In vitro virology core

体外病毒学核心

• 批准号: 10512624
• 负责人: Melanie Maria Ott
• 金额: $ 743.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10512624
• 关键词: 2019-nCoV; 3-Dimensional; A549; ACE2; Air; American; Animal Model; Animals; Antiviral Agents; Biological Assay; Bunyavirales; COVID-19 pandemic; Cell Line; Cell model; Cells; Chikungunya virus; Common Cold; Complex; Coronavirus; Coronavirus Infections; Coxsackie Viruses; Data; Dengue; Dengue Virus; Disease Outbreaks; Dose; Drug Combinations; Drug resistance; Engineering; Enterovirus; Epithelial Cells; Evaluation; Failure; Family Picornaviridae; Flavivirus; Goals; Hela Cells; Human; Human Cell Line; Human poliovirus; In Vitro; Infection; Innate Immune Response; Insecta; Intestines; Lead; Liquid substance; London; Lung; Middle East Respiratory Syndrome Coronavirus; Modeling; Molecular Cloning; Monitor; Mutation; Organoids; Paramyxovirus; Pharmaceutical Chemistry; Polymerase; Process; Proteins; Proteomics; RNA; RNA Viruses; Reagent; Recording of previous events; Reporter; Resistance; Resistance development; Rhinovirus; SARS coronavirus; Serial Passage; Serotyping; Services; Site; Testing; Togaviridae; Tonsil; Toxic effect; Triage; Universities; Variant; Vero Cells; Viral; Viral Proteins; Virus; Virus Inhibitors; Virus-like particle; Work; Zika Virus; anti-viral efficacy; antiviral drug development; bronchial epithelium; college; experience; health economics; human coronavirus; in vivo; in vivo evaluation; inhibitor; lead optimization; medical schools; member; molnupiravir; mosquito-borne; nanoluciferase; pandemic disease; programs; remdesivir; resistance mutation; response; reverse genetics; screening; social; structural biology; therapeutically effective; variants of concern; virology

CORE 6: IN VITRO VIROLOGY SUMMARY QCRG Pandemic Response Program In Vitro Virology Core leverages in-depth virology expertise from 10 groups to support Projects 1–6 in discovering and developing antivirals against coronaviruses and other RNA viruses with pandemic potential. The goal is to identify lead compounds from hits using live-virus assays and channel them, in an iterative process, through lead optimization to obtain Optimized Leads for each project. The In Vitro Virology Core is led by Melanie Ott (Gladstone, UCSF) and supported by co-Is Adolfo Garcia-Sastre, Ana Sesma (Icahn School of Medicine at Mt Sinai), Greg Towers, Clare Jolly (University College London), Luis Martinez-Sobrido (UT San Antonio), Marco Vignuzzi, Carla Saleh (Institut Pasteur), and Lorena Zuliani- Alvarez (UCSF). We will provide live virus, reverse genetics and subgenomic virological assays in cell lines and advanced primary cell models for 20 RNA viruses, including various coronaviruses, picornaviruses, togaviruses, flaviviruses, paramyxoviruses and bunyavirales. We will also provide tight organizational oversight with precisely defined and safe work- and data flows, central interpretation of results, and guidance on pan-antiviral potential of lead compounds. In Vitro Virology Core members will meet monthly and interface tightly with all projects and cores. Aim 1 will test inhibitors against coronaviruses (SARS-CoV, MERS-CoV, SARS-CoV-2, hCoV-OC43, NL63, 229E and HKU1) in a tiered approach, initially using nanoluciferase SARS-CoV-2 reporter virus assays and later parallel, multi-site IC50, IC90 and CC50 determinations with various SARS-CoV-2 isolates. Compounds will be selected for further evaluation of their: (a) pan-coronavirus inhibition, (b) efficacy in lung organoids and primary lung epithelial cells grown at the air-liquid interface, (c) mechanism of action (with the Proteomics Core), and (d) combination studies including polymerase inhibitors remdesivir and molnupiravir. Resulting leads will be serially passaged in cell lines to identify drug-resistance mutations that will be cloned into SARS-CoV-2 molecular clones and studied for lead optimization. We will also provide reagents and pre-formed virus-like particles to Projects 3 and 6 for hit identification. Aim 2 will test inhibitors against other RNA viruses with pandemic potential, including enteroviruses EV-A71 and EV-D68, chikungunya virus, coxsackie viruses, poliovirus, rhinovirus, zika and dengue viruses and paramyxovirus, in live virus assays using viral isolates and engineered molecular clones. In a tiered approach similar to Aim 1, Target Characterization, Hit-to-Lead, and Lead Optimization steps will be performed in appropriate cell lines, as well as in lung and gut organoids and tonsil histocultures. Combination and resistance studies will be performed as described above and will include the pan-serotype dengue inhibitor JNJ-A07. If appropriate, we will test lead compounds from Aim 1 for antiviral activity against viruses within Aim 2 and vice versa. We anticipate to be critical to advance Lead Compounds from both aims for testing in small animal models in the In Vivo Virology Core as well as for iterative Lead Optimization with the Medicinal Chemistry, Structural Biology and Integrative Modeling Cores.

核心6：体外病毒学 概括 QCRG大流行响应计划中的体外病毒学核心核心杠杆作用，深入的病毒学专业知识。 支持项目1-6的小组在发现和开发针对冠状病毒和其他RNA的抗病毒药 具有大流行潜力的病毒。目的是使用实时病毒测定法识别命中率的铅化合物， 在迭代过程中通过铅优化引导它们，以获取每个项目的优化潜在客户。 体外病毒学核心由Melanie Ott（UCSF Gladstone）领导，并由Co-IS Adolfo Garcia-Sastre支持 Ana Sesma（Sinai Mt Sinai伊坎医学院），Greg Towers，Clare Jolly（伦敦大学学院）， 路易斯·马丁内斯·索伯里多（UT San Antonio），Marco Vignuzzi，Carla Saleh（Carla Saleh（Castitut Pasteur））和Lorena Zuliani- Alvarez（UCSF）。我们将在细胞系和 20种RNA病毒的高级主细胞模型，包括各种冠状病毒，picornaviruses，togaviruses， 黄病毒，paramyxoviruses和bunyavirales。我们还将确切地提供紧张的组织监督 定义且安全的工作和数据流，结果的中心解释以及泛抗病毒潜力的指导 铅化合物。体外病毒学核心成员将每月与所有项目紧密互动， 内核。 AIM 1将测试针对冠状病毒的抑制剂（SARS-COV，MERS-COV，SARS-COV-2，HCOV-OC43， NL63，229E和HKU1）以分层的方法，最初使用纳米酸酯酶SARS-COV-2报告基因病毒测定法 后来平行于与各种SARS-COV-2分离株的多站点IC50，IC90和CC50测定。化合物 将选择以进一步评估其：（a）泛氧化病毒抑制作用，（b）肺器官的效率和 在空气界面生长的原发性肺上皮细胞（C）作用机理（具有蛋白质组学核心）， （d）包括聚合酶抑制剂Remdesivir和molnupiravir在内的组合研究。由此产生的线索将是 在细胞系中连续通过以识别将克隆到SARS-COV-2的药物抗性突变 分子克隆和研究铅优化的研究。我们还将提供试剂和预形成病毒样 项目3和6的粒子进行命中识别。 AIM 2将测试与其他RNA病毒的抑制剂 大流行潜力，包括肠病毒EV-A71和EV-D68，Chikungunya病毒，Coxsackie病毒， 使用病毒分离株和 工程分子克隆。在类似于目标1的分层方法中 铅优化步骤将在适当的细胞系，肺和肠道类器官中进行 扁桃体组织文化。将如上所述进行组合和抵抗研究，并将包括 Pan-Serotype Dengu抑制剂JNJ-A07。如果适当的话，我们将测试AIM 1抗病毒的铅化合物 在AIM 2中针对病毒的活动，反之亦然。我们预计对推进铅化合物至关重要 从两种目标中都在体内病毒学核心中的小动物模型中进行测试以及迭代铅 使用药物化学，结构生物学和综合建模核心进行优化。