@article{37bbdc9a1529417cb980fab26dabd4b6,
title = "Systematic analysis of SARS-CoV-2 infection of an ACE2-negative human airway cell",
abstract = "Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) variants govern transmissibility, responsiveness to vaccination, and disease severity. In a screen for new models of SARS-CoV-2 infection, we identify human H522 lung adenocarcinoma cells as naturally permissive to SARS-CoV-2 infection despite complete absence of angiotensin-converting enzyme 2 (ACE2) expression. Remarkably, H522 infection requires the E484D S variant; viruses expressing wild-type S are not infectious. Anti-S monoclonal antibodies differentially neutralize SARS-CoV-2 E484D S in H522 cells as compared to ACE2-expressing cells. Sera from vaccinated individuals block this alternative entry mechanism, whereas convalescent sera are less effective. Although the H522 receptor remains unknown, depletion of surface heparan sulfates block H522 infection. Temporally resolved transcriptomic and proteomic profiling reveal alterations in cell cycle and the antiviral host cell response, including MDA5-dependent activation of type I interferon signaling. These findings establish an alternative SARS-CoV-2 host cell receptor for the E484D SARS-CoV-2 variant, which may impact tropism of SARS-CoV-2 and consequently human disease pathogenesis.",
keywords = "ACE2-independent, COVID-19, RIG-I-like receptors, SARS-CoV-2, clathrin-mediated endocytosis, heparan sulfate, proteomics, spike variants, type I interferon, virus-host interactions",
author = "Maritza Puray-Chavez and LaPak, {Kyle M.} and Schrank, {Travis P.} and Elliott, {Jennifer L.} and Bhatt, {Dhaval P.} and Agajanian, {Megan J.} and Ria Jasuja and Lawson, {Dana Q.} and Keanu Davis and Rothlauf, {Paul W.} and Zhuoming Liu and Heejoon Jo and Nakyung Lee and Kasyap Tenneti and Eschbach, {Jenna E.} and {Shema Mugisha}, Christian and Cousins, {Emily M.} and Cloer, {Erica W.} and Vuong, {Hung R.} and VanBlargan, {Laura A.} and Bailey, {Adam L.} and Pavlo Gilchuk and Crowe, {James E.} and Diamond, {Michael S.} and Hayes, {D. Neil} and Whelan, {Sean P.J.} and Amjad Horani and Brody, {Steven L.} and Dennis Goldfarb and Major, {M. Ben} and Kutluay, {Sebla B.}",
note = "Funding Information: S.P.J.W., P.W.R., and Washington University have filed a patent application for uses of VSV-SARS-CoV-2. S.P.J.W. has received unrelated funding support in sponsored research agreements with Vir Biotechnology, Abbvie, and SAB therapeutics. M.S.D. is a consultant for Inbios, Vir Biotechnology, Fortress Biotech, and Carnival Corporation, and is on the Scientific Advisory Boards of Moderna and Immunome. The Diamond laboratory has received funding support in sponsored research agreements from Moderna, Vir Biotechnology, Kaleido, and Emergent BioSolutions. J.E.C. has served as a consultant for Luna Biologics, is a member of the Scientific Advisory Boards of Meissa Vaccines, and is Founder of IDBiologics. The Crowe laboratory has received sponsored research agreements from Takeda, AstraZeneca, and IDBiologics. Vanderbilt University has applied for patents related to antibodies described in this paper. The remaining authors declare no competing interests. Funding Information: This work was supported by Washington University funds to S.B.K., a V Foundation grant ( T2014-009 ) to M.B.M. and D.N.H., a T32 training grant ( T32CA009547-34 ) to K.M.L., a T32 training grant ( 5T32HL007106-39 ) to E.M.C., the Dorothy R. and Hubert C. Moog Professor of Medicine to S.L.B., a K08HL150223 grant to A.H., NIH AI059371 to S.P.J.W., NIH R01 AI157155 to M.S.D., NIH 75N93019C00074 , NIH R01 AI157155 , and the Defense Advanced Research Projects Agency ( HR001117S0019 ) grants to J.E.C. This study utilized samples obtained from the Washington University School of Medicine{\textquoteright}s COVID-19 biorepository supported by NIH /National Center for Advancing Translational Sciences grant UL1 TR002345. We thank Drs. Ali Ellebedy, Rachel Presti, Jane O{\textquoteright}Halloran, Daved Fremont, and Paul D. Bieniasz for key reagents utilized in the study, including neutralizing antibodies, sera from vaccine recipients, convalescent sera, Fc-ACE2, and plasmids. We additionally thank other members of the Whelan and Diamond laboratories for reagents. Funding Information: This work was supported by Washington University funds to S.B.K. a V Foundation grant (T2014-009) to M.B.M. and D.N.H. a T32 training grant (T32CA009547-34) to K.M.L. a T32 training grant (5T32HL007106-39) to E.M.C. the Dorothy R. and Hubert C. Moog Professor of Medicine to S.L.B. a K08HL150223 grant to A.H. NIH AI059371 to S.P.J.W. NIH R01 AI157155 to M.S.D. NIH 75N93019C00074, NIH R01 AI157155, and the Defense Advanced Research Projects Agency (HR001117S0019) grants to J.E.C. This study utilized samples obtained from the Washington University School of Medicine's COVID-19 biorepository supported by NIH/National Center for Advancing Translational Sciences grant UL1 TR002345. We thank Drs. Ali Ellebedy, Rachel Presti, Jane O'Halloran, Daved Fremont, and Paul D. Bieniasz for key reagents utilized in the study, including neutralizing antibodies, sera from vaccine recipients, convalescent sera, Fc-ACE2, and plasmids. We additionally thank other members of the Whelan and Diamond laboratories for reagents. M.P.-C. K.M.L. M.B.M. and S.B.K. conceptualized the study; M.P.-C. K.M.L. M.B.M. and S.B.K. designed the methodology. M.P.-C. K.M.L. J.L.E. D.P.B. M.J.A. D.Q.L. K.D. K.T. J.E.E. C.S.M. H.R.V. E.M.C. E.W.C. and S.B.K performed the experiments. T.P.S. R.J. H.J. and D.G. performed all statistical and bioinformatics analyses with help from N.L. and K.T. P.W.R. Z.L. A.L.B. L.A.V. M.S.D. P.G. and J.E.C. generated and provided key reagents. A.H. and S.L.B. generated and cultured primary basal epithelial cells. K.M.L. M.B.M. and S.B.K. wrote the manuscript with input from all of the authors. S.P.J.W. P.W.R. and Washington University have filed a patent application for uses of VSV-SARS-CoV-2. S.P.J.W. has received unrelated funding support in sponsored research agreements with Vir Biotechnology, Abbvie, and SAB therapeutics. M.S.D. is a consultant for Inbios, Vir Biotechnology, Fortress Biotech, and Carnival Corporation, and is on the Scientific Advisory Boards of Moderna and Immunome. The Diamond laboratory has received funding support in sponsored research agreements from Moderna, Vir Biotechnology, Kaleido, and Emergent BioSolutions. J.E.C. has served as a consultant for Luna Biologics, is a member of the Scientific Advisory Boards of Meissa Vaccines, and is Founder of IDBiologics. The Crowe laboratory has received sponsored research agreements from Takeda, AstraZeneca, and IDBiologics. Vanderbilt University has applied for patents related to antibodies described in this paper. The remaining authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021 The Author(s)",
year = "2021",
month = jul,
day = "13",
doi = "10.1016/j.celrep.2021.109364",
language = "English",
volume = "36",
journal = "Cell Reports",
issn = "2211-1247",
number = "2",
}