Multidrug-resistant Pseudomonas aeruginosa is predisposed to lasR mutation through up-regulated activity of efflux pumps in non-cystic fibrosis bronchiectasis patients.
Multidrug-resistant Pseudomonas aeruginosa is predisposed to lasR mutation through up-regulated activity of efflux pumps in non-cystic fibrosis bronchiectasis patients.
复制标题
DOI：
10.3389/fcimb.2022.934439
复制
发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
--
作者：
耐多药(MDR)铜绿假单胞菌是一种常见的机会致病菌，可导致非囊性纤维支气管扩张症(NCFB)患者的显著死亡。虽然群体感应(QS)系统是一个潜在的治疗靶点，但在临床分离的铜绿假单胞菌中，经常报告具有QS缺陷表型的LasR突变株。我们的目的是研究在慢性铜绿假单胞菌肺部感染过程中，抗生素耐药性是否会选择LasR突变株，并确定这种现象的机制。我们在我们机构的两个中心前瞻性地评估了NCFB患者在两年时间里发生的慢性铜绿假单胞菌肺部感染。对分离的铜绿假单胞菌进行QS表型分析和全基因组测序。通过进化实验证实临床多药耐药铜绿假单胞菌存在LasR突变体。我们分析了97名NCFB患者的铜绿假单胞菌感染情况，发现只有既往接触碳青霉烯类才能独立预测多药耐药铜绿假单胞菌的分离。与非MDR分离株相比，MDR分离株表现出明显的QS缺陷表型，这不能通过外源添加3OC12-HSL来补充。配对的菌株表明，它们的QS表型缺陷发生在MDR形成后。全基因组测序分析表明，LasR非同义突变在MDR分离株中显著增多，且LasR基因突变频率与负外排泵调节基因(NALC和mexZ)呈正相关。外排泵抑制剂PA-β-N的加入不仅可以促进这些耐药菌株的qs表型，而且在进化实验中还可以推迟突变株的早期出现。我们的数据表明，铜绿假单胞菌通过外排泵活性上调，易发生LasR突变。这些发现提示，抗QS治疗联合外排泵抑制剂可能是NCFB患者应对耐多药铜绿假单胞菌感染的一种潜在策略。
Multidrug-resistant (MDR) Pseudomonas aeruginosa is a frequent opportunistic pathogen that causes significant mortality in patients with non-cystic fibrosis bronchiectasis (NCFB). Although the quorum sensing (QS) system is a potential target for treatment, lasR mutants that present with a QS-deficient phenotype have been frequently reported among clinical P. aeruginosa isolates. We aimed to investigate whether antibiotic resistance would select for lasR mutants during chronic P. aeruginosa lung infection and determine the mechanism underlying the phenomenon. We prospectively evaluated episodes of chronic P. aeruginosa lung infections in NCFB patients over a 2-year period at two centers of our institution. QS phenotypic assessments and whole-genome sequencing (WGS) of P. aeruginosa isolates were performed. Evolution experiments were conducted to confirm the emergence of lasR mutants in clinical MDR P. aeruginosa cultures. We analyzed episodes of P. aeruginosa infection among 97 NCFB patients and found only prior carbapenem exposure independently predictive of the isolation of MDR P. aeruginosa strains. Compared with non-MDR isolates, MDR isolates presented significantly QS-deficient phenotypes, which could not be complemented by the exogenous addition of 3OC12-HSL. The paired isolates showed that their QS-phenotype deficiency occurred after MDR was developed. Whole-genome sequencing analysis revealed that lasR nonsynonymous mutations were significantly more frequent in MDR isolates, and positive correlations of mutation frequencies were observed between genes of lasR and negative-efflux-pump regulators (nalC and mexZ). The addition of the efflux pump inhibitor PAβN could not only promote QS phenotypes of these MDR isolates but also delay the early emergence of lasR mutants in evolution experiments. Our data indicated that MDR P. aeruginosa was predisposed to lasR mutation through the upregulated activity of efflux pumps. These findings suggest that anti-QS therapy combined with efflux pump inhibitors might be a potential strategy for NCFB patients in the challenge of MDR P. aeruginosa infections.
登录
查看更多内容
影响因子：
4.2
作者：
Minagawa S;Inami H;Kato T;Sawada S;Yasuki T;Miyairi S;Horikawa M;Okuda J;Gotoh N
通讯作者：
Gotoh N
影响因子：
3.2
作者：
ESSAR, DW;EBERLY, L;CRAWFORD, IP
通讯作者：
CRAWFORD, IP
影响因子：
14.2
作者：
Magiorakos, A. -P.;Srinivasan, A.;Monnet, D. L.
通讯作者：
Monnet, D. L.
影响因子：
5.1
作者：
Olivares, Jorge;Alvarez-Ortega, Carolina;Luis Martinez, Jose
通讯作者：
Luis Martinez, Jose
影响因子：
3.7
作者：
Lamarche MG;Déziel E
通讯作者：
Déziel E