@article{203a59148bb041c4a49182bc32ff5612,
title = "SARS-CoV-2 replication in airway epithelia requires motile cilia and microvillar reprogramming",
abstract = "How SARS-CoV-2 penetrates the airway barrier of mucus and periciliary mucins to infect nasal epithelium remains unclear. Using primary nasal epithelial organoid cultures, we found that the virus attaches to motile cilia via the ACE2 receptor. SARS-CoV-2 traverses the mucus layer, using motile cilia as tracks to access the cell body. Depleting cilia blocks infection for SARS-CoV-2 and other respiratory viruses. SARS-CoV-2 progeny attach to airway microvilli 24 h post-infection and trigger formation of apically extended and highly branched microvilli that organize viral egress from the microvilli back into the mucus layer, supporting a model of virus dispersion throughout airway tissue via mucociliary transport. Phosphoproteomics and kinase inhibition reveal that microvillar remodeling is regulated by p21-activated kinases (PAK). Importantly, Omicron variants bind with higher affinity to motile cilia and show accelerated viral entry. Our work suggests that motile cilia, microvilli, and mucociliary-dependent mucus flow are critical for efficient virus replication in nasal epithelia.",
keywords = "airway epithelial cells, cilia, COVID-19, microvilli, organoids, protein kinases, proteomics, respiratory synctital virus, respiratory virus, SARS-CoV-2",
author = "Wu, {Chien Ting} and Lidsky, {Peter V.} and Yinghong Xiao and Ran Cheng and Lee, {Ivan T.} and Tsuguhisa Nakayama and Sizun Jiang and Wei He and Janos Demeter and Knight, {Miguel G.} and Turn, {Rachel E.} and Rojas-Hernandez, {Laura S.} and Chengjin Ye and Kevin Chiem and Judy Shon and Luis Martinez-Sobrido and Bertozzi, {Carolyn R.} and Nolan, {Garry P.} and Nayak, {Jayakar V.} and Carlos Milla and Raul Andino and Jackson, {Peter K.}",
note = "Funding Information: We thank John Perrino and Ruth Yamawakithe (Stanford Cell Sciences Imaging Facility) for electron microscope sample preparation and image processing. This work was supported by the National Institutes of Health R01DK127665, R01HD085901, R01GM121565, and P30DK116074 to P.K.J.; R01AI149672-01 and U54-CA209971 to G.P.N.; Stanford Diabetes Research Center Pilot and Feasibility Research Grant to P.K.J; Fast Grant Funding for COVID-19 Science to P.K.J. and G.P.N.; NIH (R01 AI36178, AI40085, P01 AI091575), the Bill and Melinda Gates Foundation, and the DARPA Intercept program (Contract No. HR0011-17-2-0027) to R.A.; the Botnar Research Centre for Child Health Emergency Response to COVID-19 Grant and a Bill and Melinda Gates Foundation COVID-19 Pilot Award to S.J. and G.P.N.; the Rachford & Carlotta A. Harris Endowed Chair to G.P.N.; California Institute for Regenerative Medicine (DISC2-09637), Defense Advanced Research Projects Agency (HR001118S0037-PREPARE-FP-001), and The Operndorf Foundation to J.V.N.; Stanford Respond. Innovate. Scale. Empower (RISE) COVID-19 Crisis Response Trainee Seed Grant to C.-T.W. R.C. I.T.L. S.J. and T.N.; Stanford Translational Research and Applied Medicine (TRAM) Pilot Grant and Thrasher Research Fund Early Career Award to I.T.L.; Stanford Maternal and Child Health Research Institute (MCHRI) Clinical (MD) Trainee Support Award to I.T.L. Ernest and Amelia Gallo Endowed Postdoctoral Fellow; Leukemia & Lymphoma Society Career Development Program to S.J.; Cellular and Molecular Biology Training Grant (NIH 5 T32 GM007276) to R.C.; the Mossier Laboratories Research Fund to C.M.; and the ARRA Award Number 1S10RR026780-01 from the National Center for Research Resources., Concept and study coordination, C.-T.W. and P.K.J. Experimental design/execution, C.-T.W. P.L. Y.X. R.C. I.T.L. T.N. S.J. W.H. R.A. P.K.J. R.E.T. and M.G.K. Microscopy, C.-T.W. IHC, I.T.L. and T.N. Virus production/human nasal epithelial cell infection, C.-T.W. P.L. Y.X. M.G.K. K.C. J.S. and L.M.S. StcE enzyme purification, C.Y. and C.R.B. Patient consent, sample collection/testing, and banking, I.T.L. S.J. T.N. C.M. and J.V.N. Live-cell imaging for QD-RBD, W.H. and C.-T.W. Phosphoproteomics/data analysis, C.-T.W. R.C. and J.D. Statistical analysis, C.-T.W. T.N. and J.D. Human nasal cells, J.V.N. C.M. T.N. and L.S.R.H. Figures, C.-T.W. with help from I.T.L. S.J. and G.P.N. Funding/scientific guidance, R.A. and P.K.J. All authors reviewed the manuscript. The authors declare no competing interests. Funding Information: We thank John Perrino and Ruth Yamawakithe (Stanford Cell Sciences Imaging Facility) for electron microscope sample preparation and image processing. This work was supported by the National Institutes of Health R01DK127665 , R01HD085901 , R01GM121565 , and P30DK116074 to P.K.J.; R01AI149672-01 and U54-CA209971 to G.P.N.; Stanford Diabetes Research Center Pilot and Feasibility Research Grant to P.K.J; Fast Grant Funding for COVID-19 Science to P.K.J. and G.P.N.; NIH ( R01 AI36178 , AI40085 , P01 AI091575 ), the Bill and Melinda Gates Foundation, and the DARPA Intercept program (Contract No. HR0011-17-2-0027 ) to R.A.; the Botnar Research Centre for Child Health Emergency Response to COVID-19 Grant and a Bill and Melinda Gates Foundation COVID-19 Pilot Award to S.J. and G.P.N.; the Rachford & Carlotta A. Harris Endowed Chair to G.P.N.; California Institute for Regenerative Medicine ( DISC2-09637 ), Defense Advanced Research Projects Agency ( HR001118S0037-PREPARE-FP-001 ), and The Operndorf Foundation to J.V.N.; Stanford Respond. Innovate. Scale. Empower ( RISE ) COVID-19 Crisis Response Trainee Seed Grant to C.-T.W., R.C., I.T.L., S.J., and T.N.; Stanford Translational Research and Applied Medicine (TRAM) Pilot Grant and Thrasher Research Fund Early Career Award to I.T.L.; Stanford Maternal and Child Health Research Institute (MCHRI) Clinical (MD) Trainee Support Award to I.T.L., Ernest and Amelia Gallo Endowed Postdoctoral Fellow; Leukemia & Lymphoma Society Career Development Program to S.J.; Cellular and Molecular Biology Training Grant ( NIH 5 T32 GM007276 ) to R.C.; the Mossier Laboratories Research Fund to C.M.; and the ARRA Award Number 1S10RR026780-01 from the National Center for Research Resources. Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2023",
month = jan,
day = "5",
doi = "10.1016/j.cell.2022.11.030",
language = "English (US)",
volume = "186",
pages = "112--130.e20",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "1",
}