Table_2_Identification of a SARS-CoV-2 virus-derived vmiRNA in COVID-19 patients holding potential as a diagnostic biomarker.xls

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a lasting threat to public health. To minimize the viral spread, it is essential to develop more reliable approaches for early diagnosis of the infection and immediate suppression of the viral replication. Herein, through computational prediction of SARS-CoV-2 genome and screening analysis of specimens from covid-19 patients, we predicted 15 precursors for SARS-CoV-2-encoded miRNAs (CvmiRNAs) containing 20 mature CvmiRNAs, in which CvmiR-2 was successfully detected by quantitative analysis in both serum and nasal swab samples of patients. CvmiR-2 showed high specificity in distinguishing covid-19 patients from normal controls, and high conservation between SARS-CoV-2 and its mutants. A positive correlation was observed between the CvmiR-2 expression level and the severity of patients. The biogenesis and expression of CvmiR-2 were validated in the pre-CvmiR-2-transfected A549 cells, showing a dose-dependent pattern. The sequence of CvmiR-2 was validated by sequencing analysis of human cells infected by either SARS-CoV-2 or pre-CvmiR-2. Target gene prediction analysis suggested CvmiR-2 may be involved in the regulation of the immune response, muscle pain and/or neurological disorders in covid-19 patients. In conclusion, the current study identified a novel v-miRNA encoded by SARS-CoV-2 upon infection of human cells, which holds the potential to serve as a diagnostic biomarker or a therapeutic target in clinic.

严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）已成为对公共健康的持续威胁。为了最大限度地减少病毒传播，开发更可靠的早期诊断感染和立即抑制病毒复制的策略至关重要。本研究所述，通过对SARS-CoV-2基因组的计算预测以及对新冠患者样本的筛选分析，我们预测了15个SARS-CoV-2编码的miRNA（CvmiRNAs）的前体，其中包含20个成熟的CvmiRNAs，其中CvmiR-2在患者的血清和鼻拭子样本中通过定量分析成功检测到。CvmiR-2在区分新冠患者与正常对照者方面展现出极高的特异性，且SARS-CoV-2及其突变体之间高度保守。CvmiR-2的表达水平与患者的严重程度呈正相关。在预CvmiR-2转染的A549细胞中，CvmiR-2的生物发生和表达得到了验证，显示出剂量依赖性模式。通过感染SARS-CoV-2或预CvmiR-2的人类细胞的测序分析验证了CvmiR-2的序列。目标基因预测分析表明，CvmiR-2可能参与调节新冠患者的免疫反应、肌肉疼痛以及/或神经系统疾病。总之，本研究确定了SARS-CoV-2感染人类细胞后编码的一种新型v-miRNA，该miRNA在临床上有望作为诊断生物标志物或治疗靶点。