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

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.
Ching-Wen Chang 1,  Krishna Mohan Parsi 2,  Mohan Somasundaran 3,  Emma Vanderleeden 1,  Ping Liu 1,  John Cruz 4,  Alyssa Cousineau 2,  Robert W Finberg 1,  Evelyn A Kurt-Jones 1
Affiliations

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.
PMID: 35891350 DOI: 10.3390/v14071369
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.

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