The pro-fibrotic and senescence phenotype of old lung fibroblasts is reversed or ameliorated by genetic and pharmacological manipulation of Slc7a11 expression

2022 ◽  
Author(s):  
Jeffrey D. Ritzenthaler ◽  
Edilson Torres-Gonzalez ◽  
Yuxuan Zheng ◽  
Igor N. Zelko ◽  
Victor Van Berkel ◽  
...  
Keyword(s):  
Gene Expression ◽  
Idiopathic Pulmonary Fibrosis ◽  
Lung Fibroblasts ◽  
Pharmacological Manipulation ◽  
Cystine Transporter ◽  
Primary Fibroblasts ◽  
Old Mice ◽  
Lower Expression ◽  
Senescence Phenotype ◽  
Expression And Activity

Increased senescence and expression of pro-fibrotic genes in old lung fibroblasts contribute to disrepair responses. We reported that primary lung fibroblasts from old mice have lower expression and activity of the cystine transporter Slc7a11/xCT than cells from young mice, resulting in changes in both the intracellular and extracellular redox environments. This study examines the hypothesis that low Slc7a11 expression in old lung fibroblasts promotes senescence and pro-fibrotic gene expression. The levels of mRNA and protein of Slc7a11, senescence markers, and pro-fibrotic genes were measured in primary fibroblasts from the lungs of old (24 months) and young (3 months) mice. In addition, the effects of genetic and pharmacological manipulation of Slc7a11 were investigated. We found that decreased expression of Slc7a11 in old cells was associated with elevated markers of senescence (p21, p16, p53 and b-galactosidase) and increased expression of pro-fibrotic genes (Tgfb1, Smad3, Acta2, Fn1, Col1a1 and Col5a1). Silencing of Slc7a11 in young cells replicated the aging phenotype, whereas overexpression of Slc7a11 in old cells decreased expression of senescence and pro-fibrotic genes. Young cells were induced to express the senescence and pro-fibrotic phenotype by sulfasalazine, an Slc7a11 inhibitor, whereas treatment of old cells with sulforaphane, an Slc7a11 inducer, decreased senescence without affecting pro-fibrotic genes. Like aging cells, idiopathic pulmonary fibrosis fibroblasts show decreased Slc7a11 expression and increased pro-fibrotic markers. In short, old lung fibroblasts manifest a pro-fibrotic and senescence phenotype that is modulated by genetic or pharmacological manipulation of Slc7a11.

scholarly journals Diverse and Specific Gene Expression Responses to Stresses in Cultured Human Cells

2004 ◽  
Vol 15 (5) ◽  
pp. 2361-2374 ◽  
Author(s):  
John Isaac Murray ◽  
Michael L. Whitfield ◽  
Nathan D. Trinklein ◽  
Richard M. Myers ◽  
Patrick O. Brown ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Heat Shock ◽  
Hela Cells ◽  
Stress Responses ◽  
Cell Communication ◽  
Lung Fibroblasts ◽  
Specific Gene ◽  
Primary Fibroblasts ◽  
Stress Oxidative

We used cDNA microarrays in a systematic study of the gene expression responses of HeLa cells and primary human lung fibroblasts to heat shock, endoplasmic reticulum stress, oxidative stress, and crowding. Hierarchical clustering of the data revealed groups of genes with coherent biological themes, including genes that responded to specific stresses and others that responded to multiple types of stress. Fewer genes increased in expression after multiple stresses than in free-living yeasts, which have a large general stress response program. Most of the genes induced by multiple diverse stresses are involved in cell-cell communication and other processes specific to higher organisms. We found substantial differences between the stress responses of HeLa cells and primary fibroblasts. For example, many genes were induced by oxidative stress and dithiothreitol in fibroblasts but not HeLa cells; conversely, a group of transcription factors, including c-fos and c-jun, were induced by heat shock in HeLa cells but not in fibroblasts. The dataset is freely available for search and download at http://microarray-pubs.stanford.edu/human_stress/Home.shtml .

scholarly journals Itaconate controls the severity of pulmonary fibrosis

2020 ◽  
Vol 5 (52) ◽  
pp. eabc1884 ◽  
Author(s):  
Patricia P. Ogger ◽  
Gesa J. Albers ◽  
Richard J. Hewitt ◽  
Brendan J. O’Sullivan ◽  
Joseph E. Powell ◽  
...  
Keyword(s):  
Gene Expression ◽  
Wound Healing ◽  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Lung Fibrosis ◽  
Therapeutic Strategy ◽  
Lung Fibroblasts ◽  
Disease Phenotype ◽  
Wild Type

Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease in which airway macrophages (AMs) play a key role. Itaconate has emerged as a mediator of macrophage function, but its role during fibrosis is unknown. Here, we reveal that itaconate is an endogenous antifibrotic factor in the lung. Itaconate levels are reduced in bronchoalveolar lavage, and itaconate-synthesizing cis-aconitate decarboxylase expression (ACOD1) is reduced in AMs from patients with IPF compared with controls. In the murine bleomycin model of pulmonary fibrosis, Acod1−/− mice develop persistent fibrosis, unlike wild-type (WT) littermates. Profibrotic gene expression is increased in Acod1−/− tissue-resident AMs compared with WT, and adoptive transfer of WT monocyte-recruited AMs rescued mice from disease phenotype. Culture of lung fibroblasts with itaconate decreased proliferation and wound healing capacity, and inhaled itaconate was protective in mice in vivo. Collectively, these data identify itaconate as critical for controlling the severity of lung fibrosis, and targeting this pathway may be a viable therapeutic strategy.

scholarly journals Pan-transcriptome-based candidate therapeutic discovery for idiopathic pulmonary fibrosis

2020 ◽  
Vol 14 ◽  
pp. 175346662097114
Author(s):  
Yunguan Wang ◽  
Jaswanth K. Yella ◽  
Sudhir Ghandikota ◽  
Tejaswini C. Cherukuri ◽  
Harshavardhana H. Ediga ◽  
...  
Keyword(s):  
Gene Expression ◽  
Clinical Trials ◽  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Histone Deacetylase Inhibitors ◽  
Kinase Inhibitor ◽  
Lung Fibroblasts ◽  
Joint Analysis ◽  
Potential Candidate ◽  
Opioid Receptor Antagonists

Background: There are two US Food and Drug Administration (FDA)-approved drugs, pirfenidone and nintedanib, for treatment of patients with idiopathic pulmonary fibrosis (IPF). However, neither of these drugs provide a cure. In addition, both are associated with several drug-related adverse events. Hence, the pursuit for newer IPF therapeutics continues. Recent studies show that joint analysis of systems-biology-level information with drug–disease connectivity are effective in discovery of biologically relevant candidate therapeutics. Methods: Publicly available gene expression signatures from patients with IPF were used to query a large-scale perturbagen signature library to discover compounds that can potentially reverse dysregulated gene expression in IPF. Two methods were used to calculate IPF–compound connectivity: gene expression-based connectivity and feature-based connectivity. Identified compounds were further prioritized if their shared mechanism(s) of action were IPF-related. Results: We found 77 compounds as potential candidate therapeutics for IPF. Of these, 39 compounds are either FDA-approved for other diseases or are currently in phase II/III clinical trials suggesting their repurposing potential for IPF. Among these compounds are multiple receptor kinase inhibitors (e.g. nintedanib, currently approved for IPF, and sunitinib), aurora kinase inhibitor (barasertib), epidermal growth factor receptor inhibitors (erlotinib, gefitinib), calcium channel blocker (verapamil), phosphodiesterase inhibitors (roflumilast, sildenafil), PPAR agonists (pioglitazone), histone deacetylase inhibitors (entinostat), and opioid receptor antagonists (nalbuphine). As a proof of concept, we performed in vitro validations with verapamil using lung fibroblasts from IPF and show its potential benefits in pulmonary fibrosis. Conclusions: As about half of the candidates discovered in this study are either FDA-approved or are currently in clinical trials for other diseases, rapid translation of these compounds as potential IPF therapeutics is possible. Further, the integrative connectivity analysis framework in this study can be adapted in early phase drug discovery for other common and rare diseases with transcriptomic profiles. The reviews of this paper are available via the supplemental material section.

scholarly journals 2027 The role of lysyl oxidase in systemic sclerosis-associated lung fibrosis

2018 ◽  
Vol 2 (S1) ◽  
pp. 32-33
Author(s):  
Xinh-Xinh Nguyen ◽  
Tetsuya Nishimoto ◽  
Takahisa Takihara ◽  
Logan Mlakar ◽  
Ellen Riemer ◽  
...  
Keyword(s):  
Lung Fibrosis ◽  
Human Lung ◽  
Ex Vivo ◽  
Lysyl Oxidase ◽  
Lung Fibroblasts ◽  
Primary Fibroblasts ◽  
Expression And Activity

OBJECTIVES/SPECIFIC AIMS: Systemic sclerosis (SSc) is a connective tissue disease of unknown etiology characterized by progressive fibrosis of the skin and multiple visceral organs. Effective therapies for SSc are needed. Lysyl oxidase (LOX) is a copper-dependent amide oxidase that plays a critical role in the crosslinking of the extracellular matrix (ECM). In this study, we investigated the role of LOX in the pathophysiology of SSc. METHODS/STUDY POPULATION: LOX expression and protein levels were measured in lung tissues and primary fibroblasts from patients with SSc and healthy controls. The effects of recombinant LOX (rLOX) were measured in vitro in primary fibroblasts, ex vivo in human lung tissues and in vivo in mice given bleomycin in combination with rLOX. LOX levels and activity were evaluated in lung fibroblasts treated with an endostatin-derived peptide that ameliorates fibrosis and in mice treated with bleomycin in combination with the peptide. Further, to differentiate the crosslinking activity of LOX from other potential effects, primary human fibroblasts were cultured with rLOX in the presence of the inhibitor, beta-aminopropionitrile. The expression levels of ECM (collagen and fibronectin), pro-fibrotic factors (IL-6 and TGF-beta), and transcription factor (c-Fos) were examined by real-time PCR, ELISA, immunoblotting, or hydroxyproline assay. RESULTS/ANTICIPATED RESULTS: LOX mRNA was increased in lung tissues and matching fibroblasts of SSc patients. rLOX-induced ECM production in vitro and ex vivo in lung fibroblasts and in human lung tissues maintained in organ culture, respectively. Additionally, TGF-beta and bleomycin induced ECM production, LOX mRNA expression and activity. Endostatin peptide abrogated these effects. In vivo, rLOX synergistically exacerbated pulmonary fibrosis in bleomycin-treated mice. The inhibition of LOX catalytic activity by beta-aminopropionitrile failed to abrogate LOX-induced ECM production. LOX increased the production of IL-6. IL-6 neutralization blocked the effects of LOX. Further, LOX induced c-Fos expression and its nuclear localization. DISCUSSION/SIGNIFICANCE OF IMPACT: LOX expression and activity were increased with fibrosis in vitro, ex vivo, and in vivo. LOX induced fibrosis via increasing ECM, IL-6 and c-Fos translocation to the nucleus. These effects were independent of the crosslinking activity of LOX and mediated by IL-6. Our findings suggest that inhibition of LOX may be a viable option for the treatment of lung fibrosis. Further, the use of human lung in organ culture establishes the relevance of our findings to human disease.

scholarly journals Pan-transcriptome-based Candidate Therapeutic Discovery for Idiopathic Pulmonary Fibrosis

2019 ◽  
Author(s):  
Yunguan Wang ◽  
Jaswanth K. Yella ◽  
Sudhir Ghandikota ◽  
Tejaswini C. Cherukuri ◽  
Harshavardhana H. Ediga ◽  
...  
Keyword(s):  
Gene Expression ◽  
Clinical Trials ◽  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Large Scale ◽  
Kinase Inhibitor ◽  
Lung Fibroblasts ◽  
Joint Analysis ◽  
Potential Candidate ◽  
Opioid Receptor Antagonists

ABSTRACTBACKGROUNDAlthough the advent of two FDA-approved therapies for idiopathic pulmonary fibrosis (IPF) has energized the field, their effects are largely suppressive than pulmonary fibrosis remission- or reversion-inducing. Hence, the pursuit for newer IPF therapeutics continues. Recent studies show that joint analysis of systems biology level information with drug-disease connectivity are effective in discovery of biologically relevant candidate therapeutics.METHODSPublicly available gene expression signatures from IPF patients are used to query large scale perturbagen signature library to discover compounds that can potentially reverse dysregulated gene expression in IPF. Two methods are used to calculate IPF-compound connectivity: gene expression-based connectivity and feature-based connectivity. Identified compounds are further prioritized based on their shared mechanism(s) of action.RESULTSWe identified 77 compounds as potential candidate therapeutics for IPF. Of these 39 compounds are either FDA-approved for other diseases or are currently in phase 2/3 clinical trials suggesting their repurposing potential for IPF. Among these compounds are multiple receptor kinase inhibitors (e.g., nintedanib, currently approved for IPF, and sunitinib), aurora kinase inhibitor (barasertib), EGFR inhibitors (erlotinib, gefitinib), calcium channel blocker (verapamil), phosphodiesterase inhibitors (roflumilast, sildenafil), PPAR agonists (pioglitazone), HDAC inhibitors (entinostat), and opioid receptor antagonists (nalbuphine). As a proof-of-concept, we performed in vitro validations with verapamil using lung fibroblasts from IPF and show its potential benefits in pulmonary fibrosis.CONCLUSIONSSince about half of the candidates discovered in this study are either FDA-approved or are currently in clinical trials for other diseases, rapid translation of these compounds as potential IPF therapeutics is feasible. Further, the generalizable, integrative connectivity analysis framework in this study can be readily adapted in early phase drug discovery for other common and rare diseases with transcriptomic profiles.

Gene expression analysis of metabolic markers during bone regeneration in a rat model

2014 ◽  
Vol 23 (03) ◽  
pp. 207-211
Author(s):  
C. Kasch ◽  
A. Osterberg ◽  
Thordis Granitzka ◽  
T. Lindner ◽  
M. Haenle ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Analysis ◽  
Saline Solution ◽  
Female Rats ◽  
Synthesis Rate ◽  
Metabolic Markers ◽  
Expression Levels ◽  
Fibrotic Tissue ◽  
Intermittent Pth ◽  
Lower Expression

SummaryThe RANK/RANKL/OPG system plays an important role in the regulation of bone metabolism and bony integration around implants. The aim of this study was to analyse gene expression of OPG, RANK, and RANKL in regenerating bone during implant integration. Additionally, the effect of intermittent para - thyroid hormone (PTH) treatment was analysed. A titanium chamber was implanted in the proximal tibiae of 48 female rats. The animals received either human PTH or saline solution (NaCl). After 21 and 42 days, RNA was isolated from tissue adjacent to the implant and expression of RANK, RANKL, and OPG was analysed. After 21 days, very low expression levels of all genes were shown. In contrast, increased gene expression after 42 days was determined. Expression of RANK and RANKL was lower than that for OPG. The lower expression levels after 21 days might be due to still ossifying, fibrotic tissue around the titanium chamber. An increased OPG synthesis rate associated with decreased RANKL expression after 42 days revealed bone-forming processes. Despite significant differences in gene expression between the time points, only slight differences were observed between application of intermittent PTH and NaCl after a period of 42 days.

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