In vitro virology core

体外病毒学核心

基本信息

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
项目类别：
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

项目摘要

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 大流行应对计划体外病毒学核心利用了来自 10 个国家的深入病毒学专业知识小组支持项目 1-6 发现和开发针对冠状病毒和其他 RNA 的抗病毒药物目标是使用活病毒检测来识别具有大流行潜力的病毒。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。体外病毒学核心由 Melanie Ott（Gladstone，UCSF）领导，并得到 Adolfo Garcia-Sastre 的支持， Ana Sesma（西奈山伊坎医学院）、Greg Towers、Clare Jolly（伦敦大学学院）、 Luis Martinez-Sobrido（德克萨斯大学圣安东尼奥分校）、Marco Vignuzzi、Carla Saleh（巴斯德研究所）和 Lorena Zuliani- Alvarez (UCSF) 我们将提供细胞系和亚基因组病毒学检测的活病毒、反向遗传学和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、我们还将提供严格的组织监督和精确的黄病毒、副粘病毒和布尼亚病毒。明确且安全的工作和数据流、结果的集中解释以及泛抗病毒潜力的指导体外病毒学核心成员将每月举行一次会议，并与所有项目和人员紧密联系。目标 1 将测试冠状病毒（SARS-CoV、MERS-CoV、SARS-CoV-2、hCoV-OC43、 NL63、229E 和 HKU1）采用分层方法，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 化合物进行平行多位点 IC50、IC90 和 CC50 测定。将被选择进行进一步评估：（a）泛冠状病毒抑制，（b）肺类器官的功效和在空气-液体界面生长的原代肺上皮细胞，(c) 作用机制（使用蛋白质组学核心）， (d) 包括聚合酶抑制剂瑞德西韦和莫努匹拉韦的联合研究将产生先导化合物。在细胞系中连续传代，以确定将克隆到 SARS-CoV-2 中的耐药突变我们还将提供试剂和预制病毒样。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-D68、基孔肯雅病毒、柯萨奇病毒、脊髓灰质炎病毒、鼻病毒、寨卡病毒和登革热病毒以及副粘病毒，使用病毒分离物和病毒进行活病毒检测采用类似于目标 1、目标表征、命中先导等的分层方法。先导化合物优化步骤将在适当的细胞系以及肺和肠道类器官中进行将如上所述进行组合和耐药性研究，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 Aim 1 中的先导化合物的抗病毒作用。我们预计，Aim 2 内的抗病毒活性对于推进先导化合物至关重要。来自体内病毒学核心的小动物模型测试以及迭代领导的目标使用药物化学、结构生物学和综合建模核心进行优化。