Differentiated Human Nasopharyngeal Organoids and Air-Liquid Interface Culture of Nasopharyngeal Epithelial Cells are susceptible models to study human rhinovirus C infection
Refereed conference paper presented and published in conference proceedings


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摘要Background
Rhinovirus C (RV-C) could cause severe lower respiratory infections and is associated with childhood asthma exacerbations. However, there is not yet a consistent and susceptible model to study RV-C since it cannot be propagated in immortalized cell lines. We aim to establish novel in vitro models, with define genetic background, to access the infectivity and pathogenesis of RV-C. We hypothesized that differentiated human nasopharyngeal (NP) organoid and air-liquid interface (ALI) cultures of NP epithelial cells could function as experimental models to study RV-C. Based on the selective expression of the RV-C receptor, cadherin related family member 3 (CDHR3), on ciliated epithelial cells, we postulated that increased ciliated cell differentiation in NP cultures enhances RV-C infectivity .
Method
NP swabs were collected from patients admitted to the Prince of Wales hospital, Hong Kong. Airway basal cells were propagated from cells obtained from the swab and further differentiated into human NP organoids and ALI cultures on transwells. The Notch signaling pathway inhibitor DAPT was added in the differentiation protocol to enhance ciliated cell differentiation. NP organoids and ALI cultures, differentiated with or without DAPT, were subjected to RV-C infection. Culture supernatants and RNA lysates were harvested at 6 and 24 hours post infection (hpi) to monitor the infectivity of RV-C in these models. CXCL10 gene expression was determined as an innate immune response biomarker.
Results
ALI cultures differentiated with DAPT showed a higher gene expression of the ciliated cell marker FOXJ1 and CDHR3 compared to those without DAPT treatment. RV-C replicated with a higher efficiency in NP organoids and ALI cultures when cultured with DAPT. Moreover, RV-C infection in both culture models, induced CXCL10 gene expression. A stronger induction was observed in the infected cultures with the addition of DAPT.
Conclusion
Ciliated cell differentiation and CDHR3 expression are enhanced by inhibiting the Notch signaling pathway. Moreover, Notch inhibition improves RV-C replication and an innate immune response in differentiated human NP organoids and ALI cultures. Our findings suggested that both NP organoids and ALI cultures are promising and potential models to investigate the pathogenesis of RV-C.
Acknowledgement
The project is supported by the CUHK Research Committee's One-off Funding for Joint Laboratory / Research Collaboration (3132958) .
著者Louisa LY Chan, Gimano D Amatngalim, Kin P Tao, Sjanna Besteman, Louis Bont, Jeffrey Beekman, Renee WY Chan
會議名稱Origins of Allergic Disease: Microbial, Epithelial and Immune Interactions
會議開始日24.03.2019
會議完結日27.03.2019
會議地點Tahoe City
會議國家/地區美國
出版年份2019
月份3
語言美式英語

上次更新時間 2019-04-12 於 12:44