Porcine Nasal Organoids as a model to study the interactions between the swine nasal microbiota and the host

Bonillo-Lopez, Laura; Carmona-Vicente, Noelia; Tarrés-Freixas, Ferran; Kochanowski, Karl; Martínez, Jorge; Perez, Mònica; Sibila, Marina; Correa-Fiz, Florencia; Aragon, Virginia

doi:10.1101/2024.08.13.606910

Cited by 2 publications

References 44 publications

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In vitrometabolic interaction network of a rationally designed nasal microbiota community

Bonillo-Lopez,

Rouam-el Khatab,

Obregon-Gutierrez

et al. 2024

Preprint

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Mounting evidence suggests that metabolite exchange between microbiota members is a key driver of microbiota composition. However, we still know little about the metabolic interaction networks that occur within many microbiota. This is particularly true for the nasal microbiota, and current efforts towards this end are hampered by a lack of microbial consortia that would enable the mapping of metabolic interactions between nasal microbiota members under in vitro conditions. To tackle these issues, we developed the Porcine Nasal Consortium (PNC8), a rationally designed microbial consortium of eight strains representing the most in vivo abundant genera in the nasal microbiota of healthy piglets. We used this consortium to systematically examine the metabolic capabilities of nasal microbiota members, as well as the metabolic interactions occurring between them. We found that PNC8 strains differ substantially in their metabolic pathway repertoire and ability to grow across various in vitro conditions. Nevertheless, spent-media experiments revealed that most metabolic interactions between PNC8 strains are negative, and exometabolomics data pointed to co-depletion of sugars as a key driver of this interaction network. Finally, direct co-cultivation experiments showed that, as a result of this largely negative metabolic interaction network, competition is common among pairs of PNC8 strains and leads to a complex competition hierarchy in which only few strains are able to consistently outcompete all others. Overall, this work provides a valuable resource for studying the nasal microbiota under experimentally tractable in vitro conditions and is a key step towards mapping its metabolic interaction network.

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In vitrometabolic interaction network of a rationally designed nasal microbiota community

Bonillo-Lopez,

Rouam-el Khatab,

Obregon-Gutierrez

et al. 2024

Preprint

View full text Add to dashboard Cite

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Porcine Airway Organoid-Derived Well-Differentiated Epithelial Cultures as a Tool for the Characterization of Swine Influenza a Virus Strains

Gerhards,

Vrieling,

Dresken

et al. 2024

Viruses

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Swine influenza A viruses (IAVsw) are important causes of disease in pigs but also constitute a public health risk. IAVsw strains show remarkable differences in pathogenicity. We aimed to generate airway organoids from the porcine lower respiratory tract and use these to establish well-differentiated airway epithelial cell (WD-AEC) cultures grown at an air–liquid interface (ALI) for in vitro screening of IAVsw strain virulence. Epithelial cells were isolated from bronchus tissue of juvenile pigs, and airway organoids were cultured in an extracellular matrix in a culture medium containing human growth factors. Single-cell suspensions of these 3D organoids were seeded on Transwell filters and differentiated at ALI to form a pseudostratified epithelium containing ciliated cells, mucus-producing cells and tight junctions. Inoculation with a low dose of IAVsw in a low volume inoculum resulted in virus replication without requiring the addition of trypsin, and was quantified by the detection of viral genome loads in apical washes. Interestingly, inoculation of an H3N2 strain known to cause severe disease in pigs induced a greater reduction in trans-epithelial resistance and more damage to tight junctions than H1N2 or H1N1 strains associated with mild disease in pigs. We conclude that the porcine WD-AEC model is useful in assessing the virulence of IAVsw strains.

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Porcine Nasal Organoids as a model to study the interactions between the swine nasal microbiota and the host

Cited by 2 publications

References 44 publications

In vitrometabolic interaction network of a rationally designed nasal microbiota community

In vitrometabolic interaction network of a rationally designed nasal microbiota community

Porcine Airway Organoid-Derived Well-Differentiated Epithelial Cultures as a Tool for the Characterization of Swine Influenza a Virus Strains

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