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LubriShieldTM - a permanent urinary catheter coating that prevents uropathogen biofilm formation in vitro independent of host protein conditioning

ABSTRACT: LubriShieldTM - a novel permanent coating was invented, and evenly applied to both the internal and external surfaces of indwelling urinary Foley catheters. Without releasing active substances, it effectively prevented pathogens from producing biofilm. The coating was superhydrophilic and incorporated a proprietary anti-fouling ligand, which created a surface that significantly inhibited up to 99% of colonizing uropathogens from forming biofilm for the duration of use without any microbial killing (p< 0.001).RNA-seq analysis revealed that gene expression associated with microbial extracellular polymeric substances formation was significantly downregulated on the coated surfaces. Additionally, microorganisms adhering to LubriShieldTM coated catheters were 78% more susceptible to antibiotics compared to those on uncoated silicone catheters (p=0.004).

ORGANISM(S): Pseudomonas aeruginosa PAO1

PROVIDER: GSE299077 | GEO | 2025/06/05

REPOSITORIES: GEO

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Project description:Purpose: The goals of this study are to compare RNA-Seq transcriptomic data of P. aeruginosa PAO1 in response to chronic exposure of clindamycin and rifampicin eluted from antibiotic-impregnated catheters in a continuous biofilm culture condition. Methods: Transcriptomic profiles of P. aeruginosa PAO1 grown with catheters without antibiotic coating or with clindamycin/rifampicin coating were generated by deep sequencing, in quadruplicates, using Illumina NextSeq 550 sequencer with SE-75. FastQC and Trimmomatic (v0.39) were used for sequencing quality assessment. Raw read counts of the annotated genes were calculated using HTSeq-count. The log2 fold change and false discovery rate (FDR) were measured by edgeR. Genes with |log2FC| > 1.0 and FDR < 0.05 were considered as differential expression genes. Transcripts per million (TPM) was measured by Kallisto, which is an alignment free expression estimation tool. Results: P. aeruginosa PAO1 RNA sequencing using Illumina NextSeq 550 sequencer generated 5,205,190 of average number of reads with average read length of 65 bp and a mean coverage of 54X. Differentially regulated proteins of 877 in the planktonic cells were identified, 55 of which displayed up-regulation and 822 displayed down-regulation. On the other hand, 178 proteins were differentially expressed in the biofilm cells containing 152 up-regulated and 26 down-regulated. Conclusions: Our study represents the first detailed analysis of transcriptomes generated by RNA-seq technology after P. aeruginosa PAO1 planktonic and biofilm cells were continuously exposed to clindamycin/rifampicin-impregnated catheters. Our results show that RNA sequencing offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content of strain PAO1 cells. We conclude that RNA-seq based transcriptome characterization would help the understanding for the mechanism of antimicrobial resistance and virulence in P. aeruginosa after exposure of antimicrobial-coated catheter.

Project description:Actinotignum massiliense, a Gram-positive, facultatively anaerobic coccoid rod, is a rare human pathogen able to infect the urinary tract and belongs to the order of Actinomycetales. We identified A. massiliense as a resident of microbial biofilms growing on indwelling urethral catheter surfaces that were isolated from two patients with neurogenic bladders. These catheter biofilms (CBs) also harbored common uropathogens such as Proteus mirabilis and Aerococcus urinae, supporting the notion that A. massiliense depends on other co-colonizing microbes for survival. We isolated the bacterium from an anaerobically grown culture of a clinical sample, identified the species by 16S rRNA gene sequencing and verified this result via shotgun proteomics. Bacterial proteomes were profiled from the in vitro grown strain and four clinical ‘in vivo’ samples. The quantified proteomes allowed us to infer metabolic pathways and virulence/survival factors of importance in the CB milieu. Two putative subtilisin-like proteases, two Rib/Esp surface antigen repeat-containing proteins, a papain-like cysteine protease and a metal/heme/oligopeptide uptake system were highly expressed in vivo, but less so in vitro. We predict these proteins to be critical for adhesion and growth in CBs attacked by the host’s innate immune system or to improve bacterial fitness. Mixed acid fermentation following uptake and metabolism of xylose and glucuronate, sugars highly represented in proteoglycans and glycoglycerolipids of the urothelial mucosa and, in the case of glucuronate, shed into urine via renal xenobiotic conjugates, is inferred to be a major pathway for A. massiliense to generate energy under microaerobic conditions in CBs. The bacteria also appear to have active pathways for storage and utilization of glycogen as a carbon resource. Finally, we identified a putative polyketide synthase which may generate a secondary metabolite that interacts with either the host or co-colonizing organisms to enable A. massiliense survival in CBs.

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LubriShieldTM - a permanent urinary catheter coating that prevents uropathogen biofilm formation in vitro independent of host protein conditioning

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