biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsKazushi Matsumuratranscription profiling by arrayHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-6175Cigarette smoke (CS) is a major risk factor in the development of chronic inflammatory lung diseases such as chronic obstructive pulmonary disease. To evaluate the biological impact of CS on lung tissue, three-dimensional (3D) organotypic bronchial tissue cultures can be used to replicate in vivo conditions. We developed an original 3D human bronchial epithelial co-culture model to assess the biological impact of repeated CS exposure on cell differentiation and on the inflammatory response. We found that CS can disrupt homeostatic capacity in a dose-dependent manner, and that the activation of the EGFR pathway, which is involved in the early-stage pathogenesis of airway diseases, was predicted from transcriptomic data. We believe that our model of bronchial tissues, used for repeated CS exposure, can provide valuable information on tissue-specific alterations in biological systems.biostudies-arrayexpressHybridization - The biotin-labeled RNA was Hybridized on Affymetrix GeneChip Human Genome U133 Plus 2.0 Array using GeneChip® Hybridization, Wash and Stain Kit.Scaning - Command Console Software (Affymetrix) was used to automatically grid the DAT files and create the CEL files (probe cell intensity data).Sample Collection - We developed an original three-dimensional (3D) human bronchial epithelial co-culture model to assess the biological impact of repeated cigarette smoke (CS) exposure. The 3D co-culture model of human bronchial tissue was composed of a collagen matrix layer with human fibroblast cells (IMR-90; catalog no.CCL-186, American Type Culture Collection, Manassas, VA, USA) and an epithelial layer of normal human bronchial cells (NHBE; catalog no.CC-2540S, Lonza, Basel, Switzerland). The cells were cultured for 7 days under an air-liquid interface (ALI) culture, then exposed to mainstream 3R4F smoke using a VC 10 smoking robot and Cultex RFS module. One, two, and four 3R4F cigarettes were smoked in accordance with the ISO regime (35-mL puffs of 2 sec in duration each minute), and whole smoke was released into the mixing device in 2.8-sec exhausts followed by 1.0 L/min dilution with humidified clean air (more than 90% relative humidity). Diluted smoke was introduced into the exposure chamber with 5 mL/min flow controlled using mass flow controllers. Clean air was used as the control. Whole-cigarette smoke exposure was conducted from ALI culture day 7 to day 19 every other day, and the medium was replaced before each exposure. Forty-eight hours after the last exposure on ALI culture day 21, mRNA was extracted for transcriptomic analysis. Samples (=total mRNA) are a mix of bronchial cells and fibroblast cells.Nucleic Acid Extraction - Total RNA was isolated and purified using the Qiagen RNeasy® Mini Kit (Qiagen, Hilden, Germany).Labeling - Biotinylated cRNA was generated using GeneChip® 3′IVT PLUS Reagent Kit.MIAME ScoreRaw DataOrganizationAssays and DataMAGE-TAB FilesArray DesignsShinkichi IshikawaKazushi MatsumuraShigeaki ItofalseTranscription profiling by array to investigate effect of repeated cigarette exposure on three-dimensional human bronchial epithelial culturesCigarette smoke (CS) is a major risk factor in the development of chronic inflammatory lung diseases such as chronic obstructive pulmonary disease. To evaluate the biological impact of CS on lung tissue, three-dimensional (3D) organotypic bronchial tissue cultures can be used to replicate in vivo conditions. We developed an original 3D human bronchial epithelial co-culture model to assess the biological impact of repeated CS exposure on cell differentiation and on the inflammatory response. We found that CS can disrupt homeostatic capacity in a dose-dependent manner, and that the activation of the EGFR pathway, which is involved in the early-stage pathogenesis of airway diseases, was predicted from transcriptomic data. We believe that our model of bronchial tissues, used for repeated CS exposure, can provide valuable information on tissue-specific alterations in biological systems.2018-09-30T00:00:00Z2022-02-02T13:27:52.303Z2022-02-02T13:27:52.303ZE-MTAB-6175EFO_0002768EFO_0002944EFO_0003814EFO_0003813EFO_0005518EFO_0003815