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      High-resolution in situ transcriptomics of Pseudomonas aeruginosa unveils genotype independent patho-phenotypes in cystic fibrosis lungs

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          Abstract

          Life-long bacterial infections in cystic fibrosis (CF) airways constitute an excellent model both for persistent infections and for microbial adaptive evolution in complex dynamic environments. Using high-resolution transcriptomics applied on CF sputum, we profile transcriptional phenotypes of Pseudomonas aeruginosa populations in patho-physiological conditions. Here we show that the soft-core genome of genetically distinct populations, while maintaining transcriptional flexibility, shares a common expression program tied to the lungs environment. We identify genetically independent traits defining P. aeruginosa physiology in vivo, documenting the connection between several previously identified mutations in CF isolates and some of the convergent phenotypes known to develop in later stages of the infection. In addition, our data highlight to what extent this organism can exploit its extensive repertoire of physiological pathways to acclimate to a new niche and suggest how alternative nutrients produced in the lungs may be utilized in unexpected metabolic contexts.

          Abstract

          How genetic adaptation and phenotypic acclimation are interrelated and allow Pseudomonas aeruginosa to persist in cystic fibrosis lungs is poorly understood. Here, Rossi et al. use high-resolution transcriptomics on expectorates to link phenotypic conservation to ecological flexibility and persistence.

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          Pseudomonas aeruginosa Evolutionary Adaptation and Diversification in Cystic Fibrosis Chronic Lung Infections

          Pseudomonas aeruginosa populations undergo a characteristic evolutionary adaptation during chronic infection of the cystic fibrosis (CF) lung, including reduced production of virulence factors, transition to a biofilm-associated lifestyle, and evolution of high-level antibiotic resistance. Populations of P. aeruginosa in chronic CF lung infections typically exhibit high phenotypic diversity, including for clinically important traits such as antibiotic resistance and toxin production, and this diversity is dynamic over time, making accurate diagnosis and treatment challenging. Population genomics studies reveal extensive genetic diversity within patients, including for transmissible strains the coexistence of highly divergent lineages acquired by patient-to-patient transmission. The inherent spatial structure and spatial heterogeneity of selection in the CF lung appears to play a key role in driving P. aeruginosa diversification.
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            Adaptation of Pseudomonas aeruginosa to the cystic fibrosis airway: an evolutionary perspective.

            The airways of patients with cystic fibrosis (CF) are nearly always infected with many different microorganisms. This environment offers warm, humid and nutrient-rich conditions, but is also stressful owing to frequent antibiotic therapy and the host immune response. Pseudomonas aeruginosa is commonly isolated from the airways of patients with CF, where it most often establishes chronic infections that usually persist for the rest of the lives of the patients. This bacterium is a major cause of mortality and morbidity and has therefore been studied intensely. Here, we discuss how P. aeruginosa evolves from a state of early, recurrent intermittent colonization of the airways of patients with CF to a chronic infection state, and how this process offers opportunities to study bacterial evolution in natural environments. We believe that such studies are valuable not only for our understanding of bacterial evolution but also for the future development of new therapeutic strategies to treat severe chronic infections.
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              Nutritional cues control Pseudomonas aeruginosa multicellular behavior in cystic fibrosis sputum.

              The sputum (mucus) layer of the cystic fibrosis (CF) lung is a complex substrate that provides Pseudomonas aeruginosa with carbon and energy to support high-density growth during chronic colonization. Unfortunately, the CF lung sputum layer has been difficult to mimic in animal models of CF disease, and mechanistic studies of P. aeruginosa physiology during growth in CF sputum are hampered by its complexity. In this study, we performed chromatographic and enzymatic analyses of CF sputum to develop a defined, synthetic CF sputum medium (SCFM) that mimics the nutritional composition of CF sputum. Importantly, P. aeruginosa displays similar phenotypes during growth in CF sputum and in SCFM, including similar growth rates, gene expression profiles, carbon substrate preferences, and cell-cell signaling profiles. Using SCFM, we provide evidence that aromatic amino acids serve as nutritional cues that influence cell-cell signaling and antimicrobial activity of P. aeruginosa during growth in CF sputum.
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                Author and article information

                Contributors
                sm@bio.dtu.dk
                hkj@biosustain.dtu.dk
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                27 August 2018
                27 August 2018
                2018
                : 9
                : 3459
                Affiliations
                [1 ]GRID grid.475435.4, Department of Clinical Microbiology, , Rigshospitalet, ; DK-2100 Copenhagen Ø, Denmark
                [2 ]ISNI 0000 0001 2181 8870, GRID grid.5170.3, Novo Nordisk Foundation Center for Biosustainability, , Technical University of Denmark, ; DK-2800 Kgs. Lyngby, Denmark
                [3 ]ISNI 0000 0001 0674 042X, GRID grid.5254.6, Department of Clinical Medicine, Faculty of Health and Medical Sciences, , University of Copenhagen, ; DK-2200 Copenhagen N, Denmark
                Author information
                http://orcid.org/0000-0002-2042-608X
                http://orcid.org/0000-0001-7838-7258
                http://orcid.org/0000-0003-0268-3717
                Article
                5944
                10.1038/s41467-018-05944-5
                6110831
                30150613
                171df50b-cb16-4039-9ddc-418087e50e52
                © The Author(s) 2018

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 14 February 2018
                : 30 July 2018
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100005111, Rigshospitalet;
                Award ID: R88-A3537
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100003554, Lundbeckfonden (Lundbeck Foundation);
                Award ID: R167-2013-15229
                Award Recipient :
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