TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells
TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells
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DOI：
10.26508/lsa.202000786
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发表时间：
2020-09-01
影响因子：
4.4
通讯作者：
Boettcher-Friebertshaeuser, Eva
中科院分区：
文献类型：
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作者：
Bestle, Dorothea;Heindl, Miriam Ruth;Boettcher-Friebertshaeuser, Eva
新出现的SARS-CoV-2已在世界各地迅速传播，引起急性呼吸道感染（COVID-19），可导致严重疾病和死亡。为了使SARS-CoV-2进入细胞，其表面糖蛋白刺突（S）必须在两个不同的位点被宿主细胞蛋白酶切割，因此这是潜在的药物靶点。在本研究中，我们表明，S可以被切割的前蛋白转化酶弗林蛋白酶在S1/S2网站和跨膜丝氨酸蛋白酶2（TMPRSS 2）在S29网站。我们证明，TMPRSS 2是必不可少的激活SARS-CoV-2 S在Calu-3人气道上皮细胞通过反义介导的敲低TMPRSS 2的表达。此外，合成弗林蛋白酶抑制剂MI-1851在人气道细胞中也强烈抑制SARS-CoV-2的复制。相反，E64 d对内体组织蛋白酶的抑制并不影响病毒的复制。将各种TMPRSS 2抑制剂与弗林蛋白酶抑制剂MI-1851组合产生比等摩尔量的任何单一丝氨酸蛋白酶抑制剂更有效的抗SARS-CoV-2的抗病毒活性。因此，这种方法对于治疗COVID-19具有相当大的治疗潜力。
The novel emerged SARS-CoV-2 has rapidly spread around the world causing acute infection of the respiratory tract (COVID-19) that can result in severe disease and lethality. For SARS-CoV-2 to enter cells, its surface glycoprotein spike (S) must be cleaved at two different sites by host cell proteases, which therefore represent potential drug targets. In the present study, we show that S can be cleaved by the proprotein convertase furin at the S1/S2 site and the transmembrane serine protease 2 (TMPRSS2) at the S29 site. We demonstrate that TMPRSS2 is essential for activation of SARS-CoV-2 S in Calu-3 human airway epithelial cells through antisense-mediated knockdown of TMPRSS2 expression. Furthermore, SARS-CoV-2 replication was also strongly inhibited by the synthetic furin inhibitor MI-1851 in human airway cells. In contrast, inhibition of endosomal cathepsins by E64d did not affect virus replication. Combining various TMPRSS2 inhibitors with furin inhibitor MI-1851 produced more potent antiviral activity against SARS-CoV-2 than an equimolar amount of any single serine protease inhibitor. Therefore, this approach has considerable therapeutic potential for treatment of COVID-19.