Genes In Lung Epithelial Cells Research Articles

Protective interplay: Mycobacterium tuberculosis diminishes SARS-CoV-2 severity through innate immune priming.

At the beginning of the COVID-19 pandemic those with underlying chronic lung conditions, including tuberculosis (TB), were hypothesized to be at higher risk of severe COVID-19 disease. However, there is inconclusive clinical and preclinical data to confirm the specific risk SARS-CoV-2 poses for the millions of individuals infected with Mycobacterium tuberculosis (M.tb). We and others have found that compared to singly infected mice, mice co-infected with M.tb and SARS-CoV-2 leads to reduced SARS-CoV-2 severity compared to mice infected with SARS-CoV-2 alone. Consequently, there is a large interest in identifying the molecular mechanisms responsible for the reduced SARS-CoV-2 infection severity observed in M.tb and SARS-CoV-2 co-infection. To address this, we conducted a comprehensive characterization of a co-infection model and performed mechanistic in vitro modeling to dynamically assess how the innate immune response induced by M.tb restricts viral replication. Our study has successfully identified several cytokines that induce the upregulation of anti-viral genes in lung epithelial cells, thereby providing protection prior to challenge with SARS-CoV-2. In conclusion, our study offers a comprehensive understanding of the key pathways induced by an existing bacterial infection that effectively restricts SARS-CoV-2 activity and identifies candidate therapeutic targets for SARS-CoV-2 infection.

Carbon nanoparticles adversely affect CFTR expression and toxicologically relevant pathways

Cystic fibrosis is an autosomal recessive disorder caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) that can lead to terminal respiratory failure. Ultrafine carbonaceous particles, which are ubiquitous in ambient urban and indoor air, are increasingly considered as major contributors to the global health burden of air pollution. However, their effects on the expression of CFTR and associated genes in lung epithelial cells have not yet been investigated. We therefore evaluated the effects of carbon nanoparticles (CNP), generated by spark-ablation, on the human bronchial epithelial cell line 16HBE14o− at air–liquid interface (ALI) culture conditions. The ALI-cultured cells exhibited epithelial barrier integrity and increased CFTR expression. Following a 4-h exposure to CNP, the cells exhibited a decreased barrier integrity, as well as decreased expression of CFTR transcript and protein levels. Furthermore, transcriptomic analysis revealed that the CNP-exposed cells showed signs of oxidative stress, apoptosis and DNA damage. In conclusion, this study describes spark-ablated carbon nanoparticles in a realistic exposure of aerosols to decrease CFTR expression accompanied by transcriptomic signs of oxidative stress, apoptosis and DNA damage.

Cellular response of lung fibroblasts and epithelial cells to particulate matter10 treatment examined via comparative transcriptome analysis.

Particulate matter (PM) can be categorized by particle size (PM10, PM2.5 and PM1.0), which is an important factor affecting the biological response. Exposure to PM in the air (dust, smoke, dirt and biological contaminants) is clearly associated with lung disease (lung cancer, pneumonia and asthma). Although PM primarily affects lung epithelial cells, the specific response of related cell types to PM remains to be elucidated. The present study performed Gene Ontology (GO) analysis programs (Clustering GO and Database for Annotation, Visualization and Integrated Discovery) on differentially expressed genes in lung epithelial cells (WI‑38 VA‑13) and fibroblasts (WI‑38) following treatment with PM10 and evaluated the cell‑specific biological responses related to cell proliferation, apoptosis, adhesion and extracellular matrix production. The results suggested that short‑ or long‑term exposure to PM may affect cell condition and may consequently be related to several human diseases, including lung cancer and cardiopulmonary disease.

Particulate matter less than 10 μm (PM10) activates cancer related genes in lung epithelial cells

Introduction: Particulate matter (PM) has various systemic effects. We researched the effects of PM on lung epithelial cells with next generation sequencing (NGS) and validated this with quantitative real-time polymerase chain reaction (qRT-PCR). Methods: We cultured the group exposed to PM10 (Particulate matter less than 10 μm)-like fine dust (ERM® CZ120 fine dust) at a concentration of 50 μg/mL and the untreated group for seven days in one normal lung epithelial cell line (BEAS-2B) and four lung cancer epithelial cell lines (NCI-H358, HCC-827, A549, NCI-H292). Then, we extracted the RNA from the sample and performed NGS. As a result of NGS, various gene expressions were upregulated or downregulated. Among them, we selected the gene whose mean fold change was more than doubled and changed in the same direction in all five cell lines. Based on these genes, we selected the top 10 genes, either upregulated or downregulated, to validate with the qRT-PCR. Results: There were the four genes that matched the NGS and qRT-PCR results, all of which were upregulated genes. The four genes are CYP1A1, CYP1B1, LINC01816, and BPIFA2. All four genes that matched the two results were upregulated genes and none of the downregulated genes matched. Conclusion: CYP1A1 and CYP1B1 are known to cause lung cancer by metabolizing polycyclic aromatic hydrocarbons, and long noncoding RNA is also known to play an important role in lung cancer. Considering this, we thought PM10 might be associated with lung cancer by activating CYP1A1, CYP1B1, and LINC01816.

Expression profile analysis reveals that Aspergillus fumigatus but not Aspergillus niger makes type II epithelial lung cells less immunological alert

BackgroundAspergillus fumigatus is the main causative agent of aspergillosis. Infections rarely occur in immunocompetent individuals, indicating efficient clearance of conidia by pulmonary defense mechanisms. Other aspergilli like Aspergillus niger also cause infections but to a much lesser extent. Our previous studies showed that A. fumigatus and A. niger have different behavior in the presence of type II alveolar A549 epithelial cells. A. fumigatus conidia are more efficiently internalized by these cells and germination is delayed when compared to A. niger. In addition, hyphae that have escaped the epithelial cells grow parallel to the epithelium, while A. niger grows away from this cell layer.ResultsHere it is shown that global gene expression of A. fumigatus and A. niger is markedly different upon contact with A549 cells. A total of 545 and 473 genes of A. fumigatus and A. niger, respectively, were differentially expressed when compared to growth in the absence of A549 cells. Notably, only 53 genes (approximately 10%) were shared in these gene sets. The different response was also illustrated by the fact that only 4 out of 75 GO terms were shared that were enriched in the differentially expressed gene sets. The orthologues of A. fumigatus genes involved in hypoxia regulation and heat shock were also up-regulated in A. niger, whereas thioredoxin reductase and allergen genes were found up-regulated in A. fumigatus but down-regulated in A. niger. Infection with A. fumigatus resulted in only 62 up and 47 down-regulated genes in A549. These numbers were 17 and 34 in the case of A. niger. GO terms related with immune response were down-regulated upon exposure to A. fumigatus but not in the case of A. niger. This indicates that A. fumigatus reprograms A549 to be less immunologically alert.ConclusionsOur dual transcriptomic analysis supports earlier observations of a marked difference in life style between A. fumigatus and A. niger when grown in the presence of type II epithelial cells. The results indicate important differences in gene expression, amongst others down regulation of immune response genes in lung epithelial cells by A. fumigatus but not by A niger.

Effect of progesterone on Smad signaling and TGF-β/Smad-regulated genes in lung epithelial cells.

The effect of endogenous progesterone and/or exogenous pre- or postnatal progesterone application on lung function of preterm infants is poorly defined. While prenatal progesterone substitution may prevent preterm birth, in vitro and in vivo data suggest a benefit of postnatal progesterone replacement on the incidence and severity of bronchopulmonary dysplasia (BPD). However, the molecular mechanisms responsible for progesterone’s effects are undefined. Numerous factors are involved in lung development, airway inflammation, and airway remodeling: the transforming growth factor beta (TGF-β)/mothers against decapentaplegic homolog (Smad) signaling pathway and TGF-β-regulated genes, such as connective tissue growth factor (CTGF), transgelin (TAGLN), and plasminogen activator inhibitor-1 (PAI-1). These processes contribute to the development of BPD. The aim of the present study was to clarify whether progesterone could affect TGF-β1-activated Smad signaling and CTGF/transgelin/PAI-1 expression in lung epithelial cells. The pharmacological effect of progesterone on Smad signaling was investigated using a TGF-β1-inducible luciferase reporter and western blotting analysis of phosphorylated Smad2/3 in A549 lung epithelial cells. The regulation of CTGF, transgelin, and PAI-1 expression by progesterone was studied using a promoter-based luciferase reporter, quantitative real-time PCR, and western blotting in the same cell line. While progesterone alone had no direct effect on Smad signaling in lung epithelial cells, it dose-dependently inhibited TGF-β1-induced Smad3 phosphorylation, as shown by luciferase assays and western blotting analysis. Progesterone also antagonized the TGF-β1/Smad-induced upregulation of CTGF, transgelin, and PAI-1 at the promoter, mRNA, and/or protein levels. The present study highlights possible new molecular mechanisms involving progesterone, including inhibition of TGF-β1-activated Smad signaling and TGF-β1-regulated genes involved in BPD pathogenesis, which are likely to attenuate the development of BPD by inhibiting TGF-β1-mediated airway remodeling. Understanding these mechanisms might help to explain the effects of pre- or postnatal application of progesterone on lung diseases of preterm infants.

Foxp1/2/4-NuRD Interactions Regulate Gene Expression and Epithelial Injury Response in the Lung via Regulation of Interleukin-6

To determine the underlying mechanism of Foxp1/2/4-mediated transcriptional repression, a yeast two-hybrid screen was performed that identified p66beta, a transcriptional repressor and component of the NuRD chromatin-remodeling complex. We show that direct interactions between Foxp1/4 and p66beta are mediated by the CR2 domain within p66beta and the zinc finger/leucine zipper repression domain found in Foxp1/2/4. These direct interactions are functionally relevant as overexpression of p66beta in combination with Foxp factors cooperatively represses Foxp target gene expression, whereas loss of p66 and Foxp factors results in de-repression of endogenous Foxp target genes in lung epithelial cells. Moreover, the NuRD components HDAC1/2 associate in a macromolecular complex with Foxp proteins, and loss of expression or inhibition of HDAC1/2 activity leads to de-repression of Foxp target gene expression. Importantly, we show in vivo that Foxp1 and HDAC2 act cooperatively to regulate expression of the cytoprotective cytokine interleukin-6, which results in increased resistance to hyperoxic lung injury in Foxp1/HDAC2 compound mutant animals. These data reveal an important interaction between the Foxp transcription factors and the NuRD chromatin-remodeling complex that modulates transcriptional repression critical for the lung epithelial injury response.

Systems Biology: The Big Picture

Genomics, proteomics, and metabolomics have all vastly advanced our understanding of human biology and disease. But the functioning of even a simple system such as a single yeast cell or bacterium is much more complicated than the sum of its genes or proteins or metabolites; it’s the activity of all those components and their relationships to one another that add up to a living organism. Recognizing that complexity, the emerging field of systems biology attempts to harness the power of mathematics, engineering, and computer science to analyze and integrate data from all the “omics” and ultimately create working models of entire biological systems. “Traditionally, scientists—toxicologists included—have relied on a reductionist approach to biology,” says William Suk, director of the NIEHS Center for Risk and Integrated Sciences. Even now, many studies examine complex systems by looking at cellular components in isolation. For instance, a common experiment involves using DNA microarrays to observe the effect of a chemical exposure on thousands of genes at once. This technique can quickly tell a scientist which genes may be vulnerable to that exposure. But a systems biology approach would attempt to model not only the chemical’s effect on gene expression but also how that expression will affect protein function, and in turn how the exposure will affect cell signaling. “There’s nothing wrong with what we’ve been doing,” Suk says. “But systems biology is going to take it to another level.”

Identification of glucocorticoid- and adenovirus E1A-regulated genes in lung epithelial cells by differential display.

Adenovirus infection has been implicated in the pathogenesis of lung inflammatory diseases for which glucocorticoids provide effective antiinflammatory treatment. In this study, the differential display assay was used to identify messenger RNAs (mRNAs) differentially expressed in dexamethasone (1 microM for 24 h)-treated A549 lung epithelial cells compared to A549 cells transfected with the adenoviral E1A gene. Thirty-seven complimentary DNAs (cDNAs) (15 glucocorticoid-regulated, 22 adenovirus E1A-regulated) were isolated. DNA sequence analysis showed that 35 of these were unique, 2 were identical with each other, and 3 were common to the glucocorticoid- and E1A-regulated groups. Genes identified included those involved in transcription/translation, cytoskeletal/contractile element genes, metabolic enzyme genes, and genes associated with cell regulation/signal transduction. After further analysis of the isolated clones by Northern blotting, ribonuclease protection, and semiquantitative RT-PCR (reverse transcriptase-polymerase chain reaction), 10 of the 14 glucocorticoid-regulated and one of the three common to both the adenovirus E1A- and glucocorticoid-regulated cDNAs were confirmed for this control of their expression. We conclude that the strategy of identifying cDNAs regulated by both adenovirus E1A and glucocorticoids provides a promising approach for identifying genes that may be important in the pathogenesis of lung inflammation and therefore targets for glucocorticoid treatment.

Differential Effects of Adenoviral-Mediated Transfer of Na+/K+-ATPase Subunit Genes in Lung Epithelial Cells

Studies in rats, rabbits, sheep, and humans indicate that edema is cleared from the alveolar airspace by active ion transport.1-4 This active transport is due, in part, to the function of alveolar NA+/K+-adenosine triphosphatases (ATPases).1-3,5,6 Na+/K+-ATPase is a transmembrane heterodimer that generates the transalveolar Na+ gradient, which results in the iso-osmotic movement of water out of the airspace. In the lung, Na+/K+-ATPase is principally, but not exclusively, composed of α1 and β1 subunits and is primarily located on the basolateral surface of alveolar type II (ATII) epithelial cells.