Transcription profiling of mouse lung resistance or sensitivity to cigarette smoke
ABSTRACT: We have investigated the effects of cigarette smoke exposure in three different strains of mice. DBA/2 and C57Bl/6J are susceptible to smoke and develop different lung changes in response to chronic exposure, while ICR mice are resistant to smoke and do not develop emphysema. The present study was carried out to determine early changes in the gene expression profile of mice exposed to cigarette smoke with either a susceptible or resistant phenotype. Experiment Overall Design: Three strains of mice were exposed to smoke from three cigarette/day, 5d/wk for 4 weeks. Microarray analysis was carried out on total RNA extracted from the lung utilizing the Affymetrix platform.
Project description:We have investigated the effects of cigarette smoke exposure in three different strains of mice. DBA/2 and C57Bl/6J are susceptible to smoke and develop different lung changes in response to chronic exposure, while ICR mice are resistant to smoke and do not develop emphysema. The present study was carried out to determine early changes in the gene expression profile of mice exposed to cigarette smoke with either a susceptible or resistant phenotype. Overall design: Three strains of mice were exposed to smoke from three cigarette/day, 5d/wk for 4 weeks. Microarray analysis was carried out on total RNA extracted from the lung utilizing the Affymetrix platform.
Project description:The A/J mouse is highly susceptible to lung tumor induction and has been widely used as a screening system in carcinogenicity testing and chemoprevention studies. However, the A/J mouse model has several disadvantages. Most notably, it develops tumors spontaneously. Moreover, there is a considerable gap in our understanding of the underlying mechanisms of pulmonary chemical carcinogenesis in the A/J mouse. Therefore, we examined the differences between spontaneous and cigarette smoke-related lung tumors in the A/J mouse using transcriptomics and microRNA (miRNA) profiling. Male A/J mice were exposed whole-body to mainstream cigarette smoke (MS) for 18 months. Gene expression analysis of lung tumors and surrounding non-tumorous parenchyma samples from animals that were exposed to either 300 mg/m3 MS or sham-exposed to fresh air indicated significant differential expression of 296 genes. Ingenuity Pathway Analysis illustrated an overall suppression of the humoral immune response, which was accompanied by a disruption of sphingolipid and glycosaminoglycan metabolism in tumors of MS-exposed A/J mice. Thus, we propose that MS exposure leads to severe perturbations in pathways essential for tumor recognition by the immune system, thereby potentiating the ability of tumor cells to escape from immune surveillance. Further, exposure to MS appeared to affect expression of miRNA which have previously been implicated in carcinogenesis and are thought to contribute to tumor progression. Finally, we identified a 50-gene signature and show its utility in distinguishing between the cigarette smoke-related and spontaneous lung tumors.
Project description:Cigarette smoke is the most relevant risk factor for the development of lung cancer and chronic obstructive pulmonary disease. Many of its more than 4500 chemicals are highly reactive, thereby altering protein structure and function. Here, we used subcellular fractionation coupled to label-free quantitative MS to globally assess alterations in the proteome of different compartments of lung epithelial cells upon exposure to cigarette smoke extract. Proteomic profiling of the human alveolar derived cell line A549 revealed the most pronounced changes within the cellular secretome with preferential downregulation of proteins involved in wound healing and extracellular matrix organization. In particular, secretion of secreted protein acidic and rich in cysteine, a matricellular protein that functions in tissue response to injury, was consistently diminished by cigarette smoke extract in various pulmonary epithelial cell lines and primary cells of human and mouse origin as well as in mouse ex vivo lung tissue cultures. Our study reveals a previously unrecognized acute response of lung epithelial cells to cigarette smoke that includes altered secretion of proteins involved in extracellular matrix organization and wound healing. This may contribute to sustained alterations in tissue remodeling as observed in lung cancer and chronic obstructive pulmonary disease.
Project description:Introduction: Prenatal and postnatal cigarette smoke exposure enhances the risk of developing asthma. Despite this as well as other smoking related risks, 11% of women still smoke during pregnancy. We hypothesized that cigarette smoke exposure during prenatal development generates long lasting differential methylation altering transcriptional activity that correlates with disease. Methods: In a house dust mite (HDM) model of allergic airway disease, we measured airway hyperresponsiveness (AHR) and airway inflammation between mice exposed prenatally to cigarette smoke (CS) or filtered air (FA). DNA methylation and gene expression were then measured in lung tissue. Results: We demonstrate that HDM-treated CS mice develop a more severe allergic airway disease compared to HDM-treated FA mice including increased AHR and airway inflammation. While DNA methylation changes between the two HDM-treated groups failed to reach genome-wide significance, 99 DMRs had an uncorrected p-value < 0.001. 6 of these 99 DMRs were selected for validation, based on the immune function of adjacent genes, and only 2 of the 6 DMRs confirmed the bisulfite sequencing data. Additionally, genes near these 6 DMRs (Lif, Il27ra, Tle4, Ptk7, Nfatc2, and Runx3) are differentially expressed between HDM-treated CS mice and HDM-treated FA mice. Conclusions: Our findings confirm that prenatal exposure to cigarette smoke is sufficient to modify allergic airway disease, however, it is unlikely that specific methylation changes account for the exposure-response relationship. These findings highlight the important role in utero cigarette smoke exposure plays in the development of allergic airway disease. Lung DNA methylation profiles of mice exposed in utero to cigarette smoke (CS) then treated with house dust mite (HDM, n = 8) or saline (n = 6), or exposed in utero to filtered air (FA) then treated with HDM (n = 9) or saline (n = 6)
Project description:COPD is a disorder characterized by the progressive development of airflow limitation that is not fully reversible. Cigarette smoke has been generally accepted as the most important of many risk factors for the development of COPD. We used microarray technology to perform comprehensive gene expression profiling of smoke exposure and cessation effects in mouse lung tissue. Overall design: Mice received nose-only exposure of 4% mainstream cigarette smoke or air (sham exposure) for 2 hours/day, 5 days/week for 2, 12 or 24 weeks. Mice undergoing smoke cessation received cigarette smoke exposure for 12 weeks, and then sham exposure for 12 weeks.
Project description:Pregnant mice (Dams) were exposed to cigarette smoke or filtered air. Lung tissue and RNA were harvested from the lungs of the Dams and their offspring and underwent transcriptomic analysis. Overall design: Two-condition experiment, Cigarette smoke vs. filtered air. 3 biological replicates for each condition of Dam female mice and Prenatal offspring mice.
Project description:Lung antigen presenting cells isolated from wild type but not Spp1-/- mice induce Th1 and Th17 cells differentiation. The goal of this study is to identify the genes differentially expressed by lung antigen presenting cells from cigarette smoke exposed mice. These genes may play crucial roles in directing Th1 and Th17 cells differentiation. Overall design: Lung antigen presenting cells were isolated from lungs of two groups of wild type and Spp1-/- mice that have been exposed to cigarette smoke for 4 months. Total mRNA was extracted from these samples.
Project description:Bacterial infections and cigarette smoking have been linked to exacerbations of respiratory disease including severe asthma and chronic obstructive pulmonary disease (COPD). Epidemiological studies have also shown increased incidences of respiratory tract infections even in young smokers. We have previously shown that cigarette smoke exacerbates the inflammatory response to influenza A virus (PLoS ONE. 2010 Oct 12;5(10): e13251) and nontypeable Haemophilus influenzae (Am J Respir Crit Care Med. 2009 Apr 15;179(8):666) infection. While it is well understood that cigarette smoke impairs respiratory host defense, little is known with respect to mechanisms driving the outcome of infection in smoke-exposed mice. In this study, we attempt to characterize the antibacterial responses of lung resident cells from smoke-exposed mice to TLR agonists and live pathogens by gene expression profiling and to compare similarities and differences of inflammatory profiles between mouse and man. Overall design: C57BL/6 mice were exposed to room air or cigarette smoke for four days. Mice were exposed to twelve cigarettes (2R4 reference cigarettes; Tobacco and Health Research Institute, University of Kentucky), twice daily in a whole body smoke exposure system (SIU48, PROMECH LAB AB, Vintrie, Sweden). Mice were subsequently euthanized and the lungs inflated with a low-gelling agarose. 120 μm precision cut lung slices (PCLS) were generated. PCLS from room-air and cigarette smoke exposed mice were stimulated ex vivo with TLR agonists (Pam3CSK for TLR2, LPS for TLR4), as well as live and heat-killed versions of Hemophilus Influenza and S. Pneumoniae. Samples were collected 6h, 12h and 24h post-stimulation for gene expression profiling.
Project description:Epidemiological data clearly establishes cigarette smoking as one of the major cause for lung cancer worldwide. There is no standard screening method for lung cancer even in high-risk populations and the overall five-year survival has not changed significantly in the last decade. First-line treatment for lung cancer includes surgical resection, chemotherapy, and radiation. Recently with the advancement of systems biology, targeted therapy has become one of the most preferred modes of treatment for cancer. Though certain targeted therapies such as anti-EGFR are in clinical practice, they have shown limited success in the smokers suffering from lung cancer. This demands the discovery of alternative drug targets through systematic investigation of altered signaling mechanisms. To study dysregulated signaling pathways due to chronic cigarette smzoke exposure, we carried out SILAC-based phosphoproteome analysis of lung cell line H358 chronically exposed to cigarette smoke. We identified 1,812 phosphosites, of which 278 were hyperphosphorylated (≥ 3-fold) in the H358 cells exposed to cigarette smoke. We identified several known and some novel kinases and key signaling molecules that were hyperphosphorylated in response to chronic exposure to cigarette smoke
Project description:Despite the fact that smoking is the major cause of lung cancer, there is no clear mechanism detailing how chronic exposure of cigarette smoke alters the global signaling in lung cells. To investigate the altered signaling in lung cells, in this study, we carried out SILAC-based quantitative phosphoproteomic analysis of H292 cells chronically exposed to cigarette smoke. Using high resolution Orbitrap Velos mass spectrometer, we identified hyperphosphorylation of 493 sites which corresponds to 341 proteins and 195 hypophosphorylated sites mapping to 142 proteins, upon smoke exposure (2.0-fold). Bioinformatics analysis revealed differentially phosphotylated proteins were involved in multiple cellular processes including cellular polarity, cytoskeletal remodelling, cellular migration, protein synthesis, autophagy and apoptosis. The present study will significantly improve the current knowledge of smoke mediated signaling in lung cells, which in turn may aid in the development of targeted therapies specially for smokers.