微生物群落 细胞外聚合物物质，对抗菌剂不重要。铜绿假单胞菌是第二 这些感染的主要原因，这是医院患者死亡的最常见原因 感染。通过协助解决生物膜 - 相关感染。这是因为分散是一个活跃的，受调节的过程，细菌逃脱了 生物膜作为自由活动细胞，再次容易受到抗菌剂的影响，并具有分散性 提高抗生素的效率。但是，铜绿假单胞菌的色散提示以及这些如何感应 提示被翻译成C-Di-GMP池的调制，已经详细阐明了，鲜为人知 关于使铜绿假单胞菌从生物膜中出现的下游事件，C-DI-GMP的贡献或 通过分散细胞推动病毒转换的因素。使用RNA-seq，我们确定了独特的基因 分散细胞（称为“分散调节”），并有助于分配给分散细胞的表型 包括药物敏感性，病毒和基质降解。我们确认了矩阵中涉及的因素 降解和生物膜基质的加强促进分散。但是，没有明显的矩阵 到目前为止，降解酶的靶向矩阵EPS PSL，表明在 不同类别的基质生成铜绿假单胞菌菌株的基质降解和分散。而且，我们的 调查结果表明，C-DI-GMP的调节不足以诱导分散体，仅解释了一些 分散细胞的表型。基于这些前提，我们假设分散和分散 - 相关表型需要c-Di-GMP-（in）依赖途径，包括涉及的基因和基因功能 矩阵利用类别之间可能有所不同的矩阵降解。这个项目的目标是 阐明导致分散的信号传导和调节事件，包括基质降解，并分散 采用急性病毒表型的细胞。因此，我们将在AIM 1途径中阐明导致分散的途径 体外和体内与色散相关的表型，并在空间上评估基质的命运 AIM 2中分散期间的组件。AIM3的目的旨在确定分散体和所需因素是否 对于代表4个不同矩阵类别的非核酸临床分离株，分散株相似或不同。 我们预计我们的发现不仅有助于理解感染，而且还有助于指导 开发用于生物膜控制的辅助疗法的分散体。