Abstract
Understanding pulmonary diseases requires robust culture models that are reproducible, sustainable in long-term culture, physiologically relevant, and suitable for assessment of therapeutic interventions. Primary human lung cells are physiologically relevant but cannot be cultured in vitro long term and, although engineered organoids are an attractive choice, they do not phenotypically recapitulate the lung parenchyma; overall, these models do not allow for the generation of reliable disease models. Recently, we described a new cell culture platform based on H441 cells that are grown at the air-liquid interface to produce the SALI culture model, for studying and correcting the rare interstitial lung disease surfactant protein B (SPB) deficiency. Here, we report the characterization of the effects of SALI culture conditions on the transcriptional profile of the constituent H441 cells. We further analyze the transcriptomics of the model in the context of surfactant metabolism and the disease phenotype through SFTPB knockout SALI cultures. By comparing the gene expression profile of SALI cultures with that of human lung parenchyma obtained via single-cell RNA sequencing, we found that SALI cultures are remarkably similar to human alveolar type II cells, implying clinical relevance of the SALI culture platform as a non-diseased human lung alveolar cell model.
Highlights
Interstitial lung diseases (ILDs) comprise a variety of disorders broadly related to the lung interstitium that are characterized by inflammation and buildup of scar tissue in the lungs.[1]
We explore the transcriptomic profiles of the wild-type (WT) and KO surfactant air-liquid interface (SALI) culture models to assess how this platform relates to the primary human alveolar epithelium and how the gene expression profiles of H441 cells respond to the different culturing conditions
Through RNA sequencing (RNA-seq) analyses, we show that the Global transcriptome analysis of the four different cultures clearly demonstrated that the SALI culture condition was the major driver of the transcriptional changes in H441 cells (Figure 1A)
Summary
Interstitial lung diseases (ILDs) comprise a variety of disorders broadly related to the lung interstitium that are characterized by inflammation and buildup of scar tissue in the lungs.[1]. Further understanding the mechanistic outcomes of the disorders, as well as screening potential drug and treatment approaches, requires robust models of ILDs, based on the alveolar epithelium comprising alveolar type II (ATII) and I (ATI) cells. As primary human alveolar cells lose their functional characteristics when cultured ex vivo,[5,6] a number of approaches have been undertaken to develop such pulmonary models in vitro. A promising approach was the establishment of organoid-like spheres from isolated primary human ATII cells or stem cells,[7] but these alveolospheres failed to replicate the structure of the alveolus, lacking cells expressing markers of ATI cells. In the case of proximal lung bud organoids, these were phenotypically closer to a developing fetal lung.[8,9,10] these models do not allow for systematic manipulation of cells to reliably generate successful disease models, as they cannot be reliably expanded in culture following CRISPR-Cas9-based interventions
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Molecular Therapy - Methods & Clinical Development
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
