Analysis of Bacterial Biofilm Formation and MUC5AC and MUC5B Expression in Chronic Rhinosinusitis Patients

Popov, G.; Aleksandrov, Radoslav; Petkova, V.; Kaneva, Radka; Gergova, Raina; Kundurzhiev, Todor; Попова, Д.

doi:10.3390/jcm12051808

Cited by 5 publications

(3 citation statements)

References 72 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Biofilms are communities of bacteria encased in an extracellular matrix of protein, polysaccharides, and extracellular DNA and associated with bacterial tolerance to immune recognition, immune killing, and antibiotic treatment [1,2]. Biofilms can form on abiotic surfaces like plastic and glass but can also form in association with mucosal surfaces in many diseases, including chronic respiratory diseases such as chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), and chronic rhinosinusitis [3][4][5][6]. While much is understood about biofilm formation on abiotic surfaces, little is known about the host factors regulating biofilm growth in the host.…”

Section: Introductionmentioning

confidence: 99%

Pseudomonas aeruginosa senses and responds to epithelial potassium flux via Kdp operon to promote biofilm

Rapsinski,

Michaels,

Hill

et al. 2024

PLoS Pathog

View full text Add to dashboard Cite

Mucosa-associated biofilms are associated with many human disease states, but the host mechanisms promoting biofilm remain unclear. In chronic respiratory diseases like cystic fibrosis (CF), Pseudomonas aeruginosa establishes chronic infection through biofilm formation. P. aeruginosa can be attracted to interspecies biofilms through potassium currents emanating from the biofilms. We hypothesized that P. aeruginosa could, similarly, sense and respond to the potassium efflux from human airway epithelial cells (AECs) to promote biofilm. Using respiratory epithelial co-culture biofilm imaging assays of P. aeruginosa grown in association with CF bronchial epithelial cells (CFBE41o-), we found that P. aeruginosa biofilm was increased by potassium efflux from AECs, as examined by potentiating large conductance potassium channel, BKCa (NS19504) potassium efflux. This phenotype is driven by increased bacterial attachment and increased coalescence of bacteria into aggregates. Conversely, biofilm formation was reduced when AECs were treated with a BKCa blocker (paxilline). Using an agar-based macroscopic chemotaxis assay, we determined that P. aeruginosa chemotaxes toward potassium and screened transposon mutants to discover that disruption of the high-sensitivity potassium transporter, KdpFABC, and the two-component potassium sensing system, KdpDE, reduces P. aeruginosa potassium chemotaxis. In respiratory epithelial co-culture biofilm imaging assays, a KdpFABCDE deficient P. aeruginosa strain demonstrated reduced biofilm growth in association with AECs while maintaining biofilm formation on abiotic surfaces. Furthermore, we determined that the Kdp operon is expressed in vivo in people with CF and the genes are conserved in CF isolates. Collectively, these data suggest that P. aeruginosa biofilm formation can be increased by attracting bacteria to the mucosal surface and enhancing coalescence into microcolonies through aberrant AEC potassium efflux sensed by the KdpFABCDE system. These findings suggest host electrochemical signaling can enhance biofilm, a novel host-pathogen interaction, and potassium flux could be a therapeutic target to prevent chronic infections in diseases with mucosa-associated biofilms, like CF.

show abstract

Section: Introductionmentioning

confidence: 99%

Pseudomonas aeruginosa senses and responds to epithelial potassium flux via Kdp operon to promote biofilm

Rapsinski,

Michaels,

Hill

et al. 2024

PLoS Pathog

View full text Add to dashboard Cite

show abstract

“…Although the exact role of bacterial biofilms in CRS has not yet been fully elucidated, there is growing evidence to suggest that biofilms are an important factor in CRS pathogenesis which could help to explain resistance to conventional therapies. 3 The presence of bacterial biofilms on the mucosa of CRS patients is well documented, [4][5][6][7][8][9][10] and has been associated with undesirable clinical outcomes such as more severe disease pre-and post-operatively, greater risk of recurrence, and requirement of additional courses of antibiotics and revisional surgeries. 11 Multiple common biofilm-forming bacterial pathogens have been implicated in CRS, 3 and specifically Staphylococcus aureus biofilms have the greatest association with severely recurrent and recalcitrant cases.…”

Section: Introductionmentioning

confidence: 99%

Z‐Form Extracellular DNA in Pediatric CRS May Provide a Mechanism for Recalcitrance to Treatment

Hofer,

Jurcisek,

Elmaraghy

et al. 2023

The Laryngoscope

View full text Add to dashboard Cite

ObjectivesWe examined sinus mucosal samples recovered from pediatric chronic rhinosinusitis (CRS) patients for the presence of Z‐form extracellular DNA (eDNA) due to its recently elucidated role in pathogenesis of disease. Further, we immunolabeled these specimens for the presence of both members of the bacterial DNA‐binding DNABII protein family, integration host factor (IHF) and histone‐like protein (HU), due to their known role in converting common B‐DNA to the rare Z‐form.MethodsSinus mucosa samples recovered from 20 patients during functional endoscopic sinus surgery (FESS) were immunolabelled for B‐ and Z‐DNA, as well as for both bacterial DNABII proteins.ResultsNineteen of 20 samples (95%) included areas rich in eDNA, with the majority in the Z‐form. Areas positive for B‐DNA were restricted to the most distal regions of the mucosal specimen. Labeling for both DNABII proteins was observed on B‐ and Z‐DNA, which aligned with the role of these proteins in the B‐to‐Z DNA conversion.ConclusionsAbundant Z‐form eDNA in culture‐positive pediatric CRS samples suggested that bacterial DNABII proteins were responsible for the conversion of eukaryotic B‐DNA that had been released into the luminal space by PMNs during NETosis, to the Z‐form. The presence of both DNABII proteins on B‐DNA and Z‐DNA supported the known role of these bacterial proteins in the B‐to‐Z DNA conversion. Given that Z‐form DNA both stabilizes the bacterial biofilm and inactivates PMN NET‐mediated killing of trapped bacteria, we hypothesize that this conversion may be contributing to the chronicity and recalcitrance of CRS to treatment.Level of EvidenceNot applicable Laryngoscope, 2023

show abstract

Section: Introductionmentioning

confidence: 99%

Pseudomonas aeruginosasenses and responds to epithelial potassium flux via Kdp operon to promote biofilm biogenesis

Rapsinski

Hill

Yarrington

et al. 2023

Preprint

View full text Add to dashboard Cite

Mucosa-associated biofilms are associated with many human disease states, but the mechanisms by which the host promotes biofilm biogenesis remain unclear. In chronic respiratory diseases like cystic fibrosis (CF), Pseudomonas aeruginosa establishes chronic infection through biofilm formation. P. aeruginosa can be attracted to interspecies biofilms through potassium currents emanating from the biofilms. We hypothesized that P. aeruginosa could, similarly, sense and respond to the potassium efflux from human airway epithelial cells (AECs) to promote biofilm biogenesis. Using respiratory epithelial co-culture biofilm imaging assays of P. aeruginosa grown in association with CF bronchial epithelial cells (CFBE41o-), we found that P. aeruginosa biofilm biogenesis was increased by potassium efflux from AECs, as examined by potentiating large conductance potassium channel, BKCa (NS19504) potassium efflux. This phenotype is driven by increased bacterial attachment and increased coalescence of bacteria into aggregates. Conversely, biofilm formation was reduced when AECs were treated with a BKCa blocker (paxilline). Using an agar-based macroscopic chemotaxis assay, we determined that P. aeruginosa chemotaxes toward potassium and screened transposon mutants to discover that disruption of the high-sensitivity potassium transporter, KdpFABC, and the two-component potassium sensing system, KdpDE, reduces P. aeruginosa potassium chemotaxis. In respiratory epithelial co-culture biofilm imaging assays, a KdpFABCDE deficient P. aeruginosa strain demonstrated reduced biofilm growth in association with AECs while maintaining biofilm formation on abiotic surfaces. Collectively, these data suggest that P. aeruginosa biofilm formation can be increased by attracting bacteria to the mucosal surface via a potassium gradient and enhancing coalescence of single bacteria into microcolonies through aberrant AEC potassium efflux sensed through the bacterial KdpFABCDE system. These findings suggest that electrochemical signaling from the host can amplify biofilm biogenesis, a novel host-pathogen interaction, and that potassium flux could be a potential target for therapeutic intervention to prevent chronic bacterial infections in diseases with mucosa-associated biofilms, like CF.

show abstract

Analysis of Bacterial Biofilm Formation and MUC5AC and MUC5B Expression in Chronic Rhinosinusitis Patients

Cited by 5 publications

References 72 publications

Pseudomonas aeruginosa senses and responds to epithelial potassium flux via Kdp operon to promote biofilm

Pseudomonas aeruginosa senses and responds to epithelial potassium flux via Kdp operon to promote biofilm

Z‐Form Extracellular DNA in Pediatric CRS May Provide a Mechanism for Recalcitrance to Treatment

Pseudomonas aeruginosasenses and responds to epithelial potassium flux via Kdp operon to promote biofilm biogenesis

Contact Info

Product

Resources

About