Muscarinic receptor M3 activation promotes fibrocytes contraction

Fibrocytes are monocyte-derived cells able to differentiate into myofibroblasts-like cells. We have previously shown that they are increased in the bronchi of Chronic Obstructive Pulmonary Disease (COPD) patients and associated to worse lung function. COPD is characterized by irreversible airflow ob...

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Main Authors: Pauline Henrot, Edmée Eyraud, Elise Maurat, Sophie Point, Guillaume Cardouat, Jean-François Quignard, Pauline Esteves, Thomas Trian, Pierre-Olivier Girodet, Roger Marthan, Maéva Zysman, Patrick Berger, Isabelle Dupin
Format: Article
Language:English
Published: Frontiers Media S.A. 2022-09-01
Series:Frontiers in Pharmacology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2022.939780/full
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author Pauline Henrot
Pauline Henrot
Edmée Eyraud
Elise Maurat
Sophie Point
Guillaume Cardouat
Jean-François Quignard
Pauline Esteves
Thomas Trian
Pierre-Olivier Girodet
Pierre-Olivier Girodet
Pierre-Olivier Girodet
Roger Marthan
Roger Marthan
Maéva Zysman
Maéva Zysman
Patrick Berger
Patrick Berger
Patrick Berger
Isabelle Dupin
author_facet Pauline Henrot
Pauline Henrot
Edmée Eyraud
Elise Maurat
Sophie Point
Guillaume Cardouat
Jean-François Quignard
Pauline Esteves
Thomas Trian
Pierre-Olivier Girodet
Pierre-Olivier Girodet
Pierre-Olivier Girodet
Roger Marthan
Roger Marthan
Maéva Zysman
Maéva Zysman
Patrick Berger
Patrick Berger
Patrick Berger
Isabelle Dupin
author_sort Pauline Henrot
collection DOAJ
description Fibrocytes are monocyte-derived cells able to differentiate into myofibroblasts-like cells. We have previously shown that they are increased in the bronchi of Chronic Obstructive Pulmonary Disease (COPD) patients and associated to worse lung function. COPD is characterized by irreversible airflow obstruction, partly due to an increased cholinergic environment. Our goal was to investigate muscarinic signalling in COPD fibrocytes. Fibrocytes were isolated from 16 patients with COPD’s blood and presence of muscarinic M3 receptor was assessed at the transcriptional and protein levels. Calcium signalling and collagen gels contraction experiments were performed in presence of carbachol (cholinergic agonist) ± tiotropium bromide (antimuscarinic). Expression of M3 receptor was confirmed by Western blot and flow cytometry in differentiated fibrocytes. Immunocytochemistry showed the presence of cytoplasmic and membrane-associated pools of M3. Stimulation with carbachol elicited an intracellular calcium response in 35.7% of fibrocytes. This response was significantly blunted by the presence of tiotropium bromide: 14.6% of responding cells (p < 0.0001). Carbachol induced a significant contraction of fibrocytes embedded in collagen gels (13.6 ± 0.3% versus 2.5 ± 4.1%; p < 0.0001), which was prevented by prior tiotropium bromide addition (4.1 ± 2.7% of gel contraction; p < 0.0001). Finally, M3-expressing fibrocytes were also identified in situ in the peri-bronchial area of COPD patients’ lungs, and there was a tendency to an increased density compared to healthy patient’s lungs. In conclusion, around 1/3 of COPD patients’ fibrocytes express a functional muscarinic M3 receptor. Cholinergic-induced fibrocyte contraction might participate in airway diameter reduction and subsequent increase of airflow resistance in patients with COPD. The inhibition of these processes could participate to the beneficial effects of muscarinic antagonists for COPD treatment.
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spelling doaj.art-308ff3109e1a4a8e98be9283773d13222022-12-22T01:44:13ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-09-011310.3389/fphar.2022.939780939780Muscarinic receptor M3 activation promotes fibrocytes contractionPauline Henrot0Pauline Henrot1Edmée Eyraud2Elise Maurat3Sophie Point4Guillaume Cardouat5Jean-François Quignard6Pauline Esteves7Thomas Trian8Pierre-Olivier Girodet9Pierre-Olivier Girodet10Pierre-Olivier Girodet11Roger Marthan12Roger Marthan13Maéva Zysman14Maéva Zysman15Patrick Berger16Patrick Berger17Patrick Berger18Isabelle Dupin19Univ-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceUniv-Bordeaux, Centre de Recherche Cardio-thoracique de Bordeaux, INSERM U1045, Pessac, FranceFibrocytes are monocyte-derived cells able to differentiate into myofibroblasts-like cells. We have previously shown that they are increased in the bronchi of Chronic Obstructive Pulmonary Disease (COPD) patients and associated to worse lung function. COPD is characterized by irreversible airflow obstruction, partly due to an increased cholinergic environment. Our goal was to investigate muscarinic signalling in COPD fibrocytes. Fibrocytes were isolated from 16 patients with COPD’s blood and presence of muscarinic M3 receptor was assessed at the transcriptional and protein levels. Calcium signalling and collagen gels contraction experiments were performed in presence of carbachol (cholinergic agonist) ± tiotropium bromide (antimuscarinic). Expression of M3 receptor was confirmed by Western blot and flow cytometry in differentiated fibrocytes. Immunocytochemistry showed the presence of cytoplasmic and membrane-associated pools of M3. Stimulation with carbachol elicited an intracellular calcium response in 35.7% of fibrocytes. This response was significantly blunted by the presence of tiotropium bromide: 14.6% of responding cells (p < 0.0001). Carbachol induced a significant contraction of fibrocytes embedded in collagen gels (13.6 ± 0.3% versus 2.5 ± 4.1%; p < 0.0001), which was prevented by prior tiotropium bromide addition (4.1 ± 2.7% of gel contraction; p < 0.0001). Finally, M3-expressing fibrocytes were also identified in situ in the peri-bronchial area of COPD patients’ lungs, and there was a tendency to an increased density compared to healthy patient’s lungs. In conclusion, around 1/3 of COPD patients’ fibrocytes express a functional muscarinic M3 receptor. Cholinergic-induced fibrocyte contraction might participate in airway diameter reduction and subsequent increase of airflow resistance in patients with COPD. The inhibition of these processes could participate to the beneficial effects of muscarinic antagonists for COPD treatment.https://www.frontiersin.org/articles/10.3389/fphar.2022.939780/fullfibrocyteCOPDM3cholinergiccontraction
spellingShingle Pauline Henrot
Pauline Henrot
Edmée Eyraud
Elise Maurat
Sophie Point
Guillaume Cardouat
Jean-François Quignard
Pauline Esteves
Thomas Trian
Pierre-Olivier Girodet
Pierre-Olivier Girodet
Pierre-Olivier Girodet
Roger Marthan
Roger Marthan
Maéva Zysman
Maéva Zysman
Patrick Berger
Patrick Berger
Patrick Berger
Isabelle Dupin
Muscarinic receptor M3 activation promotes fibrocytes contraction
Frontiers in Pharmacology
fibrocyte
COPD
M3
cholinergic
contraction
title Muscarinic receptor M3 activation promotes fibrocytes contraction
title_full Muscarinic receptor M3 activation promotes fibrocytes contraction
title_fullStr Muscarinic receptor M3 activation promotes fibrocytes contraction
title_full_unstemmed Muscarinic receptor M3 activation promotes fibrocytes contraction
title_short Muscarinic receptor M3 activation promotes fibrocytes contraction
title_sort muscarinic receptor m3 activation promotes fibrocytes contraction
topic fibrocyte
COPD
M3
cholinergic
contraction
url https://www.frontiersin.org/articles/10.3389/fphar.2022.939780/full
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