Stress-induced changes in viral receptor and susceptibility gene expression were measured in embryonic stem cells (ESC) and differentiated progeny. Rex1 promoter-Red Fluorescence Protein reporter ESC were tested by RNAseq after 72hr exposures to control stress hyperosmotic sorbitol under stemness culture (NS) to quantify stress-forced differentiation (SFD) transcriptomic programs. Control ESC cultured with stemness factor removal produced normal differentiation (ND). Bulk RNAseq transcriptomic analysis showed significant upregulation of two genes involved in Covid-19 cell uptake, Vimentin (VIM) and Transmembrane Serine Protease 2 (TMPRSS2). SFD increased the hepatitis A virus receptor (Havcr1) and the transplacental Herpes simplex 1 (HSV1) virus receptor (Pvrl1) compared with ESC undergoing ND. Several other coronavirus receptors, Glutamyl Aminopeptidase (ENPEP) and Dipeptidyl Peptidase 4 (DPP4) were upregulated significantly in SFD>ND. Although stressed ESC are more susceptible to infection due to increased expression of viral receptors and decreased resistance, the necessary Covid-19 receptor, angiotensin converting enzyme (ACE)2, was not expressed in our experiments. TMPRSS2, ENPEP, and DPP4 mediate Coronavirus uptake, but are also markers of extra-embryonic endoderm (XEN), which arise from ESC undergoing ND or SFD. Mouse and human ESCs differentiated to XEN increase TMPRSS2 and other Covid-19 uptake-mediating gene expression, but only some lines express ACE2. Covid-19 susceptibility appears to be genotype-specific and not ubiquitous. Of the 30 gene ontology (GO) groups for viral susceptibility, 15 underwent significant stress-forced changes. Of these, 4 GO groups mediated negative viral regulation and most genes in these increase in ND and decrease with SFD, thus suggesting that stress increases ESC viral susceptibility. Taken together, the data suggest that a control hyperosmotic stress can increase Covid-19 susceptibility and decrease viral host resistance in mouse ESC. However, this limited pilot study should be followed with studies in human ESC, tests of environmental, hormonal, and pharmaceutical stressors and direct tests for infection of stressed, cultured ESC and embryos by Covid-19.

Graphical Abstract

中文翻译：

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压力降低宿主病毒抵抗力并增加胚胎干细胞的 Covid 易感性

在胚胎干细胞 (ESC) 和分化后代中测量了应激诱导的病毒受体和易感基因表达的变化。Rex1 启动子-红色荧光蛋白报告基因 ESC 在暴露 72 小时后通过 RNAseq 进行测试，以控制干细胞培养 (NS) 下的应激高渗山梨糖醇，以量化应激诱导分化 (SFD) 转录组学程序。去除干性因子培养的对照 ESC 产生正常分化 (ND)。Bulk RNAseq 转录组学分析显示，参与 Covid-19 细胞摄取的两个基因波形蛋白 (VIM) 和跨膜丝氨酸蛋白酶 2 (TMPRSS2) 显着上调。与进行 ND 的 ESC 相比，SFD 增加了甲型肝炎病毒受体 (Havcr1) 和经胎盘单纯疱疹病毒 1 (HSV1) 病毒受体 (Pvrl1)。其他几种冠状病毒受体，谷氨酰氨基肽酶 (ENPEP) 和二肽基肽酶 4 (DPP4) 在 SFD>ND 中显着上调。尽管受压的 ESC 由于病毒受体表达增加和抵抗力降低而更容易受到感染，但我们的实验中并未表达必要的 Covid-19 受体血管紧张素转换酶 (ACE)2。TMPRSS2、ENPEP 和 DPP4 介导冠状病毒摄取，但也是胚胎外内胚层 (XEN) 的标志物，它由经历 ND 或 SFD 的 ESC 产生。分化为 XEN 的小鼠和人类 ESC 增加了 TMPRSS2 和其他 Covid-19 摄取介导基因的表达，但只有一些细胞系表达 ACE2。Covid-19 易感性似乎是基因型特异性的，并非普遍存在。在病毒易感性的 30 个基因本体论 (GO) 组中，有 15 个经历了显着的应激变化。这些，4 个 GO 组介导了负病毒调节，其中大多数基因在 ND 中增加，在 SFD 中减少，因此表明压力增加了 ESC 病毒易感性。综上所述，数据表明，控制高渗应激可以增加 Covid-19 易感性并降低小鼠 ESC 中的病毒宿主抵抗力。然而，在这项有限的试点研究之后，应该进行人类胚胎干细胞研究、环境、激素和药物应激源测试，以及直接测试 Covid-19 对受压培养的胚胎干细胞和胚胎的感染。数据表明，控制高渗应激可以增加 Covid-19 易感性并降低小鼠 ESC 中的病毒宿主抵抗力。然而，在这项有限的试点研究之后，应该进行人类胚胎干细胞研究、环境、激素和药物应激源测试，以及直接测试 Covid-19 对受压培养的胚胎干细胞和胚胎的感染。数据表明，控制高渗应激可以增加 Covid-19 易感性并降低小鼠 ESC 中的病毒宿主抵抗力。然而，在这项有限的试点研究之后，应该进行人类胚胎干细胞研究、环境、激素和药物应激源测试，以及直接测试 Covid-19 对受压培养的胚胎干细胞和胚胎的感染。

图形概要