Transcriptional regulation of ACE2 and the adaption of SARS-CoV-2

ACE2 的转录调控和 SARS-CoV-2 的适应

• 批准号: 10590248
• 负责人: Yiping Zhu
• 金额: $ 23.1万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-11 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10590248
• 关键词: 2019-nCoV; ACE2; Amino Acid Substitution; Antibodies; Architecture; Binding; Binding Sites; COVID-19; COVID-19 treatment; Cell Line; Cells; Chromatin; Circulation; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA Binding; DNA Sequence Alteration; Data; Databases; Dependence; Deposition; Development; Epigenetic Process; Genetic Transcription; Human; India; Integration Host Factors; Kidney; Knock-out; Knowledge; Large Intestine; Lung; Maps; Mexico; Molecular; Monkeys; Mutation; Nucleocapsid; Orthologous Gene; Play; Production; Promoter Regions; Receptor Cell; Regulation; Research; Residual state; SARS-CoV-2 infection; TNF receptor-associated factor 3; Testing; Tissues; Transcriptional Activation; Transcriptional Regulation; Variant; Viral; Viral Pathogenesis; Virion; Virus; Virus Diseases; antiviral drug development; cofactor; differential expression; genome-wide; global health; hepatic nuclear factor 1; human tissue; innovation; novel; prevent; protein protein interaction; receptor; recruit; therapeutic development; therapy design; vaccine access; validation studies

ABSTRACT Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus disease 2019 (COVID-19), has caused a global health crisis. Currently effective vaccines are available to prevent SARS- CoV-2 infection, but our options for the treatment of SARS-CoV-2 infection remain limited. To inform the development of innovative interventions designed to counteract SARS-CoV-2 infection, we need a better understanding of the molecular details of SARS-CoV-2 infection, especially the host dependency factors that are required for virus infection. We have performed a genome-wide CRISPR knockout screen and identified PCBD1, TRAF3, and RAD54L2 as novel host factors required for SARS-CoV-2 infection. Our preliminary data showed that PCBD1, TRAF3, and RAD54L2 are required for the transcription of angiotensin converting enzyme 2 (ACE2). ACE2 is the primary receptor for the cellular entry of SARS-CoV-2, and plays key roles in virus infection and pathogenesis. ACE2 is differentially expressed in a wide variety of human tissues. However, the molecular basis for the transcriptional regulation of ACE2 in different tissues remains unexplored, which constitutes a clear knowledge gap in SARS-CoV-2 research. In Aim 1, we will determine the function of PCBD1, TRAF3, and RAD54L2 in transcriptional regulation of ACE2 in four cell lines derived from different tissues: Vero E6 (monkey kidney), Calu-3 (human lung), C2BBe1 (human large intestine), and HK-2 (human kidney). SARS-CoV-2 continuously evolves by genetic mutations for the adaption to the host. We identified a number of mutations in Spike (S) and Nucleocapsid (N) that may enhance the replication of SARS-CoV-2 in cells expressed low levels of ACE2. Some mutation (such as N S194L) was prevalent in India and Mexico during early circulation, suggesting these variants may contribute to the viral adaption to the host. In Aim 2, we will analyze the function of Spike (N81S, L242P, E484D, and P1079T) and Nucleocapsid (S194L) variants in virion production and cell entry of SARS-CoV-2. Our proposed studies will provide new information regarding the transcriptional regulation of ACE2 and the adaption of SARS-CoV-2. In the long term, these studies will provide new targets and strategies for the development of antiviral drugs.

抽象的 严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2)，冠状病毒病的病原体 2019 年（COVID-19）引发了全球健康危机。目前已有有效的疫苗可以预防SARS CoV-2 感染，但我们治疗 SARS-CoV-2 感染的选择仍然有限。通知 开发旨在对抗 SARS-CoV-2 感染的创新干预措施，我们需要更好的 了解 SARS-CoV-2 感染的分子细节，尤其是宿主依赖性因素 病毒感染所需。我们进行了全基因组 CRISPR 敲除筛选并鉴定出 PCBD1， TRAF3 和 RAD54L2 作为 SARS-CoV-2 感染所需的新宿主因子。我们的初步数据显示 PCBD1、TRAF3 和 RAD54L2 是血管紧张素转换酶 2 转录所必需的 （ACE2）。 ACE2是SARS-CoV-2进入细胞的主要受体，在病毒感染中发挥关键作用 和发病机制。 ACE2 在多种人体组织中差异表达。然而，分子 ACE2 在不同组织中的转录调控基础仍未被探索，这构成了一个明确的 SARS-CoV-2 研究中的知识差距。在目标 1 中，我们将确定 PCBD1、TRAF3 和 RAD54L2 在来自不同组织的四种细胞系中 ACE2 的转录调节：Vero E6（猴） 肾）、Calu-3（人肺）、C2BBe1（人大肠）和 HK-2（人肾）。 SARS-CoV-2 通过基因突变不断进化以适应宿主。我们发现了一些突变 刺突 (S) 和核衣壳 (N) 可能会增强低水平表达的细胞中 SARS-CoV-2 的复制 ACE2 的。一些突变（如N S194L）在早期流通期间在印度和墨西哥流行， 表明这些变异可能有助于病毒适应宿主。在目标 2 中，我们将分析该函数 病毒颗粒生产和细胞中的 Spike（N81S、L242P、E484D 和 P1079T）和核衣壳（S194L）变体 SARS-CoV-2 的进入。我们提出的研究将提供有关转录调控的新信息 ACE2 和 SARS-CoV-2 的适应。从长远来看，这些研究将提供新的目标和策略 用于开发抗病毒药物。