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

在英国，每年约有16万人被诊断患有哮喘，11万人患有慢性阻塞性肺疾病（COPD）， 1万人患有支气管扩张症，300人患有囊性纤维化（CF）。这些都是呼吸道疾病，患者的病情会因持续的细菌感染而恶化。多年来，研究人员一直试图在动物模型中重现这些感染，在这样做的过程中，他们几乎完全集中在一次只感染一种呼吸道病原体的动物（通常是小鼠或大鼠）上。然而，许多细菌不容易培养，在过去的十年中，16S rDNA测序技术揭示了许多慢性呼吸道感染实际上是多微生物。然而，值得注意的是，尽管有了这些启示，却很少或根本没有努力尝试在实验室中重建这种多物种群落。当前研究的主要目的是通过开发一种体外系统来纠正这一问题，该系统能够繁殖和维持来自CF肺环境的稳定的多微生物群落。多微生物呼吸道感染的动物模型开始出现，而且越来越明显的是，我们将在未来十年看到更多这样的动物模型。因此，我们有机会开发和推广一种竞争性的体外系统，以减少对这些动物模型的依赖。然而，我们也想更进一步，通过展示我们的模型系统可以揭示新的生物学。例如，多微生物群落如何应对抗生素的挑战？如果我们引入一个新物种或现有物种的突变变体会发生什么？社区架构是如何随时间变化的？因为体外模型可以在实验上被操纵和干扰，这在哺乳动物身上是不可能的，所以这些问题在实验上是可以处理的。本项目旨在建立、验证和传播一种模拟动物多微生物感染的体外模型。我们的目的是，这里开发的体外模型将成为研究多微生物感染的“去”模型，从而直接有助于减少和替代3Rs方案的“Rs”。目前，还没有基于动物的多微生物感染模型——尽管这些模型正在开发中——因此，通过在体外模型中灌输早期的“品牌忠诚度”，我们处于一个很好的位置，可以在国内和国际上推进3r。然而，在能够公开传播之前，该项目将需要大量的开发，改进和验证，使其成为3Rs博士研究生的理想选择。正如下文第3节所述，我们的目标是将学生融入当地和国家的3r培训工作中，同时提供以3r精神为基础的优秀科学培训。在这方面，值得注意的是，对于像剑桥地区这样的大型研究集群，我们接待的3rs型学生相对较少。我们希望加强这方面的能力建设，并认为香港有潜力成为未来的“3r枢纽”。例如，我们当然会热衷于举办3Rs学生会议和讲座（向全英国所有与3Rs相关的学生和研究员开放）来推广这一点，并认为剑桥将对潜在的演讲者产生“巨大的吸引力”。

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.