tetano
Editor, Senior Moderator
Sci Rep
. 2021 Jul 22;11(1):14961.
doi: 10.1038/s41598-021-94095-7.
High-throughput human primary cell-based airway model for evaluating influenza, coronavirus, or other respiratory viruses in vitro
A L Gard[SUP] 1 [/SUP], R J Luu[SUP] 1 [/SUP], C R Miller[SUP] 1 [/SUP], R Maloney[SUP] 1 [/SUP], B P Cain[SUP] 1 [/SUP], E E Marr[SUP] 1 [/SUP], D M Burns[SUP] 1 [/SUP], R Gaibler[SUP] 1 [/SUP], T J Mulhern[SUP] 1 [/SUP], C A Wong[SUP] 1 [/SUP], J Alladina[SUP] 2 [/SUP], J R Coppeta[SUP] 1 [/SUP], P Liu[SUP] 3 [/SUP], J P Wang[SUP] 3 [/SUP], H Azizgolshani[SUP] 1 [/SUP], R Fennell Fezzie[SUP] 1 [/SUP], J L Balestrini[SUP] 1 [/SUP], B C Isenberg[SUP] 1 [/SUP], B D Medoff[SUP] 2 [/SUP], R W Finberg[SUP] 3 [/SUP], J T Borenstein[SUP] 4 [/SUP]
Affiliations
Abstract
Influenza and other respiratory viruses present a significant threat to public health, national security, and the world economy, and can lead to the emergence of global pandemics such as from COVID-19. A barrier to the development of effective therapeutics is the absence of a robust and predictive preclinical model, with most studies relying on a combination of in vitro screening with immortalized cell lines and low-throughput animal models. Here, we integrate human primary airway epithelial cells into a custom-engineered 96-device platform (PREDICT96-ALI) in which tissues are cultured in an array of microchannel-based culture chambers at an air-liquid interface, in a configuration compatible with high resolution in-situ imaging and real-time sensing. We apply this platform to influenza A virus and coronavirus infections, evaluating viral infection kinetics and antiviral agent dosing across multiple strains and donor populations of human primary cells. Human coronaviruses HCoV-NL63 and SARS-CoV-2 enter host cells via ACE2 and utilize the protease TMPRSS2 for spike protein priming, and we confirm their expression, demonstrate infection across a range of multiplicities of infection, and evaluate the efficacy of camostat mesylate, a known inhibitor of HCoV-NL63 infection. This new capability can be used to address a major gap in the rapid assessment of therapeutic efficacy of small molecules and antiviral agents against influenza and other respiratory viruses including coronaviruses.
. 2021 Jul 22;11(1):14961.
doi: 10.1038/s41598-021-94095-7.
High-throughput human primary cell-based airway model for evaluating influenza, coronavirus, or other respiratory viruses in vitro
A L Gard[SUP] 1 [/SUP], R J Luu[SUP] 1 [/SUP], C R Miller[SUP] 1 [/SUP], R Maloney[SUP] 1 [/SUP], B P Cain[SUP] 1 [/SUP], E E Marr[SUP] 1 [/SUP], D M Burns[SUP] 1 [/SUP], R Gaibler[SUP] 1 [/SUP], T J Mulhern[SUP] 1 [/SUP], C A Wong[SUP] 1 [/SUP], J Alladina[SUP] 2 [/SUP], J R Coppeta[SUP] 1 [/SUP], P Liu[SUP] 3 [/SUP], J P Wang[SUP] 3 [/SUP], H Azizgolshani[SUP] 1 [/SUP], R Fennell Fezzie[SUP] 1 [/SUP], J L Balestrini[SUP] 1 [/SUP], B C Isenberg[SUP] 1 [/SUP], B D Medoff[SUP] 2 [/SUP], R W Finberg[SUP] 3 [/SUP], J T Borenstein[SUP] 4 [/SUP]
Affiliations
- PMID: 34294757
- DOI: 10.1038/s41598-021-94095-7
Abstract
Influenza and other respiratory viruses present a significant threat to public health, national security, and the world economy, and can lead to the emergence of global pandemics such as from COVID-19. A barrier to the development of effective therapeutics is the absence of a robust and predictive preclinical model, with most studies relying on a combination of in vitro screening with immortalized cell lines and low-throughput animal models. Here, we integrate human primary airway epithelial cells into a custom-engineered 96-device platform (PREDICT96-ALI) in which tissues are cultured in an array of microchannel-based culture chambers at an air-liquid interface, in a configuration compatible with high resolution in-situ imaging and real-time sensing. We apply this platform to influenza A virus and coronavirus infections, evaluating viral infection kinetics and antiviral agent dosing across multiple strains and donor populations of human primary cells. Human coronaviruses HCoV-NL63 and SARS-CoV-2 enter host cells via ACE2 and utilize the protease TMPRSS2 for spike protein priming, and we confirm their expression, demonstrate infection across a range of multiplicities of infection, and evaluate the efficacy of camostat mesylate, a known inhibitor of HCoV-NL63 infection. This new capability can be used to address a major gap in the rapid assessment of therapeutic efficacy of small molecules and antiviral agents against influenza and other respiratory viruses including coronaviruses.