Hydroxychloroquine-mediated inhibition of SARS-CoV-2 entry is attenuated by TMPRSS2.
Hydroxychloroquine-mediated inhibition of SARS-CoV-2 entry is attenuated by TMPRSS2.
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DOI：
10.1371/journal.ppat.1009212
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发表时间：
2021-01
期刊：
影响因子：
6.7
通讯作者：
Farzan M
中科院分区：
文献类型：
--
作者：
Ou T;Mou H;Zhang L;Ojha A;Choe H;Farzan M
在细胞培养研究中，用于治疗疟疾和一些自身免疫性疾病的羟氯喹能有效抑制SARS冠状病毒（SARS- cov -1）和SARS- cov -2的病毒感染。然而，羟氯喹的人体临床试验未能确定其治疗COVID-19的有效性。已知该化合物干扰组织蛋白酶蛋白水解活性所必需的内体酸化。在受体结合和内吞作用后，组织蛋白酶L可以切割SARS-CoV-1和SARS-CoV-2刺突(S)蛋白，从而激活膜融合进入细胞。质膜相关蛋白酶TMPRSS2同样可以切割这些S蛋白并激活病毒进入细胞表面。本研究表明，SARS-CoV-2的进入过程比SARS-CoV-1更依赖于TMPRSS2的表达。当SARS-CoV-2 S蛋白的糠蛋白切割位点被切除或将其引入SARS-CoV-1 S蛋白时，这种差异可以逆转。我们还发现，羟氯喹可以有效阻断由组织蛋白酶L介导的病毒进入，但不能阻断TMPRSS2介导的病毒进入，并且羟氯喹和经临床测试的TMPRSS2抑制剂联合使用比单独使用任何一种药物更有效地阻止SARS-CoV-2感染。这些研究确定了SARS-CoV-1和sars - cov -2进入过程之间的功能差异，并为羟氯喹在体内治疗COVID-19的有限效用提供了机制解释。新型致病性冠状病毒SARS-CoV-2引起COVID-19，仍然对全球公共卫生构成威胁。氯喹和羟氯喹已被证明可以预防细胞培养系统中的病毒感染，但人体临床试验未观察到使用这些化合物治疗的COVID-19患者的显着改善。本研究表明，羟氯喹仅干扰SARS-CoV-2刺突(S)蛋白激活介导感染的两条冗余途径中的一条。第一种途径依赖于内体蛋白酶组织蛋白酶L，对羟氯喹敏感，而第二种途径依赖于TMPRSS2，不受该化合物的影响。我们进一步表明，SARS- cov -2比SARS冠状病毒（SARS- cov -1）更依赖于TMPRSS2途径，这种差异是由于SARS- cov -2 S蛋白中存在furin切割位点。最后，我们发现羟氯喹和临床测试的TMPRSS2抑制剂联合使用可有效抑制SARS-CoV-2的进入。因此，TMPRSS2在生理相关的SARS-CoV-2靶细胞上的表达可能绕过羟氯喹的抗病毒活性，并解释了其缺乏体内疗效的原因。
Hydroxychloroquine, used to treat malaria and some autoimmune disorders, potently inhibits viral infection of SARS coronavirus (SARS-CoV-1) and SARS-CoV-2 in cell-culture studies. However, human clinical trials of hydroxychloroquine failed to establish its usefulness as treatment for COVID-19. This compound is known to interfere with endosomal acidification necessary to the proteolytic activity of cathepsins. Following receptor binding and endocytosis, cathepsin L can cleave the SARS-CoV-1 and SARS-CoV-2 spike (S) proteins, thereby activating membrane fusion for cell entry. The plasma membrane-associated protease TMPRSS2 can similarly cleave these S proteins and activate viral entry at the cell surface. Here we show that the SARS-CoV-2 entry process is more dependent than that of SARS-CoV-1 on TMPRSS2 expression. This difference can be reversed when the furin-cleavage site of the SARS-CoV-2 S protein is ablated or when it is introduced into the SARS-CoV-1 S protein. We also show that hydroxychloroquine efficiently blocks viral entry mediated by cathepsin L, but not by TMPRSS2, and that a combination of hydroxychloroquine and a clinically-tested TMPRSS2 inhibitor prevents SARS-CoV-2 infection more potently than either drug alone. These studies identify functional differences between SARS-CoV-1 and -2 entry processes, and provide a mechanistic explanation for the limited in vivo utility of hydroxychloroquine as a treatment for COVID-19. The novel pathogenic coronavirus SARS-CoV-2 causes COVID-19 and remains a threat to global public health. Chloroquine and hydroxychloroquine have been shown to prevent viral infection in cell-culture systems, but human clinical trials did not observe a significant improvement in COVID-19 patients treated with these compounds. Here we show that hydroxychloroquine interferes with only one of two somewhat redundant pathways by which the SARS-CoV-2 spike (S) protein is activated to mediate infection. The first pathway is dependent on the endosomal protease cathepsin L and sensitive to hydroxychloroquine, whereas the second pathway is dependent on TMPRSS2, which is unaffected by this compound. We further show that SARS-CoV-2 is more reliant than SARS coronavirus (SARS-CoV-1) on the TMPRSS2 pathway, and that this difference is due to a furin cleavage site present in the SARS-CoV-2 S protein. Finally, we show that combinations of hydroxychloroquine and a clinically tested TMPRSS2 inhibitor work together to effectively inhibit SARS-CoV-2 entry. Thus TMPRSS2 expression on physiologically relevant SARS-CoV-2 target cells may bypass the antiviral activities of hydroxychloroquine, and explain its lack of in vivo efficacy.
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影响因子：
64.8
作者：
Li W;Moore MJ;Vasilieva N;Sui J;Wong SK;Berne MA;Somasundaran M;Sullivan JL;Luzuriaga K;Greenough TC;Choe H;Farzan M
通讯作者：
Farzan M
影响因子：
158.5
作者：
Boulware, David R.;Pullen, Matthew F.;Hullsiek, Kathy H.
通讯作者：
Hullsiek, Kathy H.
影响因子：
5.4
作者：
Glowacka, Ilona;Bertram, Stephanie;Poehlmann, Stefan
通讯作者：
Poehlmann, Stefan
DOI：
10.1073/pnas.1407087111
发表时间：
2014-10-21
影响因子：
11.1
作者：
Millet, Jean Kaoru;Whittaker, Gary R.
通讯作者：
Whittaker, Gary R.
影响因子：
5.4
作者：
Shirato, Kazuya;Kawase, Miyuki;Matsuyama, Shutoku
通讯作者：
Matsuyama, Shutoku