Title : Establishment of a human lung organoid method platform for infectious disease modelling
Abstract:
Background: In recent years, organoids have advanced to prominent and versatile tools in 3R research, representing valuable alternatives for animal models and physiologically relevant human disease models at the same time. As the federal Public Health Institute, the RKI aims to establish an in-house multi-species, multi-organ organoid platform. The objective is a broad methodology in order to strengthen the Institute`s pandemic preparedness plans for novel and re-emerging pathogens, which often have to be handled in a BSL4 laboratory. So far, in our group, bronchial and nasal lung organoids have been generated from adult stem cells derived from various primary and commercial cells. Characterisation on mRNA and protein level showed that, with small variances, all organoids consist of a physiologically applicable cell composition.
Methodology: Single-cell RNA sequencing (scRNA sec) should be applied to investigate complex questions like e.g. viral cell tropism, host cell response etc. Less generalized questions regarding differential gene expression during a viral infection can be tackled with the Nanostring method. In order to study inter-organ viral dissemination, as well as inter-species transmission of pathogens, the relatively new multi-organ-chip technology is currently being established. Classic antiviral tests are also included in our method range. Once conditions will be established, these tests will be applied to the multi-organ-chip format, which allows physiologically more relevant tests. Finally, using a fluorescent labelled virus, 4D image data have been generated and with the help of deep learning, will be used for modelling viral replication kinetics.
Results: Quality control scRNA sec runs yielded a good cell viability, sufficing for future runs. Differential gene expression following infection of lung organoids with the BSL4 pathogens EBOV, MARV, NiV and LASV has been studied by a Nanostring host-response panel and is currently being analysed. Preliminary infection of organoids on organ chips with VACV-GFP have been performed and successful inter-organ transmission could be observed. Furthermore, the anti-viral effect of the two clinically licensed drugs itraconazole and fluoxetine has been tested in EBOV-infected organoids. Results are currently being analysed. Lastly, the successfully generated 4D data is currently being processed by our AI department.
