Amelioration of SARS-CoV-2 infection by ANO6 phospholipid scramblase inhibition
Authors
Ju-Ri Sim ; Dong Hoon Shin ; Pil-Gu Park ; So-Hyeon Park ; Joon-Yong Bae ; Youngchae Lee ; Dha-Yei Kang ; Ye Jin Kim ; Sowon Aum ; Shin Hye Noh ; Su Jin Hwang ; Hye-Ran Cha ; Cheong Bi Kim ; Si Hwan Ko ; Sunghoon Park ; Dongkyu Jeon ; Sungwoo Cho ; Gee Eun Lee ; Jeonghun Kim ; Young-Hye Moon ; Jae-Ouk Kim ; Jae-Sung Nam ; Chang-Hoon Kim ; Sungmin Moon ; Youn Wook Chung ; Man-Seong Park ; Ji-Hwan Ryu ; Wan Namkung ; Jae Myun Lee ; Min Goo Lee
As an enveloped virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) delivers its viral genome into host cells via fusion of the viral and cell membranes. Here, we show that ANO6/TMEM16F-mediated cell surface exposure of phosphatidylserine is critical for SARS-CoV-2 entry and that ANO6-selective inhibitors are effective against SARS-CoV-2 infections. Application of the SARS-CoV-2 Spike pseudotyped virus (SARS2-PsV) evokes a cytosolic Ca2+ elevation and ANO6-dependent phosphatidylserine externalization in ACE2/TMPRSS2-positive mammalian cells. A high-throughput screening of drug-like chemical libraries identifies three different structural classes of chemicals showing ANO6 inhibitory effects. Among them, A6-001 displays the highest potency and ANO6 selectivity and it inhibits the single-round infection of SARS2-PsV in ACE2/TMPRSS2-positive HEK 293T cells. More importantly, A6-001 strongly inhibits authentic SARS-CoV-2-induced phosphatidylserine scrambling and SARS-CoV-2 viral replications in Vero, Calu-3, and primarily cultured human nasal epithelial cells. These results provide mechanistic insights into the viral entry process and offer a potential target for pharmacological intervention to protect against coronavirus disease 2019 (COVID-19).