Dual inhibition of TMPRSS2 and Cathepsin B prevents SARS-CoV-2 infection in iPS cells

Abstract

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.

Dual Inhibition of Vacuolar-ATPase and TMPRSS2 Is Required for Complete Blockade of SARS-CoV-2 Entry into Cells

“…The SARS-CoV-2 replication cycle starts with the spike (S) protein binding to the cell-surface receptor angiotensin converting enzyme 2 (ACE2) ( 14 , 15 ). Then, a cell-surface transmembrane protease, serine 2 (TMPRSS2) cleaves the S protein at the S1/S2 junction and activates the S2 domain ( 16 – 19 ). The activated S2 domain brings the viral membrane into close proximity to the cell membrane and initiates membrane fusion, which ultimately leads to the release of the viral genome into the host cell ( 20 ).…”

GNS561 Exhibits Potent Antiviral Activity against SARS-CoV-2 through Autophagy Inhibition

“…Zaho et al showed that functional CTSL is essential to cleave the SARS-CoV-2 S protein and enhance virus entry [34]. Another study demonstrated that both CTSB and TMPRSS2 are required for SARS-CoV-2 infection in ACE2-expressing human induced pluripotent stem (iPS) cells [35]. We also know that GNS561 does not block the SARS-CoV-2 entry pathway.…”

mRNA vaccines for COVID-19 and diverse diseases

“…The S protein is primed by the TMPRSS2 (serine protease) crucial to the pathogenesis of the virus ( Fig. 4 ) [ 82 ]. The ACE2 is located mostly in the tissues of major organs, including the kidney, heart, lung, heart, testes, and brain.…”