Let-7b-5p in vesicles secreted by human airway cells reduces biofilm formation and increases antibiotic sensitivity of P. aeruginosa
Let-7b-5p in vesicles secreted by human airway cells reduces biofilm formation and increases antibiotic sensitivity of P. aeruginosa
复制标题
由人气道细胞分泌的囊泡中的Let-7 b-5 p减少生物膜形成并增加铜绿假单胞菌的抗生素敏感性
DOI：
10.1073/pnas.2105370118
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发表时间：
2021-07-13
影响因子：
11.1
通讯作者：
Stanton, Bruce A.
中科院分区：
文献类型：
--
作者：
Koeppen, Katja;Nymon, Amanda;Stanton, Bruce A.
铜绿假单胞菌是一种机会性病原体，可形成耐抗生素生物膜，促进免疫功能低下宿主的慢性感染。我们之前的研究表明，铜绿假单胞菌分泌的外膜囊泡可向人气道上皮细胞（AECs）传递小RNA，从而抑制先天免疫反应。在这里，我们证明了通过含有小rna的膜囊泡的结构域间通信是双向的，并且人类AECs分泌的细胞外囊泡（EVs）中的microRNAs （miRNAs）调节铜绿假单胞菌的蛋白表达、抗生素敏感性和生物膜形成。具体来说，人类ev将miRNA let-7b-5p传递给P. aeruginosa，从而系统性地降低了生物膜形成所需蛋白质的丰度，包括PpkA和ClpV1-3，并增加了β -内酰胺类抗生素通过靶向β -内酰胺酶AmpC来减少生物膜形成的能力。从生物信息学的角度预测，let -7b-5p不仅能靶向铜绿假单胞菌中的PpkA、ClpV1和AmpC，还能靶向另一种臭名昭著的机会性肺病原体新绿伯克氏菌中的相应同源物，这表明let -7b-5p减少生物膜形成和增加β -内酰胺敏感性的能力不仅限于铜绿假单胞菌。在这里，我们提供了真核细胞分泌的ev中的mirna转移到原核生物的直接证据，导致原核生物随后的表型改变，这是由于这种域间通信的结果。由于let-7家族mirna正处于减少炎症的临床试验中，并且由于慢性铜绿假单胞菌肺部感染与高炎症状态相关，因此在纳米颗粒或ev中使用let-7b-5p和β -内酰胺抗生素治疗可能对耐药铜绿假单胞菌感染患者有益。
Pseudomonas aeruginosa is an opportunistic pathogen that forms antibiotic-resistant biofilms, which facilitate chronic infections in immunocompromised hosts. We have previously shown that P. aeruginosa secretes outer-membrane vesicles that deliver a small RNA to human airway epithelial cells (AECs), in which it suppresses the innate immune response. Here, we demonstrate that interdomain communication through small RNA-containing membrane vesicles is bidirectional and that microRNAs (miRNAs) in extracellular vesicles (EVs) secreted by human AECs regulate protein expression, antibiotic sensitivity, and biofilm formation by P. aeruginosa. Specifically, human EVs deliver miRNA let-7b-5p to P. aeruginosa, which systematically decreases the abundance of proteins essential for biofilm formation, including PpkA and ClpV1-3, and increases the ability of beta-lactam antibiotics to reduce biofilm formation by targeting the beta-lactamase AmpC. Let-7b-5p is bioinformatically predicted to target not only PpkA, ClpV1, and AmpC in P. aeruginosa but also the corresponding orthologs in Burkholderia cenocepacia, another notorious opportunistic lung pathogen, suggesting that the ability of let 7b-5p to reduce biofilm formation and increase beta-lactam sensitivity is not limited to P. aeruginosa. Here, we provide direct evidence for transfer of miRNAs in EVs secreted by eukaryotic cells to a prokaryote, resulting in subsequent phenotypic alterations in the prokaryote as a result of this interdomain communication. Since let-7-family miRNAs are in clinical trials to reduce inflammation and because chronic P. aeruginosa lung infections are associated with a hyperinflammatory state, treatment with let-7b-5p and a beta-lactam antibiotic in nanoparticles or EVs may benefit patients with antibiotic resistant P. aeruginosa infections.