A Newly Engineered A549 Cell Line Expressing ACE2 and TMPRSS2 Is Highly Permissive to SARS-CoV-2, Including the Delta and Omicron Variants

Ching-Wen Chang; Krishna Mohan Parsi; Mohan Somasundaran; Emma Vanderleeden; Ping Liu; John Cruz; Alyssa Cousineau; Robert W Finberg; Evelyn A Kurt-Jones

doi:10.3390/v14071369

A Newly Engineered A549 Cell Line Expressing ACE2 and TMPRSS2 Is Highly Permissive to SARS-CoV-2, Including the Delta and Omicron Variants

Viruses. 2022 Jun 23;14(7):1369. doi: 10.3390/v14071369.

Authors

Ching-Wen Chang¹, Krishna Mohan Parsi², Mohan Somasundaran³, Emma Vanderleeden¹, Ping Liu¹, John Cruz⁴, Alyssa Cousineau², Robert W Finberg¹, Evelyn A Kurt-Jones¹

Affiliations

¹ Department of Medicine, University of Massachusetts Chan Medical School, Worcester, MA 01655, USA.
² Diabetes Center of Excellence and Program in Molecular Medicine, University of Massachusetts Chan Medical School, Worcester, MA 01655, USA.
³ Department of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, MA 01655, USA.
⁴ Department of Pathology, University of Massachusetts Chan Medical School, Worcester, MA 01655, USA.

Abstract

New variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to emerge, causing surges, breakthrough infections, and devastating losses-underscoring the importance of identifying SARS-CoV-2 antivirals. A simple, accessible human cell culture model permissive to SARS-CoV-2 variants is critical for identifying and assessing antivirals in a high-throughput manner. Although human alveolar A549 cells are a valuable model for studying respiratory virus infections, they lack two essential host factors for SARS-CoV-2 infection: angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2). SARS-CoV-2 uses the ACE2 receptor for viral entry and TMPRSS2 to prime the SARS-CoV-2 spike protein, both of which are negligibly expressed in A549 cells. Here, we report the generation of a suitable human cell line for SARS-CoV-2 studies by transducing human ACE2 and TMPRSS2 into A549 cells. We show that subclones highly expressing ACE2 and TMPRSS2 ("ACE2plus" and the subclone "ACE2plusC3") are susceptible to infection with SARS-CoV-2, including the delta and omicron variants. These subclones express more ACE2 and TMPRSS2 transcripts than existing commercial A549 cells engineered to express ACE2 and TMPRSS2. Additionally, the antiviral drugs EIDD-1931, remdesivir, nirmatrelvir, and nelfinavir strongly inhibit SARS-CoV-2 variants in our infection model. Our data show that ACE2plusC3 cells are highly permissive to SARS-CoV-2 infection and can be used to identify anti-SARS-CoV-2 drugs.

Keywords: A549; ACE2; EIDD-1931; SARS-CoV-2; TMPRSS2; delta and omicron variants; nirmatrelvir; remdesivir.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

A549 Cells
Angiotensin-Converting Enzyme 2* / genetics
Antiviral Agents / pharmacology
COVID-19*
Humans
Peptidyl-Dipeptidase A / metabolism
SARS-CoV-2 / genetics
Serine Endopeptidases / genetics
Spike Glycoprotein, Coronavirus / genetics
Spike Glycoprotein, Coronavirus / metabolism

Substances

Antiviral Agents
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2
Peptidyl-Dipeptidase A
Angiotensin-Converting Enzyme 2
Serine Endopeptidases
TMPRSS2 protein, human

Supplementary concepts

SARS-CoV-2 variants

Grants and funding

This work was supported by the Department of Defense Award (W81XWH2110029) “COVID-19 Expansion for AIRe” to R.W.F. and a Ferring COVID-19 Investigational Grant to C.-W.C.