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Changes in microRNA and mRNA expression with differentiation of human bronchial epithelial cells [miRNA]

ABSTRACT: Normal human bronchial epithelial (NHBE) cells cultured in an air-liquid interface (ALI) system form a polarized, pseudostratified epithelium composed of basal, ciliated and goblet cells that closely resemble the in vivo airway epithelium structure. ALI cultures of NHBE cells provide a unique in vitro system to investigate airway epithelial biology, including developmental, structural and physiologic aspects. MicroRNAs (miRNAs) are short, single-stranded, non-coding RNAs of 20-23 nucleotides that down-regulate gene expression by either inducing degradation of target mRNAs or impairing their translation. They are phylogenetically well conserved, which probably implies an important role of miRNAs in biological processes. In this way, we wanted to shed some light on miRNA specific roles and the relationship with their mRNA targets during airway epithelium differentiation. By using microarrays, we studied the changes in expression of microRNAs in normal human bronchial epithelial cells as they differentiate from an undifferentiated monolayer to a differentiated pseudostratified epithelium after 28 days of air-liquid interface (ALI) culture, when epithelial cells differentially express basal, ciliated and goblet cell markers.

ORGANISM(S): synthetic construct Homo sapiens

PROVIDER: GSE39060 | GEO | 2012/10/01

SECONDARY ACCESSION(S): PRJNA169856

REPOSITORIES: GEO

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Changes in microRNA and mRNA expression with differentiation of human bronchial epithelial cells [mRNA]

Project description:Normal human bronchial epithelial (NHBE) cells cultured in an air-liquid interface (ALI) system form a polarized, pseudostratified epithelium composed of basal, ciliated and goblet cells that closely resemble the in vivo airway epithelium structure. ALI cultures of NHBE cells provide a unique in vitro system to investigate airway epithelial biology, including developmental, structural and physiologic aspects. In this study, we wanted to investigate mRNA expression patterns during airway epithelium differentiation. By using microarrays, we studied the changes in expression of mRNAs in normal human bronchial epithelial cells as they differentiate from an undifferentiated monolayer to a differentiated pseudostratified epithelium after 28 days of air-liquid interface (ALI) culture, when epithelial cells differentially express basal, ciliated and goblet cell markers. Normal human bronchial epithelial cells were cultured in an air-liquid interface (ALI) system and harvested at three different time-points: subconfluent, confluent and day 28 of ALI. Samples were processed for total RNA extraction and hybridization on Affymetrix microarrays. All the experiments were performed by triplicate.

2012-10-01 | E-GEOD-39059 | biostudies-arrayexpress

Changes in microRNA and mRNA expression with differentiation of human bronchial epithelial cells [mRNA]

2012-10-01 | GSE39059 | GEO

Changes in microRNA and mRNA expression with differentiation of human bronchial epithelial cells [miRNA]

Project description:Normal human bronchial epithelial (NHBE) cells cultured in an air-liquid interface (ALI) system form a polarized, pseudostratified epithelium composed of basal, ciliated and goblet cells that closely resemble the in vivo airway epithelium structure. ALI cultures of NHBE cells provide a unique in vitro system to investigate airway epithelial biology, including developmental, structural and physiologic aspects. MicroRNAs (miRNAs) are short, single-stranded, non-coding RNAs of 20-23 nucleotides that down-regulate gene expression by either inducing degradation of target mRNAs or impairing their translation. They are phylogenetically well conserved, which probably implies an important role of miRNAs in biological processes. In this way, we wanted to shed some light on miRNA specific roles and the relationship with their mRNA targets during airway epithelium differentiation. By using microarrays, we studied the changes in expression of microRNAs in normal human bronchial epithelial cells as they differentiate from an undifferentiated monolayer to a differentiated pseudostratified epithelium after 28 days of air-liquid interface (ALI) culture, when epithelial cells differentially express basal, ciliated and goblet cell markers. Normal human bronchial epithelial cells were cultured in an air-liquid interface (ALI) system and harvested at three different time-points: subconfluent, confluent and day 28 of ALI. Samples were processed for total RNA (including small RNAs) extraction and hybridization on Affymetrix microarrays. All the experiments were performed by triplicate.

2012-10-01 | E-GEOD-39060 | biostudies-arrayexpress

Do Airway Epithelium Air-liquid Cultures Represent the In Vivo Airway Epithelium Transcriptome?

Project description:Human airway epithelial cells cultured in vitro at air-liquid interface (ALI) form a pseudostratified epithelium that forms tight junctions and cilia, and produces mucin, and are widely used as a model of differentiation, injury, and repair. To assess how closely the transcriptome of ALI epithelium matches that of in vivo airway epithelial cells, we used microarrays to compare the transcriptome of human large airway epithelial cells cultured at ALI with the transcriptome of large airway epithelium obtained via bronchoscopy and brushing. Gene expression profiling showed global gene expression correlated well between ALI cells and brushed cells, but there were some differences. Gene expression patterns mirrored differences in proportions of cell types (ALI have higher percentages of basal cells, brushed cells have higher percentages of ciliated cells), with ALI cells expressing higher levels of basal cell-related genes and brushed cells expressing higher levels of cilia-related genes. Pathway analysis showed ALI cells had increased expression of cell cycle and proliferation genes, while brushed cells had increased expression of cytoskeletal organization and humoral immune response genes. Overall, ALI cells are a good representation of the in vivo airway epithelial transcriptome, but for some biologic questions, the differences in the in vitro vs in vivo environments need to be considered. Affymetrix arrays were used to assess the gene expression of large airway cells cultured in vitro at air-liquid interface (12 samples) and large airway epithelial cells obtained by fiberoptic bronchoscopy of 20 healthy nonsmokers. *** Air-liquid interface Samples not provided in this Series. ***

2010-07-07 | E-GEOD-18637 | biostudies-arrayexpress

Do Airway Epithelium Air-liquid Cultures Represent the In Vivo Airway Epithelium Transcriptome?

2010-07-07 | GSE18637 | GEO

Analysis of the apical secretome of Calu-3 and NHBE cells in 3D models of the human bronchial epithelium

Project description:The mucus secreted by airway epithelial cells plays a key role in the protection against and clearance of particles and pathogens. In this project, a 3D model of the bronchial epithelium was established using Calu-3 cells grown on porous inserts at the air-liquid interface. The secreted proteins were characterized in long-term cultures and compared to the apical secretome of primary normal human bronchial epithelial (NHBE) cells. The apical secretome was collected and characterized in the Calu-3 model at day 4, day 11, and day 18 after air-liquid interface and in the NHBE model at day 4, day 12, and day 18 after air-liquid interface. "This project received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no 760928 (BIORIMA)."

2021-03-26 | PXD024242 | Pride

Global transcriptomic analyses of pluripotent stem cell-derived airway epithelial cells at 1 and 3 days after infection with SARS-CoV-2

Project description:We performed transcriptomic profiling of human iPSC-derived airway epithelial cells after infection with SARS-Co-V2. Using a recently described protocol to first generate airway basal cells from iPSCs and sunsequently differentiate those basal cells (iBCs) into a mucociliary epithelium in air-liquid interface culture. In this experiment we generated airway epithelial cultures from two different iPSC lines ("BU3 NGPT" and "1566"). In stage 6 of this protocol (>D45) iBCs are identified and purified by sorting on NGFR+ cells, and purified cells are expanded and differentiated on Transwells in dual-SMAD inhibition media then transitioned to ALI media when >80% confluent. Apical chamber media is removed to initiate ALI to recapitulate a physiologically-relevant environment and stimulate differentiation. After 14 days, iBCs form a pseudostratified airway epithelium that displays morphologic, molecular, and functional similarities to primary human airway epithelial cells and is comprised of the major airway cell types of multi-ciliated, secretory, and basal cellscomposed of the major airway epithelial cell types, that is permissive to SARS-CoV-2 infection. We studied the response of these iPSC-derived airway epithelial cells to SARS-CoV-2 infection using RNA-Sequencing 1 and 3 days post infection (DPI).

2022-02-10 | GSE196464 | GEO

MicroRNA signature during the time course of regeneration of the human airway mucociliary epithelium

Project description:The regeneration of the airway mucociliary epithelium involves several sequential events including migration, proliferation, polarization and final differentiation (i.e ciliogenesis). The airway mucociliary epithelium is consituted of three main cell types : ciliated cells, secretory cells and basal cells. We used microRNA microrrays to investigate the signature of microRNA during the four step of regeneration of the airway epithelium. Four time points (ALI-D0, ALI-D7, ALI-D14, ALI-D21) of regeneration of the airway epithelium for 3 donors.

2011-06-06 | E-GEOD-22141 | biostudies-arrayexpress

miRNAs high throughput sequencing profiling of regenerating human airway epithelial cells

2011-06-06 | E-GEOD-22144 | biostudies-arrayexpress

RSV-induced Cytoskeletal Inflammation

Project description:We studied RSV infection in an appropriate in vitro model of respiratory epithelium, a pseudostratified and fully differentiated mucociliary epithelium of normal human bronchial epithelial (NHBE) cells. RSV infection increased actin cytoskeleton without compromising adherent-, tight-, and tricellular-junctions as well as ciliary functions and epithelial tissue barrier integrity. This increased cytoskeleton depends on actin polymerization and the induction of proinflammatory cytokines and chemokines. Thus, we observed a novel signature “increased cytoskeleton” termed “cytoskeletal inflammation” in RSV-infected respiratory epithelium that presumably lacks classical antigen presenting cells, such as resident dendritic cells and macrophages. Our results suggest that RSV-induced cytoskeletal inflammation is a noncanonical earliest host response to the pathogen and contributes to airway inflammation.

2023-03-10 | GSE146795 | GEO

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