SARS-CoV-2 infection models using in vivo and in vitro hACE2-lentivirus transduction

Rawle, D. J.; Le, T. T.; Dumenil, T.; Yan, K.; Tang, B.; Bishop, C.; Suhrbier, A.

doi:10.1101/2021.02.09.430547

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bioRxiv · 10.1101/2021.02.09.430547

SARS-CoV-2 infection models using in vivo and in vitro hACE2-lentivirus transduction

Rawle, D. J.·Le, T. T.·Dumenil, T.·Yan, K.·Tang, B.·Bishop, C.·Suhrbier, A.

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Abstract

SARS-CoV-2 uses the human ACE2 (hACE2) receptor for cell attachment and entry, with mouse ACE2 (mACE2) unable to support infection. Herein we describe an ACE2-lentivirus system and illustrate its utility for in vitro and in vivo SARS-CoV-2 infection models. Transduction of non-permissive cell lines with hACE2 imparted replication competence, and transduction with mACE2 containing N30D, N31K, F83Y and H353K substitutions, to match hACE2, rescued SARS-CoV-2 replication. Intranasal hACE2-lentivirus transduction of C57BL/6J mice permitted significant virus replication in lungs. RNA-Seq analyses illustrated that the model involves an acute inflammatory disease followed by resolution and tissue repair, with a transcriptomic profile similar to that seen in COVID-19 patients. Intranasal hACE2-lentivirus transduction of IFNAR-/- and IL-28RA-/- mice lungs was used to illustrate that loss of type I or III interferon responses have no significant effect on virus replication. However, their importance in driving inflammatory responses was illustrated by RNA-Seq analyses. We also demonstrate the utility of the hACE2-lentivirus transduction system for vaccine evaluation in C57BL/6J mice. The ACE2-lentivirus system thus has broad application in SARS-CoV-2 research, providing a tool for both mutagenesis studies and mouse model development. AUTHOR SUMMARYSARS-CoV-2 uses the human ACE2 (hACE2) receptor to infect cells, but cannot infect mice because the virus cannot bind mouse ACE2 (mACE2). We use an ACE2-lentivirus system in vitro to identify four key amino acids in mACE2 that explain why SARS-CoV-2 cannot infect mice. hACE2-lentivirus was used to express hACE2 in mouse lungs in vivo, with the inflammatory responses after SARS-CoV-2 infection similar to those seen in human COVID-19. Genetically modified mice were used to show that type I and III interferon signaling is required for the inflammatory responses. We also show that the hACE2-lentivirus mouse model can be used to test vaccines. Overall this paper demonstrates that our hACE2-lentivirus system has multiple applications in SARS-CoV-2 and COVID-19 research.

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Rawle, D. J., Le, T. T., Dumenil, T., Yan, K., Tang, B., Bishop, C., Suhrbier, A.. 2021-02-10. SARS-CoV-2 infection models using in vivo and in vitro hACE2-lentivirus transduction. https://doi.org/10.1101/2021.02.09.430547

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