In vitro reconstitution of the polymicrobial community associated with cystic fibrosis (CF) airway infections.

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Each year in the UK, around 160,000 people are diagnosed with asthma, 110,000 with chronic obstructive pulmonary disorder (COPD), 10,000 with bronchiectasis and 300 with cystic fibrosis (CF). These are all respiratory diseases in which the patient's condition becomes worsened by persistent bacterial infections. Over the years, researchers have tried to recapitulate these infections in animal models, and in doing so, they have focussed almost exclusively on infecting the animals (usually mice or rats) with just one species of respiratory pathogen at a time. However, many bacteria are not easily cultured, and over the last decade 16S rDNA sequencing technologies have revealed that many of these chronic respiratory infections are in fact, polymicrobial. Remarkably though, and in spite of these revelations, there have been little or no efforts to try and reconstitute such multi-species communities in the laboratory. The main aim of the current studentship is to rectify this by developing an in vitro system that enables the propagation and maintenance of a stable polymicrobial community derived from the CF lung environment. Animal models of polymicrobial respiratory infections are beginning to appear, and it becoming increasingly apparent that we will see more of these over the next decade. Therefore, we have a window of opportunity in which to develop and disseminate a competing alternative in vitro system that will reduce reliance on these animal models. However, we also want to go further by showing that our model system can reveal novel biology. For example, how does the polymicrobial community respond to antibiotic challenge? What happens if we introduce a new species or a mutant variant of an existing species? How does the community architecture change over time? Because the in vitro model can be experimentally manipulated and perturbed in ways that are just not possible in living mammals, such questions become experimentally tractable.This project is specifically aimed at establishing, validating and disseminating an in vitro model that mimics polymicrobial infections in animals. The intention is that the in vitro model developed here will become the "go to" model for studying polymicrobial infections, thereby contributing directly towards the reduction and replacement "Rs" of the 3Rs scheme. At present, there are no animal-based polymicrobial infection models - although these *are being* developed - so we are poised in a good position to advance the 3Rs nationally and internationally by instilling early "brand loyality" in the in vitro model. However, the project will require considerable development, refinement and validation before it is able to be publicly disseminated, making it ideal for a 3Rs PhD studentship. As outlined in section 3 below, we aim to embed the student in the local and national 3Rs training effort, and at the same time, provide an outstanding scientific training underpinned by the 3Rs ethos. In this regard, it is worth noting that for a large research cluster like the Cambridge area, we host relatively few 3Rs-oriented studentships. We are keen to build capacity in this area and feel that we have the potential to become a future "3Rs hub". For example, we would certainly be keen on hosting 3Rs studentship meetings and lectures (open to all 3Rs-asociated students and fellows across the UK) to promote this, and feel that Cambridge would be a "big draw" for potential speakers.