High-throughput human primary cell-based airway model for evaluating influenza, coronavirus, or other respiratory viruses in vitro

Sci Rep. 2021 Jul 22;11(1):14961. 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.

Publication types

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

MeSH terms

  • Antiviral Agents / pharmacology*
  • Bronchi / cytology
  • Bronchi / virology
  • COVID-19 / drug therapy
  • COVID-19 / virology
  • Cell Culture Techniques / instrumentation
  • Cell Line
  • Coronavirus / drug effects
  • Coronavirus Infections / drug therapy
  • Coronavirus Infections / virology*
  • Equipment Design
  • High-Throughput Screening Assays / instrumentation
  • Humans
  • Influenza A virus / drug effects
  • Influenza, Human / drug therapy
  • Influenza, Human / virology*
  • Microbial Sensitivity Tests / instrumentation*
  • Microfluidic Analytical Techniques / instrumentation*
  • Respiratory Mucosa / cytology*
  • Respiratory Mucosa / virology
  • Respiratory Tract Infections / drug therapy
  • Respiratory Tract Infections / virology
  • SARS-CoV-2 / drug effects

Substances

  • Antiviral Agents