Contribution of altered lipid metabolism to resistance to cell envelope-targeting antimicrobials in MRSA

脂质代谢改变对 MRSA 细胞包膜靶向抗菌药物耐药性的影响

• 批准号: 9763437
• 负责人: Brian James Werth
• 金额: $ 46.38万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9763437
• 关键词: Address; Alleles; American; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Bacteremia; Bacteria; Biophysics; Categories; Cell Wall; Cell membrane; Cessation of life; Characteristics; Clinical; Clinical Management; Coupled; Daptomycin; Development; Diagnostic; Drug resistance; Failure; Foundations; Gene Expression; Gene Mutation; Genes; Genetic study; Genomics; Genotype; Glycopeptides; Goals; Health; Human; In Vitro; Individual; Infection; Lead; Libraries; Lipids; Mass Spectrum Analysis; Measures; Methicillin Resistance; Methodology; Modeling; Mutation; Outcome; Parents; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Phosphatidylglycerols; Play; Population Analysis; Predisposition; Prevention; Property; Resistance; Resistance development; Role; Staphylococcus aureus infection; Superbug; Techniques; Testing; Therapeutic; Time; Transcript; Treatment Failure; Vancomycin; Work; antimicrobial; base; beta-Lactams; biological adaptation to stress; cell envelope; clinically relevant; dalbavancin; differential expression; drug response prediction; druggable target; genome sequencing; improved; innovation; insight; lipid metabolism; lipoteichoic acid; methicillin resistant Staphylococcus aureus; mutant; novel; novel diagnostics; novel therapeutics; pathogenic bacteria; pharmacokinetics and pharmacodynamics; predicting response; resistance mechanism; resistant strain; response; small molecule; specific biomarkers; standard of care; success; synergism; transcriptome sequencing; transcriptomics; whole genome

Project Summary Invasive infections due to the “superbug” methicillin-resistant Staphylococcus aureus (MRSA) are responsible for more deaths than any other drug-resistant bacterial pathogen in the USA. The glycopeptide (GP), vanco- mycin, is the standard of care for the treatment of MRSA, but therapeutic failures are common. Vancomycin is closely related to two other classes of antibiotics that are also important for the treatment of MRSA infections, the lipopeptides (LP) and the lipoglycopeptides (LGP). Drugs from each of these classes can select for cross- resistance to the others to various degrees, but the mechanisms behind this cross-resistance are not well un- derstood. In addition, there is a critical need to develop new diagnostics that can predict drug response to GP, LP, and LGP antimicrobials, and to develop novel therapies that can modulate resistance. Recently, using an innovative lipidomic approach we have observed significant and characteristic changes in lipid metabolism as- sociated with resistance to each class of drugs (GP, LP, and LGP). We hypothesize that lipidomic signatures in the cell membrane reflect specific antibiotic resistance mechanisms and that these lipidomic signatures can be modified by other small molecules to favor a susceptible phenotype. We propose to comprehensively inter- rogate the mechanisms of resistance and cross-resistance to GP, LP, and LGP antimicrobials in MRSA by in- tegrated lipidomics, genomics, and transcriptomics. In AIM 1, our strategy will focus on elucidating the role of altered lipid metabolism in the development of resistant phenotypes in in vitro-selected strains that are re- sistant to GP, LP, and/or LGP using comprehensive lipidomics, coupled with genomics, transcriptomics, stand- ard and advanced susceptibility testing, and quantitative biophysical assessment of cell wall and cell mem- brane properties. Contribution of each gene mutation to the resistant phenotypes will be elucidated by similar characterization of single-gene mutants generated by allelic replacement. In AIM 2, we will test the hypothesis that β-lactams modulate the susceptibility of MRSA to GP, LP, and LGP, at least in part, by modifying the lipid composition of the bacteria. We will examine the synergistic interactions between the study drugs and β- lactams by the integrated omics approach, which would yield insights into the mechanisms of β-lactam synergy with these agents and the β-lactam “seesaw effect”. In AIM 3, we will evaluate the sensitivity of our lipidomic technique to identify sub-MIC differences in susceptibility and predict response to clinically relevant GP, LP, and LGP exposures. We anticipate that this strategy will yield valuable insight into the role of altered lipid me- tabolism in producing antimicrobial resistance. This work is significant because the pathways identified as crit- ical to resistance and susceptible to modulation will lay the foundation for further studies that could lead to re- sistance prevention therapeutics and advanced diagnostics, which in turn will improve human health. The in- novation of this proposal includes the novel lipidomics methodology, the integrated omics approach, and cor- relation of lipidomic signatures with clinically relevant pharmacodynamics endpoints.

项目摘要 由于“超级细菌”耐甲氧西林的金黄色葡萄球菌（MRSA）是造成的侵入性感染是负责 在美国，与任何其他耐药细菌病原体相比，死亡人数更多。糖肽（GP），Vanco- mycin是治疗MRSA的护理标准，但治疗失败很常见。万古霉素是 与其他两种类类的抗生素密切相关，这些抗生素对于治疗MRSA感染也很重要， 脂蛋白肽（LP）和脂肪肽（LGP）。这些类别的药物可以选择交叉 在各种程度上对其他人的抵抗力，但是这种交叉抗性背后的机制并不是很好 德斯特。此外，有迫切需要开发可以预测药物反应GP的新诊断， LP和LGP抗菌剂，并开发可调节抗性的新型疗法。最近，使用一个 创新的脂质组方法我们观察到脂质代谢的显着和特征变化 对每类药物（GP，LP和LGP）的抵抗力进行社交。我们假设脂质组学特征 在细胞膜中 通过其他小分子修饰以偏爱易感表型。我们建议全面间 通过IN-抗抗药性和对GP，LP和LGP抗菌药物的抗性机制，通过In- 培养的脂质组学，基因组学和转录组学。在AIM 1中，我们的战略将重点阐明 脂质代谢改变在耐药表型中的体外菌株中的发展 使用综合脂质组学与GP，LP和/或LGP相关，再加上基因组学，转录组学，独立 ARD和晚期敏感性测试，以及细胞壁和细胞内置的定量生物物理评估 Brane属性。每个基因突变对抗性表型的贡献将被相似 由等位基因替代产生的单基因突变体的表征。在AIM 2中，我们将检验假设 β-内酰胺可调节MRSA对GP，LP和LGP的敏感性，至少部分通过修饰脂质 细菌的组成。我们将研究研究药物与β- LACTAMS通过综合OMICS方法，该方法将对β-内酰胺协同的机制产生见解 这些药物和β-内酰胺的“ Seesaw效应”。在AIM 3中，我们将评估脂质组的灵敏度 识别易感性差异并预测对临床相关GP，LP的反应的技术差异 和LGP暴露。我们预计，这种策略将对改变的脂质变化的作用产生宝贵的见解。 产生抗菌抗性的破烂。这项工作很重要，因为被确定为批评的途径 具有抵抗力和容易受到调节的影响将为进一步的研究奠定基础 预防疗法和晚期诊断，这反过来将改善人类健康。内 该提案的小说包括新型脂肪组学方法论，综合法学方法和cor- 与临床相关的药效学终点的脂质组信号的关系。