TY - JOUR
T1 - Differential expression of RNA-binding proteins in bronchial epithelium of stable COPD patients
AU - Ricciardi, Luca
AU - Dal Col, Jessica
AU - Casolari, Paolo
AU - Memoli, Domenico
AU - Conti, Valeria
AU - Vatrella, Alessandro
AU - Vonakis, Becky M.
AU - Papi, Alberto
AU - Caramori, Gaetano
AU - Stellato, Cristiana
N1 - Funding Information:
This study was funded by POR Campania FESR 2007–2013– RETE DELLE BIOTECNOLOGIE IN CAMPANIA – Project “Terapie Innovative di Malattie Infiammatorie croniche, metaboliche, Neoplastiche e Geriatriche” – TIMING (University of Salerno, CS), FARB 2014/2015 (University of Salerno, CS, VA), and FAR 2015 (University of Ferrara, GC).
Funding Information:
This study was funded by POR Campania FESR 2007-2013-RETE DELLE BIOTECNOLOGIE IN CAMPANIA - Project “Terapie Innovative di Malattie Infiammatorie croniche, metaboliche, Neoplastiche e Geriatriche” - TIMING (University of Salerno, CS), FARB 2014/2015 (University of Salerno, CS, VA), and FAR 2015 (University of Ferrara, GC).
Publisher Copyright:
© 2018 Ricciardi et al.
PY - 2018
Y1 - 2018
N2 - 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.
AB - 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.
KW - AUF-1
KW - Airway epithelium
KW - COPD
KW - Inflammation
KW - Posttranscriptional gene regulation
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U2 - 10.2147/COPD.S166284
DO - 10.2147/COPD.S166284
M3 - Article
C2 - 30349226
AN - SCOPUS:85055205848
SN - 1176-9106
VL - 13
SP - 3173
EP - 3190
JO - International Journal of COPD
JF - International Journal of COPD
ER -