Molecular characterization of Calu-3 cells from submerged to Air-liquid interface to model lung infections

ABSTRACT: Air-liquid interface (ALI) model using the immortalized Calu-3 cells has been used to model lung diseases. From submerged to ALI culture condition, Calu-3 cells polarize and change to a ciliary mucus-producing cell. Polarized Calu-3 molecular and phenotypic similarity with primary cells has been proven; however, no studies have been focused on the pathways differentially expressed in ALI vs. submerged conditions. In this study, we profiled the proteome and transcriptome of Calu-3 cells from submerged (non-polarized, NP) to ALI (polarized, P) culture conditions and in the omics data we observed an increase in cell replication in the non-polarized condition while polarized Calu-3 cells presented higher activation of cellular energy production, protein maturation and recycle and expression of immune active molecules. Moreover, the omics findings showed upregulation of different biological processes related to protein quality control system and antigen processing presentation in polarized Calu-3 cells. Immunoblot and fluorescence microscopy confirmed increased expression of bronchial epithelium integrity components like mucus and tight junctions in polarized Calu-3 and revealed a characteristic protein expression and cellular organization found in normal lung epithelial tissue. Furthermore, SARS-CoV-2 infection of polarized cells revealed higher cell death due to higher viral entry associated with higher apical membrane expression of ACE2 receptor. The differences observed in this study give us a better understanding of how ALI can mimic the human bronchial-epithelial cells and its applications in different contexts of lung diseases.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Deivid Martins Santos  

LAB HEAD: Giuseppe Palmisano

PROVIDER: PXD056869 | Pride | 2026-02-23

REPOSITORIES: Pride