Amelioration of SARS-CoV-2 infection by ANO6 phospholipid scramblase inhibition

• Published in: Cell reports (Cambridge) Vol. 40; no. 3; p. 111117
• Main Authors: Sim, Ju-Ri, Shin, Dong Hoon, Park, Pil-Gu, Park, So-Hyeon, Bae, Joon-Yong, Lee, Youngchae, Kang, Dha-Yei, Kim, Ye Jin, Aum, Sowon, Noh, Shin Hye, Hwang, Su Jin, Cha, Hye-Ran, Kim, Cheong Bi, Ko, Si Hwan, Park, Sunghoon, Jeon, Dongkyu, Cho, Sungwoo, Lee, Gee Eun, Kim, Jeonghun, Moon, Young-hye, Kim, Jae-Ouk, Nam, Jae-Sung, Kim, Chang-Hoon, Moon, Sungmin, Chung, Youn Wook, Park, Man-Seong, Ryu, Ji-Hwan, Namkung, Wan, Lee, Jae Myun, Lee, Min Goo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 19.07.2022; The Author(s)
• Subjects: Angiotensin-Converting Enzyme 2; Animals; ANO6/TMEM16F; Anoctamins; COVID-19 Drug Treatment; Humans; Mammals - metabolism; phosphatidylserine; Phosphatidylserines; Phospholipid Transfer Proteins - metabolism; SARS-CoV-2; Virus Internalization; virus-cell fusion; phosphatidylserine; SARS-CoV-2; CP: Microbiology; ANO6/TMEM16F; virus-cell fusion
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2022.111117

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). [Display omitted] •A high-throughput screening of chemical libraries identifies several ANO6 inhibitors•SARS-CoV-2 Spike evokes ANO6-mediated phosphatidylserine scrambling in host cells•Phosphatidylserine scrambling promotes fusion of the viral and cell membranes•The identified ANO6 inhibitors inhibit SARS-CoV-2 viral replications Sim et al. show that ANO6/TMEM16F-mediated phosphatidylserine scrambling participates in the SARS-CoV-2 entry into host cells and ANO6 inhibitors are effective against SARS-CoV-2 infection. These findings provide mechanistic insights into the viral entry process as well as a potential target for the development of drugs to treat COVID-19.