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N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor
Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis, alteration in the microvasculature and immunologic abnormalities. It has been hypothesized that an abnormal redox state could regulate the persistent fibrotic phenotype in SSc patients. N-Formyl peptide receptors (FP...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5893650/ https://www.ncbi.nlm.nih.gov/pubmed/29670612 http://dx.doi.org/10.3389/fimmu.2018.00574 |
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author | Napolitano, Filomena Rossi, Francesca Wanda Pesapane, Ada Varricchio, Silvia Ilardi, Gennaro Mascolo, Massimo Staibano, Stefania Lavecchia, Antonio Ragno, Pia Selleri, Carmine Marone, Gianni Matucci-Cerinic, Marco de Paulis, Amato Montuori, Nunzia |
author_facet | Napolitano, Filomena Rossi, Francesca Wanda Pesapane, Ada Varricchio, Silvia Ilardi, Gennaro Mascolo, Massimo Staibano, Stefania Lavecchia, Antonio Ragno, Pia Selleri, Carmine Marone, Gianni Matucci-Cerinic, Marco de Paulis, Amato Montuori, Nunzia |
author_sort | Napolitano, Filomena |
collection | PubMed |
description | Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis, alteration in the microvasculature and immunologic abnormalities. It has been hypothesized that an abnormal redox state could regulate the persistent fibrotic phenotype in SSc patients. N-Formyl peptide receptors (FPRs) are chemotactic receptors overexpressed in fibroblasts derived from SSc patients. In this study, we demonstrated that stimulation of FPRs promotes the generation of reactive oxygen species (ROS) in skin fibroblasts. In fibroblast cells, ROS production was due to FPRs interaction with the urokinase receptor (uPAR) and to β(1) integrin engagement. FPRs cross-talk with uPAR and integrins led to Rac1 and ERKs activation. FPRs stimulation increased gp91phox and p67phox expression as well as the direct interaction between GTP-Rac1 and p67phox, thus promoting assembly and activation of the NADPH oxidase complex. FPRs functions occur through interaction with a specific domain of uPAR (residues (88)SRSRY(92)) that can be exposed on the cell membrane by protease-mediated receptor cleavage. Immunohistochemistry analysis with a specific anti-SRSRY antibody showed increased expression of uPAR in a cleaved form, which exposes the SRSRY sequence at its N-terminus (DIIDIII-uPAR88–92) in skin biopsies from SSc patients. As expected by the increased expression of both FPRs and DII-DIII-uPAR(88-92), fibroblasts derived from SSc patients showed a significantly increase in ROS generation both at a basal level than after FPRs stimulation, as compared to fibroblasts from normal subjects. C37, a small molecule blocking the interaction between FPRs and uPAR, and selumetinib, a clinically approved MAPKK/ERK inhibitor, significantly inhibited FPRs-mediated ROS production in fibroblasts derived from SSc patients. Thus, FPRs, through the interaction with the uPA/uPAR system, can induce ROS generation in fibroblasts by activating the NADPH oxidase, playing a role in the alteration of the redox state observed in SSc. |
format | Online Article Text |
id | pubmed-5893650 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-58936502018-04-18 N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor Napolitano, Filomena Rossi, Francesca Wanda Pesapane, Ada Varricchio, Silvia Ilardi, Gennaro Mascolo, Massimo Staibano, Stefania Lavecchia, Antonio Ragno, Pia Selleri, Carmine Marone, Gianni Matucci-Cerinic, Marco de Paulis, Amato Montuori, Nunzia Front Immunol Immunology Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis, alteration in the microvasculature and immunologic abnormalities. It has been hypothesized that an abnormal redox state could regulate the persistent fibrotic phenotype in SSc patients. N-Formyl peptide receptors (FPRs) are chemotactic receptors overexpressed in fibroblasts derived from SSc patients. In this study, we demonstrated that stimulation of FPRs promotes the generation of reactive oxygen species (ROS) in skin fibroblasts. In fibroblast cells, ROS production was due to FPRs interaction with the urokinase receptor (uPAR) and to β(1) integrin engagement. FPRs cross-talk with uPAR and integrins led to Rac1 and ERKs activation. FPRs stimulation increased gp91phox and p67phox expression as well as the direct interaction between GTP-Rac1 and p67phox, thus promoting assembly and activation of the NADPH oxidase complex. FPRs functions occur through interaction with a specific domain of uPAR (residues (88)SRSRY(92)) that can be exposed on the cell membrane by protease-mediated receptor cleavage. Immunohistochemistry analysis with a specific anti-SRSRY antibody showed increased expression of uPAR in a cleaved form, which exposes the SRSRY sequence at its N-terminus (DIIDIII-uPAR88–92) in skin biopsies from SSc patients. As expected by the increased expression of both FPRs and DII-DIII-uPAR(88-92), fibroblasts derived from SSc patients showed a significantly increase in ROS generation both at a basal level than after FPRs stimulation, as compared to fibroblasts from normal subjects. C37, a small molecule blocking the interaction between FPRs and uPAR, and selumetinib, a clinically approved MAPKK/ERK inhibitor, significantly inhibited FPRs-mediated ROS production in fibroblasts derived from SSc patients. Thus, FPRs, through the interaction with the uPA/uPAR system, can induce ROS generation in fibroblasts by activating the NADPH oxidase, playing a role in the alteration of the redox state observed in SSc. Frontiers Media S.A. 2018-04-04 /pmc/articles/PMC5893650/ /pubmed/29670612 http://dx.doi.org/10.3389/fimmu.2018.00574 Text en Copyright © 2018 Napolitano, Rossi, Pesapane, Varricchio, Ilardi, Mascolo, Staibano, Lavecchia, Ragno, Selleri, Marone, Matucci-Cerinic, de Paulis and Montuori. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Napolitano, Filomena Rossi, Francesca Wanda Pesapane, Ada Varricchio, Silvia Ilardi, Gennaro Mascolo, Massimo Staibano, Stefania Lavecchia, Antonio Ragno, Pia Selleri, Carmine Marone, Gianni Matucci-Cerinic, Marco de Paulis, Amato Montuori, Nunzia N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor |
title | N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor |
title_full | N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor |
title_fullStr | N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor |
title_full_unstemmed | N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor |
title_short | N-Formyl Peptide Receptors Induce Radical Oxygen Production in Fibroblasts Derived From Systemic Sclerosis by Interacting With a Cleaved Form of Urokinase Receptor |
title_sort | n-formyl peptide receptors induce radical oxygen production in fibroblasts derived from systemic sclerosis by interacting with a cleaved form of urokinase receptor |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5893650/ https://www.ncbi.nlm.nih.gov/pubmed/29670612 http://dx.doi.org/10.3389/fimmu.2018.00574 |
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