Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue
Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue
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DOI：
10.1183/13993003.01123-2020
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发表时间：
2020-09-01
影响因子：
24.3
通讯作者：
Hirota, Jeremy A.
中科院分区：
文献类型：
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作者：
Aguiar, Jennifer A.;Tremblay, Benjamin J-M;Hirota, Jeremy A.
2019年12月，严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）出现，引发2019年冠状病毒病（COVID-19）大流行。SARS- cov是导致2003年SARS爆发的病原体，它利用血管紧张素转换酶2 （ACE2）和跨膜丝氨酸蛋白酶2 （TMPRSS2）宿主分子进入病毒。ACE2和TMPRSS2最近与SARS-CoV-2病毒感染有关。其他宿主分子包括ADAM17、组织蛋白酶L、CD147和GRP78也可能作为SARS-CoV-2的受体。为了确定候选SARS-CoV-2受体在呼吸道粘膜中的表达和原位定位，我们分析了515名健康受试者气道上皮细胞的基因表达数据集、使用包含120种不同样品类型的FANTOM5数据集进行基因启动子活性分析、10名健康受试者的单细胞RNA测序（scRNAseq）、蛋白质组学数据集、多种气道上皮细胞类型的免疫印迹和98个人肺样本的免疫组织化学。我们在多种肺上皮细胞样本中发现了低ACE2启动子活性，在上皮细胞群体的微阵列和scRNAseq数据集中发现了低ACE2基因表达。与基因表达一致，ACE2蛋白在人肺气道上皮和肺泡中罕见表达，蛋白质组学证实。我们证实了TMPRSS2、CD147和GRP78蛋白在体外气道上皮细胞中存在，并证实了CD147和GRP78蛋白在呼吸道黏膜中广泛的原位表达。总之，我们的数据表明，可能在SARS-CoV-2感染期间，存在一种动态调节人肺中ACE2表达的机制，并且还表明存在其他SARS-CoV-2受体，以促进初始宿主细胞感染。
In December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged, causing the coronavirus disease 2019 (COVID-19) pandemic. SARS-CoV, the agent responsible for the 2003 SARS outbreak, utilises angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) host molecules for viral entry. ACE2 and TMPRSS2 have recently been implicated in SARS-CoV-2 viral infection. Additional host molecules including ADAM17, cathepsin L, CD147 and GRP78 may also function as receptors for SARS-CoV-2.To determine the expression and in situ localisation of candidate SARS-CoV-2 receptors in the respiratory mucosa, we analysed gene expression datasets from airway epithelial cells of 515 healthy subjects, gene promoter activity analysis using the FANTOM5 dataset containing 120 distinct sample types, single cell RNA sequencing (scRNAseq) of 10 healthy subjects, proteomic datasets, immunoblots on multiple airway epithelial cell types, and immunohistochemistry on 98 human lung samples.We demonstrate absent to low ACE2 promoter activity in a variety of lung epithelial cell samples and low ACE2 gene expression in both microarray and scRNAseq datasets of epithelial cell populations. Consistent with gene expression, rare ACE2 protein expression was observed in the airway epithelium and alveoli of human lung, confirmed with proteomics. We present confirmatory evidence for the presence of TMPRSS2, CD147 and GRP78 protein in vitro in airway epithelial cells and confirm broad in situ protein expression of CD147 and GRP78 in the respiratory mucosa.Collectively, our data suggest the presence of a mechanism dynamically regulating ACE2 expression in human lung, perhaps in periods of SARS-CoV-2 infection, and also suggest that alternative receptors for SARS-CoV-2 exist to facilitate initial host cell infection.