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

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.

Evaluation of rapid transepithelial electrical resistance (TEER) measurement as a metric of kidney toxicity in a high-throughput microfluidic culture system

“…In addition to efficient readouts of barrier function, TEER sensing technology is conducive to multiplexing with other PT toxicity indicators, such as oxygen consumption and injury marker secretion, which could enable the generation of multi-parametric datasets with more predictive power. Broader application of rapid TEER sensing technology could also improve understanding of tight junction dynamics in a variety of biological contexts including diseases of the PT, such as polycystic kidney disease 51 , or in other tissue types such as lung 52 , vascular 53 , and liver 54 , which have been previously modeled in PREDICT96.…”

Human organs-on-chips for disease modelling, drug development and personalized medicine

“…1e ). A streamlined plastic 384-well format organ chip design also was described recently and used to create an air–liquid interface 21 (Fig. 1f ).…”

“…Most organ chips are also low throughput, but high content; this can be valuable at later stages in the drug development pipeline (for example, where selection of lead compounds must be made for drug advancement), but not in the earlier discovery phase. Higher-throughput systems are needed to enable practical implementation of statistically significant studies with many replicates, which is essential for pharmaceutical validation, and indeed, the recent development of higher-throughput organ chips (that is, single devices containing many parallel culture chambers) that still retain many of the key features required for drug developers 21 , 120 (Table 1 ) suggest that this path should be pursued as well. However, this goal also can be met by developing automated instruments that can culture and analyse multiple lower-throughput organ chips simultaneously.…”

Organoid Models of SARS-CoV-2 Infection: What Have We Learned about COVID-19?

“…ALI-differentiated cultures established from bronchial lavage or lung resections in a 96-well format have recently been established as a high-throughput research platform (for example, [97]). Several respiratory tissue-derived ALI cultures are now commercially available (including nose, bronchial and alveolar).…”