Epitranscriptomic m6A profile of SARS-CoV-2-infected human lung epithelial cells

摘要 由传染性非典型肺炎冠状病毒2型引起的新冠肺炎大流行 非典冠状病毒(SARS-CoV-2)已导致全球健康和经济危机。开发有效的疫苗 和抗病毒药物预防和控制SARS-CoV-2或其他冠状病毒感染，关键是 了解SARS-CoV-2基因组复制在宿主细胞中的调控机制。在这 R21计划，我们将使用新的表位转录技术来研究SARS-CoV-2 人肺上皮细胞的复制。N6-甲基腺苷(M6A)的许多修饰 病毒在病毒复制、基因表达和基因表达的表位转录调控中起主要作用 免疫逃避。然而，目前还不清楚是否以及如何对SARS-CoV-2进行m6A修改 基因组和细胞基因影响病毒的复制和致病性。 我们的初步研究表明，SARS-CoV-2感染人肺上皮细胞 上调细胞RNA中m6A的水平。我们对13,699个全长SARS的生物信息学分析- 冠状病毒2基因组序列预测高度保守的多个m6A修饰位点 在世界范围内的SARS-CoV-2分离株中。因此，我们假设M6A修改 SARS-CoV-2基因组和细胞基因通过 表外转录调控。我们提出了两个具体的目标来检验这一假设。目标1.目标 目的1.研究SARS-CoV-2感染的肺上皮细胞m6A表位转录谱。 绘制SARS-CoV-2基因组上m6A位点的图谱，并确定病毒复制的关键位点。 确定SARS-CoV-2感染细胞的表位转录M6A谱具有重要意义 在了解新冠肺炎的发病机制和寻找新的药物靶点方面。

Abstract The COVID-19 pandemic caused by severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) has resulted in global health and economic crises. To develop effective vaccines and antivirals to prevent and control SARS-CoV-2 or other coronavirus infections, it is critical to understand the regulatory mechanisms of SARS-CoV-2 genome replication in host cells. In this R21 project, we will use novel epitranscriptomic technologies to investigate SARS-CoV-2 replication in human lung epithelial cells. N6-methyladenosine (m6A) modifications of many viruses play a major role in epitranscriptomic regulation of viral replication, gene expression, and immune evasion. However, it is unclear whether and how m6A modifications of the SARS-CoV-2 genome and cellular genes affect viral replication and pathogenicity. Our preliminary studies showed that SARS-CoV-2 infection of human lung epithelial cells upregulated m6A levels in cellular RNA. Our bioinformatic analysis of 13,699 full-length SARS- CoV-2 genome sequences predicts multiple m6A modification sites that are highly conserved among SARS-CoV-2 isolates worldwide. Thus, we hypothesize that m6A modifications of the SARS-CoV-2 genome and cellular genes enhance viral replication in cells through epitranscriptomic regulation. We propose two specific aims to test this hypothesis. Aim 1. To investigate the m6A epitranscriptomic profile of SARS-CoV-2-infected lung epithelial cells; Aim 2. To map m6A sites on the SARS-CoV-2 genome and to identify critical sites for viral replication. Defining epitranscriptomic m6A profile of SARS-CoV-2-infected cells has significant implications in understanding the COVID-19 pathogenesis and identifying novel drug targets.