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

摘要 严重急性呼吸综合征相关冠状病毒2引起的2019冠状病毒病大流行 （SARS-CoV-2）导致了全球健康和经济危机。为了研制有效的疫苗 和抗病毒药物来预防和控制SARS-CoV-2或其他冠状病毒感染， 了解SARS-CoV-2基因组在宿主细胞中复制的调控机制。在这 R21项目，我们将使用新的epitranscriptomic技术来研究SARS-CoV-2 在人肺上皮细胞中复制。N6-甲基腺苷（m6 A）修饰的许多 病毒在病毒复制、基因表达和 免疫逃避然而，目前还不清楚SARS-CoV-2的m6 A修饰是否以及如何修饰 基因组和细胞基因影响病毒复制和致病性。 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中的m6 A水平。我们对13,699例全长SARS病毒的生物信息学分析- CoV-2基因组序列预测高度保守的多个m6 A修饰位点 在全球SARS-CoV-2分离株中。因此，我们假设，m6 A修饰的 SARS-CoV-2基因组和细胞基因通过以下途径增强病毒在细胞中的复制： 表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.到 目的2.研究SARS冠状病毒2型感染肺上皮细胞的m6 A表型。 绘制SARS-CoV-2基因组m6 A位点，并确定病毒复制的关键位点。 定义SARS-CoV-2感染细胞的表观转录组m6 A谱具有重要意义 在理解COVID-19发病机制和确定新的药物靶点方面。

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.