Interleukin-1β Promotes Epithelial-Derived Alveolar Elastogenesis via αvβ6 Integrin-Dependent TGF-β Activation

Background/Aims: IL-1β creates persistent pulmonary inflammation accompanied by elevated transforming growth factor β (TGF-β levels and is associated with abnormal elastogenesis, which is observed in bronchopulmonary dysplasia (BPD). Although progress has been made in this field, the mechanisms underlying this process remain only partially understood. Methods: We assessed aberrant elastin localization-associated signaling in mouse pups exposed to 85% O2 treated with either IL-1Ra or 1D11, using morphometric analyses, quantitative RT-PCR, immunostaining, and ELISA. We also evaluated the derivation of elastin-producing cells using dual marker tracking. The regulatory mechanisms of IL-1β were investigated in vitro in lung epithelial and mesenchymal cells. Results: Elevated levels of IL-1β, αvβ6 and TGF-β1 were each associated with aberrant elastin production in O2-exposed lungs. IL-1Ra abolished TGF-β1 activation and αvβ6 upregulation, which occurred as a result of exposure to hyperoxia, whereas 1D11 had no discernible effect on the expression of either αvβ6 or IL-1β even following O2-exposure, suggesting that IL-1β was initially induced. Additionally, double staining revealed the presence of epithelium-derived elastin-producing cells, which was confirmed via in vitro IL-1β stress-induced epithelial-mesenchymal transformation (EMT) morphological and molecular marker changes, which may explain the altered lung elastin deposition and defective septation observed in BPD. Conclusions: These data support the hypothesis that IL-1β was initially induced by hyperoxia; αvβ6 subsequently interacted with and activated TGF-β1, acting as an epithelial/mesenchymal signaling molecule that contributed to excessive alveolar elastogenesis, the primary pathological feature of BPD.

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Abstract 17285: A Selective Transforming Growth Factor-β and Growth Differentiation Factor-15 Ligand Trap Attenuates Pulmonary Hypertension

Circulation ◽

10.1161/circ.130.suppl_2.17285 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

Lai-Ming Yung ◽

Samuel D Paskin-Flerlage ◽

Ivana Nikolic ◽

Scott Pearsall ◽

Ravindra Kumar ◽

...

Keyword(s):

Growth Factor ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Type I ◽

Rna Seq ◽

Differentiation Factor ◽

Group I ◽

Tgf Β1

Introduction: Excessive Transforming Growth Factor-β (TGF-β) signaling has been implicated in pulmonary arterial hypertension (PAH), based on activation of TGF-β effectors and transcriptional targets in affected lungs and the ability of TGF-β type I receptor (ALK5) inhibitors to improve experimental PAH. However, clinical use of ALK5 inhibitors has been limited by cardiovascular toxicity. Hypothesis: We tested whether or not selective blockade of TGF-β and Growth Differentiation Factor (GDF) ligands using a recombinant TGFβ type II receptor extracellular domain Fc fusion protein (TGFBRII-Fc) could impact experimental PAH. Methods: Male SD rats were injected with monocrotaline (MCT) and received vehicle or TGFBRII-Fc (15 mg/kg, twice per week, i.p.). C57BL/6 mice were treated with SU-5416 and hypoxia (SUGEN-HX) and received vehicle or TGFBRII-Fc. RNA-Seq was used to profile transcriptional changes in lungs of MCT rats. Circulating levels of GDF-15 were measured in 241 PAH patients and 41 healthy controls. Human pulmonary artery smooth muscle cells were used to examine signaling in vitro . Results: TGFBRII-Fc is a selective ligand trap, inhibiting the ability of GDF-15, TGF-β1, TGF-β3, but not TGF-β2 to activate SMAD2/3 in vitro . In MCT rats, prophylactic treatment with TGFBRII-Fc normalized expression of TGF-β transcriptional target PAI-1, attenuated PAH and vascular remodeling. Delayed administration of TGFBRII-Fc in rats with established PAH at 2.5 weeks led to improved survival, decreased PAH and remodeling at 5 weeks. Similar findings were observed in SUGEN-HX mice. No valvular abnormalities were found with TGFBRII-Fc treatment. RNA-Seq revealed GDF-15 to be the most highly upregulated TGF-β ligand in the lungs of MCT rats, with only modest increases in TGF-β1 and no change in TGF-β2/3 observed, suggesting a dominant role of GDF-15 in the pathophysiology of this model. Plasma levels of GDF-15 were significantly increased in patients with diverse etiologies of WHO Group I PAH. Conclusions: These findings demonstrate that a selective TGF-β/GDF-15 trap attenuates experimental PAH, remodeling and mortality, without causing valvulopathy. These data highlight the potential role of GDF-15 as a pathogenic molecule and therapeutic target in PAH.

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Knockdown of Long Noncoding RNA H19 Represses the Progress of Pulmonary Fibrosis through the Transforming Growth Factor β/Smad3 Pathway by Regulating MicroRNA 140

Molecular and Cellular Biology ◽

10.1128/mcb.00143-19 ◽

2019 ◽

Vol 39 (12) ◽

Cited By ~ 14

Author(s):

Xi Wang ◽

Zhe Cheng ◽

Lingling Dai ◽

Tianci Jiang ◽

Liuqun Jia ◽

...

Keyword(s):

Growth Factor ◽

Pulmonary Fibrosis ◽

Noncoding Rna ◽

Molecular Mechanisms ◽

Transforming Growth Factor ◽

Animal Experiments ◽

A549 Cells ◽

Transforming Growth Factor Β ◽

Tgf Β1

ABSTRACT Long noncoding RNAs (lncRNAs) are involved in various human diseases. Recently, H19 was reported to be upregulated in fibrotic rat lung and play a stimulative role in bleomycin (BLM)-induced pulmonary fibrosis in mice. However, its expression in human fibrotic lung tissues and mechanism of action remain unclear. Here, our observations showed that H19 expression was significantly upregulated and that of microRNA 140 (miR-140) was markedly reduced in pulmonary fibrotic tissues from idiopathic pulmonary fibrosis (IPF) patients and transforming growth factor β1 (TGF-β1)-induced HBE and A549 cells. Moreover, the expression of H19 was negatively correlated with the expression of miR-140 in IPF tissues. H19 knockdown attenuated TGF-β1-induced pulmonary fibrosis in vitro. Furthermore, animal experiments showed that H19 knockdown attenuated BLM-induced pulmonary fibrosis in mice. The study of molecular mechanisms showed that H19 functioned via reduction of miR-140 expression by binding to miR-140. The increase of miR-140 inhibited TGF-β1-induced pulmonary fibrosis, and H19 upregulation diminished the inhibitory effects of miR-140 on TGF-β1-induced pulmonary fibrosis, which was involved in the TGF-β/Smad3 pathway. Taken together, our findings showed that H19 knockdown attenuated pulmonary fibrosis via the regulatory network of lncRNA H19–miR-140–TGF-β/Smad3 signaling, and H19 and miR-140 might represent therapeutic targets and early diagnostic and prognostic biomarkers for patients with pulmonary fibrosis.

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Decorin-transforming Growth Factor-β Interaction Regulates Matrix Organization and Mechanical Characteristics of Three-dimensional Collagen Matrices

Journal of Biological Chemistry ◽

10.1074/jbc.m705180200 ◽

2007 ◽

Vol 282 (49) ◽

pp. 35887-35898 ◽

Cited By ~ 91

Author(s):

Zannatul Ferdous ◽

Victoria Mariko Wei ◽

Renato Iozzo ◽

Magnus Höök ◽

Kathryn Jane Grande-Allen

Keyword(s):

Growth Factor ◽

Transforming Growth Factor ◽

Three Dimensional ◽

Transforming Growth Factor Β ◽

Material Behavior ◽

Wild Type ◽

Collagen Matrices ◽

Matrix Organization ◽

Tgf Β1

The small leucine-rich proteoglycan decorin has been demonstrated to be a key regulator of collagen fibrillogenesis; decorin deficiencies lead to irregularly shaped collagen fibrils and weakened material behavior in postnatal murine connective tissues. In an in vitro investigation of the contributions of decorin to tissue organization and material behavior, model tissues were engineered by seeding embryonic fibroblasts, harvested from 12.5–13.5 days gestational aged decorin null (Dcn-/-) or wild-type mice, within type I collagen gels. The resulting three-dimensional collagen matrices were cultured for 4 weeks under static tension. The collagen matrices seeded with Dcn-/- cells exhibited greater contraction, cell density, ultimate tensile strength, and elastic modulus than those seeded with wild-type cells. Ultrastructurally, the matrices seeded with Dcn-/- cells contained a greater density of collagen. The decorin-null tissues contained more biglycan than control tissues, suggesting that this related proteoglycan compensated for the absence of decorin. The effect of transforming growth factor-β (TGF-β), which is normally sequestered by decorin, was also investigated in this study. The addition of TGF-β1 to the matrices seeded with wild-type cells improved their contraction and mechanical strength, whereas blocking TGF-β1 in the Dcn-/- cell-seeded matrices significantly reduced the collagen gel contraction. These results indicate that the inhibitory interaction between decorin and TGF-β1 significantly influenced the matrix organization and material behavior of these in vitro model tissues.

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NR4A1 is Involved in Fibrogenesis in Ovarian Endometriosis

Cellular Physiology and Biochemistry ◽

10.1159/000488838 ◽

2018 ◽

Vol 46 (3) ◽

pp. 1078-1090 ◽

Cited By ~ 6

Author(s):

Xinliu Zeng ◽

Zhang Yue ◽

Ying Gao ◽

Guosong Jiang ◽

Fuqing Zeng ◽

...

Keyword(s):

Stromal Cells ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Dependent Manner ◽

Ovarian Endometriosis ◽

Normal Endometrium ◽

Elevated Expression ◽

Tgf Β1

Background/Aims: Excess fibrosis may lead to chronic pain, scarring, and infertility as endometriosis develops and progresses. The pathogenesis of endometriosis has been linked to transforming growth factor-β (TGF-β), the most potent promoter of fibrosis. Methods: Levels of NR4A1 and P-NR4A1 protein in human endometrial and endometriotic tissue were assessed by western blotting and immunohistochemistry. The expression levels of fibrotic markers in stromal cells were evaluated by real-time PCR. The degree of fibrosis in mouse endometriotic lesions was detected by Masson trichrome and Sirius red staining. Results: The level of phosphorylated-NR4A1 was higher in ovarian endometriotic tissue than in normal endometrium, and long-term TGF-β1 stimulation phosphorylated NR4A1 in an AKT-dependent manner and then promoted the expression of fibrotic markers. Furthermore, inhibition of NR4A1 in stromal cells increased the TGF-β1-dependent elevated expression of fibrotic markers, and loss of NR4A1 stimulated fibrogenesis in mice with endometriosis. Additionally, Cytosporone B (Csn-B), an NR4A1 agonist, effectively decreased the TGF-β1-dependent elevated expression of fibrotic markers in vitro and significantly inhibited fibrogenesis in vivo. Conclusion: NR4A1 can regulate fibrosis in endometriosis and may serve as a new target for the treatment of endometriosis.

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Mechanism of Dandelion Sterol in Treating Pulmonary Fibrosis Through Transforming Growth Factor-β Signaling Pathway

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2568 ◽

2021 ◽

Vol 11 (4) ◽

pp. 612-618

Author(s):

Qun Lv ◽

Jianjun Wang ◽

Zhaoyang Ruan

Keyword(s):

Epithelial Cells ◽

Pulmonary Fibrosis ◽

Signaling Pathway ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Cellular Level ◽

Lung Epithelial Cells ◽

Migration And Invasion ◽

Lung Epithelial ◽

Tgf Β1

Background: The paper aimed to elucidate the molecular mechanism of Dandelion sterol in the treatment of pulmonary fibrosis, to study its effect on EMT of lung epithelial cells, and to find its target and downstream signaling pathways. Material and methods: The effects of Dandelion sterol on parathyroid (PQ)-induced EMT in lung epithelial cells were studied by immunofluorescence method. Immunohistochemistry and western-blot methods were used to verify that Dandelion sterol inhibited TGF-β1-induced EMT at the cellular level in animals, demonstrating that Dandelion sterol targets TGF-β1 to exert an anti-pulmonary fibrosis effect. Results: Dandelion sterol significantly inhibited PQ-induced migration and invasion of lung epithelial cells, and also inhibited the induced EMT. Dandelion sterol had a proper binding activity with the lung fibrosis-inducing factor TGF-β1. Dandelion sterol inhibited the TGF-β1-induced EMT process, and acted to treat pulmonary fibrosis by inhibiting the TGF-β1/Smad3 signaling pathway. Conclusion: Dandelion sterol can inhibit the pulmonary fibrosis by inhibiting the EMT process of lung epithelial cells through targeting the TGF- β1/Smad signaling pathway.

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Myc Downregulation by Transforming Growth Factor β Required for Activation of the p15Ink4b G1 Arrest Pathway

Molecular and Cellular Biology ◽

10.1128/mcb.19.9.5913 ◽

1999 ◽

Vol 19 (9) ◽

pp. 5913-5922 ◽

Cited By ~ 151

Author(s):

Beverley J. Warner ◽

Stacy W. Blain ◽

Joan Seoane ◽

Joan Massagué

Keyword(s):

Growth Factor ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

G1 Arrest ◽

Cyclin D ◽

Oligomeric State ◽

Proliferating Cells ◽

Lung Epithelial ◽

Epithelial Cell Lines

ABSTRACT The antimitogenic action of transforming growth factor β (TGF-β) in epithelial cells involves cyclin-dependent kinase (cdk) inhibitory gene responses and downregulation of c-Myc expression. Although the cdk inhibitory responses are sufficient for G1arrest, enforced expression of c-Myc prevents G1 arrest by TGF-β. We investigated the basis of this antagonism by using Mv1Lu lung epithelial cell lines that conditionally express levels of human c-Myc. We show that c-Myc prevents induction of the cdk4 inhibitor p15Ink4b and the subsequent inhibition of G1cdks by TGF-β. We assessed the significance of this effect by analyzing the oligomeric state of cdk4 in these cells. In proliferating cells, endogenous cdk4 is distributed among three populations: an abundant high-molecular-mass (>400-kDa) pool of latent cdk4 that serves as a source of cdk4 for cyclin D, a low-abundance pool containing active cyclin D-cdk4 complexes, and an inactive population of monomeric cdk4. Cell stimulation with TGF-β converts the latent and active cdk4 pools into inactive cdk4, an effect that is specifically mimicked by overexpression of p15 but not by other forms of G1 arrest. This process of TGF-β-induced cdk4 inactivation is completely blocked by expression of c-Myc, even though the latent and active cdk4 complexes from c-Myc-expressing cells remain sensitive to dissociation by p15 in vitro. c-Myc causes a small increase in cyclin D levels, but this effect contributes little to the loss of TGF-β responses in these cells. The evidence suggests that c-Myc interferes with TGF-β activation of the p15 G1arrest pathway. TGF-β must therefore downregulate c-Myc in order to activate this pathway.

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Dermatopontin interacts with transforming growth factor β and enhances its biological activity

Biochemical Journal ◽

10.1042/bj3370537 ◽

1999 ◽

Vol 337 (3) ◽

pp. 537-541 ◽

Cited By ~ 50

Author(s):

Osamu OKAMOTO ◽

Sakuhei FUJIWARA ◽

Mayumi ABE ◽

Yasufumi SATO

Keyword(s):

Extracellular Matrix ◽

Biological Activity ◽

Growth Factor ◽

Epithelial Cells ◽

Inhibitory Activity ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Lung Epithelial Cells ◽

Lung Epithelial ◽

Tgf Β1

Dermatopontin, a recently found low-molecular-mass component of the extracellular matrix, was studied for its interaction with decorin and transforming growth factor β (TGF-β) and its influence on TGF-β bioactivity. Dermatopontin reacted with decorin with an apparent Kd of 100 nM in a solid-phase assay. Dermatopontin inhibited the formation of the decorin–TGF-β1 complex. Decorin also competed with dermatopontin for the binding of this cytokine. The dermatopontin–decorin complex bound 3-fold more TGF-β1 than did each component individually, and binding was inhibited more strongly by decorin preincubated with dermatopontin than by dermatopontin or decorin alone. Dermatopontin augmented the biological activity of TGF-β1, as analysed by the expression of luciferase in mink lung epithelial cells transfected with a plasminogen activator inhibitor–promoter–luciferase construct, although dermatopontin itself did not show apparent induction of luciferase. Dermatopontin showed weak inhibitory activity on the proliferation of mink lung epithelial cells, and it enhanced the growth-inhibitory activity of TGF-β on these cells. Thus dermatopontin increases the cellular response to TGF-β. These findings strongly suggest that dermatopontin modifies the behaviour of TGF-β through interaction with decorin in the microenvironment of the extracellular matrix in vivo.

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Expression of Transforming Growth Factor-β by Human Islets: Impact on Islet Viability and Function

Cell Transplantation ◽

10.3727/000000007783465217 ◽

2007 ◽

Vol 16 (8) ◽

pp. 775-785 ◽

Cited By ~ 6

Author(s):

Omaima M. Sabek ◽

Daniel W. Fraga ◽

James Henry ◽

Lillian W. Gaber ◽

Malak Kotb ◽

...

Keyword(s):

Growth Factor ◽

Matrix Protein ◽

Transforming Growth Factor ◽

Active Form ◽

Transforming Growth Factor Β ◽

Human Islets ◽

Extracellular Matrix Protein ◽

Tgf Β1

Transforming growth factor-β1 (TGF-β1) is a pleotropic cytokine that promotes angiogenesis and extracellular matrix protein synthesis in addition to its immunosuppressive effects. The purpose of this study is to identify optimal conditions for in vivo expression of TGF-β1 by human islets to exploit the possible beneficial effects and minimize undesirable side effects. We transduced human islets with adenoviral vectors encoding the active form of Ad-TGF-β1 or Ad-LacZ to test the effects of TGF-β1 gene expression on islet in vivo function following their transplantation into a NOD-SCID mouse model. Islets were transduced with multiplicity of infection (MOI) of 20, 10, 5, and 2.5 per islet cell. At a MOI ranging from 2.5 to 20, expression of TGF-β1 in islet supernatant persisted for 1–2 months and ranged from 153 ± 5 to 2574 ± 1299 pg/ml, respectively. Transduction with the lowest MOI (2.5) did not compromise the in vivo production of human C-peptide. We conclude that TGF-β1 expression in transplanted islets does not compromise viability and that adenoviral transduction with the TGF-β1 gene has a dose-dependent effect, with larger MOIs being deleterious. The data also indicate that in vitro culture system and the in vivo NOD-SCID model could be used successfully to evaluate the nonimmune effects of gene transduction.

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Vardenafil Activity in Lung Fibrosis and In Vitro Synergy with Nintedanib

Cells ◽

10.3390/cells10123502 ◽

2021 ◽

Vol 10 (12) ◽

pp. 3502

Author(s):

Michael H. Bourne ◽

Theodore J. Kottom ◽

Deanne M. Hebrink ◽

Malay Choudhury ◽

Edward B. Leof ◽

...

Keyword(s):

Extracellular Matrix ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Enzyme Linked Immunosorbent Assay ◽

Vascular Effects ◽

Phosphodiesterase Type 5 Inhibitors ◽

Concurrent Use ◽

Mesenchymal Transformation ◽

Tgf Β1

Idiopathic pulmonary fibrosis (IPF) remains an intractably fatal disorder, despite the recent advent of anti-fibrotic medication. Successful treatment of IPF, like many chronic diseases, may benefit from the concurrent use of multiple agents that exhibit synergistic benefit. In this light, phosphodiesterase type 5 inhibitors (PDE5-Is), have been studied in IPF primarily for their established pulmonary vascular effects. However, recent data suggest certain PDE5-Is, particularly vardenafil, may also reduce transforming growth factor beta 1 (TGF-β1) activation and extracellular matrix (ECM) accumulation, making them a potential target for therapy for IPF. We evaluated fibroblast TGF-β1-driven extracellular matrix (ECM) generation and signaling as well as epithelial mesenchymal transformation (EMT) with pretreatment using the PDE5-I vardenafil. In addition, combinations of vardenafil and nintedanib were evaluated for synergistic suppression of EMC using a fibronectin enzyme-linked immunosorbent assay (ELISA). Finally, the effects of vardenafil on fibrosis were investigated in a bleomycin mouse model. Our findings demonstrate that vardenafil suppresses ECM generation alone and also exhibits significant synergistic suppression of ECM in combination with nintedanib in vitro. Interestingly, vardenafil was shown to improve fibrosis markers and increase survival in bleomycin-treated mice. Vardenafil may represent a potential treatment for IPF alone or in combination with nintedanib. However, additional studies will be required.

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Inhibition of lysyl oxidase-like 2 ameliorates folic acid-induced renal tubulointerstitial fibrosis

10.1101/2020.12.01.406009 ◽

2020 ◽

Author(s):

Sung-Eun Choi ◽

Nara Jeon ◽

Hoon Young Choi ◽

Hyeon Joo Jeong ◽

Beom Jin Lim

Keyword(s):

Folic Acid ◽

Cancer Metastasis ◽

Transforming Growth Factor ◽

Lysyl Oxidase ◽

Transforming Growth Factor Β ◽

Treated Group ◽

Tubulointerstitial Fibrosis ◽

Renal Tubulointerstitial Fibrosis ◽

Mesenchymal Transformation

AbstractTubulointerstitial fibrosis is characterized by accumulation of the extracellular matrix in the interstitium. Lysyl oxidase-like 2 (LOXL2), a member of the lysyl oxidase family, is known for promoting cancer metastasis, invasion, and stromal fibrosis in various organs. Our previous study demonstrated expression of LOXL2 in kidney podocytes and tubular epithelial cells, and the association between elevated LOXL2 and tubulointerstitial fibrosis. The present study evaluated the effect of LOXL2 inhibition using an inhibitory monoclonal antibody (AB0023) on tubulointerstitial fibrosis in a folic acid-induced tubulointerstitial fibrosis mouse model. We also evaluated the association of LOXL2 with epithelial-mesenchymal transformation related molecules in vitro using HK-2 cells. Our data demonstrate that AB0023 prevented the progression of tubulointerstitial fibrosis significantly, as determined by trichrome and picro-sirius red staining, as well as the total collagen assay. The mean expression of phosphorylated Smad2 and Smad4 was lower in the AB0023-treated group although it was not statistically significant. Following transforming growth factor-β (TGF-β) challenge, LOXL2-deficient HK-2 cells exhibited significantly lower expression of the mesenchymal markers vimentin and fibronectin than control HK-2 cells. In conclusion, LOXL2 inhibition ameliorates renal fibrosis through the TGF-β/Smad signalling pathway.

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