Development of an in vitro model of the ruminant intestine for studies of host-parasite-microbe interactions

BBSRC战略主题：生物科学促进可持续农业和食品胃肠道蠕虫是家畜最重要的感染源之一；对新的、可持续的寄生虫控制策略的需求很高。已有证据表明肠道微生物区系在宿主-寄生虫串扰中所起的作用，这使得开发以微生物群为靶标的抗寄生虫成为一种合理的选择。然而，需要更好地了解支撑这种相互作用的机制。通常，对这种相互作用的分析依赖于从活体动物身上收集的肠道样本。然而，这种方法(1)往往涉及数十到数百只动物，长期暴露在寄生虫下；(2)由于影响肠道微生物区系组成的各种因素，数据高度可变；(3)影响大小的计算以及反过来的统计能力是具有挑战性的。为了解决这些问题，我们的实验室开发了脊椎动物肠道免疫轴的3D体外模型原型，作为实地研究和实验感染GI蠕虫的绵羊模型的替代方案。这项建议旨在将该系统优化为一种适应性设备，以适应对宿主-寄生虫-微生物区系关系的纵向、机械性研究，从而减少对蠕虫感染的体内模型和不一致的粪便采样的依赖。该项目的具体目标将是：1)优化设备的设计，以允许肠道微生物组的厌氧共培养；2)调整设备以包括来自关键养殖动物的肠道上皮和免疫细胞；3)将复杂的动物微生物组整合到设备中；4)将优化的3D模型用于宿主-蠕虫-微生物组相互作用的概念验证研究。

BBSRC strategic theme: Bioscience for sustainable agriculture and foodGastrointestinal (GI) helminths are amongst the most important infectious agents of livestock; demand for novel, sustainable strategies for parasite control is high. Evidence has emerged of the role of the gut microbiota in host-parasite crosstalk, which makes the development of microbiome-targeting anti-parasitics a plausible option. An improved understanding of the mechanisms underpinning this interplay is however needed. Typically, the analysis of such interactions relies on gut samples collected from live animals. However, this approach (i) often involves tens to hundreds of animals continually exposed to parasites for prolonged periods; (ii) data is highly variable because of a myriad of factors that shape the composition of the gut microbiota; (iii) calculations of effect size and, in turn, statistical power, are challenging. To address these issues, our laboratories have developed a prototype 3D in vitro model of the vertebrate gut-immune axis, as an alternative to both field studies and experimental infections of sheep models with GI worms. This proposal aims to optimise this system as an adaptable device to accommodate longitudinal, mechanistic studies of host-parasite-microbiota relationships, therefore reducing the reliance on in vivo models of worm infections and inconsistent faecal sampling.The specific aims of this project will be to:1) Optimise the design of the device to allow anaerobic co-cultures of gut microbiome;2) Adapt the device to include gut epithelial and immune cells from key farmed animals;3) Incorporate a complex animal microbiome into the device;4) Employ the optimised 3D model in proof-of-concept studies of host-helminth-microbiome interactions.