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

细胞包膜代谢改变对 MRSA 细胞包膜靶向抗菌药物耐药性的贡献

• 批准号: 10733982
• 负责人: Brian James Werth
• 金额: $ 55.02万
• 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 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10733982
• 关键词: Affect; Alleles; Antimetabolites; Bacteremia; Biochemical Pathway; Biophysics; CRISPR interference; Cell Wall; Clinical; Combined Modality Therapy; Coupled; Daptomycin; Dose; Drug Kinetics; Drug Targeting; Drug resistance; Endocarditis; Exhibits; Exposure to; Folic Acid Antagonists; Fosfomycin; Frequencies; Functional disorder; Gene Expression; Genetic; Glycopeptides; Goals; Grant; Half-Life; Hospitals; In Vitro; Individual; Infection; Lead; Lipids; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolism; Methicillin Resistance; Modification; Mutate; Mutation; Nucleotides; Outcome; Outpatients; Parents; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Play; Predisposition; Prevention; Property; Proteomics; Publishing; Reporting; Resistance; Role; Serial Passage; Staphylococcus aureus; Staphylococcus aureus infection; Sulfamethoxazole; Superbug; System; Techniques; Testing; Trimethoprim; Vancomycin; Vancomycin Resistance; Walking; Water; Work; antimicrobial; beta-Lactams; biophysical analysis; cell envelope; cost; dalbavancin; drug preservation; experimental study; genome sequencing; infection management; inhibitor; innovation; knock-down; lipid metabolism; lipidomics; metabolic phenotype; metabolomics; methicillin resistant Staphylococcus aureus; mortality; mutant; nucleotide metabolism; optimal treatments; pharmacodynamic model; pharmacokinetics and pharmacodynamics; prevent; resistant strain; response; standard care; synergism; targeted agent; transcriptome sequencing; transcriptomics; treatment strategy; Affect; Alleles; Antimetabolites; Bacteremia; Biochemical Pathway; Biophysics; CRISPR interference; Cell Wall; Clinical; Combined Modality Therapy; Coupled; Daptomycin; Dose; Drug Kinetics; Drug Targeting; Drug resistance; Endocarditis; Exhibits; Exposure to; Folic Acid Antagonists; Fosfomycin; Frequencies; Functional disorder; Gene Expression; Genetic; Glycopeptides; Goals; Grant; Half-Life; Hospitals; In Vitro; Individual; Infection; Lead; Lipids; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolism; Methicillin Resistance; Modification; Mutate; Mutation; Nucleotides; Outcome; Outpatients; Parents; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Play; Predisposition; Prevention; Property; Proteomics; Publishing; Reporting; Resistance; Role; Serial Passage; Staphylococcus aureus; Staphylococcus aureus infection; Sulfamethoxazole; Superbug; System; Techniques; Testing; Trimethoprim; Vancomycin; Vancomycin Resistance; Walking; Water; Work; antimicrobial; beta-Lactams; biophysical analysis; cell envelope; cost; dalbavancin; drug preservation; experimental study; genome sequencing; infection management; inhibitor; innovation; knock-down; lipid metabolism; lipidomics; metabolic phenotype; metabolomics; methicillin resistant Staphylococcus aureus; mortality; mutant; nucleotide metabolism; optimal treatments; pharmacodynamic model; pharmacokinetics and pharmacodynamics; prevent; resistant strain; response; standard care; synergism; targeted agent; transcriptome sequencing; transcriptomics; treatment strategy

Invasive infections due to the “superbug” methicillin-resistant Staphylococcus aureus (MRSA) have poor outcomes that are worsened by reduced susceptibility to first-line agents, vancomycin (glycopeptide; GP), and daptomycin (lipopeptide; LP). The long-acting lipoglycopeptide (LGP), dalbavancin, is an alternative that can be given weekly or as a single dose, which can facilitate discharge and reduce costs. However, we have shown that its long half-life may increase its resistance selection potential and can select for cross-resistance to vancomycin and daptomycin. Thus, there is a critical need to understand the mechanism(s) of cross-resistance among cell envelope-targeting drugs in MRSA and to investigate strategies to mitigate or overcome such resistance. Our work from the previous grant periods found that 75% of GP/LP/LGP non-susceptible isolates from in vitro PK/PD models simulating dalbavancin exposures acquired mutations related to the essential two-component regulatory system walKR. Furthermore, we recently published a case in which dalbavancin treatment selected for GP/LP/LGP-resistant MRSA in a patient with endocarditis, via a walK mutation. These recent findings led to the goal for this renewal: to elucidate the multiple mechanisms through which walKR mutations lead to GP/LP/LGP cross-resistance and reveal how beta-lactams and other metabolic modulators interact with WalKR-regulated metabolic networks to synergize with GP/LP/LGP and prevent resistance. We hypothesize that walKR mutations underlie GP/LP/LGP cross-resistance phenotypes through modulation of both cell envelope and nucleotide metabolism, and metabolic modulators can re-sensitize these strains to GP/LP/LGP or prevent resistance by further altering cell envelope or nucleotide metabolism. In AIM 1, we will measure the contribution of reduced WalKR function to cross-resistance phenotypes in MRSA using genetic, lipidomic, metabolomic, transcriptomic, and proteomic approaches in combination with susceptibility testing and quantitative biophysical assessment of the cell envelope properties. In AIM 2, we will test the hypothesis that beta-lactams and other metabolic modulators can re-sensitize walK-knockdown strains to GP/LP/LGP. We will examine the synergistic effects of cell wall inhibitors beta-lactams and fosfomycin, lipid synthesis inhibitors, and anti-folate drugs, trimethoprim/sulfamethoxazole, which will inform metabolic pathways that are important for walKR mutation- caused resistance. In AIM 3, we will evaluate the potential of beta-lactams and other metabolic modulators to prevent the selection of GP/LP/LGP resistance by dalbavancin in vitro using serial passage and PK/PD models. This work is significant because dalbavancin exposures readily select for vancomycin and daptomycin-resistant S. aureus and a strategy to prevent resistance and/or re-sensitize MRSA to GP/LG/LGP is critical to preserve these drugs, especially in the current context of increasing dalbavancin use. The innovation of this proposal includes leveraging metabolic phenotypes to inform and modulate resistance phenotypes and the application of high-throughput lipidomics and metabolomics to characterize drug resistance and responses.

由于“超级细菌”耐甲氧西林金黄色葡萄球菌（MRSA）引起的侵入性感染较差 通过降低对一线药物的敏感性，万古霉素（糖肽； GP）和 daptomycin（脂肽； LP）。长效脂肪糖肽（LGP）Dalbavancin是一种可以是的替代方法 每周或单剂量给予，这可以促进放电并降低成本。但是，我们已经表明 其长期寿命可能会增加其阻力选择潜力，并可以选择对万古霉素的抗性 和Daptomycin。这是迫切需要了解细胞之间的交叉抗性机制 MRSA中的靶向药物，并调查减轻或克服这种耐药性的策略。我们的 从前赠款期的工作发现，来自体外PK/PD的GP/LP/LGP不可启示的分离株的75％ 模拟Dalbavancin暴露的模型获得了与必需的两部分调节有关的突变 系统Walkr。此外，我们最近发表了一个案例，在其中选择了达尔巴万林治疗 通过行走突变，患有心内膜炎患者的GP/LP/LGP耐药性MRSA。这些最近的发现导致了 续约的目标：阐明Wallr突变导致GP/LP/LGP的多种机制 交叉抗性并揭示β-内酰胺和其他代谢调节剂如何与WalkR调节的相互作用 代谢网络与GP/LP/LGP协同作用并防止阻力。我们假设Walkr突变 GP/LP/LGP基础通过调节细胞包膜和核苷酸的跨抗性表型 代谢和代谢调节剂可以将这些菌株重新敏感到GP/LP/LGP或防止抗性 进一步改变细胞膜或核苷酸代谢。在AIM 1中，我们将衡量减少的贡献 使用遗传学，脂质组，代谢组，转录组，WALKR功能可在MRSA中进行抗抗性表型 和蛋白质组学方法结合了易感性测试和定量生物物理评估 细胞包络属性。在AIM 2中，我们将测试β-内酰胺和其他代谢的假设 调节器可以将敏感性的敲低应变重新敏感到GP/LP/LGP。我们将研究 细胞壁抑制剂β-内酰胺和Fosfomycin，脂质合成抑制剂和抗叶酸药物， 甲氧苄啶/磺胺甲恶唑，这将为Walkr突变至关重要的代谢途径提供信息 引起阻力。在AIM 3中，我们将评估β-内酰胺和其他代谢调节剂的潜力 防止使用串行通道和PK/PD模型在体外选择Dalbavancin在体外选择GP/LP/LGP。 这项工作很重要，因为达尔巴万氏素很容易选择万古霉素和耐达霉素 S.金黄色葡萄球菌和防止抗性和/或重新敏感的MRSA对GP/LG/LGP的策略对于保存至关重要 这些药物，尤其是在达尔巴氏蛋白使用增加的当前情况下。该提议的创新 包括利用代谢表型告知和调节抗药性表型以及应用 高通量脂质组学和代谢组学以表征耐药性和反应。