Stanford/UNC Biomimetic U19 Research Center

斯坦福大学/斯坦福大学仿生U19研究中心 项目总结/摘要-中心概述 传染病继续普遍影响全球健康和社会经济稳定，尽管 预防和治疗措施。呼吸道和胃肠道病原体是最常见的 难治性传染病，特别是人畜共患冠状病毒的反复出现和最近的 由SARS-CoV-2引起的COVID-19大流行。总的来说，迫切需要改进的体外 人类疾病的实验模型，以研究发病机制和验证治疗方法。中心使命 因此，斯坦福大学/斯坦福大学仿生U19研究中心的研究人员正在部署新的三维类器官培养 用于阐明呼吸道和胃肠道感染病原体的生物学和治疗的模型。 我们的申请是我们之前的斯坦福大学NAMSED U19研究中心的更新，包括 两个核心和三个研究项目，利用我们研究人员的互补和协同专业知识 在斯坦福大学和北卡罗来纳州大学。该中心继续由多个PI领导， 卡尔文郭和曼努埃尔阿米耶娃，谁也共同领导核心A（行政核心）。引导核心B（类器官核心） 由卡尔文郭，并提供了新的能力，肺和胃肠道类器官培养，基因编辑和多路复用 筛选这三个项目广泛利用类器官仿生学来探索GI和呼吸系统 病原体项目1（PI，Manuel Amieva）研究H.幽门螺杆菌和沙门氏菌定植，竞争和 胃肠道中的入侵，而项目2（PI，Harry Greenberg）研究轮状病毒宿主范围，中和， 和M细胞的相互作用。项目3（PI，Ralph Baric）是一个新的补充，广泛使用 类器官模型SARS-CoV-2，其他密切相关的流行和流行前的新冠病毒， 1918年H1N1流感揭示了与严重肺部结局相关的常见和独特的宿主网络。 斯坦福大学/斯坦福大学仿生U19研究中心的活动主要集中在三个方面： 目标。在目标1中，我们的中心进行上皮-病原体界面的类器官建模，以研究 致病性、易感性和宿主范围限制。这采用了强大的反向遗传学和CRISPR 筛选系统地操纵宿主与病毒/细菌隔室，在新的顶部-底部极性内 调节远端肺/肺泡、鼻窦、胃和肠类器官系统。目标2定义了SARS- CoV-2、流行前冠状病毒、轮状病毒和沙门氏菌可干扰组织之间的相互串扰 上皮和常驻免疫细胞。这利用了一种独特的3D空气-液体界面类器官方法， 胃肠道和肺上皮整体，具有多种内源性浸润免疫细胞类型，无人工 重组最后，Aim 3对SARS-CoV的治疗候选物进行了基于类器官的评估。 2、中高通量形式的流行前冠状病毒和轮状病毒，包括上皮和/或 免疫细胞总体而言，斯坦福大学/斯坦福大学仿生U19研究中心的探索直接适用于 先进的类器官系统的调查和治疗的寄生虫感染性病原体。

Stanford/UNC Biomimetic U19 Research Center PROJECT SUMMARY/ABSTRACT – CENTER OVERVIEW Infectious diseases continue to pervasively afflict global health and socioeconomic stability despite substantial prevention and treatment initiatives. Respiratory and gastrointestinal pathogens rank amongst the most intractable infectious diseases, particularly notable for recurrent waves of zoonotic coronaviruses and the recent COVID-19 pandemic, engendered by SARS-CoV-2. Overall, an urgent need exists for improved in vitro experimental models of human disease to study pathogenesis and to validate therapeutics. The central mission of the Stanford/UNC Biomimetic U19 Research Center is thus to deploy novel 3-dimensional organoid culture models to elucidate the biology and therapy of respiratory and gastrointestinal infectious pathogens. Our application is a renewal of our prior Stanford NAMSED U19 Research Center and is comprised of two Cores and three research Projects, leveraging complementary and synergistic expertise of our investigators at Stanford University and the University of North Carolina. The Center continues to be led by the Multi-PIs, Calvin Kuo and Manuel Amieva, who also co-lead Core A (Administrative Core). Core B (Organoid Core) is led by Calvin Kuo and provides novel capabilities for lung and GI organoid culture, gene editing and multiplexed screening. The three Projects extensively utilize organoid biomimetics for exploration of GI and respiratory pathogens. Project 1, (PI, Manuel Amieva) investigates H. pylori and Salmonella colonization, competition and invasion in the GI tract, while Project 2 (PI, Harry Greenberg) investigates rotavirus host range, neutralization, and M cell interactions in enteric biomimetics. Project 3 (PI, Ralph Baric) is a new addition and extensively uses organoids to model SARS-CoV-2, other closely related epidemic and pre-epidemic emerging coronaviruses and 1918 H1N1 influenza to reveal common and unique host networks associated with severe pulmonary outcomes. The activities of the Stanford/UNC Biomimetic U19 Research Center reside within three overarching Aims. In Aim 1, our Center performs organoid modeling of the epithelium-pathogen interface to investigate pathogenesis, susceptibility and host range restriction. This employs robust reverse genetics and CRISPR screens to systematically manipulate host versus viral/bacterial compartments, within novel apical-basal polarity modulated distal lung/alveolar, nasal sinus, stomach and intestinal organoid systems. Aim 2 defines how SARS- CoV-2, pre-epidemic coronaviruses, rotavirus and Salmonella can perturb reciprocal cross-talk between tissue epithelium and resident immune cells. This exploits a unique 3D air-liquid interface organoid method preserving GI and lung epithelium en bloc with diverse endogenous infiltrating immune cell types without artificial reconstitution. Lastly, Aim 3 performs organoid-based evaluation of therapeutic candidates against SARS-CoV- 2, pre-epidemic coronaviruses and rotavirus in medium- to high-throughput formats including epithelium and/or immune cells. Overall, the explorations of the Stanford/UNC Biomimetic U19 Research Center directly apply advanced organoid systems to the investigation and therapy of recalcitrant infectious pathogens.