Hashimoto Rina, Sakamoto Ayaka, Deguchi Sayaka, Yi Renxing, Sano Emi, Hotta Akitsu, Takahashi Kazutoshi, Yamanaka Shinya, Takayama Kazuo
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Mol Ther Nucleic Acids. 2021 Dec 3;26:1107-1114. doi: 10.1016/j.omtn.2021.10.016. Epub 2021 Oct 20.
It has been reported that many receptors and proteases are required for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. Although angiotensin-converting enzyme 2 (ACE2) is the most important of these receptors, little is known about the contribution of other genes. In this study, we examined the roles of neuropilin-1, basigin, transmembrane serine proteases (TMPRSSs), and cathepsins (CTSs) in SARS-CoV-2 infection using the CRISPR interference system and ACE2-expressing human induced pluripotent stem (iPS) cells. Double knockdown of TMPRSS2 and cathepsin B (CTSB) reduced the viral load to 0.036% ± 0.021%. Consistently, the combination of the CTPB inhibitor CA-074 methyl ester and the TMPRSS2 inhibitor camostat reduced the viral load to 0.0078% ± 0.0057%. This result was confirmed using four SARS-CoV-2 variants (B.1.3, B.1.1.7, B.1.351, and B.1.1.248). The simultaneous use of these two drugs reduced viral load to less than 0.01% in both female and male iPS cells. These findings suggest that compounds targeting TMPRSS2 and CTSB exhibit highly efficient antiviral effects independent of gender and SARS-CoV-2 variant.
据报道，严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染需要多种受体和蛋白酶。尽管血管紧张素转换酶2（ACE2）是这些受体中最重要的，但对于其他基因的作用知之甚少。在本研究中，我们使用CRISPR干扰系统和表达ACE2的人类诱导多能干细胞（iPS细胞），研究了神经纤毛蛋白-1、基底膜蛋白、跨膜丝氨酸蛋白酶（TMPRSSs）和组织蛋白酶（CTSs）在SARS-CoV-2感染中的作用。TMPRSS2和组织蛋白酶B（CTSB）的双重敲低将病毒载量降低至0.036%±0.021%。同样，CTPB抑制剂CA-074甲酯和TMPRSS2抑制剂卡莫司他的联合使用将病毒载量降低至0.0078%±0.0057%。使用四种SARS-CoV-2变体（B.1.3、B.1.1.7、B.1.351和B.1.1.248）证实了这一结果。这两种药物同时使用在雌性和雄性iPS细胞中均将病毒载量降低至低于0.01%。这些发现表明，靶向TMPRSS2和CTSB的化合物表现出高效的抗病毒作用，且与性别和SARS-CoV-2变体无关。
