Pseudomonas Quinolone Signal-Induced Outer Membrane Vesicles Enhance Biofilm Dispersion in Pseudomonas aeruginosa.
Pseudomonas Quinolone Signal-Induced Outer Membrane Vesicles Enhance Biofilm Dispersion in Pseudomonas aeruginosa.
复制标题
假单胞菌喹诺酮信号诱导的外膜囊泡增强铜绿假单胞菌的生物膜分散。
DOI：
10.1128/msphere.01109-20
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发表时间：
2020-11-25
期刊：
影响因子：
4.8
通讯作者：
Schertzer JW
中科院分区：
文献类型：
--
作者：
Cooke AC;Florez C;Dunshee EB;Lieber AD;Terry ML;Light CJ;Schertzer JW
操纵生物膜分散的治疗有可能将慢性耐药生物膜感染从受保护的固着群落转化为对抗菌治疗更敏感的数量级的释放群体。然而，分散的细胞往往表现出增加的急性毒力和传播表型。细菌生物膜是人类慢性感染的主要贡献者。由于它们对常规治疗无效，因此它们提出了特别困难的治疗挑战。确定参与生物膜发展的因素可以帮助发现新的目标，并指导生物膜战略的发展。铜绿假单胞菌引起手术部位、烧伤伤口和医院获得性感染，并且还与囊性纤维化患者肺中的侵袭性生物膜形成相关。铜绿假单胞菌毒力的一个有效但知之甚少的贡献者是产生外膜囊泡（OMV）的能力。OMV的运输与细胞间通讯、毒力因子传递和抗生素抗性基因的转移有关。由于OMV几乎完全是使用无菌培养物进行研究的，因此对其在生物膜中的生物起源和功能知之甚少。几个研究小组已经表明，假单胞菌喹诺酮信号（PQS）诱导铜绿假单胞菌中OMV的形成。我们的小组描述了一种生物物理机制，最近表明它在生物膜中起作用。在这里，我们表明，PQS诱导的OMV生产是高度动态的生物膜的发展过程中。有趣的是，与附着和成熟阶段相比，PQS和OMV合成在分散期间显著升高。PQS生物合成和受体突变体生物膜的分散能力显着受损，但这种表型被拯救的遗传互补或外源添加PQS。最后，我们表明，纯化的OMV可以主动降解细胞外蛋白质，脂质和DNA。因此，我们建议，提高生产的PQS诱导的OMV在生物膜分散促进细胞逃逸协调生物膜基质成分的控制降解。重要性操纵生物膜分散的治疗有可能将慢性耐药生物膜感染从受保护的固着群落转化为对抗菌治疗更敏感的数量级的释放群体。然而，分散的细胞往往表现出增加的急性毒力和传播表型。因此，在有效地采用这种有前途的策略之前，对分散过程的透彻理解至关重要。假单胞菌喹诺酮信号（PQS）已牵连在早期生物膜的发展，但我们假设，其功能作为外膜囊泡（OMV）诱导剂可能有助于在多个阶段。在这里，我们证明，PQS和OMV的差异产生在铜绿假单胞菌生物膜的发展，并提供证据表明，有效的生物膜分散是依赖于生产的PQS诱导的OMV，这可能作为交付车辆的基质降解酶。这些研究结果奠定了基础，了解OMV的贡献，生物膜的发展，并提出了一个模型来解释控制基质降解，伴随着生物膜分散在许多物种。
Treatments that manipulate biofilm dispersion hold the potential to convert chronic drug-tolerant biofilm infections from protected sessile communities into released populations that are orders-of-magnitude more susceptible to antimicrobial treatment. However, dispersed cells often exhibit increased acute virulence and dissemination phenotypes. Bacterial biofilms are major contributors to chronic infections in humans. Because they are recalcitrant to conventional therapy, they present a particularly difficult treatment challenge. Identifying factors involved in biofilm development can help uncover novel targets and guide the development of antibiofilm strategies. Pseudomonas aeruginosa causes surgical site, burn wound, and hospital-acquired infections and is also associated with aggressive biofilm formation in the lungs of cystic fibrosis patients. A potent but poorly understood contributor to P. aeruginosa virulence is the ability to produce outer membrane vesicles (OMVs). OMV trafficking has been associated with cell-cell communication, virulence factor delivery, and transfer of antibiotic resistance genes. Because OMVs have almost exclusively been studied using planktonic cultures, little is known about their biogenesis and function in biofilms. Several groups have shown that Pseudomonas quinolone signal (PQS) induces OMV formation in P. aeruginosa. Our group described a biophysical mechanism for this and recently showed it is operative in biofilms. Here, we demonstrate that PQS-induced OMV production is highly dynamic during biofilm development. Interestingly, PQS and OMV synthesis are significantly elevated during dispersion compared to attachment and maturation stages. PQS biosynthetic and receptor mutant biofilms were significantly impaired in their ability to disperse, but this phenotype was rescued by genetic complementation or exogenous addition of PQS. Finally, we show that purified OMVs can actively degrade extracellular protein, lipid, and DNA. We therefore propose that enhanced production of PQS-induced OMVs during biofilm dispersion facilitates cell escape by coordinating the controlled degradation of biofilm matrix components. IMPORTANCE Treatments that manipulate biofilm dispersion hold the potential to convert chronic drug-tolerant biofilm infections from protected sessile communities into released populations that are orders-of-magnitude more susceptible to antimicrobial treatment. However, dispersed cells often exhibit increased acute virulence and dissemination phenotypes. A thorough understanding of the dispersion process is therefore critical before this promising strategy can be effectively employed. Pseudomonas quinolone signal (PQS) has been implicated in early biofilm development, but we hypothesized that its function as an outer membrane vesicle (OMV) inducer may contribute at multiple stages. Here, we demonstrate that PQS and OMVs are differentially produced during Pseudomonas aeruginosa biofilm development and provide evidence that effective biofilm dispersion is dependent on the production of PQS-induced OMVs, which likely act as delivery vehicles for matrix-degrading enzymes. These findings lay the groundwork for understanding OMV contributions to biofilm development and suggest a model to explain the controlled matrix degradation that accompanies biofilm dispersion in many species.