Effects of envelope- or membrane-protein segments of SARS-CoV-2 on gene expression of HUVEC cells. Effects of envelope- or membrane-protein segments of SARS-CoV-2 on gene expression of HUVEC cells

Exterior segments of E-proteins bound onto and modulated gene expression in human vascular endothelial cells in vitro The finding that EC38 segment laid out of SARS-CoV-2 virus envelope prompted us to wonder if this segment would bind onto and impact the biology of host cells. Bearing the viremia during severe COVID-19 in mind, more attention was paid to the circulation system. Cultured human umbilical vein endothelial cells (HUVECs) were used to test the binding of EC38 and MN19 peptides and the potential consequence in terms of gene expression upon such binding. To check if the binding of interest peptides onto HUVEC concurred functional consequences in the cells, HUVECs were treated with EC38 and MN19 peptides (5μM) for 24 hours, and their transcriptome was measured via RNA sequencing. Overall design: To check if the interested peptides corresponding to segments of envelope (E-) or membrane (M-) proteins of SARS-CoV-2 would stimulate changes of transcriptome in HUVEC cells. HUVECs were treated with EC38 and MN19 peptides (5μM) for 24 hours, and their transcriptome was measured via RNA sequencing.

结合于人血管内皮细胞并调控其基因表达的E蛋白胞外区段（体外实验）。此前研究发现，严重急性呼吸综合征冠状病毒2（SARS-CoV-2）病毒包膜的EC38区段外露于病毒外，这引发我们思考该区段是否可结合宿主细胞并影响其生物学功能。鉴于重症新型冠状病毒肺炎（COVID-19）患者存在病毒血症，我们将研究重点聚焦于循环系统。我们采用培养的人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVECs），检测EC38与MN19肽段的结合情况，以及该结合对基因表达的潜在影响。为验证目标肽段结合HUVECs后是否会引发细胞内的功能变化，我们以5μM浓度的EC38与MN19肽段处理HUVECs 24小时，随后通过RNA测序检测其转录组（transcriptome）。实验总体设计：为明确对应SARS-CoV-2包膜（E）或膜（M）蛋白区段的目标肽段是否会诱导HUVEC细胞的转录组发生改变，我们以5μM浓度的EC38与MN19肽段处理HUVECs 24小时，随后通过RNA测序检测其转录组。