tetano
Editor, Senior Moderator
Respir Res
. 2025 Jan 16;26(1):18.
doi: 10.1186/s12931-025-03095-y. Optimization of a micro-scale air-liquid-interface model of human proximal airway epithelium for moderate throughput drug screening for SARS-CoV-2
Chandani Sen[SUP] 1 [/SUP], Tammy M Rickabaugh[SUP] 1 [/SUP], Arjit Vijey Jeyachandran[SUP] 2 [/SUP], Constance Yuen[SUP] 2 3 [/SUP], Maisam Ghannam[SUP] 1 [/SUP], Abdo Durra[SUP] 1 [/SUP], Adam Aziz[SUP] 1 [/SUP], Kristen Castillo[SUP] 1 [/SUP], Gustavo Garcia Jr[SUP] 2 [/SUP], Arunima Purkayastha[SUP] 1 [/SUP], Brandon Han[SUP] 2 3 [/SUP], Felix W Boulton[SUP] 1 [/SUP], Eugene Chekler[SUP] 4 [/SUP], Robert Garces[SUP] 4 [/SUP], Karen C Wolff[SUP] 5 [/SUP], Laura Riva[SUP] 5 [/SUP], Melanie G Kirkpatrick[SUP] 5 [/SUP], Amal Gebara-Lamb[SUP] 5 [/SUP], Case W McNamara[SUP] 5 [/SUP], Ulrich A K Betz[SUP] #[/SUP][SUP] 6 [/SUP], Vaithilingaraja Arumugaswami[SUP] #[/SUP][SUP] 7 [/SUP], Robert Damoiseaux[SUP] #[/SUP][SUP] 8 9 10 [/SUP], Brigitte N Gomperts[SUP] #[/SUP][SUP] 11 12 13 14 15 [/SUP]
Affiliations
Background: Many respiratory viruses attack the airway epithelium and cause a wide spectrum of diseases for which we have limited therapies. To date, a few primary human stem cell-based models of the proximal airway have been reported for drug discovery but scaling them up to a higher throughput platform remains a significant challenge. As a result, most of the drug screening assays for respiratory viruses are performed on commercial cell line-based 2D cultures that provide limited translational ability.
Methods: We optimized a primary human stem cell-based mucociliary airway epithelium model of SARS-CoV-2 infection, in 96-well air-liquid-interface (ALI) format, which is amenable to moderate throughput drug screening. We tested the model against SARS-CoV-2 parental strain (Wuhan) and variants Beta, Delta, and Omicron. We applied this model to screen 2100 compounds from targeted drug libraries using a high throughput-high content image-based quantification method.
Results: The model recapitulated the heterogeneity of infection among patients with SARS-CoV-2 parental strain and variants. While there were heterogeneous responses across variants for host factor targeting compounds, the two direct-acting antivirals we tested, Remdesivir and Paxlovid, showed consistent efficacy in reducing infection across all variants and donors. Using the model, we characterized a new antiviral drug effective against both the parental strain and the Omicron variant.
Conclusion: This study demonstrates that the 96-well ALI model of primary human mucociliary epithelium can recapitulate the heterogeneity of infection among different donors and SARS-CoV-2 variants and can be used for moderate throughput screening. Compounds that target host factors showed variability among patients in response to SARS-CoV-2, while direct-acting antivirals were effective against SARS-CoV-2 despite the heterogeneity of patients tested.
Keywords: Air–liquid-interface; Anti-viral screening; Heterogeneity; High throughput drug screening; Human mucociliary epithelium; Image quantification; RNA sequencing; Respiratory viral infections; SARS-CoV-2; Small-molecules.
. 2025 Jan 16;26(1):18.
doi: 10.1186/s12931-025-03095-y. Optimization of a micro-scale air-liquid-interface model of human proximal airway epithelium for moderate throughput drug screening for SARS-CoV-2
Chandani Sen[SUP] 1 [/SUP], Tammy M Rickabaugh[SUP] 1 [/SUP], Arjit Vijey Jeyachandran[SUP] 2 [/SUP], Constance Yuen[SUP] 2 3 [/SUP], Maisam Ghannam[SUP] 1 [/SUP], Abdo Durra[SUP] 1 [/SUP], Adam Aziz[SUP] 1 [/SUP], Kristen Castillo[SUP] 1 [/SUP], Gustavo Garcia Jr[SUP] 2 [/SUP], Arunima Purkayastha[SUP] 1 [/SUP], Brandon Han[SUP] 2 3 [/SUP], Felix W Boulton[SUP] 1 [/SUP], Eugene Chekler[SUP] 4 [/SUP], Robert Garces[SUP] 4 [/SUP], Karen C Wolff[SUP] 5 [/SUP], Laura Riva[SUP] 5 [/SUP], Melanie G Kirkpatrick[SUP] 5 [/SUP], Amal Gebara-Lamb[SUP] 5 [/SUP], Case W McNamara[SUP] 5 [/SUP], Ulrich A K Betz[SUP] #[/SUP][SUP] 6 [/SUP], Vaithilingaraja Arumugaswami[SUP] #[/SUP][SUP] 7 [/SUP], Robert Damoiseaux[SUP] #[/SUP][SUP] 8 9 10 [/SUP], Brigitte N Gomperts[SUP] #[/SUP][SUP] 11 12 13 14 15 [/SUP]
Affiliations
- PMID: 39819574
- PMCID: PMC11740480
- DOI: 10.1186/s12931-025-03095-y
Background: Many respiratory viruses attack the airway epithelium and cause a wide spectrum of diseases for which we have limited therapies. To date, a few primary human stem cell-based models of the proximal airway have been reported for drug discovery but scaling them up to a higher throughput platform remains a significant challenge. As a result, most of the drug screening assays for respiratory viruses are performed on commercial cell line-based 2D cultures that provide limited translational ability.
Methods: We optimized a primary human stem cell-based mucociliary airway epithelium model of SARS-CoV-2 infection, in 96-well air-liquid-interface (ALI) format, which is amenable to moderate throughput drug screening. We tested the model against SARS-CoV-2 parental strain (Wuhan) and variants Beta, Delta, and Omicron. We applied this model to screen 2100 compounds from targeted drug libraries using a high throughput-high content image-based quantification method.
Results: The model recapitulated the heterogeneity of infection among patients with SARS-CoV-2 parental strain and variants. While there were heterogeneous responses across variants for host factor targeting compounds, the two direct-acting antivirals we tested, Remdesivir and Paxlovid, showed consistent efficacy in reducing infection across all variants and donors. Using the model, we characterized a new antiviral drug effective against both the parental strain and the Omicron variant.
Conclusion: This study demonstrates that the 96-well ALI model of primary human mucociliary epithelium can recapitulate the heterogeneity of infection among different donors and SARS-CoV-2 variants and can be used for moderate throughput screening. Compounds that target host factors showed variability among patients in response to SARS-CoV-2, while direct-acting antivirals were effective against SARS-CoV-2 despite the heterogeneity of patients tested.
Keywords: Air–liquid-interface; Anti-viral screening; Heterogeneity; High throughput drug screening; Human mucociliary epithelium; Image quantification; RNA sequencing; Respiratory viral infections; SARS-CoV-2; Small-molecules.