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: 10570255
• 负责人: MATTHEW R. PARSEK
• 金额: $ 71.31万
• 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/10570255
• 关键词: Acute; Adopted; Animal Model; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Automobile Driving; Bacteremia; Bacteria; Bacterial Adhesins; Cells; Classification; Clinic; Clinical; Communities; Complex; Cues; Deoxyribonucleases; Development; Enzymes; Event; Excision; Extracellular Matrix; Future; Gene Expression Profile; Genes; Glycoside Hydrolases; Goals; Hospitalization; Human; Immune system; In Vitro; Individual; Infection; Knowledge; Laboratories; Microbial Biofilms; Mus; Names; Nature; Nosocomial Infections; Operative Surgical Procedures; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Polymers; Predisposition; Process; Proteins; Pseudomonas aeruginosa; Psychological reinforcement; Regulon; Research; Role; Signal Transduction; Societies; Swimming; Testing; Translating; Virulence; World Health Organization; Wound models; antimicrobial; burn wound; cell motility; chronic infection; gene function; global health; human pathogen; in vivo; microorganism; novel; resilience; response; spatiotemporal; 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.

项目总结 抗菌素耐药性、慢性感染是当今社会面临的最大挑战之一。这 这一问题如此严重，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年对全球健康的威胁。超过80%的这些感染被归因于引人注目的 细菌形成生物膜的能力，包裹在保护性基质中的微生物群落 对抗菌剂不敏感的胞外聚合物。铜绿假单胞菌居第二位 这些感染的主要原因，通常也是医院感染患者死亡的最常见原因 感染。分散体被认为是控制生物膜的一条很有前途的途径，因为它有助于分解生物膜。 相关感染。这是因为扩散是细菌逃逸的一个活跃的、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，因此具有分散性 提高抗生素的疗效。然而，虽然铜绿假单胞菌对弥散线索的感知以及这些 信号被翻译成c-di-GMP池的调制已经被非常详细地阐明，鲜为人知 关于下游事件使铜绿假单胞菌能够离开生物膜，c-di-GMP的贡献，或 驱动毒力的因素通过分散的细胞进行切换。使用rna-seq，我们已经确定了独一无二的基因。 导致分散的细胞(称为“分散调节基因”)，并导致分散的细胞的表型 包括药物敏感性、毒性和基质降解。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。然而，没有明显的矩阵 到目前为止，已经确定了针对基质EPS PSL的降解酶，这表明在 不同类型产生基质的铜绿假单胞菌对基质的降解和分散。而且，我们的 研究结果表明，c-di-GMP的调节不足以引起弥散，只能解释一些 分散细胞的表型。基于这些前提，我们假设分散和分散- 相关的表型需要c-di-GMP(In)依赖的途径，包括参与 在不同的矩阵利用类别中可能有所不同的矩阵退化。这个项目的目标是 阐明导致分散的信号和调节事件，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。 以及体内和体外分散相关的表型，以及时空评估基质的命运 目标2中分散过程中的组件。目标3旨在确定分散和所需的因素 在代表4个不同基质类别的非粘液性临床分离株中，对分散性的影响相似或不同。 我们期待我们的发现不仅有助于了解感染，还有助于指导 分散体向生物被膜控制的辅助疗法的发展。