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 摘要 QCRG大流行应对计划体外病毒学核心利用来自10个国家的深入病毒学专业知识 支持项目1-6发现和开发针对冠状病毒和其他RNA的抗病毒药物的小组 具有大流行潜力的病毒。其目标是使用活病毒检测从HITS中识别先导化合物和 在迭代过程中引导他们，通过线索优化来获得每个项目的最佳线索。这个 体外病毒学核心由Melanie Ott(Gladstone，加州大学旧金山分校)领导，并由共同IS Adolfo Garcia-Sastre支持， Ana Sesma(西奈山伊坎医学院)，Greg Towers，Clare Jolly(伦敦大学学院)， 路易斯·马丁内斯-索布里多(德克萨斯大学圣安东尼奥分校)、马尔科·维格努齐、卡拉·萨利赫(巴斯德研究所)和洛雷娜·祖利安尼- 阿尔瓦雷斯(加州大学旧金山分校)。我们将提供活病毒、反向遗传学和亚基因组病毒学检测 20种RNA病毒的高级原代细胞模型，包括各种冠状病毒、小核糖核酸病毒、TOGA病毒、 黄病毒、副粘病毒和布尼亚病毒。我们还将提供严格的组织监督，准确地 明确和安全的工作和数据流、对结果的集中解释以及泛抗病毒潜力的指导 先导化合物。在体外，病毒学核心成员将每月开会，并与所有项目和 核心。AIM 1将测试针对冠状病毒(SARS-CoV，MERS-CoV，SARS-CoV-2，hCoV-OC43， NL63、229E和HKU1)，最初使用纳米纤维素酶SARS-CoV-2报告病毒检测 随后对不同的SARS-CoV-2分离株进行平行、多点IC50、IC90和CC50测定。化合物 将被选择用于进一步评估它们：(A)抑制泛冠状病毒，(B)在肺器官中的疗效和 原代肺上皮细胞在气液界面生长，(C)作用机制(以蛋白质组学为核心)， 和(D)包括聚合酶抑制剂瑞拜韦和莫那普拉韦在内的联合研究。最终的销售线索将是 在细胞系中连续传代以识别将被克隆到SARS-CoV-2中的耐药突变 分子克隆，并进行了先导优化研究。我们还将提供试剂和预制病毒样 粒子到项目3和6进行命中识别。AIM 2将测试针对其他RNA病毒的抑制剂 大流行的可能性，包括肠道病毒EV-A71和EV-D68，基孔肯雅病毒，柯萨奇病毒， 脊髓灰质炎病毒、鼻病毒、寨卡病毒、登革热病毒和副粘病毒，在使用病毒分离株和 基因工程的分子克隆。在类似于目标1的分层方法中，目标特征、点击到领先，以及 铅优化步骤将在适当的细胞系以及肺和肠道器官中执行，并 扁桃体组织培养。将如上所述进行组合和抗性研究，并将包括 泛血清型登革热抑制剂JNJ-A07。如果合适，我们将测试目标1的先导化合物是否具有抗病毒作用。 在AIM 2内对抗病毒的活动，反之亦然。我们预计对推动先导化合物的发展至关重要 这两个目标都是为了在体内病毒学核心的小动物模型中测试以及迭代领导 以药物化学、结构生物学和综合建模为核心的优化。