Stanford/UNC Biomimetic U19 Research Center

斯坦福大学/北卡罗来纳大学仿生 U19 研究中心

• 批准号: 10614385
• 负责人: MANUEL R AMIEVA
• 金额: $ 150.1万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614385
• 关键词: 2019-nCoV; 3-Dimensional; Acute; Acute Respiratory Distress Syndrome; Air; Alveolar; Alveolus; Apical; Bacteria; Bacterial Infections; Basic Science; Binding; Biology; Biomimetics; COVID-19; COVID-19 pandemic; COVID-19 test; CRISPR interference; CRISPR screen; CRISPR-mediated transcriptional activation; Cell Communication; Cells; Chiroptera; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Communicable Diseases; Coronavirus; Digestion; Disease; Distal; Enteral; Epidemic; Epithelium; Epitopes; Escherichia coli; Etiology; Exhibits; Experimental Models; Functional disorder; Gastrointestinal Diseases; Gastrointestinal tract structure; Genes; Genetic Screening; Glycoproteins; Goals; Growth; Helicobacter pylori; Human; Immune; Immune response; Immunity; In Vitro; Incubators; Infection; Influenza; Innate Immune Response; Intestines; Invaded; Investigation; Lead; Liquid substance; Lung; Lung infections; M cell; Mediating; Methods; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Mission; Modeling; Monoclonal Antibodies; Morbidity - disease rate; Myelogenous; National Institute of Allergy and Infectious Disease; Norovirus; North Carolina; Organoids; Outcome; Pathogenesis; Pathogenicity; Play; Pneumonia; Population; Predisposition; Prevention; Recurrence; Research; Research Personnel; Research Project Grants; Resistance; Respiratory Failure; Respiratory Mucosa; Respiratory Therapy; Respiratory Tract Infections; Role; Rotavirus; Route; SARS coronavirus; SARS-CoV-2 inhibitor; Salmonella; Salmonella typhi; Salmonella typhimurium; Severe Acute Respiratory Syndrome; Sinus; Spanish flu; Stomach; Surface; System; Terminal Bronchiole; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Universities; Viral; Virulence; Virus; Virus Diseases; Zoonoses; absorption; adaptive immune response; airway epithelium; cell type; comparative; cytokine release syndrome; emerging pathogen; enteric infection; gastric organoids; gastrointestinal; gastrointestinal epithelium; genetic manipulation; global health; health economics; human coronavirus; human disease; human model; immune cell infiltrate; improved; mortality; multidisciplinary; multiplexed imaging; mutant; neutralizing monoclonal antibodies; new epidemic; novel; novel coronavirus; pathogen; pathogenic virus; permissiveness; preservation; reconstitution; respiratory; respiratory pathogen; reverse genetics; screening; single-cell RNA sequencing; small molecule inhibitor; socioeconomics; synergism; therapeutic candidate; therapeutic development; therapeutic evaluation; three dimensional cell culture; translational goal; vaccine development; virus tropism; zoonotic coronavirus

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.

斯坦福/北卡罗来纳大学仿生U19研究中心 项目摘要/摘要-中心概述 传染病继续普遍困扰着全球卫生和社会经济稳定，尽管 预防和治疗举措。呼吸道和胃肠道病原体的数量最多。 顽固性传染病，特别是反复出现的人畜共患冠状病毒浪潮和最近 新冠肺炎大流行，由SARS-CoV-2引发。总体而言，迫切需要改进体外培养。 人类疾病的实验模型，用于研究发病机制和验证治疗方法。中心使命 因此，斯坦福大学/北卡罗来纳大学仿生U19研究中心的研究人员将部署新的三维有机培养 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NAMSED U19研究中心的续订，包括 两个核心和三个研究项目，利用我们调查人员的互补和协同专业知识 在斯坦福大学和北卡罗来纳大学。该中心继续由多绩效指标领导， 卡尔文·郭和曼努埃尔·阿米耶娃，他们也是核心A(行政核心)的联合负责人。核心B(有机核心)是领先的 由Calvin Kuo开发，并为肺和胃肠道器官培养、基因编辑和多路传输提供了新功能 放映。这三个项目广泛利用有机仿生技术探索胃肠道和呼吸系统。 病原体。项目1，(Pi，Manuel Amieva)调查幽门螺杆菌和沙门氏菌的定植、竞争和 在项目2(Pi，Harry Greenberg)调查轮状病毒宿主范围、中和、 肠道仿生学中的M细胞相互作用。项目3(Pi，Ralph Baric)是新增加的，用途广泛 用于模拟SARS-CoV-2、其他密切相关的流行和流行前新出现的冠状病毒的有机化合物以及 1918年H1N1流感揭示与严重肺部后果相关的共同和独特的宿主网络。 斯坦福大学/北卡罗来纳大学仿生U19研究中心的活动位于三个总体范围内 目标。在目标1中，我们的中心对上皮-病原体界面进行器官模拟以研究 发病机制、敏感性和寄主范围限制。这采用了强大的反向遗传学和CRISPR 在新的顶端-基底极性内，系统地操纵宿主与病毒/细菌隔间的筛网 调节远端肺/肺泡、鼻窦、胃和肠道器官系统。目标2定义了SARS是如何-- CoV-2、流行前冠状病毒、轮状病毒和沙门氏菌可以扰乱组织之间的相互串扰 上皮细胞和常驻免疫细胞。这利用了一种独特的3D气液界面有机物方法来保存 GI和肺上皮不同内源性浸润性免疫细胞类型的块 重建。最后，AIM 3对针对SARS-CoV的治疗候选进行基于有机物的评估- 2、流行前冠状病毒和轮状病毒，中高通量形式，包括上皮和/或 免疫细胞。总体而言，斯坦福/北卡罗来纳大学仿生U19研究中心的探索直接适用于 先进的有机系统，用于顽固性感染病原体的调查和治疗。