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Ex vivo transcriptional profiling reveals a common set of genes important for the adaptation of Pseudomonas aeruginosa to chronically infected host sites

ABSTRACT: The opportunistic bacterium Pseudomonas aeruginosa is a major nosocomial pathogen causing both devastating acute and chronic persistent infections. During the course of an infection, P. aeruginosa rapidly adapts to the specific conditions within the host. In the present study, we aimed at the identification of genes that are highly expressed during biofilm infections such as in chronically infected lungs of patients with cystic fibrosis (CF), burn wounds and subcutaneous mouse tumours. We found a common subset of differentially regulated genes in all three in vivo habitats and evaluated whether their inactivation impacts on the bacterial capability to form biofilms in vitro and to establish biofilm-associated infections in a murine model. Additive effects on biofilm formation and host colonization were discovered by the combined inactivation of several highly expressed genes. However, even combined inactivation was not sufficient to abolish the establishment of an infection completely. These findings can be interpreted as evidence that either redundant traits encode functions that are essential for in vivo survival and chronic biofilm infections and/or bacterial adaptation is considerably achieved independently of transcription levels. Supplemental screens, will have to be applied in order to identify the minimal set of key genes essential for the establishment of chronic infectious diseases.

ORGANISM(S): Pseudomonas aeruginosa

PROVIDER: GSE25945 | GEO | 2012/11/14

SECONDARY ACCESSION(S): PRJNA135703

REPOSITORIES: GEO

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Ex vivo transcriptional profiling reveals a common set of genes important for the adaptation of Pseudomonas aeruginosa to chronically infected host sites

Project description:The opportunistic bacterium Pseudomonas aeruginosa is a major nosocomial pathogen causing both devastating acute and chronic persistent infections. During the course of an infection, P. aeruginosa rapidly adapts to the specific conditions within the host. In the present study, we aimed at the identification of genes that are highly expressed during biofilm infections such as in chronically infected lungs of patients with cystic fibrosis (CF), burn wounds and subcutaneous mouse tumours. We found a common subset of differentially regulated genes in all three in vivo habitats and evaluated whether their inactivation impacts on the bacterial capability to form biofilms in vitro and to establish biofilm-associated infections in a murine model. Additive effects on biofilm formation and host colonization were discovered by the combined inactivation of several highly expressed genes. However, even combined inactivation was not sufficient to abolish the establishment of an infection completely. These findings can be interpreted as evidence that either redundant traits encode functions that are essential for in vivo survival and chronic biofilm infections and/or bacterial adaptation is considerably achieved independently of transcription levels. Supplemental screens, will have to be applied in order to identify the minimal set of key genes essential for the establishment of chronic infectious diseases. Ex-vivo samples were collected from burn wound, cystic fibrosis patients mucous and mice tumor and stabilized for the RNA extraction. Recovered clinical strains were cultivated in rich medium control conditions untill the early stationary phase and RNA was extracted. The murine tumors were infected with burn wound isolated strains

2012-11-14 | E-GEOD-25945 | biostudies-arrayexpress

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In-Vivo Expression Profiling of Pseudomonas aeruginosa Infections Reveals Niche-Specific and Strain-Independent Transcriptional Programs

Project description:Pseudomonas aeruginosa is a threatening, opportunistic pathogen causing disease in immunocompromised individuals. The hallmark of P. aeruginosa virulence is its multi-factorial and combinatorial nature. It renders such bacteria infectious for many organisms and it is often resistant to antibiotics. To gain insights into the physiology of P. aeruginosa during infection, we assessed the transcriptional programs of three different P. aeruginosa strains directly after isolation from burn wounds of humans. We compared the programs to those of the same strains using two infection models: a plant model, which consisted of the infection of the midrib of lettuce leaves, and a murine tumor model, which was obtained by infection of mice with an induced tumor in the abdomen. All control conditions of P. aeruginosa cells growing in suspension and as a biofilm were added to the analysis. We found that these different P. aeruginosa strains express a pool of distinct genetic traits that are activated under particular infection conditions regardless of their genetic variability. The knowledge herein generated will advance our understanding of P. aeruginosa virulence and provide valuable cues for the definition of prospective targets to develop novel intervention strategies. Samples collected from human burn wounds were stabilized and used for P. aeruginosa RNA extraction. The strains from burn wound infection were later used for infection models namely plant and murine tumor infection as well as rich medium controls with planctonic and biofilm growth. The RNA was treated for bacterial RNA enrichment and amplification. All samples were treated in the same way.

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rG4-seq for P. aeruginosa and E. coli

Project description:Escherichia coli is the most widely studied strains, which has irreplaceable position in medicine and biology research. Pseudomonas aeruginosa, an opportunistic human pathogen, tends to cause potentially lethal acute or chronic infections in patients with cystic fibrosis (CF), immunocompromised individuals and burn victims. However, it is little know about the effect of the special secondary structure rG4 (G-quadruplex) in the mRNA on virulence regulation. Here, we aim to reveal the new and important post-transcriptional regulatory roles of rG4 in bacterial pathogenicity and metabolic pathways.

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Project description:Bacterial interspecies interactions shape the function and structural dynamics of microbial communities and affect disease progression of polymicrobial infections. Here, we present data suggesting that the FemA-FemR-FemI (Fem) cell surface signaling system in P. aeruginosa serves as an interspecies signaling pathway between P. aeruginosa and Mycobacterium species. The Fem system is regulated by the type three secretion system (T3SS) regulator ExsA. Fem system significantly influenced virulence factors in P. aeruginosa, including the quorum sensing systems, pyocyanin production, biofilm formation and the type six secretion systems (T6SSs). Our study using a Galleria mellonella infection model indicates femA deletion significantly increased the host survival rate while femI over-expression decreased it, demonstrating that the Fem system’s role in bacterial pathogenicity in vivo. We propose that the Fem system functions as an interspecies signaling pathway enabling P. aeruginosa to alter its behaviours in response to the presence of mycobacteria.

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Circular RNA Expression Profiles Following Negative Pressure Wound Therapy in Burn Wounds with Experimental Pseudomonas aeruginosa Infection

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