GED-0507 attenuates lung fibrosis by counteracting myofibroblast transdifferentiation in vivo and in vitro

The development of more effective, better tolerated drug treatments for progressive pulmonary fibrosis (of which idiopathic pulmonary fibrosis is the most common and severe form) is a research priority. The peroxisome proliferator-activated receptor gamma (PPAR-γ) is a key regulator of inflammation and fibrosis and therefore represents a potential therapeutic target. However, the use of synthetic PPAR-γ agonists may be limited by their potentially severe adverse effects. In a mouse model of bleomycin (BLM)-induced pulmonary fibrosis, we have demonstrated that the non-racemic selective PPAR-γ modulator GED-0507 is able to reduce body weight loss, ameliorate clinical and histological features of pulmonary fibrosis, and increase survival rate without any safety concerns. Here, we focused on the biomolecular effects of GED-0507 on various inflammatory/fibrotic pathways. We demonstrated that preventive and therapeutic administration of GED-0507 reduced the BLM-induced mRNA expression of several markers of fibrosis, including transforming growth factor (TGF)-β, alpha-smooth muscle actin, collagen and fibronectin as well as epithelial-to-mesenchymal transition (EMT) and expression of mucin 5B. The beneficial effect of GED-0507 on pulmonary fibrosis was confirmed in vitro by its ability to control TGFβ-induced myofibroblast activation in the A549 human alveolar epithelial cell line, the MRC-5 lung fibroblast line, and primary human lung fibroblasts. Compared with the US Food and Drug Administration-approved antifibrotic drugs pirfenidone and nintedanib, GED-0507 displayed greater antifibrotic activity by controlling alveolar epithelial cell dysfunction, EMT, and extracellular matrix remodeling. In conclusion, GED-0507 demonstrated potent antifibrotic properties and might be a promising drug candidate for the treatment of pulmonary fibrosis.

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Effect of CDH1 gene methylation on transforming growth factor (TGF-β)-induced epithelial-mesenchymal transition in alveolar epithelial cell line A549

Genetics and Molecular Research ◽

10.4238/2014.february.13.13 ◽

2014 ◽

Vol 13 (4) ◽

pp. 8568-8576 ◽

Cited By ~ 5

Author(s):

Y.Y. Liu ◽

J.Y. Han ◽

S.C. Lin ◽

Z.Y. Liu ◽

W.T. Jiang

Keyword(s):

Growth Factor ◽

Epithelial Cell ◽

Transforming Growth Factor ◽

Epithelial Mesenchymal Transition ◽

Alveolar Epithelial Cell ◽

Epithelial Cell Line ◽

Gene Methylation ◽

Cdh1 Gene ◽

Mesenchymal Transition ◽

Alveolar Epithelial

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Transforming growth factor-alpha enhances alveolar epithelial cell repair in a new in vitro model

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1994.267.6.l728 ◽

1994 ◽

Vol 267 (6) ◽

pp. L728-L738 ◽

Cited By ~ 37

Author(s):

F. Kheradmand ◽

H. G. Folkesson ◽

L. Shum ◽

R. Derynk ◽

R. Pytela ◽

...

Keyword(s):

Growth Factor ◽

Epithelial Cell ◽

Wound Repair ◽

Wound Closure ◽

Transforming Growth Factor ◽

Alveolar Epithelial Cell ◽

Alveolar Epithelial ◽

Factor Alpha ◽

Vitro Model

Alveolar epithelial type II cells are essential for regenerating an intact alveolar barrier after destruction of type I cells in vivo. The first objective of these experimental studies was to develop an in vitro model to quantify alveolar epithelial cell wound repair. The second objective was to investigate mechanisms of alveolar epithelial cell wound healing by studying the effects of serum and transforming growth factor-alpha (TGF-alpha) on wound closure. Primary cultures of rat alveolar type II cells were prepared by standard methods and grown to form confluent monolayers in 48 h. Then a wound was made by denuding an area (mean initial area of 2.1 +/- 0.6 mm2) of the monolayer. Re-epithelialization of the denuded area over time in the presence or absence of serum was measured using quantitative measurements from time-lapse video microscopy. The half time of wound healing was significantly enhanced in the presence of serum compared with serum-free conditions (2.4 +/- 0.2 vs. 17.4 +/- 0.8 h, P < 0.001). We then tested the hypothesis that TGF-alpha is an important growth factor for stimulating wound repair of alveolar epithelial cells. Exogenous addition of TGF-alpha in serum-free medium resulted in a significantly more rapid wound closure, and, furthermore, the addition of a monoclonal antibody to TGF-alpha in the presence of serum significantly decreased fourfold the rate of wound closure. Measurement of internuclear cell distance confirmed that both cell motility and cell spreading were responsible for closure of the wound. These data demonstrate that 1) the mechanisms of alveolar cell repair can be studied in vitro and that 2) TGF-alpha is a potent growth factor that enhances in vitro alveolar epithelial cell wound closure.

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Protective Effect of Remdesivir Against Pulmonary Fibrosis in Mice

Frontiers in Pharmacology ◽

10.3389/fphar.2021.692346 ◽

2021 ◽

Vol 12 ◽

Author(s):

Xiaohe Li ◽

Rui Liu ◽

Yunyao Cui ◽

Jingjing Liang ◽

Zhun Bi ◽

...

Keyword(s):

Pulmonary Fibrosis ◽

Antiviral Drug ◽

Alveolar Epithelial Cells ◽

Lung Damage ◽

Lung Fibroblasts ◽

Mesenchymal Transition ◽

Alveolar Epithelial ◽

Tgf Β1

Pulmonary fibrosis is a known sequela of severe or persistent lung damage. Existing clinical, imaging and autopsy studies have shown that the lungs exhibit a pathological pulmonary fibrosis phenotype after infection with coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Pulmonary fibrosis may be one of the most serious sequelae associated with coronavirus disease 2019 (COVID-19). In this study, we aimed to examine the preventative effects of the antiviral drug remdesivir on pulmonary fibrosis. We used a mouse model of bleomycin-induced pulmonary fibrosis to evaluate the effects of remdesivir on pulmonary fibrosis in vivo and further explored the potential pharmacological mechanisms of remdesivir in lung fibroblasts and alveolar epithelial cells in vitro. The preventive remdesivir treatment was started on the day of bleomycin installation, and the results showed that remdesivir significantly alleviated bleomycin-induced collagen deposition and improved pulmonary function. In vitro experiments showed that remdesivir dose-dependently suppressed TGF-β1-induced lung fibroblast activation and improved TGF-β1-induced alveolar epithelial to mesenchymal transition. Our results indicate that remdesivir can preventatively alleviate the severity of pulmonary fibrosis and provide some reference for the prevention of pulmonary fibrosis in patients with COVID-19.

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β-catenin induces A549 alveolar epithelial cell mesenchymal transition during pulmonary fibrosis

Molecular Medicine Reports ◽

10.3892/mmr.2014.3013 ◽

2014 ◽

Vol 11 (4) ◽

pp. 2703-2710 ◽

Cited By ~ 9

Author(s):

PING SONG ◽

JIN-XU ZHENG ◽

JIAO XU ◽

JI-ZHU LIU ◽

LI-YAN WU ◽

...

Keyword(s):

Epithelial Cell ◽

Pulmonary Fibrosis ◽

Alveolar Epithelial Cell ◽

Mesenchymal Transition ◽

Alveolar Epithelial

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M2 macrophages have unique transcriptomes but conditioned media does not promote profibrotic responses in lung fibroblasts or alveolar epithelial cells in vitro

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00107.2021 ◽

2021 ◽

Author(s):

Elissa M Hult ◽

Stephen James Gurczynski ◽

Bethany B Moore

Keyword(s):

Pulmonary Fibrosis ◽

Alveolar Epithelial Cell ◽

Conditioned Media ◽

Alveolar Epithelial Cells ◽

Lung Fibroblasts ◽

M2 Macrophage ◽

M2 Macrophages ◽

Alveolar Epithelial ◽

Fibroblast Migration

Macrophages are critical regulators of pulmonary fibrosis. Their plasticity, proximity, and ability to crosstalk with structural cells of the lung make them a key cell type of interest in the regulation of lung fibrosis. Macrophages can express a variety of phenotypes which have been historically represented through an "M1-like" to "M2-like" delineation. In this classification, M1-like macrophages are proinflammatory and have increased phagocytic capacity compared to alternatively activated M2-like macrophages that are profibrotic and are associated with wound healing. Extensive evidence in the field in both patients and animal models align pulmonary fibrosis with M2 macrophages. In this paper, we performed RNAseq to fully characterize M1 vs. M2-skewed bone marrow-derived macrophages (BMDMs) and investigated the profibrotic abilities of M2 BMDM conditioned media (CM) to promote fibroblast migration, proliferation, alveolar epithelial cell (AEC) apoptosis, and mRNA expression of key fibrotic genes in both fibroblasts and in AECs. Although M2 CM-treated fibroblasts had increased migration and M2 CM-treated fibroblasts and AECs had increased expression of profibrotic proteins over M1 CM-treated cells, all differences can be attributed to M2 polarization reagents IL-4 and IL-13 also present in the CM. Collectively, these data suggest that the profibrotic effects associated with M2 macrophage CM in vitro are attributable to effects of polarization cytokines rather than additional factors secreted in response to those polarizing cytokines.

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Mode of action of nintedanib in the treatment of idiopathic pulmonary fibrosis

European Respiratory Journal ◽

10.1183/09031936.00174914 ◽

2015 ◽

Vol 45 (5) ◽

pp. 1434-1445 ◽

Cited By ~ 284

Author(s):

Lutz Wollin ◽

Eva Wex ◽

Alexander Pautsch ◽

Gisela Schnapp ◽

Katrin E. Hostettler ◽

...

Keyword(s):

Growth Factor ◽

Idiopathic Pulmonary Fibrosis ◽

Pulmonary Fibrosis ◽

Tyrosine Kinases ◽

Cell Injury ◽

Transforming Growth Factor ◽

Alveolar Epithelial Cell ◽

Phase Iii ◽

Lung Fibroblasts

Idiopathic pulmonary fibrosis (IPF) is a progressive and ultimately fatal disease characterised by fibrosis of the lung parenchyma and loss of lung function. Although the pathogenic pathways involved in IPF have not been fully elucidated, IPF is believed to be caused by repetitive alveolar epithelial cell injury and dysregulated repair, in which there is uncontrolled proliferation of lung fibroblasts and differentiation of fibroblasts into myofibroblasts, which excessively deposit extracellular matrix (ECM) proteins in the interstitial space. A number of profibrotic mediators including platelet-derived growth factor (PDGF), fibroblast growth factor (FGF) and transforming growth factor-β are believed to play important roles in the pathogenesis of IPF. Nintedanib is a potent small molecule inhibitor of the receptor tyrosine kinases PDGF receptor, FGF receptor and vascular endothelial growth factor receptor. Data from in vitro studies have shown that nintedanib interferes with processes active in fibrosis such as fibroblast proliferation, migration and differentiation, and the secretion of ECM. In addition, nintedanib has shown consistent anti-fibrotic and anti-inflammatory activity in animal models of lung fibrosis. These data provide a strong rationale for the clinical efficacy of nintedanib in patients with IPF, which has recently been demonstrated in phase III clinical trials.

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Transcriptional and Proteomic Characterization of Telomere-Induced Senescence in a Human Alveolar Epithelial Cell Line

Frontiers in Medicine ◽

10.3389/fmed.2021.600626 ◽

2021 ◽

Vol 8 ◽

Author(s):

Daniel I. Sullivan ◽

Mao Jiang ◽

Angela M. Hinchie ◽

Mark G. Roth ◽

Harinath Bahudhanapati ◽

...

Keyword(s):

Epithelial Cell ◽

Cell Line ◽

Alveolar Epithelial Cell ◽

Epithelial Cell Line ◽

Telomere Dysfunction ◽

Alveolar Epithelial ◽

Novel Proteins ◽

Human Alveolar Epithelial Cell ◽

Species Specific

Cellular senescence due to telomere dysfunction has been hypothesized to play a role in age-associated diseases including idiopathic pulmonary fibrosis (IPF). It has been postulated that paracrine mediators originating from senescent alveolar epithelia signal to surrounding mesenchymal cells and contribute to disease pathogenesis. However, murine models of telomere-induced alveolar epithelial senescence fail to display the canonical senescence-associated secretory phenotype (SASP) that is observed in senescent human cells. In an effort to understand human-specific responses to telomere dysfunction, we modeled telomere dysfunction-induced senescence in a human alveolar epithelial cell line. We hypothesized that this system would enable us to probe for differences in transcriptional and proteomic senescence pathways in vitro and to identify novel secreted protein (secretome) changes that potentially contribute to the pathogenesis of IPF. Following induction of telomere dysfunction, a robust senescence phenotype was observed. RNA-seq analysis of the senescent cells revealed the SASP and comparisons to previous murine data highlighted differences in response to telomere dysfunction. We conducted a proteomic analysis of the senescent cells using a novel biotin ligase capable of labeling secreted proteins. Candidate biomarkers selected from our transcriptional and secretome data were then evaluated in IPF and control patient plasma. Four novel proteins were found to be differentially expressed between the patient groups: stanniocalcin-1, contactin-1, tenascin C, and total inhibin. Our data show that human telomere-induced, alveolar epithelial senescence results in a transcriptional SASP that is distinct from that seen in analogous murine cells. Our findings suggest that studies in animal models should be carefully validated given the possibility of species-specific responses to telomere dysfunction. We also describe a pragmatic approach for the study of the consequences of telomere-induced alveolar epithelial cell senescence in humans.

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Protective effect of IL-6 on alveolar epithelial cell death induced by hydrogen peroxide

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00016.2004 ◽

2005 ◽

Vol 288 (2) ◽

pp. L342-L349 ◽

Cited By ~ 32

Author(s):

Hiroshi Kida ◽

Mitsuhiro Yoshida ◽

Shigenori Hoshino ◽

Koji Inoue ◽

Yukihiro Yano ◽

...

Keyword(s):

Gene Expression ◽

Cell Death ◽

Epithelial Cell ◽

Epithelial Cells ◽

Alveolar Epithelial Cell ◽

Alveolar Epithelial Cells ◽

Lung Fibroblasts ◽

Alveolar Epithelial ◽

Epithelial Cell Death

The goal of this study was to examine whether IL-6 could directly protect lung resident cells, especially alveolar epithelial cells, from reactive oxygen species (ROS)-induced cell death. ROS induced IL-6 gene expression in organotypic lung slices of wild-type (WT) mice. ROS also induced IL-6 gene expression in mouse primary lung fibroblasts, dose dependently. The organotypic lung slices of WT were more resistant to ROS-induced DNA fragmentation than those of IL-6-deficient (IL-6−/−) mice. WT resistance against ROS was abrogated by treatment with anti-IL-6 antibody. TdT-mediated dUTP nick end labeling stain and electron microscopy revealed that DNA fragmented cells in the IL-6−/− slice included alveolar epithelial cells and endothelial cells. In vitro studies demonstrated that IL-6 reduced ROS-induced A549 alveolar epithelial cell death. Together, these data suggest that IL-6 played an antioxidant role in the lung by protecting lung resident cells, especially alveolar epithelial cells, from ROS-induced cell death.

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