Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways

Cristina Isabel Amador Hierro, Lars Jelsbak

    Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsResearchpeer-review

    Abstract

    Introduction: In previous studies from our laboratory, a Pseudomonas aeruginosa lineage, named DK2, has been identified and characterized as highly successful, transmissible and persistent over four decades in cystic fibrosis (CF) patients. This lineage underwent substantial phenotypic and genetic changes over time and therefore provides a unique opportunity to explore the impact of those adaptational pathways on its ability to interact with other pathogenic bacteria such as Staphylococcus aureus, a pathogen frequently co-infecting the CF airways. Methods: We have used a novel method to study interspecies interactions between a CF isolate (2003) from the DK2 lineage and a wild-type S. aureus JE2. We grew both strains in mono or co-culture on LB agar, harvested RNA from the colonies after a 24-hour period. Subsequently we performed RNA-seq for the different samples. The data were then compared in a pairwise mode to isolate the transcriptomic profiles for each species. The most differentially expressed genes from both species were validated using real-time quantitative PCR. Results: Interestingly, the greatest expression change was observed in S. aureus, where large clusters of genes associated with virulence were differentially expressed, compared with the monoculture condition, while the P. aeruginosa DK2 response was much more discrete with isolated genes differentially regulated rather than whole operons or clusters. Conclusions: According to our data, S. aureus would display reduced virulence in the presence of an adapted P. aeruginosa DK2 clone, possibly as a consequence of the multiple hostile forces DK2 encountered over time during its long-term adaptation to the CF airways.
    Original languageEnglish
    Title of host publicationThe Danish Microbiological Society Annual Congress 2015 : Programme & Abstracts
    Place of PublicationCopenhagen
    Publication date2015
    Pages68-68
    Publication statusPublished - 2015
    EventThe Danish Microbiological Society Annual Congress 2015 - Eigtved's Pakhus, Copenhagen, Denmark
    Duration: 9 Nov 20159 Nov 2015

    Conference

    ConferenceThe Danish Microbiological Society Annual Congress 2015
    LocationEigtved's Pakhus
    CountryDenmark
    CityCopenhagen
    Period09/11/201509/11/2015

    Cite this

    Amador Hierro, C. I., & Jelsbak, L. (2015). Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways. In The Danish Microbiological Society Annual Congress 2015: Programme & Abstracts (pp. 68-68). Copenhagen.
    Amador Hierro, Cristina Isabel ; Jelsbak, Lars. / Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways. The Danish Microbiological Society Annual Congress 2015: Programme & Abstracts. Copenhagen, 2015. pp. 68-68
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    Amador Hierro, CI & Jelsbak, L 2015, Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways. in The Danish Microbiological Society Annual Congress 2015: Programme & Abstracts. Copenhagen, pp. 68-68, The Danish Microbiological Society Annual Congress 2015, Copenhagen, Denmark, 09/11/2015.

    Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways. / Amador Hierro, Cristina Isabel; Jelsbak, Lars.

    The Danish Microbiological Society Annual Congress 2015: Programme & Abstracts. Copenhagen, 2015. p. 68-68.

    Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsResearchpeer-review

    TY - ABST

    T1 - Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways

    AU - Amador Hierro, Cristina Isabel

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    N2 - Introduction: In previous studies from our laboratory, a Pseudomonas aeruginosa lineage, named DK2, has been identified and characterized as highly successful, transmissible and persistent over four decades in cystic fibrosis (CF) patients. This lineage underwent substantial phenotypic and genetic changes over time and therefore provides a unique opportunity to explore the impact of those adaptational pathways on its ability to interact with other pathogenic bacteria such as Staphylococcus aureus, a pathogen frequently co-infecting the CF airways. Methods: We have used a novel method to study interspecies interactions between a CF isolate (2003) from the DK2 lineage and a wild-type S. aureus JE2. We grew both strains in mono or co-culture on LB agar, harvested RNA from the colonies after a 24-hour period. Subsequently we performed RNA-seq for the different samples. The data were then compared in a pairwise mode to isolate the transcriptomic profiles for each species. The most differentially expressed genes from both species were validated using real-time quantitative PCR. Results: Interestingly, the greatest expression change was observed in S. aureus, where large clusters of genes associated with virulence were differentially expressed, compared with the monoculture condition, while the P. aeruginosa DK2 response was much more discrete with isolated genes differentially regulated rather than whole operons or clusters. Conclusions: According to our data, S. aureus would display reduced virulence in the presence of an adapted P. aeruginosa DK2 clone, possibly as a consequence of the multiple hostile forces DK2 encountered over time during its long-term adaptation to the CF airways.

    AB - Introduction: In previous studies from our laboratory, a Pseudomonas aeruginosa lineage, named DK2, has been identified and characterized as highly successful, transmissible and persistent over four decades in cystic fibrosis (CF) patients. This lineage underwent substantial phenotypic and genetic changes over time and therefore provides a unique opportunity to explore the impact of those adaptational pathways on its ability to interact with other pathogenic bacteria such as Staphylococcus aureus, a pathogen frequently co-infecting the CF airways. Methods: We have used a novel method to study interspecies interactions between a CF isolate (2003) from the DK2 lineage and a wild-type S. aureus JE2. We grew both strains in mono or co-culture on LB agar, harvested RNA from the colonies after a 24-hour period. Subsequently we performed RNA-seq for the different samples. The data were then compared in a pairwise mode to isolate the transcriptomic profiles for each species. The most differentially expressed genes from both species were validated using real-time quantitative PCR. Results: Interestingly, the greatest expression change was observed in S. aureus, where large clusters of genes associated with virulence were differentially expressed, compared with the monoculture condition, while the P. aeruginosa DK2 response was much more discrete with isolated genes differentially regulated rather than whole operons or clusters. Conclusions: According to our data, S. aureus would display reduced virulence in the presence of an adapted P. aeruginosa DK2 clone, possibly as a consequence of the multiple hostile forces DK2 encountered over time during its long-term adaptation to the CF airways.

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    BT - The Danish Microbiological Society Annual Congress 2015

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    Amador Hierro CI, Jelsbak L. Rnaseq As A Method To Study Microbial Interactions Arising In The Cystic Fibrosis Airways. In The Danish Microbiological Society Annual Congress 2015: Programme & Abstracts. Copenhagen. 2015. p. 68-68