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.

项目概要/摘要 革兰氏阴性细菌具有独特的能力来抵抗抗生素，它们的最外层，即细胞膜，是。 天然渗透屏障，包含一系列能够中和大多数现有的耐药蛋白 因此，它的存在给耐药性感染的治疗带来了重大障碍。 以及新抗生素的开发。 细胞膜也是许多保守途径的所在地，可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方面发挥着核心作用，但它们与耐药性的相互作用 尚未检查定位于细胞被膜的蛋白质。我们注意到细胞被膜的活性。 折叠催化剂可能对耐药性决定因素的功能很重要，我们在以下方面测试了这一假设： 我们发现氧化蛋白质折叠是蛋白质稳态的关键参与者。 该通路的活性对于一些与流行病学和临床最相关的功能至关重要 具有挑战性的耐药蛋白，即β-内酰胺酶、粘菌素耐药酶和外排泵。 通过从模型实验室菌株和临床分离株获得的强有力的初步数据，我们提出了一个in- 深入研究细胞包膜蛋白质稳态系统在抗生素耐药性中的作用。 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 疾病建模以实现三个具体目标：1）识别依赖于氧化的抗性组成分 蛋白质折叠，通过评估数百个临床上重要的二硫键形成的要求 2) 通过测试我们的方法来评估氧化蛋白折叠对耐药感染的影响。 临床分离株和相关小鼠慢性感染模型中的生化结果 3) 探索其作用。 其他细胞膜折叠催化剂在抗生素耐药性中的作用，通过探索它们在多重耐药临床中的功能 致病菌菌株并验证我们在模型实验室菌株中的结果。 我们预计，涵盖多个阻力决定因素和折叠催化剂的整体方法将突破 这一知识将为我们理解细胞包膜蛋白稳态在抵抗中的作用提供新的基础。 适用于许多高度优先的革兰氏阴性病原体，从长远来看，可能会激发新的广泛作用 克服抗生素耐药性的策略。