Table_9_Identification of a SARS-CoV-2 virus-derived vmiRNA in COVID-19 patients holding potential as a diagnostic biomarker.xlsx

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（SARS-CoV-2）已成为持续威胁全球公共卫生安全的病原体。为最大限度降低病毒传播风险，开发更为可靠的感染早期诊断方法以及快速抑制病毒复制的手段至关重要。本研究通过对SARS-CoV-2基因组开展计算预测，并对新型冠状病毒肺炎（COVID-19）患者的临床标本进行筛选分析，共预测得到15个SARS-CoV-2编码的微RNA（miRNA）前体（CvmiRNAs），对应20条成熟CvmiRNA；其中CvmiR-2可通过定量分析在患者的血清及鼻咽拭子样本中成功检出。CvmiR-2在区分COVID-19患者与健康对照人群时表现出极高的特异性，且在SARS-CoV-2及其变异株之间具有高度保守性。研究发现CvmiR-2的表达水平与患者的病情严重程度呈正相关。通过对转染CvmiR-2前体的A549细胞进行验证，证实了CvmiR-2的生物发生与表达呈现剂量依赖性模式。通过对感染SARS-CoV-2或转染CvmiR-2前体的人类细胞进行测序分析，进一步验证了CvmiR-2的序列正确性。靶基因预测分析显示，CvmiR-2可能参与调控COVID-19患者的免疫应答、肌肉疼痛及/或神经系统疾病。综上所述，本研究鉴定出一种人类细胞感染SARS-CoV-2后由病毒编码的新型病毒微RNA（v-miRNA），该分子具备成为临床诊断生物标志物或治疗靶点的潜力。