Project description:Transcriptional profiling of human bronchial epithelial cell lines and primary cells. Parental normal 16HBE14o- cells were stably transfected with beta/gammaENaC to generate beta/gammaENaC-16HBE14o- cells. NHBE and DHBE-CF were obtained from TaKaRa.
Project description:Chronic obstructive pulmonary disease (COPD) is a serious global health problem characterized by chronic airway inflammation, progressive airflow limitation and destruction of lung parenchyma. Remodeling of the bronchial airways in COPD includes changes in both the bronchial epithelium and the subepithelial extracellular matrix (ECM). To explore the impact of an aberrant ECM on epithelial cell phenotype in COPD we developed a new ex vivo model, in which normal human bronchial epithelial (NHBE) cells repopulate and differentiate on decellularized human bronchial scaffolds derived from COPD patients and healthy individuals. By using transcriptomics, we show that bronchial ECM from COPD patients induces differential gene expression in primary NHBE cells when compared to normal bronchial ECM. The gene expression profile indicated altered activity of upstream mediators associated with COPD pathophysiology, including hepatocyte growth factor, transforming growth factor beta 1 and platelet-derived growth factor B, which suggests that COPD-related changes in the bronchial ECM contribute to the defective regenerative ability in the airways of COPD patients.
Project description:Analysis of 2 cultured normal lung cell lines, Normal Human Bronchial Epithelial (NHBE) and Human Small Airway Epithelial (SAEC) cells (Lonza, Walkersville, MD), following treatment with 5-aza-dC to induce DNA demethylation. These results provide insight into the role of epigenetic alterations, specifically demethylation, in differential gene expression in various lung neoplasms.
Project description:Gene expression was determined for normal human bronchial epithelial (NHBE) cells incubated with DMSO or TG immediately prior to infection with USSR H1N1 virus for 12 hours. All of the NHBE cell stocks were obtained from QIAGEN.
Project description:Gene expression microarrays were used to compare gene alterations induced by exposure to equitoxic doses of crocidolite asbestos and cristobalite silica in an isolate of normal human bronchial epithelial cells. Primary human bronchial epithelial cells (NHBE) were exposed to crocidolite asbestos, cristobalite silica, and saline as control for 24 hours. Total RNA was extracted from cell cultures, 3 separate cultures were used for each condition. The isolated RNA was used to analyze mRNA expression via Affymetix GeneChip Human U133A 2.0 arrays.
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)."
Project description:We studied miRNAs and their gene targets affecting SARS-CoV-2 pathogenesis in CF airway epithelial cell models in response to TGF-β1. Small RNAseq in CF human bronchial epithelial cell line treated with TGF-β1 and miRNA profiling characterized TGF-β1 effects on the SARS-CoV-2 pathogenesis pathways. Among the effectors, we identified and validated two miRNAs targeting ACE2 mRNA using different CF and non-CF human bronchial epithelial cell models. We have shown that TGF-β1 inhibits ACE2 expression by miR-136-3p and miR-369-5p. ACE2 levels were higher in cells expressing F508del-CFTR, compared to wild-type(WT)-CFTR and TGF-β1 inhibited ACE2 in both cell types. The ACE2 protein levels were still higher in CF, compared to non-CF cells after TGF-β1 treatment. TGF-β1 prevented the functional rescue of F508del-CFTR by ETI in primary human bronchial epithelial cells while ETI did not prevent the TGF-β1 inhibition of ACE2 protein. Finally, TGF-β1 reduced binding of ACE2 to the recombinant monomeric spike RBD. Our results may help to explain, at least in part, the role of TGF-β1 on the SARS-CoV-2 entry via ACE2 in the CF and non-CF airway.
Project description:Temporal analysis of the effect of cigarette smoke on normal human bronchial epithelial cells (NHBE), and S9 toxicity. Keywords: other
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.
