Human iPSC-derived alveolar and airway epithelial cells can be cultured at air-liquid interface and express SARS-CoV-2 host factors
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Abstract
Development of an anti-SARS-CoV-2 therapeutic is hindered by the lack of physiologically relevant model systems that can recapitulate host-viral interactions in human cell types, specifically the epithelium of the lung. Here, we compare induced pluripotent stem cell (iPSC)-derived alveolar and airway epithelial cells to primary lung epithelial cell controls, focusing on expression levels of genes relevant for COVID-19 disease modeling. iPSC-derived alveolar epithelial type II-like cells (iAT2s) and iPSC-derived airway epithelial lineages express key transcripts associated with lung identity in the majority of cells produced in culture. They express ACE2 and TMPRSS2 , transcripts encoding essential host factors required for SARS-CoV-2 infection, in a minor subset of each cell sub-lineage, similar to frequencies observed in primary cells. In order to prepare human culture systems that are amenable to modeling viral infection of both the proximal and distal lung epithelium, we adapt iPSC-derived alveolar and airway epithelial cells to two-dimensional air-liquid interface cultures. These engineered human lung cell systems represent sharable, physiologically relevant platforms for SARS-CoV-2 infection modeling and may therefore expedite the development of an effective pharmacologic intervention for COVID-19.
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SciScore for 10.1101/2020.06.03.132639: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Cells were incubated with anti-pro-SFTPC (Santa Cruz sc-518029, 1:500) primary antibody overnight at 4°C. anti-pro-SFTPCsuggested: NoneSoftware and Algorithms Sentences Resources RNA-Seq Comparison to Existing Human Lung Cell Line Models: RNA-Seq data from 150 lung cell lines were downloaded from the ArrayExpress repository with accession E-MTAB-2706 (https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-2706/) characterized by Klijn et al. (2015). ArrayExpresssuggested: (ArrayExpress, RRID:SCR_00…SciScore for 10.1101/2020.06.03.132639: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Cells were incubated with anti-pro-SFTPC (Santa Cruz sc-518029, 1:500) primary antibody overnight at 4°C. anti-pro-SFTPCsuggested: NoneSoftware and Algorithms Sentences Resources RNA-Seq Comparison to Existing Human Lung Cell Line Models: RNA-Seq data from 150 lung cell lines were downloaded from the ArrayExpress repository with accession E-MTAB-2706 (https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-2706/) characterized by Klijn et al. (2015). ArrayExpresssuggested: (ArrayExpress, RRID:SCR_002964)Clusters identified with the Louvain method were annotated based on their enrichment score for molecular signatures taken from the top 20 differentially expressed genes (computed using hurdle models for sparse single cell data, as implemented in the MAST package) in the dataset from Habermann et al. Generation of HIOs, digital gene expression, and qRT-PCR: The iPSC line BU1CG was differentiated to human intestinal organoids (HIOs) and profiled by digital gene expression as previously described (Mithal 2020). MASTsuggested: (MAST, RRID:SCR_016340)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:Despite the potential advantages of iPSC-based systems discussed above, we also recognize the limitations. Directed differentiation of human iPSCs to alveolar or airway epithelial-like cells is a time-consuming process (usually more than 40 days) and is technically challenging to master in a short period of time. This is especially inconvenient given the urgency to study SARS-CoV-2 in the midst of the current pandemic. To address this issue, we have developed approaches to facilitate sharing of iAT2s and iBCs as live cell cultures or frozen vials with other researchers. This information is listed under the Resource Sharing section of this manuscript. iAT2s and iBCs can be thawed and expanded in 3D organoid culture and are simple to maintain as spheres or culture at air-liquid interface for experimentation. In summary, we identify iPSC-derived alveolar and airway epithelial-like cells as a physiologically relevant model system with the potential to model components of SARS-CoV-2 infection such as viral entry, cellular response to pathogen, and viral replication. iAT2s and iBC-derived ALI cultures express the relevant viral entry genes and are an immediately sharable resource. We believe that iAT2s and iBCs can be used to provide insight into lung epithelial cell infection by SARS-CoV-2 and test therapeutic candidates. Resource Sharing:
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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