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

BBSRC战略主题：促进可持续农业和食品的生物科学胃肠道蠕虫是家畜最重要的传染性病原体之一；对新的、可持续的寄生虫控制策略的需求很高。有证据表明肠道微生物群在宿主-寄生虫串扰中的作用，这使得开发以微生物群为目标的抗寄生虫药成为一种可行的选择。然而，需要更好地理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)通常涉及数十至数百只动物长期暴露于寄生虫；（ii）由于塑造肠道菌群组成的无数因素，数据变化很大；（iii）效应大小的计算以及统计效力的计算具有挑战性。为了解决这些问题，我们的实验室开发了一个脊椎动物肠道免疫轴的原型3D体外模型，作为实地研究和胃肠道蠕虫实验感染绵羊模型的替代方案。本提案旨在优化该系统，使其成为一种适应性强的设备，以适应宿主-寄生虫-微生物群关系的纵向、机制研究，从而减少对体内蠕虫感染模型的依赖和不一致的粪便采样。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)对该装置进行改造，纳入关键养殖动物的肠道上皮细胞和免疫细胞；3)将复杂的动物微生物组纳入装置；4)将优化后的3D模型用于宿主-蠕虫-微生物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.