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.

概括 在持续对抗耐药细菌感染的斗争中，识别新型抗生素至关重要 不太可能诱导耐药性的化学型。我们使用了合成分子进化（SME）、迭代 文库设计和筛选，鉴定具有强效广谱灭菌作用的新型抗菌肽 对所有经过测试的 ESKAPE 病原体（包括耐药病原体和生物膜形成病原体）的体外活性 和体内。这些肽具有高度可溶性、蛋白酶抗性、低细胞毒性，并且在 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. 铜绿假单胞菌在快速产生对常规抗生素和某些抗生素耐药性的条件下 AMP。据报道，对其他一些 AMP 具有抗性，并且已通过实验选择了它。然而， 也有报道称避免或延迟对 AMP 产生耐药性。在这里我们试图了解其中的机制 的抵抗-回避。迄今为止，最常见观察到的 AMP 抗性机制是修饰 具有阳离子部分的脂质 A 磷酸盐，这大大降低了 AMP 与 外膜（OM）。我们之前已经证明一些 AMP 会强烈结合并积累 OM LPS 显着。在这项工作中，我们将检验以下假设：肽需要大规模积累 外膜上的 LPS（包括脂质 A）在穿过之前能够驱动选择 使用这种机制进行抵抗。可以绕过 OM 到达内膜目标的 AMP，无需 需要在 OM 上进行大规模积累，速度会较慢或无法引起抵抗。为了检验这个假设， 我们已经确定了两种相关的 AMP，一种能够快速引发铜绿假单胞菌的耐药性，另一种能够快速引起铜绿假单胞菌的耐药性 在相同条件下的多个段落中根本不会引起任何阻力。我们将使用这两个 肽等，以研究铜绿假单胞菌和其他细菌的耐药性和耐药性避免机制 革兰氏阴性细菌使用 i) 直接测量 AMP-细菌结合，ii) 遗传评估 耐药途径，iii) 直接评估脂质 A 修饰。我们还将测试抵抗避免 一组其他革兰氏阴性病原体来测试该机制的普遍性。

项目成果

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

我们如何理解脂质双层膜的“混乱的化学异质性”。

• 发表时间: 2020
• 作者: Wimley; William C
• 通讯作者: William C

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

伯克霍尔德杆菌对阳离子抗菌肽的显着先天耐药性：深入了解 AMP 耐药机制。

• 发表时间: 2022-10
• 作者: Ghimire, Jenisha;Guha, Shantanu;Nelson, Benjamin J;Morici, Lisa A;Wimley, William C
• 通讯作者: Wimley, William C

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

蜂毒肽（典型的膜活性肽）的应用和进化。

• 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.