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Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients

PURPOSE: Inflammatory gene expression is modulated by posttranscriptional regulation via RNA-binding proteins (RBPs), which regulate mRNA turnover and translation by binding to conserved mRNA sequences. Their role in COPD is only partially defined. This study evaluated RBPs tristetraprolin (TTP), hu...

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Autores principales: Ricciardi, Luca, Col, Jessica Dal, Casolari, Paolo, Memoli, Domenico, Conti, Valeria, Vatrella, Alessandro, Vonakis, Becky M, Papi, Alberto, Caramori, Gaetano, Stellato, Cristiana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6190813/
https://www.ncbi.nlm.nih.gov/pubmed/30349226
http://dx.doi.org/10.2147/COPD.S166284
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author Ricciardi, Luca
Col, Jessica Dal
Casolari, Paolo
Memoli, Domenico
Conti, Valeria
Vatrella, Alessandro
Vonakis, Becky M
Papi, Alberto
Caramori, Gaetano
Stellato, Cristiana
author_facet Ricciardi, Luca
Col, Jessica Dal
Casolari, Paolo
Memoli, Domenico
Conti, Valeria
Vatrella, Alessandro
Vonakis, Becky M
Papi, Alberto
Caramori, Gaetano
Stellato, Cristiana
author_sort Ricciardi, Luca
collection PubMed
description PURPOSE: Inflammatory gene expression is modulated by posttranscriptional regulation via RNA-binding proteins (RBPs), which regulate mRNA turnover and translation by binding to conserved mRNA sequences. Their role in COPD is only partially defined. This study evaluated RBPs tristetraprolin (TTP), human antigen R (HuR), and AU-rich element-binding factor 1 (AUF-1) expression using lung tissue from COPD patients and control subjects and probed their function in epithelial responses in vitro. PATIENTS AND METHODS: RBPs were detected by immunohistochemistry in bronchial and peripheral lung samples from mild-to-moderate stable COPD patients and age/smoking history-matched controls; RBPs and RBP-regulated genes were evaluated by Western blot, ELISA, protein array, and real-time PCR in human airway epithelial BEAS-2B cell line stimulated with hydrogen peroxide, cytokine combination (cytomix), cigarette smoke extract (CSE), and following siRNA-mediated silencing. Results were verified in a microarray database from bronchial brushings of COPD patients and controls. RBP transcripts were measured in peripheral blood mononuclear cell samples from additional stable COPD patients and controls. RESULTS: Specific, primarily nuclear immunostaining for the RBPs was detected in structural and inflammatory cells in bronchial and lung tissues. Immunostaining for AUF-1, but not TTP or HuR, was significantly decreased in bronchial epithelium of COPD samples vs controls. In BEAS-2B cells, cytomix and CSE stimulation reproduced the RBP pattern while increasing expression of AUF-1-regulated genes, interleukin-6, CCL2, CXCL1, and CXCL8. Silencing expression of AUF-1 reproduced, but not enhanced, target upregulation induced by cytomix compared to controls. Analysis of bronchial brushing-derived transcriptomic confirmed the selective decrease of AUF-1 in COPD vs controls and revealed significant changes in AUF-1-regulated genes by genome ontology. CONCLUSION: Downregulated AUF-1 may be pathogenic in stable COPD by altering posttranscriptional control of epithelial gene expression.
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spelling pubmed-61908132018-10-22 Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients Ricciardi, Luca Col, Jessica Dal Casolari, Paolo Memoli, Domenico Conti, Valeria Vatrella, Alessandro Vonakis, Becky M Papi, Alberto Caramori, Gaetano Stellato, Cristiana Int J Chron Obstruct Pulmon Dis Original Research PURPOSE: Inflammatory gene expression is modulated by posttranscriptional regulation via RNA-binding proteins (RBPs), which regulate mRNA turnover and translation by binding to conserved mRNA sequences. Their role in COPD is only partially defined. This study evaluated RBPs tristetraprolin (TTP), human antigen R (HuR), and AU-rich element-binding factor 1 (AUF-1) expression using lung tissue from COPD patients and control subjects and probed their function in epithelial responses in vitro. PATIENTS AND METHODS: RBPs were detected by immunohistochemistry in bronchial and peripheral lung samples from mild-to-moderate stable COPD patients and age/smoking history-matched controls; RBPs and RBP-regulated genes were evaluated by Western blot, ELISA, protein array, and real-time PCR in human airway epithelial BEAS-2B cell line stimulated with hydrogen peroxide, cytokine combination (cytomix), cigarette smoke extract (CSE), and following siRNA-mediated silencing. Results were verified in a microarray database from bronchial brushings of COPD patients and controls. RBP transcripts were measured in peripheral blood mononuclear cell samples from additional stable COPD patients and controls. RESULTS: Specific, primarily nuclear immunostaining for the RBPs was detected in structural and inflammatory cells in bronchial and lung tissues. Immunostaining for AUF-1, but not TTP or HuR, was significantly decreased in bronchial epithelium of COPD samples vs controls. In BEAS-2B cells, cytomix and CSE stimulation reproduced the RBP pattern while increasing expression of AUF-1-regulated genes, interleukin-6, CCL2, CXCL1, and CXCL8. Silencing expression of AUF-1 reproduced, but not enhanced, target upregulation induced by cytomix compared to controls. Analysis of bronchial brushing-derived transcriptomic confirmed the selective decrease of AUF-1 in COPD vs controls and revealed significant changes in AUF-1-regulated genes by genome ontology. CONCLUSION: Downregulated AUF-1 may be pathogenic in stable COPD by altering posttranscriptional control of epithelial gene expression. Dove Medical Press 2018-10-12 /pmc/articles/PMC6190813/ /pubmed/30349226 http://dx.doi.org/10.2147/COPD.S166284 Text en © 2018 Ricciardi et al. This work is published and licensed by Dove Medical Press Limited The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.
spellingShingle Original Research
Ricciardi, Luca
Col, Jessica Dal
Casolari, Paolo
Memoli, Domenico
Conti, Valeria
Vatrella, Alessandro
Vonakis, Becky M
Papi, Alberto
Caramori, Gaetano
Stellato, Cristiana
Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
title Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
title_full Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
title_fullStr Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
title_full_unstemmed Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
title_short Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
title_sort differential expression of rna-binding proteins in bronchial epithelium of stable copd patients
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6190813/
https://www.ncbi.nlm.nih.gov/pubmed/30349226
http://dx.doi.org/10.2147/COPD.S166284
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