Bacterial respiratory inhibition triggers dispersal of Pseudomonas aeruginosa biofilms.
Bacterial respiratory inhibition triggers dispersal of Pseudomonas aeruginosa biofilms.
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DOI：
10.1128/aem.01101-23
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发表时间：
2023-10-31
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
作者：
铜绿假单胞菌在许多环境条件下生长为生物膜，并且细菌可以通过高度调控的动态过程从生物膜中分散。然而，生物膜分散的生理触发因素仍然知之甚少。根据描述饥饿引发的扩散的先前文献，我们在两个类似于慢性气道感染的模型中测试了细菌呼吸抑制剂对生物膜扩散的影响。我们的基本假设是呼吸抑制剂可以作为饥饿下游影响的模型。我们使用了两种实验条件。在第一种条件下，生物膜从气道上皮细胞表面生长并分散，第二种条件是生物膜在补充有宿主相关铁源的细胞培养基中的玻璃上生长的模型。在两种生物膜模型中，呼吸抑制剂氰化钾和叠氮化钠均引发生物膜扩散。我们假设氰化物诱导的扩散是由于呼吸抑制而不是通过替代机制发出信号，并且事实上，如果通过对氰化物不敏感的氧化酶的过度表达来支持呼吸，则可以阻止扩散。分散需要环二 GMP 调节的蛋白酶 LapG 的活性，从而增强基质降解在分散中的作用。最后，我们检查了单个磷酸二酯酶的作用，之前发现这些磷酸二酯酶与特定触发物的扩散有关，并发现信号传导高度冗余。需要联合删除磷酸二酯酶dipA、bifA和rbdA来减弱分散表型。总之，这项工作增加了对细菌呼吸突然受限的环境条件下生物膜分散生理学的深入了解。铜绿假单胞菌生长在生物膜群落中，在人类感染中很难治疗。生长成生物膜可以保护细菌免受抗生素和免疫系统的侵害。细菌可以通过称为“扩散”的过程留下生物膜。分散的细菌会产生新的生长区域，并且更容易被抗生素杀死。生物膜扩散的触发因素尚不清楚，如果我们更好地了解扩散，可能会导致新的感染治疗方法的开发。在本文中，我们发现抑制铜绿假单胞菌的呼吸（产生能量）的能力可以引发与培养中的人类呼吸道上皮细胞相关的生物膜的扩散。分散过程需要蛋白酶，此前已知该蛋白酶可降解生物膜基质。这些发现让我们更好地了解生物膜分散过程的工作原理，以便未来的研究能够发现更好的方法来清除生物膜中生长的细菌。
Pseudomonas aeruginosa grows as a biofilm under many environmental conditions, and the bacterium can disperse from biofilms via highly regulated, dynamic processes. However, physiologic triggers of biofilm dispersal remain poorly understood. Based on prior literature describing dispersal triggered by forms of starvation, we tested bacterial respiratory inhibitors for biofilm dispersal in two models resembling chronic airway infections. Our underlying hypothesis was that respiratory inhibitors could serve as a model for the downstream effects of starvation. We used two experimental conditions. In the first condition, biofilms were grown and dispersed from the surface of airway epithelial cells, and the second condition was a model where biofilms were grown on glass in cell culture media supplemented with host-relevant iron sources. In both biofilm models, the respiratory inhibitors potassium cyanide and sodium azide each triggered biofilm dispersal. We hypothesized that cyanide-induced dispersal was due to respiratory inhibition rather than signaling via an alternative mechanism, and, indeed, if respiration was supported by overexpression of cyanide-insensitive oxidase, dispersal was prevented. Dispersal required the activity of the cyclic-di-GMP regulated protease LapG, reinforcing the role of matrix degradation in dispersal. Finally, we examined the roles of individual phosphodiesterases, previously implicated in dispersal to specific triggers, and found signaling to be highly redundant. Combined deletion of the phosphodiesterases dipA, bifA, and rbdA was required to attenuate the dispersal phenotype. In summary, this work adds insight into the physiology of biofilm dispersal under environmental conditions in which bacterial respiration is abruptly limited. The bacterium Pseudomonas aeruginosa grows in biofilm communities that are very difficult to treat in human infections. Growing as a biofilm can protect bacteria from antibiotics and the immune system. Bacteria can leave a biofilm through a process called “dispersal.” Dispersed bacteria seed new growth areas and are more susceptible to killing by antibiotics. The triggers for biofilm dispersal are not well understood, and if we understood dispersal better it might lead to the development of new treatments for infection. In this paper, we find that inhibiting P. aeurginosa’s ability to respire (generate energy) can trigger dispersal from a biofilm grown in association with human respiratory epithelial cells in culture. The dispersal process requires a protease which is previously known to degrade the biofilm matrix. These findings give us a better understanding of how the biofilm dispersal process works so that future research can discover better ways of clearing bacteria growing in biofilms.
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影响因子：
3.7
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Harrison JJ
影响因子：
5.2
作者：
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通讯作者：
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影响因子：
4.6
作者：
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