Mechanism of Resistance Avoidance in Synthetically Evolved Antibacterial Peptides

合成进化抗菌肽避免耐药性的机制

• 批准号: 10412134
• 负责人: WILLIAM C WIMLEY
• 金额: $ 22.8万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10412134
• 关键词: Anti-Bacterial Agents; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacteria; Bacterial Infections; Binding; Binding Proteins; Burn injury; Bypass; Cations; Ceftazidime; Cell Membrane Permeability; Cells; Cephalosporins; Charge; Daughter; Drug resistance; ESKAPE pathogens; Escherichia coli; Family; Genetic; Genomics; Gram-Negative Bacteria; In Vitro; Infection Control; Klebsiella pneumoniae; Label; Learning; Libraries; Lipid A; Location; Measurement; Measures; Membrane; Microbial Biofilms; Modification; Molecular Evolution; Operon; Parents; Pathway interactions; Pattern; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Property; Pseudomonas aeruginosa; Publications; Reporting; Research Personnel; Residual state; Resistance; Resistance development; Serum Proteins; Site; Skin Tissue; Soft Tissue Infections; Solubility; Structure; Surface; Testing; Time; Toxic effect; Work; antimicrobial peptide; bactericide; clinical application; cytotoxicity; design; drug resistant bacteria; fight against; in vivo; inorganic phosphate; insight; medical implant; novel; novel strategies; pathogen; prevent; resistance mechanism; screening; small molecule; wound

Summary In the ongoing fight against drug resistant bacterial infections it will be critical to identify novel antibiotic chemotypes that are less likely to induce resistance. We have used synthetic molecular evolution (SME), iterative library design and screening, to identify novel antimicrobial peptides that have potent broad-spectrum sterilizing activity against all ESKAPE pathogens tested, including drug-resistant and biofilm-forming pathogens, in vitro and in vivo. These peptides are highly soluble, protease resistant, have low cytotoxicity, and are active in the presence of concentrated host cells and/or serum proteins. Most importantly, they do not induce resistance in P. aeruginosa under conditions that rapidly enable development of resistance to conventional antibiotics and some AMPs. Resistance to some other AMPs has been reported, and it has been experimentally selected. However, avoidance or delay in resistance to AMPs have also been reported. Here we seek to understand the mechanism of resistance-avoidance. The most commonly observed AMP resistance mechanism, by far, is the modification of Lipid A phosphates with cationic moieties, which dramatically reduces the potential for AMP interactions with the outer membrane(OM). We have previously shown that some AMPs bind strongly and accumulate significantly on OM LPS. In this work we will test the hypothesis that peptides requiring large scale accumulation on the outer membrane LPS, including Lipid A, before they can cross it will be able to drive selection for resistance using this mechanism. AMPs that can bypass the OM to reach the inner membrane target without requiring large scale accumulation on the OM will be slower or unable to elicit resistance. To test this hypothesis, we have identified two related AMPs, one that rapidly invokes resistance in Pseudomonas aeruginosa, and one that does not invoke any resistance at all in multiple passages under the same conditions. We will use these two peptides, and others, to study resistance and resistance-avoidance mechanisms in P. aeruginosa and other Gram-negative bacteria using i) direct measurements of AMP-bacteria binding, ii) genetic assessment of resistance pathways, iii) direct assessment of Lipid A modifications. We will also test for resistance avoidance in a panel of other Gram-negative pathogens to test for the generality of the mechanism.

总结 在与耐药细菌感染的持续斗争中，识别新的抗生素至关重要 不太可能诱导耐药性的化学型。我们已经使用合成分子进化（SME），迭代 文库设计和筛选，以鉴定具有强效广谱杀菌作用的新型抗菌肽 体外对所有测试的ESKAPE病原体（包括耐药性和生物膜形成病原体）的活性 和体内。这些肽是高度可溶的，具有蛋白酶抗性，具有低细胞毒性，并且在免疫组织化学中具有活性。 存在浓缩的宿主细胞和/或血清蛋白。最重要的是，它们不会诱导P的抗性。 铜绿假单胞菌的条件下，迅速使发展耐药性的常规抗生素和一些 AMPs.对其他一些AMP的耐药性也有报道，并已通过实验选择。然而，在这方面， 还报道了避免或延迟对AMP的抗性。在这里，我们试图了解的机制 抵抗-回避。到目前为止，最常见的AMP耐药机制是修饰 脂质A磷酸盐与阳离子部分，这大大降低了AMP与 外膜（OM）。我们以前已经表明，一些AMP强烈结合和积累， 对OM LPS有显著影响。在这项工作中，我们将测试的假设，肽需要大规模积累 外膜上的LPS，包括脂质A，在它们可以穿过之前，它将能够驱动选择， 利用这种机制。AMP可以绕过OM到达内膜靶点， 需要在OM上大规模积累的药物将更慢或不能引起抗性。为了验证这个假设， 我们已经鉴定了两种相关的AMP，一种在铜绿假单胞菌中迅速引起耐药性， 在相同条件下的多次传代中根本不引起任何抗性。我们将使用这两个 肽等，以研究铜绿假单胞菌和其他 使用i）AMP-细菌结合的直接测量，ii）AMP-细菌结合的遗传评估， 抗性途径，iii）脂质A修饰的直接评估。我们还将测试在 一组其他革兰氏阴性病原体，以测试该机制的一般性。

项目成果

How We Came to Understand the "Tumultuous Chemical Heterogeneity" of the Lipid Bilayer Membrane.

• DOI: 10.1007/s00232-020-00126-1
• 发表时间: 2020-06
• 期刊: The Journal of membrane biology

The Remarkable Innate Resistance of Burkholderia bacteria to Cationic Antimicrobial Peptides: Insights into the Mechanism of AMP Resistance.

• DOI: 10.1007/s00232-022-00232-2
• 发表时间: 2022-10
• 期刊: The Journal of membrane biology

Applications and evolution of melittin, the quintessential membrane active peptide.

• DOI: 10.1016/j.bcp.2021.114769
• 发表时间: 2021-11
• 期刊: BIOCHEMICAL PHARMACOLOGY
• 影响因子: 5.8
• 作者: Guha, Shantanu;Ferrie, Ryan P.;Ghimire, Jenisha;Ventura, Cristina R.;Wu, Eric;Sun, Leisheng;Kim, Sarah Y.;Wiedman, Gregory R.;Hristova, Kalina;Wimley, Wimley C.
• 通讯作者: Wimley, Wimley C.