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The Effect of Repeated Whole Cigarette Smoke Challenge on Human Air-Liquid Interface Lung Epithelial Cultures

ABSTRACT: Airway epithelial cells are the first cells of the lungs to be exposed to the toxic agents contained within cigarette smoke. Accordingly, the response of these cells to this challenge is of considerable interest in the context of diseases in which cigarette smoke is a major aetiological factor. We used Affymetrix microarrays to study the effects of repeated cigarette smoke challenge on three-dimensional human lung airway epithelial cultures.

ORGANISM(S): Homo sapiens

PROVIDER: GSE30660 | GEO | 2011/08/31

SECONDARY ACCESSION(S): PRJNA144587

REPOSITORIES: GEO

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The Effect of Repeated Whole Cigarette Smoke Challenge on Human Air-Liquid Interface Lung Epithelial Cultures

Project description:Airway epithelial cells are the first cells of the lungs to be exposed to the toxic agents contained within cigarette smoke. Accordingly, the response of these cells to this challenge is of considerable interest in the context of diseases in which cigarette smoke is a major aetiological factor. We used Affymetrix microarrays to study the effects of repeated cigarette smoke challenge on three-dimensional human lung airway epithelial cultures. We have used cultures from four individual donors, and compared the effects of repeated cigarette smoke challenge (30mins exposure on four separate days), with no challenge.

2011-08-30 | E-GEOD-30660 | biostudies-arrayexpress

Intermittent exposure to whole cigarette smoke alters the differentiation of primary small airway epithelial cells in the air-liquid interface culture

Project description:Cigarette smoke (CS) is the leading cause to develop COPD. Therefore, we investigated the pathologic effects of whole CS on the differentiation of primary small airway epithelial cells (SAEC), from three healthy donors and three COPD patients, cultured under ALI (air-liquid interface) conditions. The analysis of the epithelial physiology demonstrated that CS impaired barrier formation and reduced cilia beat activity. Although, COPD-derived ALI cultures preserved some features known from COPD patients, the CS-induced effects were similarly pronounced in ALI cultures from patients compared to healthy controls. A RNA sequencing analysis revealed the deregulation for marker genes for basal cells and secretory cells upon CS exposure. The comparison between gene signatures obtained from our in vitro model (CS vs. air) with a published data set from human epithelial brushes (smoker vs. non-smoker) reveals a high degree of similarity between the deregulated genes and pathways. ALI culture has been generated utilising cells obtained from 3 COPD and 3 healthy subjects. Subsequently cultures have been treated with CS and air respectively for 33 days. Gene expression profiles were generated using next generation sequencing.

2019-12-31 | GSE135188 | GEO

miRNA expression in differentiating primary human bronchial epithelial cells cultured at air-liquid-interface

Project description:We performed microarray analysis of miRNA expression in differentiating primary human bronchial epithelial cells. The goal was to identify miRNAs that are dynamically expressed under airway epithelial development. Cells were cultured at air-liquid-interface and were harvested day 4, 6, 8, 11, 13, 15, 18, 20 and 22 for analysis.

2021-01-09 | GSE164448 | GEO

Single cell transcriptomic profiling of pluripotent stem cell-derived alveolar type 2 cells expressing SFTPC-tdTomato cultured in 3D spheres, 2D submerged culture, and air-liquid interface culture with and without cigarette smoke exposure

Project description:We performed transcriptomic profiling of cells derived from human induced pluripotent stem cells (iPSCs) using previously described distal lung directed differentiation protocol to generate alveolar type 2 cells (iAT2s). We used the SPC2-ST-B2 (“SPC2”) line containing a SFTPC-tdTomato knock in reporter. iAT2s were cultured as 3D spheres (3D), on Transwells in 2D submerged conditions (2D), or at ALI for a total of 10 days post passaging. iAT2s in ALI culture were exposed to cigarette smoke 12 hours prior to cell collection (Smoke) or air (ALI). On day 10, and at 12 hours post cigarette smoke exposure, cultures were dissociated to single cells and live cells were sorted on calcein blue on a Mo-Flo Astrios Cell Sorter. Capture and library preparation was performed using a 10X Chromium system. Libraries were sequenced using a NextSeq 500. We find that ALI culture promotes maturation of iAT2s compared to 3D, and that iAT2s mount an inflammatory and xenobiotic metabolism response to cigarette smoke exposure at ALI.

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Mouse Tracheal Epithelial Cell Air-Liquid Interface Primary Culture Time Course (ALI d0, d2, d7)

Project description:Mouse tracheal epithelial cells were cultured at air-liquid interface (ALI), RNA was harvested at days 0, 2, and 7 post-ALI, and hybridized to two-channel MEEBO arrays. The experiment was designed to allow investigators to identify genes differentially expressed during airway epithelial cell differentiation and development, including ciliogenesis.

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Development of a Novel Air-Liquid Interface Airway Tissue Equivalent Model for In Vitro Respiratory Modeling Studies

Project description:The human airways are complex structures with important interactions between cells, extracellular matrix proteins and the biomechanical microenvironment. A robust, well-differentiated in vitro culture system that accurately models these interactions would provide a useful tool for studying normal and pathological airway biology. Here, we report the analysis of a physiologically relevant air-liquid interface (ALI) 3D airway ‘organ tissue equivalent’ (OTE) model with three novel features: native pulmonary fibroblasts, solubilized lung extracellular matrix (ECM), and hydrogel substrate with tunable stiffness and porosity. We demonstrate the versatility of the OTE model by evaluating the impact of these features on HBE cell phenotype. Variations of this model were analyzed during 28 days of ALI culture by evaluating epithelial confluence, trans-epithelial resistance (TEER), and epithelial phenotype via multispectral immuno-histochemistry and next-generation sequencing (NGS). Cultures that included both solubilized lung ECM and native pulmonary fibroblasts within the hydrogel substrate formed well differentiated ALI cultures that maintained a barrier function and expressed similar levels of goblet, club and ciliated markers to native airway tissue. Modulation of hydrogel stiffness did not negatively impact HBE differentiation and could be a valuable variable to alter epithelial phenotype. This study highlights the capability and versatility of a 3D airway OTE model to model the multiple components of the human airway 3D microenvironment.

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Regional heterogeneity in response of airway epithelial cells to cigarette smoke

Project description:To investigate the biochemical and genetic alterations that occur in response to cigarette smoke exposure among airway epithelial cells from different sites in the lungs, we performed microarray-based analysis using small airway epithelial cells (SAEC) and normal human bronchial epithelial cells (NHBE) following 24 h of cigarette smoke extract (CSE). In microarray-based analysis, the small airway showed higher susceptibility to CS compared to the large airway, such as enhanced expression of inflammatory-related pathways including the TNF signaling pathway. Among the TNF-related genes, PTGS2, also known as COX-2, showed the greatest difference in expression levels, with higher CSE-induced increments of both mRNA and protein expression in SAEC compared to NHBE.

2018-01-16 | GSE107200 | GEO

Early-life exposure to tobacco smoke alters airway signaling pathways in D. melanogaster and later mortality in D. melanogaster

Project description:To investigate the effect of cigarette smoke exposure on gene expression in airway epithelial cells of Canton S Drosophila melanogaster larvae, we isolated the airways of cigarette smoke exposed larvae and air controls. We then performed gene expression profiling analysis using data obtained from RNA-seq of smoke-exposed males, smoke-exposed females, air-control males and air-control females. For each group 4 biological replicates were prepared, representing 40-50 larval airways.

2022-09-30 | GSE205093 | GEO

Direct exposure to SARS-CoV-2 and cigarette smoke increases infection severity and alters the stem cell-derived airway repair response

Project description:To study the effect of cigarette smoke exposure on Sars-Cov2 infection, we directly exposed mucociliary air-liquid interface (ALI) cultures derived from primary human nonsmoker airway basal stem cells (ABSCs) to short term cigarette smoke and infected them with live SARS-CoV-2. We set out to examine the underlying mechanisms governing the increased susceptibility of cigarette smoke exposed ALI cultures to SARS-CoV-2 infection by usingle cell profiling of the cultures, which showed that interferon response genes were induced in SARS-CoV-2 infected airway epithelial cells in ALI cultures but smoking exposure together with SARS-CoV-2 infection reduced the interferon response.

2020-11-17 | GSE161089 | GEO

Intermittent exposure to whole cigarette smoke alters the differentiation of primary small airway epithelial cells in the air-liquid interface culture

Project description:Intermittent exposure to whole cigarette smoke alters the differentiation of primary small airway epithelial cells in the air-liquid interface culture

| PRJNA557685 | ENA

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