Core B: Organoid Core

项目总结(核心B：有机核心) 传染病研究因缺乏必要的实验模型而严重受阻。 病原体和靶组织之间的相互作用；最近的新冠肺炎放大了一个明显的缺陷 大流行。最近出现的3D有机化合物方法体现了一种更具生理性的体外感染性疾病 站台。在过去的5年里，斯坦福NAMSED U19中心运营了一个有机核心，由 Calvin Kuo，已经发布了大约74条有机类线来模拟胃肠道感染性 病原体，从而产生了许多合作出版物。有机核心还产生了新的空气- 液体界面(ALI)有机类物质，维持上皮细胞和渗透的免疫群体在没有 重建(细胞，2018)，并为长期、无饲养器、化学定义的人类肺创建了新的方案 远端器官培养，以及新的人类鼻窦器官培养(细胞干细胞，2020)。 在我们的斯坦福NAMSED U19续订申请中，核心B(有机核心)的总体目标是 斯坦福大学/北卡罗来纳大学仿生U19研究中心将在我们先前成功的基础上扩展并提供新颖的 研究胃肠道和呼吸道病原体的体外模型。因此，核心B将作为一个中心 通过生成和分发下一代人类胃肠道和呼吸系统，为我们的中心建立BioHub 支持项目1-3的有机培养。除了这些核心职责外，郭实验室还将优化 创新的有机化合物化验，将为项目1-3提供额外支持，并为 全球研究界。在目标1中，来自胃、小肠、远端的人“仅限于上皮”的有机体 将为项目1(幽门螺杆菌、伤寒沙门氏菌、沙门氏菌、沙门氏菌)生成、表征和分布肺部和鼻腔呼吸道。 项目2(轮状病毒)和项目3(SARS-CoV-2、中东呼吸综合征、流行前冠状病毒和1918年 H1N1流感病毒)。尤其缺乏结合内源性免疫的有机方法。 没有重组的种群。因此，Aim 2开发了创新的ALI有机类保存上皮的方法 以及内源性浸润性免疫细胞，用于定义上皮细胞和免疫细胞之间的串扰 胃肠道感染和肺部感染时的隔室。最后，Aim 3优化了多路复用的有机平台 病原体感染和药物筛选为评估感染、治疗和治疗提供新的方法 免疫反应。 总体而言，斯坦福大学/北卡罗来纳大学仿生U19研究中心的核心B利用了领先的专业知识 在原代3D人体器官培养中，用于研究肠道和呼吸道病原体。此外，我们 充分利用新的有机体方法，既保留附属物又保留免疫成分，以提供 创新的整体工具，剖析病原体感染过程中涉及的途径，包括基础和 翻译含义。

PROJECT SUMMARY (Core B: Organoid Core) Infectious disease research is substantially hampered by lack of experimental models necessary to study interactions between pathogens and target tissues; a glaring deficiency amplified by the recent COVID-19 pandemic. The recent advent of 3D organoid methods embodies a more physiologic in vitro infectious disease platform. Over the past 5 years, the Stanford NAMSED U19 Center has operated an Organoid Core, led by Calvin Kuo, which has distributed approximately 74 organoid lines for modeling infectivity of gastrointestinal pathogens, resulting in numerous collaborative publications. The Organoid Core has also produced new air- liquid interface (ALI) organoids that maintain epithelium and infiltrating immune populations en bloc without reconstitution (Cell, 2018) and created new protocols for long-term, feeder-free, chemically defined human lung distal organoid culture, as well as novel human nasal sinus organoids (Cell Stem Cell, 2020). The overall goal for Core B (Organoid Core) in our renewal application of the Stanford NAMSED U19 for a Stanford/UNC Biomimetic U19 Research Center is to expand upon our previous success and provide novel ex vivo models for studying gastrointestinal and respiratory pathogens. Thus, Core B will serve as a central biohub for our Center by generating and distributing next-generation human gastrointestinal and respiratory organoid cultures to support Projects 1-3. In addition to these core responsibilities, the Kuo lab will optimize innovative organoid-based assays that will provide additional support for Projects 1-3 and resources for the global research community. In Aim 1, human “epithelial-only” organoids from the stomach, small intestine, distal lung and nasal airways will be generated, characterized, and distributed for Project 1 (H. pylori, S. Typhi, S. Typhimurium), Project 2 (rotavirus), and Project 3 (SARS-CoV-2, MERS, pre-epidemic coronaviruses and 1918 influenza H1N1 virus). There has been a particular lack of organoid methods incorporating endogenous immune populations without reconstitution. Thus, Aim 2 develops innovative ALI organoid methods preserving epithelial and endogenous infiltrating immune cells en bloc for defining cross-talk between epithelial and immune compartments during GI and pulmonary infections. Lastly, Aim 3 optimizes multiplexed organoid platforms for pathogen infection and drug screening to provide novel methods for evaluating infection, therapeutics and immune responses. Overall, Core B of the Stanford/UNC Biomimetic U19 Research Center leverages leading-edge expertise in primary 3D human organoid culture for the study of enteric and respiratory pathogens. Additionally, we capitalize on novel organoid methods preserving both the epitheilal and immune components to provide innovative holistic tools to dissect pathways involved during pathogenic infection, with both basic and translational implications.