An Antipersister Strategy for Treatment of Chronic Pseudomonas aeruginosa Infections
An Antipersister Strategy for Treatment of Chronic Pseudomonas aeruginosa Infections
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DOI：
10.1128/aac.00987-17
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发表时间：
2017-12-01
影响因子：
4.9
通讯作者：
Joseph-McCarthy, Diane
中科院分区：
文献类型：
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作者：
Koeva, Martina;Gutu, Alina D.;Joseph-McCarthy, Diane
细菌存留细胞是对抗生素治疗具有耐受性的准休眠细胞亚群。氨基糖苷类妥布霉素与富马酸盐的组合作为抗菌增强剂，采用抗持久性策略，旨在通过增强对铜绿假单胞菌持久性的杀灭来减少铜绿假单胞菌的复发感染。铜绿假单胞菌的固定相培养物用于产生持续细胞。使用一系列代谢物浓度测试了一系列妥布霉素浓度，以确定代谢物在各种条件下的增强作用，包括一系列 pH 值以及阿奇霉素或囊性纤维化 (CF) 患者痰液中存在的情况。此外，还进行了 96 孔培养皿生物膜和菌落生物膜测定，并利用乳酸脱氢酶 (LDH) 测定测定了妥布霉素-富马酸盐组合的细胞毒性。与单独使用妥布霉素相比，妥布霉素-富马酸盐组合对一系列 CF 分离株（包括粘液样菌株和非粘液样菌株）的铜绿假单胞菌持久性杀灭作用增强高达 6 个数量级。此外，在阿奇霉素或 CF 患者痰液存在的情况下，观察到显着的富马酸盐介导的增强作用。富马酸盐还降低了妥布霉素处理的铜绿假单胞菌对人上皮气道细胞的细胞毒性。最后，在粘液样和非粘液样 CF 分离株中，由于富马酸盐增强作用，在菌落生物膜测定中观察到铜绿假单胞菌生物膜被完全根除。这些数据表明，妥布霉素与富马酸盐的组合作为抗菌增强剂可能是一种有吸引力的治疗方法，可以通过根除细菌持续存在来消除 CF 患者的复发性铜绿假单胞菌感染。
Bacterial persisters are a quasidormant subpopulation of cells that are tolerant to antibiotic treatment. The combination of the aminoglycoside tobramycin with fumarate as an antibacterial potentiator utilizes an antipersister strategy that is aimed at reducing recurrent Pseudomonas aeruginosa infections by enhancing the killing of P. aeruginosa persisters. Stationary-phase cultures of P. aeruginosa were used to generate persister cells. A range of tobramycin concentrations was tested with a range of metabolite concentrations to determine the potentiation effect of the metabolite under a variety of conditions, including a range of pH values and in the presence of azithromycin or cystic fibrosis (CF) patient sputum. In addition, 96-well dish biofilm and colony biofilm assays were performed, and the cytotoxicity of the tobramycin-fumarate combination was determined utilizing a lactate dehydrogenase (LDH) assay. Enhanced killing of up to 6 orders of magnitude of P. aeruginosa persisters over a range of CF isolates, including mucoid and nonmucoid strains, was observed for the tobramycin-fumarate combination compared to killing with tobramycin alone. Furthermore, significant fumarate-mediated potentiation was seen in the presence of azithromycin or CF patient sputum. Fumarate also reduced the cytotoxicity of tobramycin-treated P. aeruginosa to human epithelial airway cells. Finally, in mucoid and nonmucoid CF isolates, complete eradication of P. aeruginosa biofilm was observed in the colony biofilm assay due to fumarate potentiation. These data suggest that a combination of tobramycin with fumarate as an antibacterial potentiator may be an attractive therapeutic for eliminating recurrent P. aeruginosa infections in CF patients through the eradication of bacterial persisters.