ACE2-lentiviral transduction enables mouse SARS-CoV-2 infection and mapping of receptor interactions

PLoS Pathog. 2021 Jul 2;17(7):e1009723. doi: 10.1371/journal.ppat.1009723. eCollection 2021 Jul.

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. Intrapulmonary hACE2-lentivirus transduction of C57BL/6J mice permitted significant virus replication in lung epithelium. RNA-Seq and histological analyses illustrated that this model involved an acute inflammatory disease followed by resolution and tissue repair, with a transcriptomic profile similar to that seen in COVID-19 patients. hACE2-lentivirus transduction of IFNAR-/- and IL-28RA-/- mouse 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin-Converting Enzyme 2* / biosynthesis
  • Angiotensin-Converting Enzyme 2* / genetics
  • Animals
  • COVID-19* / genetics
  • COVID-19* / metabolism
  • Chlorocebus aethiops
  • Disease Models, Animal
  • Gene Expression Profiling*
  • Humans
  • Lentivirus*
  • Mice
  • Mice, Knockout
  • SARS-CoV-2* / genetics
  • SARS-CoV-2* / metabolism
  • Transduction, Genetic*
  • Vero Cells

Substances

  • ACE2 protein, human
  • Angiotensin-Converting Enzyme 2

Grants and funding

We thank Clive Berghofer and the Brazil Family Foundation (and many others) for their generous philanthropic donations to support SARS-CoV-2 research at QIMR Berghofer MRI (https://www.qimrberghofer.edu.au/) (intramural grants awarded to A.S. and D.J.R). The project was also partly funded by an intramural seed grant from the Australian Infectious Diseases Research Centre (https://aidrc.org.au/) awarded to A.S. A.S. holds an Investigator grant from the National Health and Medical Research Council (NHMRC, https://www.nhmrc.gov.au/) of Australia (APP1173880). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.