The interplay between cell envelope protein homeostasis and antibiotic resistance in Gram-negative bacteria

革兰氏阴性菌细胞包膜蛋白稳态与抗生素耐药性之间的相互作用

• 批准号: 10514634
• 负责人: Despoina Mavridou
• 金额: $ 38.95万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2026-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10514634
• 关键词: Acinetobacter baumannii; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacterial Antibiotic Resistance; Bacterial Infections; Biochemical; Biochemical Pathway; Biochemistry; Biogenesis; CRISPR interference; Cell Membrane Permeability; Cessation of life; Chemicals; Clinical; Complex; Cytoplasm; Data; Development; Disease model; Enzymes; Epidemiology; Escherichia coli; Evolution; Foundations; Future; Genetic Transcription; Goals; Gram-Negative Bacteria; Healthcare Systems; Home; Impairment; Infection; Investigation; Klebsiella pneumoniae; Knowledge; Laboratories; Light; Link; Mechanical Stress; Mediating; Microbiology; Modeling; Modern Medicine; Multi-Drug Resistance; Mus; Natural regeneration; Nature; Organism; Pathway interactions; Performance; Permeability; Process; Proteins; Proteome; Proteomics; Pseudomonas aeruginosa; Research; Resistance; Role; Stress; System; Testing; antimicrobial; bacterial genetics; beta-Lactamase; catalyst; cell envelope; chronic infection; clinically relevant; colistin resistance; comparative; disulfide bond; efflux pump; functional disability; global health; holistic approach; human disease; innovation; multidrug-resistant Pseudomonas aeruginosa; mutant; next generation; novel; novel therapeutics; pathogen; pathogenic bacteria; periplasm; programs; protein folding; proteostasis; resistance mechanism

PROJECT SUMMARY / ABSTRACT Gram-negative bacteria are uniquely equipped to defeat antibiotics. Their outermost layer, the cell envelope, is a natural permeability barrier that contains an array of resistance proteins capable of neutralizing most existing antimicrobials. As a result, its presence creates a major obstacle both for the treatment of resistant infections and the development of new antibiotics. The cell envelope is also home to numerous conserved pathways that safeguard the integrity of its proteome. Despite the central role of these systems in maintaining protein homeostasis, their interaction with resistance proteins localizing in the cell envelope has not been examined. We hypothesized that the activity of cell envelope folding catalysts may be important for the function of resistance determinants, and we tested this hypothesis on a key proteostasis player, the disulfide bond formation system. We discovered that the oxidative-protein-folding activity of this pathway is essential for the function of some of the most epidemiologically relevant and clinically challenging resistance proteins, namely β-lactamases, colistin resistance enzymes, and efflux pumps. Guided by strong preliminary data obtained from model laboratory strains and from clinical isolates, we propose an in- depth investigation of the role of cell envelope proteostasis systems in antibiotic resistance. We will use a combination of bacterial genetics, microbiology, biochemistry, proteomics, experimental evolution, and human disease modeling to pursue three specific aims: 1) Identify the components of the resistome that rely on oxidative protein folding, by assessing the requirement for disulfide bond formation on hundreds of clinically important resistance proteins. 2) Evaluate the impact of oxidative protein folding on resistant infections, by testing our biochemical findings in clinical isolates and in a relevant murine chronic infection model. 3) Explore the role of other cell envelope folding catalysts in antibiotic resistance, by probing their function in multidrug-resistant clinical strains of pathogenic bacteria and validating our results in model laboratory strains. We expect that our holistic approach, spanning multiple resistance determinants and folding catalysts, will break new ground in our understanding of the role of cell envelope proteostasis in resistance. This knowledge will be applicable to many high-priority Gram-negative pathogens and, in the long term, may inspire novel broad-acting strategies for overcoming antibiotic resistance.

项目摘要 /摘要 革兰氏阴性细菌与击败抗生素是独特的。它们的最外层，即细胞信封，是 一种自然的渗透性屏障，其中包含一系列能够中和大多数现有的抗性蛋白 抗菌剂。结果，它的存在既有用于治疗抗性感染的主要障碍 以及新抗生素的开发。 细胞包络也是众多构成途径的所在地，这些途径可以保护其蛋白质组的完整性。 尽管这些系统在维持蛋白质稳态中的核心作用，但它们与抗性的相互作用 尚未检查位于细胞包膜中的蛋白质。我们假设细胞包膜的活性 折叠催化剂对于电阻确定剂的功能可能很重要，我们对此假设进行了测试。 一个关键的Proteostasis播放器，二硫键形成系统。我们发现氧化蛋白折叠 该途径的活动对于某些最流行病学和临床上最相关的功能至关重要 具有挑战性的抗性蛋白，即β-内酰胺酶，结肠蛋白抗性酶和外排泵。指导 通过从模型实验室菌株和临床分离株中获得的强大初步数据，我们提出了一个In- 细胞包膜蛋白抑制系统在抗生素耐药性中的作用的深度研究。我们将使用一个 细菌遗传学，微生物学，生物化学，蛋白质组学，实验进化和人类的结合 追求三个特定目的的疾病模型：1）确定依赖氧化的抵抗组的成分 蛋白质折叠，通过评估在数百个临床上重要的二硫键形成的需求 抗性蛋白。 2）通过测试我们的氧化蛋白折叠对抗性感染的影响 临床分离株和相关的鼠慢性感染模型中的生化发现。 3）探索角色 抗生素耐药性中的其他细胞包膜折叠催化剂，通过探测其在多药耐药性临床中的功能 致病细菌的菌株并在模型实验室菌株中验证了我们的结果。 我们期望我们的整体方法跨越多种阻力决定和折叠催化剂，将破裂 我们了解细胞包膜蛋白抑制在抗性中的作用的新基础。这些知识将是 适用于许多高优先级革兰氏阴性病原体，从长远来看，可能会激发新型的广泛作用 克服抗生素耐药性的策略。