Core B: Organoid Core

项目总结（核心B：类器官核心） 传染病研究因缺乏研究所需的实验模型而受到严重阻碍 病原体和靶组织之间的相互作用;最近的COVID-19放大了一个明显的缺陷 流行病最近出现的3D类器官方法体现了一种更具生理性的体外感染性疾病 平台在过去的5年里，斯坦福大学NAMSED U19中心运营了一个类器官核心，由 卡尔文郭，其中分布了大约74个类器官线，用于模拟胃肠道感染性 病原体，导致许多合作出版物。类器官核心也产生了新的空气 液体界面（ALI）类器官，其整体维持上皮和浸润免疫群体， 重建（Cell，2018），并为长期，无饲养层，化学定义的人肺创建了新的方案 远端类器官培养，以及新的人类鼻窦类器官（细胞干细胞，2020）。 核心B（类器官核心）在我们的斯坦福大学NAMSED U19更新申请中的总体目标是 一个斯坦福大学/斯坦福大学仿生U19研究中心是扩大我们以前的成功，并提供新的 用于研究胃肠道和呼吸道病原体的离体模型。因此，核心B将作为中心 通过生成和分发下一代人类胃肠道和呼吸道的生物中心， 类器官培养支持项目1-3。除了这些核心职责外，Kuo实验室还将优化 创新的基于类器官的检测方法，将为项目1-3提供额外的支持，并为 全球研究界。在目标1中，从胃、小肠、远端和结肠中提取人“仅上皮”类器官。 肺和鼻气道将为项目1（H。pylori、S.伤寒沙门氏菌S. 项目2（轮状病毒）和项目3（SARS-CoV-2、MERS、流行前冠状病毒和1918年 H1N1流感病毒）。特别缺乏将内源性免疫结合到细胞内的类器官方法。 人口不重组。因此，Aim 2开发了创新的ALI类器官方法， 和内源性浸润性免疫细胞，用于定义上皮和免疫细胞之间的串扰。 胃肠道和肺部感染期间的隔室。最后，Aim 3优化了多重类器官平台， 病原体感染和药物筛选，以提供用于评价感染、治疗和 免疫反应。 总体而言，斯坦福大学/斯坦福大学仿生U19研究中心的核心B利用了领先的专业知识 用于肠道和呼吸道病原体研究的原代3D人类类器官培养。另外我们 利用保留上皮和免疫组分的新型类器官方法， 创新的整体工具，以解剖致病性感染过程中涉及的途径， 翻译的含义。

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.