Revue des maladies respiratoires最新文献

{"title":"Role of CRACR2A in the development of pulmonary arterial hypertension","authors":"A. Saint-Martin Willer ,&nbsp;B. Masson ,&nbsp;M. Dutheil ,&nbsp;Y. Ruchon ,&nbsp;M. Humbert ,&nbsp;D. Montani ,&nbsp;V. Capuano ,&nbsp;F. Antigny","doi":"10.1016/j.rmr.2025.02.018","DOIUrl":"10.1016/j.rmr.2025.02.018","url":null,"abstract":"<div><h3>Introduction</h3><div>Pulmonary arterial hypertension (PAH) is a rare, progressive, and devastating disease resulting from the obstruction of distal pulmonary arteries. The pathobiology of PAH is multifactorial, involving all the cell spectrum of pulmonary circulation, including pulmonary arterial (PA) smooth muscle cells (PASMCs), endothelial cells (PECs), and PA adventitial fibroblasts (PAAFs). These cellular dysfunctions are characterized by enhanced migration, proliferation and apoptosis resistance. Moreover, in PAH-hPASMCs, calcium signaling is exacerbated contributing to hPASMCs abnormalities. A mutation of CRACR2A, a regulatory partner of the calcium (Ca²⁺) channel Orai1 has been identified in an Italian PAH patients’ cohort suggesting its involvement in PAH development. Orai1, the main Ca²⁺ channel of the Store-Operated Ca²⁺ Entry (SOCE) is increased in expression and function in PAH-hPASMCs and participate to PAH pathogenesis. SOCE is triggered by STIM1, a calcium sensor of the endoplasmic reticulum (ER) that activates Orai1 after ER calcium depletion. CRACR2A is known to stabilize Orai1/STIM1 interaction.</div></div><div><h3>Methods</h3><div>We used a whole exome sequencing approach to identify <em>CRACR2A</em> variants. We used a siRNA strategy against CRACR2A in hPASMCs, hPECs, and hPAAF isolated from control and PAH patients. We then measured the consequences of the CRACR2A knockdown on SOCE by Ca²⁺ imaging measurement, on cell proliferation, migration and <em>in vitro</em> angiogenesis. Finally, we measured the consequence on SOCE and Orai1 protein expression in PAH-hPASMCs overexpressing CRACR2A.</div></div><div><h3>Results</h3><div>We identified additional variants of <em>CRACR2A</em> in PAH patients strengthening CRACR2A involvement in PAH development. Using immunofluorescence staining on human control and PAH lungs, we found that CRACR2A is expressed in control and PAH. By immunoblot, we showed that CRACR2A protein expression is unchanged in control and PAH-hPECs and hPAAFs, whereas its expression is reduced in PAH-hPASMCs. By the siRNA strategy, we found that SOCE, migration, and proliferation were decreased in control-hPASMCs transfected with siCRACR2A. Surprisingly, in PAH-hPASMCs, the knockdown of CRACR2A strongly increased SOCE and Orai1 expression. In contrast, CRACR2A overexpression in PAH-hPASMCs reduced SOCE. In PAH-hPAAFs, the knockdown of CRACR2A increases SOCE as in PAH-hPASMCs.</div></div><div><h3>Conclusion</h3><div>These results indicate that CRACR2A plays an essential role in pulmonary vascular cells from control and PAH patients by its involvement in SOCE, proliferation, and migration. The reduced CRACR2A expression in PAH-hPASMCs could partly explain increased Orai1 expression and function in PAH-hPASMCs. These results suggest that CRACR2A dysfunction is involved in PAH pathogenesis.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 190-191"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Jacketed respiratory inductive plethysmography in rats: A translational non-invasive method for cardiorespiratory exercise monitoring","authors":"A. Cambier ,&nbsp;S. Tanguy ,&nbsp;T. Flenet ,&nbsp;F. Boucher","doi":"10.1016/j.rmr.2025.02.090","DOIUrl":"10.1016/j.rmr.2025.02.090","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;div&gt;Respiratory inductive plethysmography (RIP) is one of the least restrictive methods for evaluating pulmonary ventilation. It has been used in humans since the 1980s as one of the clinical tests for functional exploration of the respiratory system under different conditions (sleep, rest or exercise) in healthy and in respiratory pathology patients. This method has been adapted and integrated into jacketed telemetry systems used in preclinical large animal research (safety pharmacology, fundamental or applied research, etc.). Thanks to the recent miniaturisation of jacketed telemetric systems, a new solution is available for assessing respiratory function combined with ECG in small mammals &lt;span&gt;&lt;span&gt;[1]&lt;/span&gt;&lt;/span&gt; (i.e. rats). As exercise was initially used to validate the measurement accuracy of dual-band respiratory inductance plethysmography in humans, it was of interest to qualify this solution in a similar way in rats. The aim is to assess the feasibility of a translational model using this technology to monitor physiological changes associated with exercise on a treadmill.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;div&gt;For this purpose, male Wistar rats (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;10.8-9 weeks old) were exposed to an incremental treadmill exercise protocol with speed levels from 5 to 45&lt;!--&gt; &lt;!--&gt;cm. s-1. Respiratory parameters (respiratory rate (RespR), minute ventilation (VM), Tidal volume (VT)) and peak inspiratory and expiratory flow (PIF, PEF, PENH) as well as cardiac (heart rate (HR)) and activity level (AL) were continuously monitored and analyzed over the last 30&lt;!--&gt; &lt;!--&gt;seconds of each level.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;div&gt;All animals were successfully fitted with the device and returned to their home cages. The control values measured after stabilization were for RespR 165&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;8&lt;!--&gt; &lt;!--&gt;brpm, HR 407&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;9&lt;!--&gt; &lt;!--&gt;bpm and AL 24&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;4&lt;!--&gt; &lt;!--&gt;mg. At least all animals ran successfully on the treadmill with the vest up to 35&lt;!--&gt; &lt;!--&gt;cm/sec (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;10), some reaching 45&lt;!--&gt; &lt;!--&gt;cm/sec (&lt;em&gt;n&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;6). For the maximum plateau, exercise induced a significant physiological increase (&lt;span&gt;&lt;span&gt;Fig. 1&lt;/span&gt;&lt;/span&gt;) in respiratory parameters (RespR +74%, 116&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;26 brpm, &lt;em&gt;P&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.01**//MV +334%, 1099&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;197&lt;!--&gt; &lt;!--&gt;mL. min-1, &lt;em&gt;P&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.01**//VT +197%, 4&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;1&lt;!--&gt; &lt;!--&gt;mL, &lt;em&gt;P&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.01**), as well as HR (+36%, 140&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;13 bpm, &lt;em&gt;P&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.05*) and AL (+2358%, 504&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;57 bpm, &lt;em&gt;P&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.05*). PIF and PEF showed a significant increase of 86&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;9&lt;!--&gt; &lt;!--&gt;ml. s-1 and 52&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;11&lt;!--&gt; &lt;!--&gt;mL. s- respectively. enhanced Pause (PENH) remained stable during exercise indicating ","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 227"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143792058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Airway-delivered propionate boost immune tolerance during allergic asthma in mice","authors":"HL. Mai ,&nbsp;B. Misme-Aucouturier ,&nbsp;M. Decarvalho ,&nbsp;J. Marteau ,&nbsp;E. Martin ,&nbsp;E. Nguyen ,&nbsp;S. Brouard ,&nbsp;G. Bouchaud","doi":"10.1016/j.rmr.2025.02.036","DOIUrl":"10.1016/j.rmr.2025.02.036","url":null,"abstract":"<div><h3>Background</h3><div>Allergic Asthma's pathomechanism is essentially a disruption of bronchial homeostasis characterized by an increase in proinflammatory mediators such as T and B effector cells, over immune regulatory cell signaling. Recent work has brought to the forefront the pivotal role of gut microbial metabolites, SCFAs, in abrogating the cardinal features of asthma, such as airway inflammation and AHR, via induction of tolerogenic pathways, tipping the balance towards homeostasis recovery.</div></div><div><h3>Objective</h3><div>The goal was to investigate on a broader scale the immune-regulatory role of propionate in allergic airway- inflammation, in vivo, in an asthma mouse model, and in vitro on human B cell behavior.</div></div><div><h3>Material and Methods</h3><div>The preventive effect of propionate was tested in vivo by treating (or not) BALB/c mice with airway-delivered propionate during house dust mite (HDM) allergen induction of asthma, at the indicated time points. Asthma-related features such as airway function and inflammation were assessed through airway hyperresponsiveness (AHR) measurement, and immune cell profile analysis performed on blood, bronchoalveolar lavage fluid, lungs and spleen, respectively. In vitro, proliferation, survival and intracellular IL-10 expression of isolated purified human B cells was determined upon propionate treatment or not, by flow cytometry.</div></div><div><h3>Results</h3><div>In vivo, Propionate treatment ameliorated AHR after allergic challenge and reduced eosinophil, neutrophil and T lymphocyte but not macrophages extraversion to the lungs. Lung tissue from propionate treated allergic mice showed increased Treg cells, decreased Th2 and Th17 cells and increased B regulatory IL-10 producing cells and Granzyme B cells compared to asthmatic controls. In vitro, propionate dose dependently reversed CPG, CD₄₀L induced B cell proliferation and IL-10 secretion.</div></div><div><h3>Conclusion</h3><div>Direct lung delivery of propionate improved allergen-induced airway inflammation by attenuating lung eosinophilia, neutrophilia and AHR. As well as Treg and B reg over effector cell differentiation in vivo and inhibited B cell Proliferation in vitro. Authors do not have conflict of interest to declare.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 199-200"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application de la microscopie à force atomique dans l’analyse fonctionnelle des éosinophiles","authors":"V. Spasovski ,&nbsp;A. Ecrement ,&nbsp;T. Gasser ,&nbsp;C. Elie-Caille ,&nbsp;G. Rolin ,&nbsp;C. Barnig","doi":"10.1016/j.rmr.2025.02.041","DOIUrl":"10.1016/j.rmr.2025.02.041","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;div&gt;Les éosinophiles sont des cellules du système immunitaire appartenant à la famille des granulocytes. Ces cellules possèdent de nombreuses fonctions comme l’élimination des pathogènes, tout en jouant un rôle dans l’homéostasie tissulaire et l’immunorégulation. Les éosinophiles peuvent changer de morphologie en réponse à une activation, une adhésion, une migration, ou des interactions cellulaires. La microscopie à force atomique (AFM ou Atomic Force Microscope) a émergé ces dernières années comme un outil précieux pour diverses applications physiques et biologiques. En biologie ultrastructurale, l’AFM permet de détecter des caractéristiques morphologiques sub-nanométriques. Grace à l’AFM, il est ainsi possible d’observer des modifications morphologiques et de mieux comprendre les mécanismes sous-jacents à l’activation et à la fonction des éosinophiles.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Méthodes&lt;/h3&gt;&lt;div&gt;Nous avons développé différents modèles fonctionnels des éosinophiles. Tout d’abord, un modèle d’adhésion utilisant de la fibronectine a été étudié en plaçant les éosinophiles dans des puits coatés préalablement avec de la fibronectine (20&lt;!--&gt; &lt;!--&gt;μg/mL pendant 24&lt;!--&gt; &lt;!--&gt;h à 4&lt;!--&gt; &lt;!--&gt;°C). L’adhésion des éosinophiles a été mesurée au cours du temps. Nous avons également mis au point un modèle de libération de pièges extracellulaires par les éosinophiles (EETosis ou eosinophil extracellular trap). L’induction de l’EETosis par le Phorbol myristate acetate (PMA) (50&lt;!--&gt; &lt;!--&gt;ng/mL) a été suivie en temps réel avec un microscope (Incucyte S3, Sartorius) en utilisant un colorant fluorescent vert spécifique aux acides nucléiques. Enfin, l’imagerie par AFM a été réalisée en mode contact (pointe DI 0,3&lt;!--&gt; &lt;!--&gt;N/m, Nanowizard 3, Bruker, USA) sur des cellules Eol-1 différenciées par action d’acide butyrique (0,5&lt;!--&gt; &lt;!--&gt;mM) et après fixation s au glutaraldéhyde (0,5 %) sur des lames en verre.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Résultats&lt;/h3&gt;&lt;div&gt;Nous avons observé une augmentation significative de l’adhésion des éosinophiles en présence de fibronectine, avec un pic à 1 heure (8,1 % &lt;em&gt;vs.&lt;/em&gt; 65,58 %). L’induction de l’EETosis était très nette lorsque les éosinophiles sont activés avec un pic à 10&lt;!--&gt; &lt;!--&gt;heures (1,81 % &lt;em&gt;vs.&lt;/em&gt; 38,2 %). L’imagerie par AFM a révélé des différences morphologiques et physiques notables entre les cellules Eol-1 différenciées et non différenciées, démontrant la faisabilité de cette technique pour observer des changements morphologiques des éosinophiles.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;div&gt;Nos modèles fonctionnels et les techniques d’imagerie par AFM développés dans cette étude ouvrent de nouvelles perspectives pour l’analyse détaillée des propriétés morphologiques et mécaniques des éosinophiles. Cette approche novatrice nous permettra de suivre avec précision les modifications induites par l’activation, l’adhésion et autres stimulations des éosinophiles, améliorant ainsi notre compréhension de leurs fonc","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 202"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791876","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fine-tuning levels of filamins A and B as a specific mechanism sustaining Th2 lymphocyte functions","authors":"K. Maire ,&nbsp;L. Chamy ,&nbsp;S. Ghazali ,&nbsp;M. Carratala-Lasserre ,&nbsp;M. Zahm ,&nbsp;C. Bouisset ,&nbsp;A. Métais ,&nbsp;L. Combes-Soia ,&nbsp;L. de la Fuente-Vizuete ,&nbsp;H. Trad ,&nbsp;A. Chaubet ,&nbsp;M. Savignac ,&nbsp;A. Gonzalez de Peredo ,&nbsp;A. Subramaniam ,&nbsp;O. Joffre ,&nbsp;P. Lutz ,&nbsp;I. Lamsoul","doi":"10.1016/j.rmr.2025.02.034","DOIUrl":"10.1016/j.rmr.2025.02.034","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;div&gt;Asthma is a chronic inflammatory disease of lower airways that affects nearly 400 million people worldwide. Nearly half of asthmatic patients present a high type-2 inflammation characterized by a massive recruitment of T helper 2 (Th2) lymphocytes producing type 2 cytokines, eosinophilia and high levels of IgE. Treatments targeting type 2 cytokines and their receptors but also targeting IgE are already used in clinics. An attractive therapy would consist in targeting the recruitment of Th2 lymphocytes to inflammatory sites. We previously showed that the &lt;em&gt;ASB2&lt;/em&gt; gene that encodes the specificity subunit of an E3 ubiquitin ligase belongs to the core set of Th2-specific genes (Cancer Immunol Res 2019.7: 1332–1344).&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;div&gt;&lt;em&gt;ASB2&lt;/em&gt; knockout mice and small molecules were used to modulate the levels of ASB2α and its substrates, filamins a and b (FLNa/b), in mouse primary Th2 lymphocytes and/or PBMC from healthy donors. In vitro experiments including global transcriptomic and proteomic approaches, high content and live imaging, as well as several immunological techniques were used to decipher the role of ASB2α in Th2 lymphocytes. In vivo mouse models of airway inflammation and asthma were used to investigate the role of the ASB2α in type 2 inflammation and determine whether the ASB2α-FLNa/b axis represents a therapeutic opportunity in Th2-driven diseases.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;div&gt;Here, we show that ASB2α triggers ubiquitylation and proteasomal degradation of FLNa/b in Th2 lymphocytes. As a consequence, Th2 lymphocytes exhibit lower levels of FLNa/b proteins compared to naïve CD&lt;sub&gt;4&lt;/sub&gt; &lt;sup&gt;+&lt;/sup&gt; T lymphocytes or other T helper subsets, associated with an elongated shape and a specific dynamic migration pattern depending on integrin αVß3 activation. Furthermore, we show decreased recruitment of Th2 lymphocytes in inflamed lungs of ASB2α-deficient mice submitted to ovalbumin or house dust mite induced airway inflammation, associated with decreased eosinophilia and decreased type 2 cytokine production. Using a model of passive asthma that relies on the injection of ovalbumin-specific Th2 lymphocytes and ovalbumin inhalations, we show that mice that received ASB2α-deficient or accumulating FLNa/b Th2 lymphocytes exhibit decreased airway inflammation but also mucus secretion and remodeling of the airways compared to mice receiving control Th2 lymphocytes.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;div&gt;In conclusion, our study highlights the key role of a fine regulation of FLNa/b levels in Th2 lymphocyte functions as a driver of Th2-dependent airway inflammation. Our results show that augmenting FLNa/b levels in Th2 lymphocytes benefits in asthma features, indicating that ASB2α and its substrates FLNa/b may represent novel pharmacological targets in type 2 pathologies (Preprint (v1) Res. Square, [&lt;span&gt;&lt;span&gt;https://doi.org/10.21203/rs.3.rs-3878460/v1&lt;/span&gt;&lt;svg&gt;&lt;path&gt;&lt;/path&gt;","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 198-199"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Is the Cx43-specific inhibitor 43Gap26 a relevant therapeutic target in BPD and BPD-PH induced by hyperoxia?","authors":"C.M. Pilard ,&nbsp;L. Gassiat ,&nbsp;C. Cardouat ,&nbsp;I. Gauthereau ,&nbsp;P. Robillard ,&nbsp;E. Dumas-de-la-Roque ,&nbsp;F. Sauvestre ,&nbsp;F. Pelluard ,&nbsp;S. Berenguer ,&nbsp;L. Sentilhes ,&nbsp;F. Coatleven ,&nbsp;M. Vincienne ,&nbsp;R. Marthan ,&nbsp;P. Berger ,&nbsp;V. Freund-Michel ,&nbsp;C. Guibert","doi":"10.1016/j.rmr.2025.02.080","DOIUrl":"10.1016/j.rmr.2025.02.080","url":null,"abstract":"<div><h3>Introduction</h3><div>Premature newborns exposed to oxygen are at risk for bronchopulmonary dysplasia (BPD), characterized by lung growth arrest and inflammation. Connexin 43-gap junction (Cx43-GJ) plays a central role in lung development and macrophages-induced inflammation, and is increased by hyperoxia. We thus explored the role of CX43-GJ in experimental BPD.</div></div><div><h3>Methods</h3><div>We used neonatal rats exposed to normoxia or hyperoxia (90% O₂) and daily treated or not with a specific Cx43 inhibitor (43Gap26) during 14 days. Some experiments were also performed in human fetal pulmonary artery smooth muscle cells (HfPA-SMC) exposed to normoxia or hyperoxia (60% O₂) and treated or not with 43Gap26 for 2 days.</div></div><div><h3>Results</h3><div>In vivo, we showed that 43Gap26 decreased hyperoxia-induced increased Cx43 expression in lung and improved hypo-alveolarization. However, 43Gap26 did not prevent 1) the hyperoxia-induced alteration of lung function (plethysmographie), 2) the decreased SP-B expression, 3) the disruption of extracellular matrix components (fibronectin and collagen I/III), 4) the increased macrophagic infiltration and M2 polarization, and 5) the increased secretion of pro-inflammatory cytokines (Tissue Inhibitor of Metalloproteinases-1). Furthermore, 43Gap26 blocked the hyperoxia-induced proliferation of type II alveolar epithelial cells, and did not prevent BPD-associated pulmonary hypertension (PH-BPD) or the increased mortality. In vitro, on HfPA-SMC, we confirmed that although an increased Cx43 expression was induced by hyperoxia, 43Gap26 had no effect on the hyperoxia-induced increase of pro-inflammatory cytokines such as IL-6 and Macrophage Inhibitory Factor (MIF). Finally, although we did not demonstrate an increase in oxidative stress in our 2 models, 43Gap26 decreased the hyperoxia-induced increase in heme oxygenase-1 expression in pulmonary arteries.</div></div><div><h3>Conclusion</h3><div>Despite an improved alveolarization in an animal model of hyperoxia-induced BPD, specific inhibition of Cx43 does not appear to be a new potential therapeutic strategy in BPD and PH-BPD as it did not prevent mortality as well as the main hallmarks of BPD and PH-BPD (altered lung function, pulmonary vascular growth arrest and remodeling, inflammation).</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 221"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impaired lung epithelial-associated phagocytes recruitment during sepsis: A pivotal role of TLR2 signalling","authors":"N. Jebnoun ,&nbsp;L. Boulant ,&nbsp;C. Rousseau ,&nbsp;L. Regard ,&nbsp;C. Martin ,&nbsp;P.R. Burgel ,&nbsp;F. Pène ,&nbsp;M.Z.L. Ladjemi","doi":"10.1016/j.rmr.2025.02.065","DOIUrl":"10.1016/j.rmr.2025.02.065","url":null,"abstract":"<div><h3>Introduction</h3><div>Post-septic hosts exhibit altered lung immunity, leading to increased susceptibility to bacterial pneumonia. Epithelial cells can attract immune cells to the lung upon injury. Phagocytic cells are essential to epithelial wound healing by clearing debris, regulating inflammation, and promoting tissue repair. Our team showed that TLR2 acts as a susceptibility factor to secondary bacterial pneumonia in a murine model of polymicrobial peritonitis <span><span>[1]</span></span>. We also demonstrated that sepsis induces TLR2-dependent bronchial epithelial morphological and functional alterations including dysplasia <span><span>[2]</span></span>. Hence, we hypothesized that sepsis might induce dysfunction of the bronchial epithelium ability to recruit and activate phagocytic immune cells.</div></div><div><h3>Methods</h3><div>C57BL/6<!--> <!-->J female mice were subjected to polymicrobial peritonitis induced by cecal ligation and puncture (CLP) or control sham surgery. Quantity and localisation of polynuclear neutrophils and macrophages were studied by immunohistochemistry at day 7 after surgery, prior to any secondary bacterial challenge.</div></div><div><h3>Results</h3><div>As compared to sham-operated mice, post-septic mice showed a significant decrease of whole lung and of epithelial-associated macrophages (median epithelial-associated macrophages staining ratio was 5.3% for septic mice compared to 2,2% for the controls, <em>P</em> <!-->=<!--> <!-->0,048). No increase in epithelial-associated polynuclear neutrophils was observed despite a significant overall lung infiltration (median epithelial-associated neutrophils staining ratio 1,484% for septic mice compared to 0,7151% for the controls, <em>P</em> <!-->=<!--> <!-->0,2571). Conversely, tlr2-/- post septic mice exhibited a significant increase of whole lung and epithelial-associated macrophages and polynuclear neutrophils in comparison with tlr2-/- control mice (median epithelial-associated macrophages staining ratio 6.726% for septic mice compared to 3,270% for the controls, <em>P</em> <!-->=<!--> <!-->0,0159; median epithelial associated neutrophils staining ratio 6,530% for septic mice compared to 2,665% for the controls, <em>P</em> <!-->=<!--> <!-->0.0159).</div></div><div><h3>Conclusion</h3><div>Our data show a TLR2-dependent defect in epithelial-recruitment of phagocytes post sepsis. These results suggest a crucial role of phagocytes in previously observed sepsis-induced alteration in epithelial morphology and functions (e.g. epithelial dysplasia) which may contribute to the increased susceptibility of post-septic hosts to secondary pneumonia. Further ex vivo experiments using primary air-liquid interface epithelial cultures from WT and tlr2-/- post-septic mice will allow us to identify the contribution of epithelial and immune TLR2 signalling in these alterations.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 214"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rhinovirus and severe asthma: Decoding epithelial dysfunction and inflammatory responses","authors":"K. Valette ,&nbsp;L. Moreno ,&nbsp;E. Redman ,&nbsp;A.S. Payet ,&nbsp;S. Ambard ,&nbsp;L.E. Zaragosi ,&nbsp;P. Chanez ,&nbsp;D. Gras","doi":"10.1016/j.rmr.2025.02.056","DOIUrl":"10.1016/j.rmr.2025.02.056","url":null,"abstract":"<div><h3>Introduction</h3><div>Alteration of bronchial epithelium is one of the main features of severe asthma. Bronchial epithelial cells promote and perpetuate airway inflammation and structural changes leading to persistent symptoms, bronchial obstruction and hyperresponsiveness. Bronchial epithelial cells are the first barriers in viral infections, which are the major contributor to asthma exacerbations. However, the consequences of the persisting long-term responses to viral infection of bronchial epithelium in severe asthma are not fully understood. Therefore, we investigated the in vitro response to rhinovirus of epithelial cells obtained from control donors or severe asthma patients and performed a long-term follow-up.</div></div><div><h3>Methods</h3><div>Primary human bronchial epithelial cells (HBECs) from control (C) (<em>n</em> <!-->=<!--> <!-->6) and severe asthmatics (SA) (<em>n</em> <!-->=<!--> <!-->6), cultured in air-liquid interface (ALI), were infected with Rhinovirus A16 (RV-A16). Differences in term of viral replication, epithelium cohesion (TEER measurement), antiviral and inflammatory responses (protein secretion) were assessed at the acute phase and at the late phase. Moreover, a single-cell RNA-sequencing analysis was performed at 3-, 7- and 14-days post-infection.</div></div><div><h3>Results</h3><div>RV-A16 replication increased during the acute phase then decreased during the late phase, without any significative difference between C and SA. Epithelium cohesion was damaged by RV-A16 in SA at the late phase whereas it was strengthened in C. RV-A16 induced the secretion of various antiviral defence peptides (IFN type I and III) and inflammatory mediators (CXCL10, IL 33, TSLP) over the first week post-infection, with different kinetics and intensities between C and SA. For example, IL-33, TSLP, IFNλ2-3 and IFNα2 secretions were significantly more induced in SA than in C. By contrast, MUC5AC secretion was significantly less important in SA than in C. Single-cell RNA sequencing data revealed that SA epithelia displayed fewer genes up-regulated after RV-A16 infection than C epithelia for all cell types (<em>max 202</em> genes upregulated in multiciliated cells vs <em>max 1262</em> genes upregulated in goblet cells respectively at 3 dpi). NF-kB pathway (e.g: RIPK1, TNFAIP3, NFKBIA, MYD₈₈) was enriched only in C at 3 dpi, not in SA.</div></div><div><h3>Conclusion</h3><div>The antiviral response is different between control and severe asthmatic, both in the acute and late phases. The virus induces alteration of epithelium cohesion in severe asthmatics, suggesting a defect in cellular events normally induced by injury. Then, an altered response of HBECs in severe asthma, illustrated by the increased secretion of epithelial alarmins and the deficiency of NF-kB pathway, could contribute to increased disease susceptibility and an enhanced persistent inflammatory response to rhinovirus infection.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 209-210"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bénéfices de la réparation épithéliale bronchique par des iPSC dans la BPCO","authors":"M. Nadaud ,&nbsp;C. Bourdais ,&nbsp;A. Fort-Petit ,&nbsp;E. Ahmed ,&nbsp;K. Hireche ,&nbsp;I. Vachier ,&nbsp;J. Vos (de) ,&nbsp;A. Bourdin","doi":"10.1016/j.rmr.2025.02.048","DOIUrl":"10.1016/j.rmr.2025.02.048","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;div&gt;La BPCO est une maladie influencée par l’environnement et les modifications épigénétiques qu’il génère. Cette pathologie pourrait être le résultat d’un profond dysfonctionnement épithélial, probablement au niveau des sous-types de cellules Club, en réponse à un environnement toxique. Notre objectif est d’améliorer la réparation épithéliale des voies respiratoires de patients atteints de le BPCO, en greffant des cellules souches pluripotentes induites (iPSC) prédifférenciées, provenant du même patient.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Méthodes&lt;/h3&gt;&lt;div&gt;Des cellules épithéliales provenant de biopsies bronchiques ont été cultivées en interface air-liquide (ALI). Les épithéliums bronchiques natifs pseudostratifiés ont été lésés mécaniquement. Les cellules épithéliales dérivées d’iPSC provenant du sang de patients &lt;span&gt;&lt;span&gt;[1]&lt;/span&gt;&lt;/span&gt; ont été modifiées pour exprimer la GFP. Les cellules issues d’iPSC reprogrammées ont été différenciées jusqu’au stade de maturation précoce vAFE (ventral anterior foregut endodermis) et ont été ajoutées à l’épithélium natif endommagé. Le ratio utilisé pour la quantité de cellules issues d’IPSC ajoutée est de 2 cellules issues d’iPSC au stade vAFE pour 1 cellule épithéliale bronchique. L’ensemencement de cellules issues d’iPSC directement après la lésion de l’épithélium bronchique s’est avéré être la condition la plus appropriée pour simuler un protocole de thérapie cellulaire. L’expression de la GFP a été contrôlée et quantifiée par microscopie à fluorescence à partir du jour de la greffe et pendant 42&lt;!--&gt; &lt;!--&gt;jours pour surveiller la réparation des lésions et évaluer la différenciation des cellules greffées. La TEER et la fermeture de la lésion ont été quantifiées. Les puits de culture ont été fixés au formol pour réaliser des marquages par immunofluorescence (GFP, epcam, eCAD, p63, tubIV).&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Résultats&lt;/h3&gt;&lt;div&gt;Les cellules GFP dérivées d’iPSC au stade vAFE ont été greffées avec succès au niveau du site de la lésion épithéliale de l’ALI. Cela a été confirmé par l’observation de la GFP au microscope optique à fluorescence. La détection de la fluorescence au jour 3, qui s’est intensifiée jusqu’au jour 7, était toujours présente au jour 42, soulignant la viabilité et la stabilité à long terme des cultures greffées. La quantification de la fluorescence indique une augmentation de l’aire marquée par la fluorescence comparée à l’aire totale, au cours du temps. La fermeture de la lésion est plus rapide au cours des premiers jours sur un épithélium greffé, comparé à un épithélium non greffé. Les expériences ont été réalisées sur des épithéliums bronchiques de 14 patients, plus de 120 puits de culture ont été greffés et la greffe a été un succès à chaque fois. La coloration par immunofluorescence au jour 7 a également démontré que les cellules GFP+ étaient épithéliales (epCAM+) et formaient des jonctions intercellulaires (eCAD) avec les cellules GFP-. Certaines cellules GFP+ se sont","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 205-206"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification de nouveaux candidats moléculaires dans les altérations associées aux cils dans l’asthme sévère","authors":"M.A. Devilliers ,&nbsp;A. Brisebarre ,&nbsp;L.M.G. Petit ,&nbsp;C. Taillé ,&nbsp;M. Polette ,&nbsp;G. Deslée ,&nbsp;V. Dormoy ,&nbsp;J.M. Perotin","doi":"10.1016/j.rmr.2025.02.050","DOIUrl":"10.1016/j.rmr.2025.02.050","url":null,"abstract":"<div><h3>Introduction</h3><div>L’asthme est caractérisé par une inflammation chronique et un remodelage des voies aériennes. La fonctionnalité de l’épithélium respiratoire y est altérée suggérant un rôle central dans la physiopathologie de la maladie. Des anomalies associées aux cils ont été mises en évidence dans l’épithélium des voies aériennes de patients asthmatiques mais les mécanismes restent encore peu étudiés. Compte tenu de l’importance des cils primaires et moteurs dans l’intégrité structurelle et fonctionnelle de l’épithélium, l’objectif de ce projet est d’étudier les dérégulations associées aux cils dans de larges cohortes de patients asthmatiques sévères (SA) afin d’identifier de potentiels acteurs moléculaires du remodelage des voies aériennes.</div></div><div><h3>Méthodes</h3><div>Les données transcriptomiques de trois bases de données publiques U-BIOPRED (GSE76226, n<!--> <!-->=<!--> <!-->105), SARP (GSE63142, n<!--> <!-->=<!--> <!-->81) et IMSA (GSE158752, n<!--> <!-->=<!--> <!-->42), obtenues à partir de brossages bronchiques de patients non-asthmatiques et SA, ont été analysées dans le but de mettre en évidence des modifications du niveau d’expression des gènes associés aux cils (879 gènes décrits) dans l’asthme sévère et d’identifier des gènes d’intérêt parmi les plus dérégulés. L’expression des trois gènes les plus dérégulés a été étudiée <em>ex vivo</em> sur biopsies bronchiques par immunofluorescence.</div></div><div><h3>Résultats</h3><div>L’analyse bio-informatique permet de mettre en évidence 17 % de l’ensemble des gènes associés aux cils dérégulés de manière significative chez les patients SA dans chacune des cohortes (144<!--> <!-->±<!--> <!-->74 gènes dérégulés représentant 17<!--> <!-->±<!--> <!-->9 %, <em>p</em> &lt; 0,05). 17 gènes sont dérégulés de manière concordante entre les trois bases de données analysées, dont PHLDB2 (13<!--> <!-->±<!--> <!-->9 %), NEK6 (8<!--> <!-->±<!--> <!-->4 %) et SCNN1G (−7<!--> <!-->±<!--> <!-->3 %) sont les plus dérégulés. L’expression protéique de NEK6 et SCNN1G est significativement augmentée dans l’épithélium bronchique des patients SA comparativement aux patients non-asthmatiques (respectivement 4120<!--> <!-->±<!--> <!-->1157 niveaux de gris moyens (ngm) <em>vs.</em> 1980<!--> <!-->±<!--> <!-->789 ngm, <em>p</em> &lt; 0,001, 7465<!--> <!-->±<!--> <!-->2143 ngm <em>vs.</em> 3492<!--> <!-->±<!--> <!-->1169 ngm, <em>p</em> &lt; 0,001)).</div></div><div><h3>Conclusion</h3><div>Cette étude permet l’identification de trois acteurs moléculaires pouvant être impliqués dans le remodelage épithélial associé à l’asthme sévère. Ces résultats ouvrent des perspectives expérimentales visant à déterminer le rôle des acteurs identifiés dans la différenciation des cellules épithéliales des voies aériennes, afin de mettre en évidence de potentiels biomarqueurs et cibles thérapeutiques dans l’asthme sévère pour restaurer la clairance mucociliaire.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 206"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}