Sequence-Specific Targeting of Bacterial Resistance Genes Increases Antibiotic Efficacy.
Sequence-Specific Targeting of Bacterial Resistance Genes Increases Antibiotic Efficacy.
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DOI：
10.1371/journal.pbio.1002552
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发表时间：
2016-09
期刊：
影响因子：
9.8
通讯作者：
Toprak E
中科院分区：
文献类型：
--
作者：
Ayhan DH;Tamer YT;Akbar M;Bailey SM;Wong M;Daly SM;Greenberg DE;Toprak E
缺乏有效且耐受性良好的抗耐药细菌疗法是一个全球公共卫生问题，导致治疗延长和死亡率增加。为了提高现有抗生素化合物的疗效，我们介绍了一种新的方法，用于在病原菌中策略性地诱导抗生素超敏反应。在系统地证实AcrAB-TolC外排系统是大肠杆菌内在抗生素抗性水平的主要决定因素之一之后，我们开发了一种设计用于抑制acrA表达并增加大肠杆菌抗生素敏感性的短反义寡聚体。杆菌采用这种策略，我们可以抑制E。大肠杆菌的生长使用2至40倍的抗生素剂量，这取决于所使用的抗生素化合物。反义寡聚体的致敏作用对靶基因的序列是高度特异性的，所述靶基因的序列在几种细菌属中是保守的，并且寡聚体对人细胞没有任何可检测的毒性。最后，我们证明了反义寡聚体提高了抗生素组合的功效，允许联合使用甚至拮抗性抗生素对，这些抗生素对通常由于其活性降低而不受欢迎。通过反义介导的翻译抑制减少大肠杆菌AcrAB-TolC外排系统的表达增加了细菌对多种抗生素的敏感性。抗生素耐药性是一个全球性的健康威胁。虽然基因组测序和遗传操作工具已经阐明了许多耐药机制，但这些工具尚未开发成成功的治疗方法。具有这种潜力的一种工具是肽缀合的磷酰二胺吗啉代寡聚物（PPM 0），其是作为反义mRNA翻译抑制剂起作用的合成DNA/RNA模拟物。在本文中，我们使用PMOs来增加细菌的抗生素敏感性。首先，我们确定AcrAB-TolC外排系统是大肠杆菌的主要内在耐药机制。杆菌然后通过用PPMO靶向该外排系统的每个组分的mRNA，我们确定acrA-PPMO为最有效的反义分子。用acrA-PPMO处理细菌导致抗生素功效增加2至40倍，阻止AcrA蛋白的翻译，并抑制抗生素分子的外排，而对人类细胞没有细胞毒性。最后，我们证明了acrA-PPMO在几种病原菌属中是有效的，并且当一起使用时增强了协同和拮抗抗生素对的活性。这项工作确定了PPMO可以潜在地用于治疗由抗生素耐药细菌引起的感染。
The lack of effective and well-tolerated therapies against antibiotic-resistant bacteria is a global public health problem leading to prolonged treatment and increased mortality. To improve the efficacy of existing antibiotic compounds, we introduce a new method for strategically inducing antibiotic hypersensitivity in pathogenic bacteria. Following the systematic verification that the AcrAB-TolC efflux system is one of the major determinants of the intrinsic antibiotic resistance levels in Escherichia coli, we have developed a short antisense oligomer designed to inhibit the expression of acrA and increase antibiotic susceptibility in E. coli. By employing this strategy, we can inhibit E. coli growth using 2- to 40-fold lower antibiotic doses, depending on the antibiotic compound utilized. The sensitizing effect of the antisense oligomer is highly specific to the targeted gene’s sequence, which is conserved in several bacterial genera, and the oligomer does not have any detectable toxicity against human cells. Finally, we demonstrate that antisense oligomers improve the efficacy of antibiotic combinations, allowing the combined use of even antagonistic antibiotic pairs that are typically not favored due to their reduced activities. Reducing expression of the AcrAB-TolC efflux system of Escherichia coli by antisense-mediated translation inhibition increases the susceptibility of bacteria to a wide range of antibiotics. Antibiotic resistance is a global health threat. While genome sequencing and genetic manipulation tools have elucidated many resistance mechanisms, these tools have not yet been developed into successful therapeutics. One tool with such potential are peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs), which are synthetic DNA/RNA mimics that function as antisense mRNA translation inhibitors. In this paper we use PPMOs to increase antibiotic susceptibility of bacteria. First, we identify the AcrAB-TolC efflux system as a major intrinsic resistance mechanism in E. coli. Then by targeting the mRNA of each component of this efflux system with PPMOs, we identify an acrA-PPMO as the most effective antisense molecule. Treatment of bacteria with acrA-PPMO resulted in a 2- to 40-fold increase in antibiotic efficacy, prevented translation of the AcrA protein, and inhibited efflux of antibiotic molecules without being cytotoxic to human cells. Finally, we demonstrate that acrA-PPMO is efficacious in several pathogenic bacterial genera and enhances activity of both synergistic and antagonistic antibiotic pairs when used together. This work establishes that PPMOs can potentially be used to treat infections caused by antibiotic resistant bacteria.
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DOI：
10.1086/652807
发表时间：
2010-06-15
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
Greenberg DE;Marshall-Batty KR;Brinster LR;Zarember KA;Shaw PA;Mellbye BL;Iversen PL;Holland SM;Geller BL
通讯作者：
Geller BL
影响因子：
46.9
作者：
Bikard, David;Euler, Chad W.;Jiang, Wenyan;Nussenzweig, Philip M.;Goldberg, Gregory W.;Duportet, Xavier;Fischetti, Vincent A.;Marraffini, Luciano A.
通讯作者：
Marraffini, Luciano A.
影响因子：
64.8
作者：
Chait, Remy;Craney, Allison;Kishony, Roy
通讯作者：
Kishony, Roy
大肠杆菌K-12的构造框架，单基因敲除突变体：Keio Collection。
影响因子：
9.9
作者：
通讯作者：
--
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL