@article{9a013f3c430c44d4b5ff79e5a58061a8,
title = "Amelioration of SARS-CoV-2 infection by ANO6 phospholipid scramblase inhibition",
abstract = "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).",
keywords = "ANO6/TMEM16F, CP: Microbiology, SARS-CoV-2, phosphatidylserine, virus-cell fusion",
author = "Sim, {Ju Ri} and Shin, {Dong Hoon} and Park, {Pil Gu} and Park, {So Hyeon} and Bae, {Joon Yong} and Youngchae Lee and Kang, {Dha Yei} and Kim, {Ye Jin} and Sowon Aum and Noh, {Shin Hye} and Hwang, {Su Jin} and Cha, {Hye Ran} and Kim, {Cheong Bi} and Ko, {Si Hwan} and Sunghoon Park and Dongkyu Jeon and Sungwoo Cho and Lee, {Gee Eun} and Jeonghun Kim and Moon, {Young hye} and Kim, {Jae Ouk} and Nam, {Jae Sung} and Kim, {Chang Hoon} and Sungmin Moon and Chung, {Youn Wook} and Park, {Man Seong} and Ryu, {Ji Hwan} and Wan Namkung and Lee, {Jae Myun} and Lee, {Min Goo}",
note = "Funding Information: The authors thank the Medical Illustration & Design team at Yonsei University College of Medicine for providing support with medical illustrations, Benjamin J. Latimer for his editorial assistance, and the Yonsei-Carl Zeiss Advanced Imaging Center for their technical assistance. This work was supported by grants 2013R1A3A2042197 , 2022R1A2C3002917 (M.G.L.), 2018R1A6A1A03023718 (W.N.), 2022M3A9I2082293 , and 2019R1A6A1A03032869 (J.M.L.) from the National Research Foundation , the Ministry of Science and ICT , and the Ministry of Education , and grant HI22C0984 (M.G.L.) from the Korea Health Industry Development Institute , the Ministry of Health and Welfare, Republic of Korea . M.G.L. was supported by the Heung Ki Kim Memorial Fund from Yonsei University, Seoul, Korea . Funding Information: The authors thank the Medical Illustration & Design team at Yonsei University College of Medicine for providing support with medical illustrations, Benjamin J. Latimer for his editorial assistance, and the Yonsei-Carl Zeiss Advanced Imaging Center for their technical assistance. This work was supported by grants 2013R1A3A2042197, 2022R1A2C3002917 (M.G.L.), 2018R1A6A1A03023718 (W.N.), 2022M3A9I2082293, and 2019R1A6A1A03032869 (J.M.L.) from the National Research Foundation, the Ministry of Science and ICT, and the Ministry of Education, and grant HI22C0984 (M.G.L.) from the Korea Health Industry Development Institute, the Ministry of Health and Welfare, Republic of Korea. M.G.L. was supported by the Heung Ki Kim Memorial Fund from Yonsei University, Seoul, Korea. J.-R.S. designed and carried out the molecular experiments of SARS-CoV-2 and analyzed the data. D.H.S. performed patch-clamp and intracellular calcium measurements. P.-G.P. performed experiments using an infectious virus. S.-H.P. performed the high-throughput screening of chemicals. J.-Y.B. performed authentic virus isolation. Y.L. D.-Y.K. Y.J.K. and S.A. generated pseudotyped virus and immunochemistry. S.H.N. and H.-R.C. performed the four-quadrant dot blot FACS analysis. S.J.H. C.B.K. S.H.K. and S.P. performed experiments using an infectious virus. D.J. and S.C. performed YFP quenching assays. G.E.L. and J.K. performed experiments with authentic virus. Y.M. and J.-O.K. generated pseudotyped virus. J.-S.N. C.-H.K. S.M. and Y.W.C. performed experiments in HNE cells, and M.-S.P. designed and supervised experiments with authentic virus. J.-H.R. designed and supervised experiments in HNE cells. W.N. and J.M.L. designed and conceived experiments involving high-throughput screening and in vitro antiviral assays and edited the manuscript. M.G.L. designed and conceived all experiments and wrote the manuscript. M.G.L. J.M.L. and W.N. are inventors on a pending patent related to this work filed by Yonsei University Office of Research Affairs/University Industry Foundation. The authors declare no other competing interests. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = jul,
day = "19",
doi = "10.1016/j.celrep.2022.111117",
language = "English",
volume = "40",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "3",
}
