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

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“超级细菌”耐甲氧西林金黄色葡萄球菌 (MRSA) 引起的侵袭性感染的效果很差 由于对一线药物、万古霉素（糖肽；GP）的敏感性降低而恶化的结果，以及 达托霉素（脂肽；LP） 长效脂糖肽（LGP）达巴万星是一种替代品。 每周或单次给药，这可以促进出院并降低成本。但是，我们已经证明这一点。 其半衰期长可能会增加其耐药性选择潜力，并可以选择与万古霉素的交叉耐药性 因此，迫切需要了解细胞间交叉耐药的机制。 MRSA 中的包膜靶向药物，并研究减轻或克服这种耐药性的策略。 之前资助期间的工作发现，75% 的 GP/LP/LGP 非敏感分离株来自体外 PK/PD 模拟达巴万星暴露的模型获得了与基本两部分调节相关的突变 此外，我们最近发表了一个选择达巴万星治疗的病例。 心内膜炎患者通过 walK 突变发现 GP/LP/LGP 耐药的 MRSA。 此次更新的目标：阐明 walKR 突变导致 GP/LP/LGP 的多种机制 交叉耐药性并揭示 β-内酰胺和其他代谢调节剂如何与 WalKR 调节相互作用 代谢网络与 GP/LP/LGP 协同作用并防止耐药性。 通过调节细胞包膜和核苷酸形成 GP/LP/LGP 交叉耐药表型 代谢，代谢调节剂可以使这些菌株对 GP/LP/LGP 重新敏感，或通过以下方式预防耐药性： 在 AIM 1 中，我们将测量减少的贡献。 WalKR 使用遗传、脂质组学、代谢组学、转录组学等方法对 MRSA 中的交叉耐药表型发挥作用 蛋白质组学方法与药敏试验和定量生物物理评估相结合 在 AIM 2 中，我们将测试 β-内酰胺和其他代谢物质的假设。 调节剂可以使 walK 敲低菌株对 GP/LP/LGP 重新敏感。我们将检查 GP/LP/LGP 的协同效应。 细胞壁抑制剂β-内酰胺和磷霉素、脂质合成抑制剂和抗叶酸药物， 甲氧苄啶/磺胺甲恶唑，它将告知对 walKR 突变很重要的代谢途径 - 在 AIM 3 中，我们将评估 β-内酰胺和其他代谢调节剂的潜力。 使用连续传代和 PK/PD 模型在体外阻止达巴万星对 GP/LP/LGP 耐药性的选择。 这项工作意义重大，因为达巴万星暴露很容易选择万古霉素和达托霉素耐药性 金黄色葡萄球菌和防止 MRSA 对 GP/LG/LGP 产生耐药性和/或重新敏感的策略对于保存至关重要 这些药物，特别是在目前达巴万星使用量不断增加的背景下，这一提议具有创新性。 包括利用代谢表型来告知和调节耐药表型以及应用 高通量脂质组学和代谢组学来表征耐药性和反应。

项目成果

Antimicrobial use across six referral hospitals in Tanzania: a point prevalence survey.

• DOI: 10.1136/bmjopen-2020-042819
• 发表时间: 2020-12-15
• 期刊: BMJ open
• 影响因子: 2.9
• 作者: Seni J;Mapunjo SG;Wittenauer R;Valimba R;Stergachis A;Werth BJ;Saitoti S;Mhadu NH;Lusaya E;Konduri N
• 通讯作者: Konduri N

Identification of a novel tedizolid resistance mutation in rpoB of MRSA after in vitro serial passage.

• DOI: 10.1093/jac/dkaa422
• 发表时间: 2020-10
• 期刊: The Journal of antimicrobial chemotherapy
• 作者: Tianwei Shen;K. Penewit;A. Waalkes;Libin Xu;S. Salipante;Abhinav Nath;Brian J. Werth

Dalbavancin exposure in vitro selects for dalbavancin-non-susceptible and vancomycin-intermediate strains of methicillin-resistant Staphylococcus aureus.

• DOI: 10.1016/j.cmi.2020.08.025
• 发表时间: 2021-06
• 期刊: Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases
• 作者: Werth BJ;Ashford NK;Penewit K;Waalkes A;Holmes EA;Ross DH;Shen T;Hines KM;Salipante SJ;Xu L
• 通讯作者: Xu L