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Table S1. Cell samples used for CAGE profiling. Table S2. Public datasets used in this study. Table S3. Differentially expressed promoters in FANTOM5 NSCLC cell lines. Table S4. Up-regulated/hypomethylated promoters in NSCLC cell lines. Table S5. Epi-markers in NSCLC. Table S6. Extended information for Table 2. Table S7. Univariate and multivariate Cox regression models of TCGA LUAD and LUSC datasets. Table S8. Gene expression profiling of SAECs treated with 5-aza-dC and TSA. Table S9. Up-regulated/hypomethylated genes in NSCLC cell lines that show up-regulation by 5-aza-dC and TSA in SAECs. Table S10. Promoters and methylation array probes that overlap nine families of repetitive elements. Table S11. Promoters and methylation array probes that overlap REP522 repetitive elements. Table S12. Transcripts commonly down-regulated by two different MYEOV siRNAs in A549 cells. Table S13. The relationship between expression of 22 epi-markers and EGFR/KRAS mutation in TCGA LUAD dataset.

Lung cancer is the leading cause of cancer-related deaths worldwide. The majority of cancer driver mutations have been identified; however, relevant epigenetic regulation involved in tumorigenesis has only been fragmentarily analyzed. Epigenetically regulated genes have a great theranostic potential, especially in tumors with no apparent driver mutations. Here, epigenetically regulated genes were identified in lung cancer by an integrative analysis of promoter-level expression profiles from Cap Analysis of Gene Expression (CAGE) of 16 non–small cell lung cancer (NSCLC) cell lines and 16 normal lung primary cell specimens with DNA methylation data of 69 NSCLC cell lines and 6 normal lung epithelial cells. A core set of 49 coding genes and 10 long noncoding RNAs (lncRNA), which are upregulated in NSCLC cell lines due to promoter hypomethylation, was uncovered. Twenty-two epigenetically regulated genes were validated (upregulated genes with hypomethylated promoters) in the adenocarcinoma and squamous cell cancer subtypes of lung cancer using The Cancer Genome Atlas data. Furthermore, it was demonstrated that multiple copies of the REP522 DNA repeat family are prominently upregulated due to hypomethylation in NSCLC cell lines, which leads to cancer-specific expression of lncRNAs, such as RP1-90G24.10, AL022344.4, and PCAT7. Finally, Myeloma Overexpressed (MYEOV) was identified as the most promising candidate. Functional studies demonstrated that MYEOV promotes cell proliferation, survival, and invasion. Moreover, high MYEOV expression levels were associated with poor prognosis.Implications: This report identifies a robust list of 22 candidate driver genes that are epigenetically regulated in lung cancer; such genes may complement the known mutational drivers.Visual Overview: http://mcr.aacrjournals.org/content/molcanres/15/10/1354/F1.large.jpg. Mol Cancer Res; 15(10); 1354–65. ©2017 AACR.

Figure S1. Methylation and expression levels of genes in lung cancer and normal lung tissues. Figure S2. Expression levels of epi-markers in TCGA LUAD dataset. Figure S3. Expression levels of epi-markers in TCGA LUSC dataset. Figure S4. The signature of 22 epigenetically regulated genes in non-small cell lung cancer. Figure S5. ZENBU genome browser view of RP1-90G24.10 and AL022344.4. Figure S6. ZENBU genome browser view of MYEOV. Figure S7. Higher MYEOV expression is associated with poor prognosis in NSCLC.

Chronic obstructive pulmonary disease (COPD) is primarily caused by inhalation of cigarette smoke and is the third leading cause of death worldwide. Pulmonary surfactant, a complex of phospholipids and proteins, plays an essential role in respiration by reducing the surface tension in the alveoli. Lysophosphatidylcholine acyltransferase 1 (LPCAT1) is an enzyme that catalyzes the biosynthesis of surfactant lipids and is expressed in type 2 alveolar epithelial cells. Its dysfunction is suggested to be involved in various lung diseases; however, the relationship between LPCAT1 and COPD remains unclear. To investigate the role of LPCAT1 in the pathology of COPD, we analyzed an elastase-induced emphysema model using Lpcat1 knockout (KO) mice. In Lpcat1 KO mice, elastase-induced emphysema was significantly exacerbated with increased apoptotic cells, which was not ameliorated by supplementation with dipalmitoylphosphatidylcholine, which is a major component of the surfactant synthesized by LPCAT1. We subsequently evaluated the effects of cigarette smoking on primary human type 2 alveolar epithelial cells (hAEC2s) and found that cigarette smoke extract (CSE) downregulated the expression of Lpcat1. Furthermore, RNA sequencing analysis revealed that the apoptosis pathway was significantly enriched in CSE-treated primary hAEC2s. Finally, we downregulated the expression of Lpcat1 using small interfering RNA, which resulted in enhanced CSE-induced apoptosis in A549 cells. Taken together, cigarette smoke-induced downregulation of LPCAT1 can promote the exacerbation of pulmonary emphysema by increasing the susceptibility of alveolar epithelial cells to apoptosis, thereby suggesting that Lpcat1 is a novel therapeutic target for irreversible emphysema.

Chronic obstructive pulmonary disease (COPD) is a systemic inflammatory disorder. It often causes weight loss, which is considered a poor prognostic factor. A Japanese herbal Kampo medicine, Hochuekkito (TJ-41), has been reported to prevent systemic inflammation and weight loss in COPD patients, but the underlying biological mechanisms remain unknown. In the present study, we investigated the role of TJ-41 in vivo using a mouse model of lung emphysema. We used lung epithelium-specific Taz conditional knockout mice (Taz CKO mice) as the lung emphysema model mimicking the chronic pulmonary inflammation in COPD. Acute inflammation was induced by intratracheal lipopolysaccharide administration, simulating COPD exacerbation. Mice were fed a diet containing 2% TJ-41 or a control diet. Taz CKO mice showed increased numbers of inflammatory cells in the bronchoalveolar lavage fluid compared to control mice. This effect was reduced by TJ-41 treatment. In the acute exacerbation model, TJ-41 mitigated the increased numbers of inflammatory cells in the bronchoalveolar lavage fluid and attenuated lung inflammation in histopathological studies. Additional in vitro experiments using the human macrophage cell line U-937 demonstrated that lipopolysaccharide-induced tumor necrosis factor-alpha expression was significantly downregulated by TJ-41. These results suggest that TJ-41 has anti-inflammatory effects in lung emphysema both in the chronic phase and during an acute exacerbation. In conclusion, our study sheds light on the anti-inflammatory effects of TJ-41 in lung emphysema. This establishes its potential as a new anti-inflammatory therapy and a preventive medicine for exacerbations during the long-time maintenance of COPD patients.

We report a case of atypical type A thymoma variant manifesting polymyalgia rheumatica. A 68-year-old man underwent extended thymectomy with concomitant resection of the pericardium and right lung for an anterior mediastinal tumor. He was diagnosed with atypical type A thymoma variant with pericardial invasion. He developed pain in his extremities 1 year and 2 months after surgery. Detailed examinations resulted in a diagnosis of polymyalgia rheumatica and bone metastasis of thymoma. He was treated with oral prednisolone for polymyalgia rheumatica. His symptoms and bone lesion have been stable up to the present time of 3.5 years post-surgery.

Background Obstructive lung disease (OLD) is a risk factor for postoperative pulmonary complications (PPC) and is incidentally discovered during preoperative evaluation. The key treatments for OLD are inhaled long-acting bronchodilators (LAB). However, the advantage of preoperative bronchodilator treatment for patients with OLD remains unclear. The aim of this study is to elucidate the effect of preoperative LAB treatment in patients with untreated OLD on postoperative outcomes. Methods In this propensity-matched cohort study, we included patients who were referred to the pulmonologists for untreated OLD. The patients were either treated with LAB or left untreated. The primary outcome was the incidence of prolonged oxygen therapy (>3 days) in the postoperative period. We evaluated patients' characteristics with and without the use of LAB using propensity score (PS) matching weight. Subsequently, the outcomes in the two groups were compared. Results We analysed 614 patients; 132 patients were part of the LAB group and 482 were included in the control group. In the crude analysis, the incidence of prolonged oxygen therapy was higher in the LAB group than in the control group (odds ratio [OR] = 1.35; P = 0.04). However, after PS matching weight, no statistically significant differences in prolonged oxygen therapy (OR = 1.15), incidence of prolonged intensive care unit stay, endotracheal re-intubation postoperatively and in-hospital death between the groups were identified. Conclusion There is a limited benefit of preoperative treatment with inhaled LAB for the reduction of PPC in patients with untreated OLD.

Naftopidil, an α‐1 adrenoceptor antagonist with few adverse effects, is prescribed for prostate hyperplasia. Naftopidil inhibits prostate fibroblast proliferation; however, its effects on lung fibroblasts and fibrosis remain largely unknown. Two normal and one idiopathic pulmonary fibrosis human lung fibroblast lines were cultured with various naftopidil concentrations with or without phenoxybenzamine, an irreversible α‐1 adrenoceptor inhibitor. We examined the incorporation of 5‐bromo‐2ʹ‐deoxyuridine into DNA and lactic acid dehydrogenase release by enzyme‐linked immunosorbent assay, cell cycle analysis by flow cytometry, scratch wound‐healing assay, and mRNA expressions of type IV collagen and α‐smooth muscle actin by polymerase chain reaction. Effects of naftopidil on bleomycin‐induced lung fibrosis in mice were evaluated using histology, micro‐computed tomography, and surfactant protein‐D levels in serum. Naftopidil, dose‐dependently but independently of phenoxybenzamine, inhibited 5‐bromo‐2ʹ‐deoxyuridine incorporation in lung fibroblasts. Naftopidil induced G1 cell cycle arrest, but lactic acid dehydrogenase release and migration ability of lung fibroblasts were unaffected. Naftopidil decreased mRNA expressions of type IV collagen and α‐smooth muscle actin in one normal lung fibroblast line. Histological and micro‐computed tomography examination revealed that naftopidil attenuated lung fibrosis and decreased serum surfactant protein‐D levels in bleomycin‐induced lung fibrosis in mice. In conclusion, naftopidil may have therapeutic effects on lung fibrosis.

TitleA mouse model of asthma­–chronic obstructive pulmonary disease overlap induced by intratracheal papain

The master regulator of neuroendocrine differentiation, achaete-scute complex homolog 1 (ASCL1) defines a subgroup of lung adenocarcinoma. However, the mechanistic role of ASCL1 in lung tumorigenesis and its relation to the immune microenvironment is principally unknown. Here, the immune landscape of ASCL1-positive lung adenocarcinomas was characterized by immunohistochemistry. Furthermore, ASCL1 was transduced in mouse lung adenocarcinoma cell lines and comparative RNA-sequencing and secretome analyses were performed. The effects of ASCL1 on tumorigenesis were explored in an orthotopic syngeneic transplantation model. ASCL1-positive lung adenocarcinomas revealed lower infiltration of CD8+, CD4+, CD20+, and FOXP3+ lymphocytes and CD163+ macrophages indicating an immune desert phenotype. Ectopic ASCL1 upregulated cyclin transcript levels, stimulated cell proliferation, and enhanced tumor growth in mice. ASCL1 suppressed secretion of chemokines, including CCL20, CXCL2, CXCL10, and CXCL16, indicating effects on immune cell trafficking. In accordance with lower lymphocytes infiltration, ASCL1-positive lung adenocarcinomas demonstrated lower abundance of CXCR3-and CCR6-expressing cells. In conclusion, ASCL1 mediates its tumor-promoting effect not only through cell-autonomous signaling but also by modulating chemokine production and immune responses. These findings suggest that ASCL1-positive tumors represent a clinically relevant lung cancer entity.

Lung cancer is the leading cause of cancer-related deaths worldwide. The majority of cancer driver mutations have been identified; however, relevant epigenetic regulation involved in tumorigenesis has only been fragmentarily analyzed. Epigenetically regulated genes have a great theranostic potential, especially in tumors with no apparent driver mutations. Here, epigenetically regulated genes were identified in lung cancer by an integrative analysis of promoter-level expression profiles from Cap Analysis of Gene Expression (CAGE) of 16 non–small cell lung cancer (NSCLC) cell lines and 16 normal lung primary cell specimens with DNA methylation data of 69 NSCLC cell lines and 6 normal lung epithelial cells. A core set of 49 coding genes and 10 long noncoding RNAs (lncRNA), which are upregulated in NSCLC cell lines due to promoter hypomethylation, was uncovered. Twenty-two epigenetically regulated genes were validated (upregulated genes with hypomethylated promoters) in the adenocarcinoma and squamous cell cancer subtypes of lung cancer using The Cancer Genome Atlas data. Furthermore, it was demonstrated that multiple copies of the REP522 DNA repeat family are prominently upregulated due to hypomethylation in NSCLC cell lines, which leads to cancer-specific expression of lncRNAs, such as RP1-90G24.10, AL022344.4, and PCAT7. Finally, Myeloma Overexpressed (MYEOV) was identified as the most promising candidate. Functional studies demonstrated that MYEOV promotes cell proliferation, survival, and invasion. Moreover, high MYEOV expression levels were associated with poor prognosis. Implications: This report identifies a robust list of 22 candidate driver genes that are epigenetically regulated in lung cancer; such genes may complement the known mutational drivers. Visual Overview: http://mcr.aacrjournals.org/content/molcanres/15/10/1354/F1.large.jpg. Mol Cancer Res; 15(10); 1354–65. ©2017 AACR.

Introduction Patients with haematological malignancies usually have a plethora of respiratory complications. Bronchoscopy is one of the most important procedures used to diagnose respiratory complications. Despite enormous benefit, patients should be carefully selected for bronchoscopy as the process is invasive; however, there are only few reports evaluating the contributing factors of bronchoscopy that result in the definitive diagnosis of respiratory complications in these patients. Objective This study aimed to elucidate and identify the contributing factors of bronchoscopy for definitive diagnosis in patients with haematological malignancies. Methods We retrospectively analysed 275 patients with haematological malignancies who later showed respiratory complications, requiring consultation with pulmonologists. We found that 62 patients underwent bronchoscopy. Our data analysis focused on this particular subset of patients to identify the factors crucial for definitive diagnosis via bronchoscopy. Results Bronchoscopy provided definitive diagnosis for 25 patients (diagnostic yield = 40.3%). We determined that nodular shadow was associated with high diagnostic yields by multivariate logistic regression [odds ratio (OR): 6.6 (2.1-23)]. Furthermore, in several bronchoscopic procedures, biopsy also contributed to definitive diagnosis of patients with nodular shadow [OR: 17 (1.5-180)]. Life-threatening complications were not observed due to bronchoscopy in our study. Conclusions Our study demonstrated that patients with haematological malignancies who showed lung nodular shadows are more likely to be definitively diagnosed by bronchoscopy, whereas transbronchial biopsy may also be beneficial for these patients.

Background: Periostin, a biomarker that reflects Th2-driven inflammatory diseases, play important roles in the asthmatic airway. Although periostin is mainly produced in Bronchial Smooth Muscle (BSM) cells after IL-13 stimulation, the mechanism of periostin production remains unclear. Methods: In order to investigate the intracellular signaling pathway of periostin production under stimulation with IL-13, we performed RNA sequence analysis. BSM cells were stimulated with IL-13 for 0, 12, 24, 48hours. We evaluated the phosphorylation of signal transducer and activator of transcription factor 6 (STAT6), extracellular signal-regulated kinase (ERK) 1/2, and Akt after IL-13 stimulation. Furthermore, the influence of IL-13 receptor α2 knockdown and several phosphorylation inhibitors on periostin production was evaluated by using qPCR and ELISA. Results: Periostin increased in a time-dependent manner in the culture supernatant. The results of RNA sequence suggested that the involvement of JAK/STAT, ERK 1/2, PI3K/Akt pathways and IL13 receptor α2. IL-13-induced periostin was attenuated by inhibiting STAT6 phosphorylation and was strongly suppressed by inhibiting MEK 1/2 phosphorylation or PI3K phosphorylation. IL-13 receptor α2 is highly expressed during IL-13 stimulation and knockdown of IL-13 receptor α2 caused increase of periostin production. Conclusions: Our study demonstrates that BSM cells produce periostin after IL-13 stimulation through the JAK/STAT6, ERK1/2, and PI3K/Akt pathways and the relation of IL-13 receptor α2. These suggest that the mechanism of periostin production in BSM cells might help clarify asthma pathogenesis and discover new therapeutic targets for managing asthma.

Background and Aim: Recently, it has been recognized that asthma and COPD can coexist as asthma-COPD overlap (ACO). Due to the difference of clinical course, it is thought the pathogenesis of ACO is different from that of asthma or COPD; however, its animal models have not yet been established. In animal experiments, papain (a cysteine protease enzyme present in papaya) is known to cause asthmatic response by daily inhalation, and lung emphysema by intermittent inhalation. Here we developed ACO mice model by weekly intratracheal administrations of papain, and we examined whether the mice model well reflects the features of ACO, comparing its features with elastase induced COPD model. Methods: PBS, papain, or porcine pancreatic elastase (PPE) was intratracheally administered to female C57BL/6N mice (5-6 weeks old) on days 0, 7, 14, 21, while PPE on day 0 only. The mice were sacrificed on day 25. Lung mechanics, airway responsiveness to methacholine, bronchoalveolar lavage fluid (BALF), lung tissue, and mRNA in lung homogenates were analyzed. Results: In lung mechanics tests, both papain and PPE group showed significantly higher compliance and inspiratory capacity than PBS group, while the airway responsiveness to methacholine was significantly higher in papain group than in other groups. Papain increased eosinophils in BALF. Both papain and PPE induced alveolar air-space enlargement. In addition, type 2 inflammatory cytokines such as IL-13 and IL-25 were significantly higher in papain group than in other groups. Conclusions: Weekly papain administrations induced ACO like features in mice. Clarification of the underlying pathogenesis and development of novel treatment might be expected using this single-agent simple model.

Background Infections are an important cause of morbidity and mortality in patients with rheumatoid arthritis. Patients receiving immunosuppressive or anti-tumor necrosis factor (TNF) agents are vulnerable to fungal infections, including those derived from Aspergillus species. Detection of the Aspergillus galactomannan antigen in serum is useful for the early diagnosis of invasive aspergillosis in patients with hematological malignancies. However, its usefulness for detecting early invasive aspergillosis in rheumatoid arthritis patients remains unestablished. Methods Galactomannan antigen levels were measured in 340 patients (311 female patients). For patients who exhibited galactomannan antigen levels ≥0.5 during the initial examination, a second examination was performed 3–6 months later. Conventional blood tests and chest radiography were also performed. Results Elevated galactomannan antigen levels (≥0.5) were observed in 62 (18.2%) of 340 patients during the initial examination. A second examination was performed in 56 of 62 patients, 50 of whom exhibited elevated antigen levels. Elevated antigen levels were not associated with the use of any drug including anti-TNF agents. Serum galactomannan antigen levels were correlated with the albumin/globulin ratio ( r =−0.19, p r =0.17, p =0.001), and hemoglobin concentration ( r =−0.15, p =0.005). No patient was clinically diagnosed with invasive aspergillosis during the study period. Conclusions Serum galactomannan antigen levels are frequently elevated in a nonspecific manner in patients with rheumatoid arthritis.

Background: Bronchial asthma is a chronic airway disease characterized by eosinophilic airway inflammation. Lung fibroblasts activated by IL-13 serve as important sources of chemokines, such as eotaxins, contributing to persistent eosinophilic inflammation. Src-homology 2-containing protein (CISH), belonging to the suppressor of cytokine signaling (SOCS) family, acts as a negative regulator of cytokine induction. The aim of this study was to elucidate the role of CISH in the production of eosinophil chemotactic chemokines in human lung fibroblasts. Methods: Normal human lung fibroblasts were stimulated by IL-13, and global gene expression profile was assessed by cDNA microarray. Expression changes and downstream of IL-13 signaling were evaluated by quantitative RT-PCR, ELISA or western blotting. Loss- and gain-of-function analyses of CISH were performed by small interfering RNA and vector overexpression, respectively. Results: Ingenuity pathway analysis revealed that IL-13 induced chemokine signaling, including the eotaxin family, while significantly suppressing IFN-α/β signaling. Among eight SOCS family members, CISH was most strongly induced by IL-13 via phosphorylation of signal transducer and activator of transcription 6 (STAT6). Loss- and gain-of-function studies demonstrated that CISH negatively regulated the expression of CCL26. Conclusions: These findings suggest that CISH plays a key role in the eosinophilic inflammation associated with bronchial asthma by regulating IL-13-induced CCL26 production. Augmentation of CISH function could be a novel approach for treating eosinophilic inflammation in severe asthma. Keywords: Asthma, Eotaxin-3, Fibroblast, Interleukin-13, Transcriptome

Introduction: Naftopidil, a selective α1 adrenoceptor antagonist, inhibits the growth of prostate fibroblasts by inducing cell apoptosis without the blocking effects of α1 adrenoceptor. However, the effects of naftopidil are little known on the growth of lung fibroblast and lung fibrosis. Methods: We examined the effects of naftopidil on the growth of lung fibroblasts using 5-Bromo-29-deoxyuridine (BrdU) incorporation assay. We also examined whether naftopidil induced the apoptosis of lung fibroblasts by analyzing the levels of cleaved caspase-3 by Western blotting. The effects of naftopidil on bleomycin-induced lung fibrosis in mice were examined. Results: Naftopidil inhibited the growth of lung fibroblasts in a dose-dependent manner. This inhibitory effect was weakened in lung fibroblast derived from a patient with idiopathic pulmonary fibrosis in comparison to normal human lung fibroblasts. Western blotting revealed that naftopidil induced the apoptosis of lung fibroblasts similarly to prostate fibroblasts. Naftopidil also attenuated bleomycin-induced lung fibrosis in mice. Conclusions: Naftopidil, a selective α1 adrenoceptor antagonist, inhibits the growth of lung fibroblasts by inducing cell apoptosis and attenuates bleomycin-induced lung fibrosis in mice.

Background: COPD and Bronchial asthma (BA) are implicated in increased risk of postoperative pulmonary complications (PPC). Risks of PPC in patients with FEV1/FVC Aims and Objectives: The aim was to elucidate the risk of PPC in patients with FEV1/FVC Methods: We retrospectively evaluated patients consulted to the Department of Respiratory Medicine for FEV1/FVC Results: Out of 408 incidentally detected FEV1/FVC 3 days (P=0.03) and oxygen therapy >4 days (P=0.02) after surgery. Whereas, patients without CADs fulfilling FEV1%pred 3 days (P=0.04). Conclusions: Depending on severity of airway obstruction, patients with FEV1/FVC

Introduction: Acute exacerbation of interstitial pneumonia often cause in inpatients with non-respiratory disease. The diagnosis and treatment are usually difficult and in-hospital mortality is high. However, there are only few studies demonstrating the factors of the disease prognosis. Aims: To evaluate clinical features and the predictive factors of in-hospital mortality of the exacerbation of interstitial pneumonia in patients hospitalized for non-respiratory diseases. Methods: Medical charts of inpatients referred to the Pulmonary Department for the exacerbation of interstitial pneumonia between 2010 and 2016 were retrospectively reviewed. Results: In total, 33 patients (25 male and 8 female) were analyzed in this study. Average age of the patients was 68.4 (42-86). The types of the interstitial pneumonia were drug-induced pneumonia in 15 patients (45%), acute exacerbation of idiopathic interstitial pneumonia in 9 patients (27%), and acute exacerbation of interstitial lung disease associated with collagen-vascular disease in 4 patients (12%). In-hospital morality rate was 36%. Serum LDH elevations to more than 500 U/L at the onset of exacerbation was significantly associated with in-hospital mortality (p Conclusions: Serum LDH elevations may be a surrogate marker for in-hospital mortality of exacerbation of interstitial pneumonia in patients hospitalized for non-respiratory diseases.

Background: Asthma is a chronic airway inflammatory disease characterized by airway remodeling, in which the bronchial smooth muscle (BSM) cells play an important role. Periostin, a biomarker that reflects Th2-driven inflammatory diseases such as asthma, may play an important role in the asthmatic airway. Although periostin is mainly produced in airway epithelial cells and fibroblasts after interleukin (IL)-13 stimulation, whether BSM cells produce periostin remains unclear. Therefore, we investigated periostin production in BSM cells and the mechanisms involved. Methods: Human BSM cells were cultured, and the effect of IL-13 stimulation on periostin production was evaluated using quantitative polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA). We evaluated the phosphorylation of signal transducer and activator of transcription factor 6 (STAT6), extracellular signal-regulated kinase (ERK)1/2, and Akt after IL-13 stimulation. Furthermore, using ELISA, we evaluated the influence of several phosphorylation inhibitors on periostin production. Results: Periostin mRNA expression increased in a dose- and time-dependent manner after IL-13 stimulation; periostin production was induced 24 and 48 h after stimulation. IL-13 stimulation induced the phosphorylation of STAT6, ERK1/2, and Akt. IL-13-induced periostin production was attenuated by inhibiting STAT6 phosphorylation and strongly suppressed by inhibiting mitogen-activated protein kinase kinase 1/2 phosphorylation or phosphatidylinositol 3-kinase (PI3K) phosphorylation. Conclusions: BSM cells produced periostin after IL-13 stimulation, via the JAK/STAT6, ERK1/2, and PI3K/Akt pathways. Understanding the mechanism of periostin production in BSM cells may help to clarify asthma pathogenesis.

Transcriptional coactivator with PDZ-binding motif (TAZ) regulates a variety of biological processes. Nuclear translocation and activation of TAZ are regulated by multiple mechanisms, including actin cytoskeleton and mechanical forces. TAZ is involved in lung alveolarization during lung development and Taz-heterozygous mice are resistant to bleomycin-induced lung fibrosis. In this study, we explored the roles of TAZ in the pathogenesis of idiopathic pulmonary fibrosis (IPF) through histological analyses of human lung tissues and cell culture experiments. TAZ was highly expressed in the fibroblastic foci of lungs from patients with IPF. TAZ controlled myofibroblast marker expression, proliferation, migration, and matrix contraction in cultured lung fibroblasts. Importantly, actin stress fibers and nuclear accumulation of TAZ were more evident when cultured on a stiff matrix, suggesting a feedback mechanism to accelerate fibrotic responses. Gene expression profiling revealed TAZ-mediated regulation of connective tissue growth factor (CTGF) and type I collagen. Clinical relevance of TAZ-regulated gene signature was further assessed using publicly available transcriptome data. These findings suggest that TAZ is involved in the pathogenesis of IPF through multifaceted effects on lung fibroblasts.

Background: Lung cancer (LC) is often incidentally found in inpatients with non-respiratory disease. Diagnosis and treatment are often difficult due to diverse backgrounds. Aim: To evaluate clinical features and analyze contributing factors for lung cancer diagnosis in patients hospitalized for non-respiratory diseases. Methods: Medical charts of inpatients referred to the Pulmonary Department for abnormal chest radiographs between 2010 and 2015 were retrospectively reviewed. Results: Of 356 patients analyzed in this study, 101 patients (67 men and 34 women) were suspected of LC. LC was diagnosed in 55 patients including 22 patients with another concurrent primary cancer, and LC was ruled out in 37 patients. Bronchoscopy was performed in 27 patients, CT-guided biopsy was performed in 5 patients, and thoracoscopic biopsy was performed in 12 patients. Twenty one cases were stage 1 and 25 cases were stage 4. Thirty four patients were treated and 21 patients had best supportive care. All patients received CT scans, and average tumor size was larger (32 mm) in the LC group than in the non-LC group (16 mm). Positron emission tomography (PET) was performed in 49 patients and maximum uptake was significantly higher in the LC group (7.6) than in the non-LC group (2.7). Serum tumor markers CEA, CYFRA, and NSE were significantly increased in LC patients. Conclusion: Patients with large nodules, chest shadows with high uptake on PET scans, and higher serum tumor markers were likely to be diagnosed as LC. Although referred inpatients had poor prognosis in general, many patients underwent curative resection. Diagnosis and treatment should be made promptly.

BACKGROUND: The viral infection is one of the main causes of bronchial asthma attack.It has been reported that the serum CCL5 levels during attack increase as compared to stable periods. The airway epithelial cell has a potential to release various chemical mediators as well as a barrier role from the outside world. CCL5, one of chemical mediators, is known to have strong chemotactic activity for eosinophils, and thus might be involved in asthma exacerbation. AIMS: To determine regulation of CCL5 production from the airway epithelial cell at the time of the viral infection. METHODS: BEAS-2B cells were stimulated by poly I:C. CCL5 mRNA levels were evaluated using RT-PCR method and concentrations of CCL5 in the supernatant were measured by ELISA method. A variety of endogenous agents such as IL-4, IL-13, IL-33, IL-37 and Clara cell 16 kD protein (CC16) were co-stimulated to investigate the roles of these factors in endogenouscontrol of CCL5 production. RESULTS: CCL5 production and mRNA levels were significantly increased by poly I:C stimulation (∼50-fold increase and∼80-fold increase, p DISCUSSION: Airway epithelial cells act as potent sources of CCL5 upon viral infection, and IL-13 was suggested to play a role in airway inflammatory exacerbation in coordinated fashion during viral infection. Therefore, targeting IL-13 might be useful for the management of the asthma exacerbation at the time of the viral infection.

Fibrosis is often involved in the pathogenesis of various chronic progressive diseases such as interstitial pulmonary disease. Pathological hallmark is the formation of fibroblastic foci, which is associated with the disease severity. Mesenchymal cells consisting of the fibroblastic foci are proposed to be derived from several cell sources, including originally resident intrapulmonary fibroblasts and circulating fibrocytes from bone marrow. Recently, mesenchymal cells that underwent epithelial-mesenchymal transition (EMT) have been also supposed to contribute to the pathogenesis of fibrosis. In addition, EMT can be induced by transforming growth factor β, and EMT can be enhanced by pro-inflammatory cytokines like tumor necrosis factor α. The gel contraction assay is an ideal in vitro model for the evaluation of contractility, which is one of the characteristic functions of fibroblasts and contributes to wound repair and fibrosis. Here, the development of a gel contraction assay is demonstrated for evaluating contractile ability of mesenchymal cells that underwent EMT.

Background and Purpose Bronchial asthma (BA) is a chronic airway disease characterized by airway hyperresponsiveness and remodeling, which are intimately linked to chronic airway inflammation. Reactive oxygen species (ROS) such as hydrogen peroxide are generated by inflammatory cells that are involved in the pathogenesis of BA. However, the role of ROS in the management of BA patients is not yet clear. We attempted to determine the role of ROS as a biomarker in the clinical setting of BA. Subjects and Methods We enrolled patients with BA from 2013 through 2015 and studied the degrees of asthma control, anti-asthma treatment, pulmonary function test results, fractional exhaled nitric oxide (FeNO), serum reactive oxygen metabolite (ROM) levels, and serum levels of interleukin (IL)-6 and IL-8. Results We recruited 110 patients with BA. Serum ROM levels correlated with white blood cell (WBC) count (rs = 0.273, p = 0.004), neutrophil count (rs = 0.235, p = 0.014), CRP (rs = 0.403, p < 0.001), and IL-6 (rs = 0.339, p < 0.001). Serum ROM levels and IL-8 and CRP levels negatively correlated with %FEV1 (rs = -0.240, p = 0.012, rs = -0.362, p < 0.001, rs = -0.197, p = 0.039, respectively). Serum ROM levels were significantly higher in patients who experienced severe exacerbation within 3 months than in patients who did not (339 [302–381] vs. 376 [352–414] CARR U, p < 0.025). Receiver-operating characteristics analysis showed that ROM levels correlated significantly with the occurrence of severe exacerbation (area under the curve: 0.699, 95% CI: 0.597–0.801, p = 0.025). Conclusions Serum levels of ROM were significantly associated with the degrees of airway obstruction, WBC counts, neutrophil counts, IL-6, and severe exacerbations. This biomarker may be useful in predicting severe exacerbations of BA.

Lung metastasis showing radiographic findings of air-space consolidation is considered to be rare. This report describes the case of a man with progressive left lower lobe air-space consolidation with a history of hepatocellular carcinoma. The pulmonary lesion was initially suspected to be infection and later clinically diagnosed as primary adenocarcinoma of the lung. Although the patient was treated with systemic chemotherapy, the disease progressed very rapidly. A postmortem examination revealed that the alveolar spaces were filled with neoplastic cells subsequently proven to be metastases of combined hepatocellular and cholangiocarcinoma.

Chronic inflammatory airway diseases, such as bronchial asthma and chronic obstructive pulmonary disease, are common respiratory disorders worldwide. Exacerbations of these diseases are frequent and worsen patients' respiratory condition and overall health. However, the mechanisms of exacerbation have not been fully elucidated. Recently, it was reported that interleukin (IL)-17A might play an important role in neutrophilic inflammation, which is characteristic of such exacerbations, through increased production of neutrophil chemoattractants. Therefore, we hypothesized that IL-17A was involved in the pathogenesis of acute exacerbation, due to viral infection in chronic inflammatory airway diseases. In this study, we assessed chemokine production by bronchial epithelial cells and investigated the underlying mechanisms. Comprehensive chemokine analysis showed that, compared with poly(I:C) alone, co-stimulation of BEAS-2B cells with IL-17A and poly(I:C) strongly induced production of such neutrophil chemoattractants as CXC chemokine ligand (CXCL)8, growth-related oncogene (GRO), and CXCL1. Co-stimulation synergistically induced CXCL8 and CXCL1 mRNA and protein production by BEAS-2B cells and normal human bronchial epithelial cells. Poly(I:C) induced chemokine expression by BEAS-2B cells mainly via Toll-like receptor 3/TIR-domain-containing adapter-inducing interferon-β-mediated signals. The co-stimulation with IL-17A and poly(I:C) markedly activated the p38 and extracellular-signal-regulated kinase 1/2 pathway, compared with poly(I:C), although there was little change in nuclear factor-κB translocation into the nucleus or the transcriptional activities of nuclear factor-κB and activator protein 1. IL-17A promoted stabilization of CXCL8 mRNA in BEAS-2B cells treated with poly(I:C). In conclusion, IL-17A appears to be involved in the pathogenesis of chronic inflammatory airway disease exacerbation, due to viral infection by promoting release of neutrophil chemoattractants from bronchial epithelial cells.

Asthma-related mortality has been decreasing due to inhaled corticosteroid use, but severe asthma remains a major clinical problem. One characteristic of severe asthma is resistance to steroid therapy, which is related to neutrophilic inflammation. Recently, the tumor necrosis factor superfamily member (TNFSF) 14/LIGHT has been recognized as a key mediator in severe asthmatic airway inflammation. However, the profiles and intracellular mechanisms of cytokine/chemokine production induced in cells by LIGHT are poorly understood. We aimed to elucidate the molecular mechanism of LIGHT-induced cytokine/chemokine production by bronchial epithelial cells. Human bronchial epithelial cells express lymphotoxin β receptor (LTβR), but not herpesvirus entry mediator, which are receptors for LIGHT. LIGHT induced various cytokines/chemokines, such as interleukin (IL)-6, oncostatin M, monocyte chemotactic protein-1, growth-regulated protein α and IL-8. Specific siRNA for LTβR attenuated IL-6 and IL-8 production by BEAS-2B and normal human bronchial epithelial cells. LIGHT activated intracellular signaling, such as mitogen-activated protein kinase and nuclear factor-κB (NF-κB) signaling. LIGHT also induced luciferase activity of NF-κB response element, but not of activator protein-1 or serum response element. Specific inhibitors of phosphorylation of extracellular signal-regulated kinase (Erk) and that of inhibitor κB attenuated IL-8 production, suggesting that LIGHT-LTβR signaling induces IL-8 production via the Erk and NF-κB pathways. LIGHT, via LTβR signaling, may contribute to exacerbation of airway neutrophilic inflammation through cytokine and chemokine production by bronchial epithelial cells.