Defining therapeutic drug targets for SARS-CoV-2-specific and pan-coronavirus inhibition

定义 SARS-CoV-2 特异性和泛冠状病毒抑制的治疗药物靶点

• 批准号: 10238377
• 负责人: Charles M Rice
• 金额: $ 48.35万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-02 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238377
• 关键词: 2019-nCoV; Antiviral Agents; Biological; Biological Assay; COVID-19; COVID-19 pandemic; COVID-19 treatment; CRISPR library; CRISPR screen; CRISPR/Cas technology; Categories; Cells; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Coronavirus; Databases; Development; Disease; Disease Outbreaks; Drug Compounding; Drug Targeting; Environment; Epithelial Cells; Future; Genes; Genomic Library; Goals; Guide RNA; Human; Infection; Integration Host Factors; Intervention; Knock-out; Laboratories; Libraries; Life; Liver; Lower respiratory tract structure; Lung; Lung diseases; Mechanical ventilation; Microscopy; Molecular; Outcome; Pharmaceutical Preparations; Phenotype; Population; Production; Proteins; Readiness; Resistance; Respiratory distress; Role; Source; Symptoms; Temperature; Testing; Therapeutic; Tissues; Tropism; Vaccines; Viral Antigens; Virion; Virus; Western Blotting; base; bronchial epithelium; cell killing; combat; coronavirus therapeutics; cytotoxicity; drug development; drug repurposing; expectation; falls; fighting; genome-wide; human coronavirus; human disease; insight; pandemic disease; respiratory distress syndrome; response; small molecule; therapeutic target; vaccine candidate; validation studies; whole genome

Project Summary COVID-19 disease, caused by severe respiratory distress syndrome coronavirus 2 (SARS-CoV-2), is currently ravaging the world. Infection of humans causes symptoms spanning the spectrum from asymptomatic infection to severe respiratory distress and death. Despite the fact that vaccines and repurposed drugs are currently under study, it is uncertain whether they will be efficacious and so efforts to develop additional treatment and preventative options are crucial. This study will leverage results from multiple bulk CRISPR knockout screens already performed in the lab that identify host factors that SARS-CoV-2 and other human coronaviruses require for replication. These screens utilized cells from two tissue sources, lung and liver, four human coronaviruses including SARS-CoV-2, two temperatures that mimic the upper and lower airway, and five different CRISPR libraries including three druggable genome libraries, a library focused on human factors recently discovered to interact with proteins of SARS-CoV-2 and a full genome library. In the first aim, factors identified as hits in each of the screens will be cross compared and prioritized into three categories: 1) specific to SARS-CoV-2, 2) common to SARS-CoV-2 and at least one other coronavirus, or 3) specific to the lung for any of the viruses (Aim 1a). Hits from these categories will be subjected to refinement using a semi-arrayed CRISPR approach (Aim 1b) as well as drug and small molecule inhibition assays (Aim 1c) in the context of a panel of coronaviruses. In the second aim, gene disruption of the prioritized targets will be performed in primary human lung cells (Aim 2a) and the effect on the replication and cell killing by a panel of coronaviruses, including SARS-CoV-2 will be determined (Aim 2b). Drugs and compounds found as antiviral in Aim 1c will be tested in the primary human lung cells for their antiviral activity against the coronavirus panel. Validated factors, which could be SARS-CoV-2-specific, or possibly factors required generally for coronaviruses, would be targets for future in depth mechanistic studies and drug development.

项目摘要 由严重呼吸窘迫综合征2（SARS-COV-2）引起的Covid-19疾病是 目前正在肆虐世界。人类感染引起跨越频谱的症状 无症状感染严重的呼吸窘迫和死亡。尽管事实是疫苗和 重新使用的药物目前正在研究中，尚不确定它们是否有效，因此努力 开发额外的治疗方法和预防选择至关重要。这项研究将利用结果 从实验室中已经执行的多个散装CRISPR淘汰屏幕，可以识别宿主因素 SARS-COV-2和其他人类冠状病毒需要复制。这些屏幕利用了细胞 两个组织来源，肺和肝脏，四个人冠状病毒，包括SARS-COV-2，两个温度 模仿上下气道，以及五个不同的CRISPR库，包括三个可吸毒 基因组库，一个库的图书馆，重点是最近发现的人类因素，以与蛋白质相互作用 SARS-COV-2和一个完整的基因组库。在第一个目标中，在每个屏幕中都确定为命中的因素 将交叉比较并优先分为三类：1）特定于SARS-COV-2，2） SARS-COV-2和至少一个其他冠状病毒，或3）特定于肺部的任何病毒（AIM 1A）。这些类别的命中将使用半阵列的CRISPR方法进行改进 （AIM 1B）以及在一个面板的背景下，药物和小分子抑制测定（AIM 1C） 冠状病毒。在第二个目标中，将在主要目标中执行优先级目标的基因破坏 人肺细胞（AIM 2A）以及对冠状病毒的复制和细胞杀死的影响， 将确定包括SARS-COV-2（AIM 2B）。在AIM 1C中被发现是抗病毒的药物和化合物 将在原发性人肺细胞中测试其针对冠状病毒面板的抗病毒活性。 经过验证的因素，这可能是SARS-COV-2特异性的，也可能是通常所需的因素 冠状病毒将是未来深度机械研究和药物开发的目标。