Epithelial interactions with indigenous and pathogenic microbes

项目摘要/摘要 肠上皮由具有吸收和分泌功能的单层上皮层组成。 负责营养吸收。此外，它还与丰富多样的原住民保持着持续的联系 生活在肠腔内的微生物区系。上皮和微生物群以一种非常稳定的、平衡的方式存在。 更高级的共生。这种共生关系负责肠道生态系统的动态平衡，并扰乱了 这种关系与多种疾病状态有关。虽然动物模型和使用 无菌小鼠和诺生菌小鼠在理解宿主-微生物相互作用方面取得了显著进展。 然而，物种特有的差异使许多人类肠道病原体的研究变得困难，并适用于 目前还严重缺乏能够准确代表人体生理的体外细胞培养模型。我们最近取消了- 描述了一种准确反映复杂细胞组成和拓扑结构的体外系统 人类肠道的。使用人类多能干细胞(HPSCs)，我们已经产生了三维(3D)界面-- 称为人类肠道器官(HIO)的组织单位。HIO含有肠上皮和间充质。 由在小肠中发现的所有不同类型的细胞组成。HIO已经被证明是一种 研究人类肠道发育的强大系统。我们的初步结果表明，HIO也是一种 这是研究肠道病原体如何影响宿主上皮的极佳系统。因此，最重要的目标是 这个项目是为了确定特定的肠道病原体和正常的微生物区系如何影响宿主的HIO上皮 在细胞和分子水平上，并确定如何建立复杂的宿主-微生物区系-病原体关系- 在HIO内放贷和维护。我们建议1)研究共生微生物之间的相互作用 和HIO上皮影响这种共生中的每个伙伴，2)确定接触病原体如何影响 HIO上皮的生理学和3)确定与共生微生物接触是否会改变病理- 基因/上皮细胞相互作用。为了实现这些具体目标，我们已经建立了一个多学科团队，将 检查HIO上皮、本地微生物区系与病毒和细菌之间的复杂关系 肠道病原体。

PROJECT SUMMARY/ABSTRACT The intestinal epithelium is comprised of a single epithelial layer with both absorptive and secretory functions and is responsible for nutrient absorption. Additionally it is in constant contact with the abundant and diverse indigenous microbiota that inhabits the intestinal lumen. The epithelium and the microbiota exists in a remarkably stable, bal- anced symbiosis. This symbiosis is responsible for homeostasis of the intestinal ecosystem, and disturbances in this relationship has been associated with a wide variety of disease states. While animal models and the use of germ-free and gnotobiotic mice have led to remarkable advances in our understanding of host-microbial interac- tions, species-specific differences have made it difficult to study many human enteric pathogens, and suitable in vitro cell culture models that accurately represent human physiology are severely lacking. We have recently de- scribed an in vitro system that accurately reflects both the complex cellular makeup and the topological organization of the human intestine. Using human pluripotent stem cells (hPSCs), we have generated 3-dimensional (3D) intes- tinal units called Human Intestinal Organoids (HIOs). HIOs contain both intestinal epithelium and mesenchyme and are comprised of all of the different cell types found in the small intestine. HIOs have already proven to be a powerful system to study human intestinal development. Our preliminary results demonstrate that HIOs are also an excellent system to study how enteric pathogens affect the host epithelium. Therefore, the overarching goal of this project is to determine how specific enteric pathogens and normal microbiota affect the host HIO epithelium at the cellular and molecular level, and to determine how complex host-microbiota-pathogen relationships are estab- lished and maintained within the HIO. We propose to 1) investigate how interaction between commensal microbes and the HIO epithelium affects each partner in this symbiosis, 2) determine how contact pathogens affects the physiology of the HIO epithelium and 3) determine if contact with commensal microbes will alter the patho- gen/epithelium interaction. To accomplish these specific aims, we have established a multidisciplinary team that will examine the complex relationship between the HIO epithelium, the indigenous microbiota and viral and bacterial enteric pathogens.