Transcriptomics,Genomics

Dataset Information

161

The glutathione-disrupted biofilm of clinical Pseudomonas aeruginosa strains: enhanced antibiotic affect and a novel biofilm transcriptome


ABSTRACT: Pseudomonas aeruginosa infections for individuals with Cystic Fibrosis (CF), result in high morbidity and mortality, with premature death often occurring. These infections are complicated by the formation of biofilms in the sputum. Antibiotic therapy is stymied by antibiotic resistance of the biofilm matrix, making novel anti-biofilm strategies highly desirable. Within the P. aeruginosa biofilm, the redox factor pyocyanin enhances biofilm integrity by intercalating with extracellular DNA. The antioxidant glutathione (GSH) reacts with pyocyanin to disrupt intercalation. This study investigated GSH disruption by assaying the physiological effects of GSH and DNase I on biofilms of clinical CF isolates grown in artificial CF sputum media (ASMDM+). Confocal scanning laser microscopy showed that 2mM GSH alone or combined with DNase I significantly disrupted the immature (24 hour) biofilms of Australian Epidemic Strain (AES) isogens AES-1R and AES-1M. GSH alone greatly disrupted the mature (72 hour) biofilm of AES-1R, resulting in significant differential expression of 587 genes, as evidenced by RNA-sequencing. Upregulated systems included cyclic diguanylate and pyoverdine biosynthesis, the Type VI secretion system, nitrate metabolism and translational machinery. Physiochemical biofilm disruption with GSH revealed a metabolically active cellular physiology distinct from either mature or dispersed biofilm physiology. RNA-seq results were validated by biochemical assay and qPCR. Biofilms of a range of CF isolates disrupted with GSH and DNase I were significantly more susceptible to ciprofloxacin, and increased antibiotic effectiveness was achieved at 10mM GSH. This study demonstrated that GSH alone or with DNase I represent effective anti-biofilm treatments when combined with appropriate antibiotics. Overall design: Examination of the effects of 2mM glutathione (GSH) on the mature (72 hour) biofilms of P. aeruginosa strain AES-1R. 2 replicates control (1× PBS treated) 2 replicates 2mM GSH treated. RNA harvest 8 hours post-treatment.

INSTRUMENT(S): Illumina NextSeq 500 (Pseudomonas aeruginosa AES-1R)

SUBMITTER: Jim Manos  

PROVIDER: GSE75334 | GEO | 2016-02-01

SECONDARY ACCESSION(S): PRJNA303970

REPOSITORIES: GEO

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Publications

Glutathione-Disrupted Biofilms of Clinical Pseudomonas aeruginosa Strains Exhibit an Enhanced Antibiotic Effect and a Novel Biofilm Transcriptome.

Klare William W   Das Theerthankar T   Ibugo Amaye A   Buckle Edwina E   Manefield Mike M   Manos Jim J  

Antimicrobial agents and chemotherapy 20160722 8


Pseudomonas aeruginosa infections result in high morbidity and mortality rates for individuals with cystic fibrosis (CF), with premature death often occurring. These infections are complicated by the formation of biofilms in the sputum. Antibiotic therapy is stymied by antibiotic resistance of the biofilm matrix, making novel antibiofilm strategies highly desirable. Within P. aeruginosa biofilms, the redox factor pyocyanin enhances biofilm integrity by intercalating with extracellular DNA. The a  ...[more]

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