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 总结 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 具有大流行潜力的病毒。目标是使用活病毒检测从命中中识别出先导化合物， 在迭代过程中，通过潜在客户优化来引导他们，以获得每个项目的优化潜在客户。的 体外病毒学核心由Melanie Ott（Gladstone，UCSF）领导，并由共同的Adolfo Garcia-Sastre支持， Ana Sesma（西奈山伊坎医学院），Greg Towers，克莱尔乔利（伦敦大学学院）， Luis Martinez-Sobrido（UT圣安东尼奥）、Marco Vignuzzi、Carla Saleh（巴斯德研究所）和洛雷纳Zuliani- 阿尔瓦雷斯（加州大学旧金山分校）。我们将提供活病毒、反向遗传学和细胞系亚基因组病毒学检测， 20种RNA病毒的先进原代细胞模型，包括各种冠状病毒、小核糖核酸病毒、披膜病毒， 黄病毒、副粘病毒和布尼亚病毒目。我们还将提供严格的组织监督， 明确和安全的工作流程和数据流，集中解释结果，以及对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举行一次会议，并与所有项目紧密联系， 丹目标1将测试针对冠状病毒（SARS-CoV、MERS-CoV、SARS-CoV-2、hCoV-OC 43、 NL 63，229 E和HKU 1），最初使用纳米荧光素酶SARS-CoV-2报告病毒检测 随后对各种SARS-CoV-2分离株进行平行、多位点IC 50、IC 90和CC 50测定。化合物 将被选择用于进一步评价其：（a）泛冠状病毒抑制，（B）在肺类器官中的功效， 在空气-液体界面生长的原代肺上皮细胞，（c）作用机制（使用Proteomics Core）， 和（d）包括聚合酶抑制剂瑞姆德西韦和莫努匹韦的联合研究。产生的线索将是 在细胞系中连续传代，以确定将克隆到SARS-CoV-2中的耐药突变 分子克隆并进行先导优化研究。我们还将提供试剂和预成型的病毒样 粒子到项目3和6进行命中识别。目标2将测试针对其他RNA病毒的抑制剂， 大流行潜力，包括肠道病毒EV-A71和EV-D 68，基孔肯雅病毒，科萨基病毒， 脊髓灰质炎病毒、鼻病毒、寨卡病毒和登革热病毒以及副粘病毒，在使用病毒分离株的活病毒测定中， 工程分子克隆。在一个类似于目标1的分层方法中，目标表征、命中到引导和 将在适当的细胞系以及肺和肠类器官中进行电极导线优化步骤， 扁桃体组织培养。将如上所述进行联合用药和耐药性研究，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中的先导化合物的抗病毒活性 目标2内的抗病毒活性，反之亦然。我们期待着成为推动铅化合物发展的关键 来自体内病毒学核心的小动物模型试验以及迭代电极导线的两个目标 优化与药物化学，结构生物学和综合建模核心。