Revue des maladies respiratoires最新文献

{"title":"Proteomic analysis of pulmonary fibrosis associated with lung cancer reveals dysregulation of the apoptotic pathway","authors":"C. Guillois ,&nbsp;H. Cazier ,&nbsp;E. Guenzi ,&nbsp;A. Chassac ,&nbsp;P. Mordant ,&nbsp;G. Zalcman ,&nbsp;A. Mailleux ,&nbsp;B. Crestani ,&nbsp;A. Cazes ,&nbsp;G. Chevreux ,&nbsp;N. Poté","doi":"10.1016/j.rmr.2025.02.072","DOIUrl":"10.1016/j.rmr.2025.02.072","url":null,"abstract":"<div><h3>Introduction</h3><div>Pulmonary fibrosis (PF), especially idiopathic pulmonary fibrosis, is associated with a high risk of lung cancer (LC), but the mechanisms of carcinogenesis remains poorly understood. Combining Liquid Chromatography Mass Spectrometry (LC-MS/MS) and Mass Spectrometry Imaging (MSI), we compared the proteome of fibrotic areas from PF tissue samples with and without LC to identify altered signaling pathways.</div></div><div><h3>Methods</h3><div>34 Formalin Fixed Paraffin-Embedded (FFPE) surgical samples from patients with PF with LC (LC+, <em>n</em> <!-->=<!--> <!-->17) and without LC (LC−, <em>n</em> <!-->=<!--> <!-->17) were included. For each case, a representative FFPE block was selected for downstream analyses. After macrodissection of fibrotic areas, whole protein extracts were analyzed by LC-MS/MS. MSI was performed on tissue section after tryptic digestion, on selected regions of interest within fibrotic areas. <em>In silico</em> tryptic digestion of peptides of interest was further performed, and spatial localization of these peptides was visualized on ion maps.</div></div><div><h3>Results</h3><div>By LC-MS/MS analysis, we identified 72 out of 4901 proteins significantly differentially expressed between fibrotic areas of LC- and LC+ samples (<em>P</em> <!-->&lt;<!--> <!-->0.001). Pathway analysis suggested the involvement of oxidative stress metabolism and apoptosis, with significant downregulation of two mitochondrial proteins (BAX, Frataxin), known to activate apoptosis, in fibrotic areas adjacent to LC. After <em>in silico</em> tryptic digestion of BAX protein, MSI analysis revealed at least 2 unique peptides per protein that were significantly underexpressed in LC+ fibrotic areas. Complementary ion map analysis showed a specific expression of these mitochondrial peptides within metaplastic epithelial cells ligning honeycomb cysts.</div></div><div><h3>Conclusion</h3><div>Using an original proteomic approach integrating LC-MS/MS and MSI, we show that, in patients with PF, fibrotic areas have distinct proteomic profiles depending on the presence of LC. Fibrotic areas adjacent to LC exhibit a significant downregulation of pro-apoptotic mitochondrial proteins. Interestingly, spatial analysis by MSI show that these proteins are expressed by metaplastic epithelial cells within fibrotic areas, suggesting that dysregulation of apoptosis in these cells might be involved in lung carcinogenesis in patients with PF.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 217-218"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791521","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":"Combined cellular and gene therapy to treat primary ciliary dyskinesia","authors":"C. Bourdais ,&nbsp;M. Nadaud ,&nbsp;A. Coeur ,&nbsp;F. Foisset ,&nbsp;E. Ahmed ,&nbsp;I. Vachier ,&nbsp;A. Bourdin ,&nbsp;S. Assou ,&nbsp;J. De Vos","doi":"10.1016/j.rmr.2025.02.026","DOIUrl":"10.1016/j.rmr.2025.02.026","url":null,"abstract":"<div><div>Primary Ciliary Dyskinesia (PCD) is a genetic disease caused by mutations that alter cilia beating, including in the respiratory airways. This results in poor mucus clearance, severe morbidity, and increased mortality. We hypothesized that bronchial cilia beating can be restored using genetically corrected iPSC differentiated into air - liquid interface bronchial epithelium model (iALI) for autologous cell therapy. We have previously demonstrated that corrected cells derived from a PCD patient iPS line can be differentiated into iALI with functional ciliary beating. The current aim of the project is to evaluate the engraftment potential of these corrected cells and their ability to repair the pathological model post-engraftment. Several key issues need to be addressed identifying competent cells for bronchial engraftment, exploring various strategies to pre-treat the bronchial epithelium, and assessing the recovery of the ciliary beating recovery to assure bronchial repair.</div><div>Our team has established the differentiation of iPSCs into iALI. We have generated a PCD patient iPSC line using Sendai viruses, a corresponding CRISPR/Cas9 corrected cell line, as well a wild-type iPSC line and its CRISPR/Cas9 mutated counterpart. We also generated a GFP-iPSC line that expresses the fluorescent GFP protein under the human elongation factor 1 alpha promoter (EF1a), which allows us to track the engraftment of GFP-labeled bronchial stem cells in both control and mutated iALI models. Our results suggest that lung progenitors at the ventralized anterior foregut endoderm stage could be the most efficient cells for engraftment. Their self-renewal capability and ability to differentiate into various cell types of the bronchial epithelium are promising for developing a long-term and effective therapy. Regarding bronchial erosion, we found that promoting cell engraftment is crucial due to the barrier function of the intact bronchial epithelium and the lack of selective advantage of corrected cells. Various chemical and enzymatic strategies have shown potential, but their safety for in-vivo use needs further assessment. Furthermore, GFP-expressing engrafted cells displaying cilia suggest successful differentiation into ciliated cells, though functional recovery still requires confirmation.</div><div>In conclusion, the engraftment of corrected lung progenitors into eroded bronchial epithelium appears to be a promising therapeutic strategy to PCD. Next step of the project involves developing this therapy for in-vivo application, assessing its safety and efficacy in an immunodeficient mini-pig model.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 194-195"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791516","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":"Impact de la matrice extracellulaire sur le canal mécanosensible PIEZO1 dans les cellules musculaires lisses pulmonaires","authors":"A. Leriche ,&nbsp;A. Gaubert ,&nbsp;C. Guibert ,&nbsp;T. Ducret ,&nbsp;JF. Quignard","doi":"10.1016/j.rmr.2025.02.021","DOIUrl":"10.1016/j.rmr.2025.02.021","url":null,"abstract":"<div><h3>Introduction</h3><div>L’hypertension pulmonaire est une pathologie caractérisée par un remodelage important des artères intra-pulmonaires ayant pour conséquence d’augmenter le travail cardiaque conduisant à la mort. Dans cette pathologie, les contraintes mécaniques exercées sur les cellules musculaires lisses vasculaires sont modifiées. Ces contraintes mécaniques sont perçues par des canaux ioniques membranaires mécano-sensibles tels que Piezo1 qui convertissent les différents stimuli mécaniques en influx calcique et en réponses biologiques. La rigidité de la matrice extracellulaire est une contrainte mécanique majeure et elle augmente au cours de la pathologie. Nous avons investigué les effets de la rigidité matricielle sur l’activité de Piezo1.</div></div><div><h3>Méthodes</h3><div>Des cellules musculaires lisses d’artères intra-pulmonaires ont été cultivées sur des hydrogels Bola Bis Urée (BBU) de différentes rigidités (1 à 10 KPa). Cette matrice synthétique ne contient pas de protéines pouvant interagir avec des récepteurs membranaires. La concentration calcique intracellulaire et le potentiel transmembranaire ont été mesurés grâce aux sondes fluorescentes Cal520® et FLIPR®. La prolifération cellulaire a été estimé avec CyQUANT®.</div></div><div><h3>Résultats</h3><div>Une forte rigidité matricielle (10<!--> <!-->kPa) augmente l’expression de Piezo1. Elle induit une diminution l’amplitude et un ralentissement de la cinétique des signaux calciques intracellulaires après stimulation de Piezo1 par son agoniste Yoda1. Par contre, cette forte rigidité augmente l’activité basale de Piezo1 induisant une augmentation de la concentration calcique intracellulaire basale. Elle induit aussi une dépolarisation cellulaire liée à Piezo1. L’augmentation de la rigidité induit une augmentation de la prolifération cellulaire mais qui est indépendante de Piezo1.</div></div><div><h3>Conclusion</h3><div>Ainsi la rigidité matricielle pourrait jouer un rôle de stimulus mécanique sur Piezo1 entraînant une altération de son activité en adéquation avec le contexte d’hypertension pulmonaire.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 192"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791499","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":"Kcnk3 deficiency aggravates right ventricular dysfunction induced by pulmonary artery banding","authors":"K. El Jekmek ,&nbsp;M. Gourmelon ,&nbsp;A. Saint-Martin Willer ,&nbsp;M. Dutheil ,&nbsp;V. Capuano ,&nbsp;O. Mercier ,&nbsp;D. Montani ,&nbsp;F. Antigny","doi":"10.1016/j.rmr.2025.02.020","DOIUrl":"10.1016/j.rmr.2025.02.020","url":null,"abstract":"<div><h3>Introduction</h3><div>Right ventricular failure (RVF) is characterized by the inability of the right ventricle (RV) to maintain adequate blood flow and is the major predictor of mortality in pulmonary arterial hypertension (PAH). PAH is defined by a mean pulmonary arterial pressure greater than 20<!--> <!-->mmHg at rest and is characterized by increased pulmonary vascular resistance, leading to right heart failure. Currently, no treatments specifically target RVF in PAH. However, slowing down the progression of RVF could delay the need for double lung transplantation and reduce PAH-related mortality. Loss-of-function mutations in the KCNK3 (Potassium channel subfamily K⁺ member 3) gene, which encodes the potassium channel known as TASK-1 (TWIK-related acid-sensitive K⁺ channel 1), have been identified in PAH patients. Our team has recently shown that KCNK3 dysfunction is involved in pulmonary vascular remodeling and contributes to the development of PAH <span><span>[1]</span></span>. They also demonstrated that reduced function and expression of KCNK3 is a hallmark of RV hypertrophy and dysfunction associated with pulmonary hypertension <span><span>[2]</span></span>.</div><div>Our objective was to investigate whether the Kcnk3 deficiency exacerbates RV remodeling in a rat model of RV dysfunction induced independently of pulmonary vascular remodeling.</div></div><div><h3>Methods</h3><div>To address this, we subjected male and female Kcnk3-deficient rats (WT, heterozygous, and homozygous) to chronic RV pressure overload by pulmonary artery banding (PAB). 4 weeks after surgery, we analyzed the consequence of Kcnk3-deficiency on RV remodeling by performing echocardiography, right heart catheterization, and RV histology.</div></div><div><h3>Results</h3><div>Our results show that rats (males and females) subjected to PAB developed RV hypertrophy and RV dysfunction. In male homozygous Kcnk3-deficient rats, PAB led to an exacerbation of RV hypertrophy, RV dilatation, and a more pronounced increase in RV systolic pressure compared to WT-PAB rats. In contrast, no difference was observed in male heterozygous Kcnk3-deficient rats compared to WT-PAB male rats. Interestingly, RV dysfunction and hypertrophy were unchanged in female homozygous Kcnk3-deficient rats subjected to PAB compared to WT-PAB female rats.</div></div><div><h3>Conclusion</h3><div>In conclusion, our results suggest that KCNK3 dysfunction is a crucial event in the physiopathology of RV failure. Further studies are needed to understand underlying molecular and cellular mechanisms and to investigate sex differences linked to Kcnk3-deficiency in these experimental conditions.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 191-192"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791498","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":"Impact de l’activation de la GTPase Rac dans la dégranulation des éosinophiles au cours de l’asthme sévère","authors":"H. Bergereau ,&nbsp;D. Hassoun ,&nbsp;Q. Marquant ,&nbsp;M. Rousselle ,&nbsp;A. Magnan ,&nbsp;G. Loirand ,&nbsp;V. Sauzeau","doi":"10.1016/j.rmr.2025.02.032","DOIUrl":"10.1016/j.rmr.2025.02.032","url":null,"abstract":"<div><h3>Introduction</h3><div>L’asthme est une pathologie chronique caractérisée par une inflammation exacerbée, une hyperréactivité bronchique et un remodelage des voies aériennes. Récemment, nous avons démontré que la GTPase Rac est activée au sein des éosinophiles dans un modèle murin d’asthme sévère. Notre objectif a été d’identifier le rôle de Rac dans la fonction principale des éosinophiles au cours de l’asthme : la dégranulation.</div></div><div><h3>Méthodes</h3><div>Les cellules progénitrices hématopoïétiques provenant de la moelle osseuse de souris sont mises en culture en présence de Stem Cell Factor (SCF) et de Fms-Like Tyrosine kinase 3 (FLT3-L) pour induire leur expansion. Au 4^e jour de culture, l’ajout d’interleukine 5 permet la différenciation des cellules progénitrices en éosinophiles. In vitro, la dégranulation des éosinophiles est induite par le Platelet Activating Factor (PAF). Le trafic des granules de sécrétion a été analysé par immunofluorescence et par microscopie électronique. Les voies de signalisation intracellulaire ont été analysées par western-blot.</div></div><div><h3>Résultats</h3><div>Nous avons observé que le traitement des éosinophiles au PAF induit une libération d’éosinophile peroxydase (EPX) mais également une activation de la protéine Rac. L’inhibition pharmacologique de Rac (A41, 10-5<!--> <!-->M) prévient la libération d’EPX des éosinophiles murins et également des éosinophiles humains. Par immunofluorescence et microscopie électronique, nous avons observé que l’activation de Rac par le PAF joue un rôle majeur dans la migration des granules de sécrétion du cytoplasme vers la membrane plasmique des éosinophiles, aboutissant à la libération des médiateurs cytotoxiques. Par une approche biochimique, nous avons démontré que l’activation de Rac par le PAF favorise la formation du complexe protéique Rac/PLCß2 (phospholipase<!--> <!-->C ß2) permettant la production d’IP3 et ainsi l’augmentation de la concentration calcique intracellulaire nécessaire à la dégranulation. Pour valider in vivo le rôle de Rac dans les éosinophiles, nous avons développé un modèle expérimental d’asthme sévère allergique. Nous avons observé que la libération d’EPX par les cellules inflammatoires présentes dans les poumons des souris asthmatiques est inhibée par l’A41.</div></div><div><h3>Conclusions</h3><div>Nos résultats démontrent que la dégranulation des éosinophiles induite par le PAF au cours de l’asthme est dépendante de l’activation de Rac. Ainsi, la GTPase apparaît comme une nouvelle cible thérapeutique d’intérêt pour lutter contre l’inflammation au cours de l’asthme allergique.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 197-198"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791452","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":"Sommaire","authors":"","doi":"10.1016/S0761-8425(25)00157-3","DOIUrl":"10.1016/S0761-8425(25)00157-3","url":null,"abstract":"","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page iii"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143790207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploration du role de la h-caldesmon dans la modulation de la contraction prolongée du muscle lisse","authors":"M. Schultz ,&nbsp;A. Bai ,&nbsp;L.H. Kachmar ,&nbsp;A.-M. Lauzon","doi":"10.1016/j.rmr.2025.02.037","DOIUrl":"10.1016/j.rmr.2025.02.037","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;div&gt;Le muscle lisse (ML) est le principal effecteur de l’hyperréactivité bronchique dans l’asthme. La contraction du ML est produite par l’interaction cyclique entre les protéines motrices appelées « myosine » et les filaments d’actine. Cette interaction est régulée par l’activation de la myosine via la phosphorylation de sa chaîne légère régulatrice (LC20) et qui est renversée par la phosphatase de la chaîne légère (MLCP). Cependant, le ML a la capacité de maintenir la contraction même après la désactivation de la majorité des myosines (ce qu’on appelle l’état latch). Cette propriété, si dérégulée, pourrait contribuer à la physiopathologie de l’asthme. Le modèle classique de l’état latch stipule qu’il se produit lorsque les myosines sont désactivées tout en restant attachées à l’actine. Toutefois, ce modèle ne considère pas d’autres éléments régulateurs de la contraction du ML, comme la protéine régulatrice de l’actine h-caldesmon (hCaD), qui est supposée jouer un rôle important dans la régulation du ML. Ainsi, dans cette étude nous cherchons à comprendre comment l’hCaD influence la capacité des myosines à maintenir la force après leur inactivation.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Objectif&lt;/h3&gt;&lt;div&gt;Déterminer les effets de l’hCaD sur l’interaction entre la myosine et l’actine lors de la désactivation des molécules de myosine.&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Méthodes&lt;/h3&gt;&lt;div&gt;Nous avons récemment développé une technique nous permettant de mesurer la génération de forces moléculaires tout en modifiant les conditions in vitro, de façon dynamique, pour simuler l’état latch. Des pinces optiques ainsi que des méthodes d’analyse d’image ont été utilisées pour mesurer la force et le temps de maintien de la force par des myosines qui tirent sur un filament unique d’actine, pendant l’injection de MLCP, en présence et en absence de l’hCaD (400&lt;!--&gt; &lt;!--&gt;nM). De plus, l’essai de motilité in-vitro a été utilisé pour examiner les interactions mécaniques entre les myosines actives et inactives en mesurant la vitesse moyenne (vavg) et la fraction (fmot) de filaments d’actine mobiles pendant la déphosphorylation de la myosine par injection de MLCP, en présence ou en absence de hCaD (400&lt;!--&gt; &lt;!--&gt;nM). vavg et fmot ont également été mesurés à quatre niveaux de phosphorylation de la myosine, en présence et en absence de hCaD (400&lt;!--&gt; &lt;!--&gt;nM).&lt;/div&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Résultats&lt;/h3&gt;&lt;div&gt;Des régressions sigmoïdales des données d’injection de MLCP montrent une diminution plus rapide et plus précoce de fmot en présence de hCaD. Une tendance similaire est observée pour vavg. Les régressions des données de tests de motilité in vitro à différents niveaux de phosphorylation de LC20 avec un modèle mathématique de l’interaction mécanique entre les myosines montre que l’hCaD augmente la concavité ascendante de la relation entre la vitesse et le pourcentage de phosphorylation, une caractéristique corrélée à une augmentation de la charge subie par les molécules de myosine phos","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 200"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791873","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":"Modeling COPD with a complex tubular model of distal airways integrating the epithelial and mesenchymal compartments","authors":"K. Raasch ,&nbsp;E. Maurat ,&nbsp;A.E. Leipold ,&nbsp;P. Henrot ,&nbsp;M. Zysman ,&nbsp;R. Prevel ,&nbsp;T. Trian ,&nbsp;T. Krammer ,&nbsp;V. Bergeron ,&nbsp;M. Thumerel ,&nbsp;P. Nassoy ,&nbsp;P. Berger ,&nbsp;A.E. Saliba ,&nbsp;L. Andrique ,&nbsp;G. Recher ,&nbsp;I. Dupin","doi":"10.1016/j.rmr.2025.02.040","DOIUrl":"10.1016/j.rmr.2025.02.040","url":null,"abstract":"<div><div>Chronic Obstructive Pulmonary Disease (COPD) is characterized by progressive irreversible limitation of expiratory airflow, primarily affecting distal airways, which show early signs of remodelling, inflammation and obliteration. The limited relevance of 2D cell models and lack of 3D models mimicking small airway features hinder early pathophysiological understanding and drug discovery. We have previously developed a 3D mesenchyme-free “bronchioid” model using an innovative tubuloid cell-based assay and human bronchial epithelial adult stem cells derived from clinical samples <span><span>[1]</span></span>. Here, we aimed at integrating mesenchymal cells into the bronchioid model to investigate epithelial-mesenchymal interactions and their role in COPD development. Fine-tuning the composition of the scaffold is required to obtain a bronchioid with mesenchymal cells. Epithelial and mesenchymal cells were viable and organized from lumen to periphery, akin to bronchial topology. Adding this mesenchymal component improved the robustness of the system, highlighting their pivotal role in adhesion and contraction balance. Flow cytometry analysis and 3D imaging shows interactions between both compartments, strongly affecting epithelial differentiation. We conclude that the bi-component bronchioid enables modeling of the distal lung's epithelial–mesenchymal unit and represents a powerful tool for a comprehensive grasp of the disease processes in COPD.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 201-202"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791976","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":"Chronic and intermittent hypoxias lead to disrupted clock genes expression in mouse lung tissues: Implications for lung cell senescence","authors":"V. Gros ,&nbsp;E. arcos ,&nbsp;J. Jacquet ,&nbsp;E. Born ,&nbsp;J.A. De Freitas ,&nbsp;S. Abid ,&nbsp;D. Beaulieu ,&nbsp;N. Vienney ,&nbsp;A. Houssaini ,&nbsp;L. Lipskaia ,&nbsp;C. Jourdan Le Saux ,&nbsp;O. Bischof ,&nbsp;H. Duez ,&nbsp;B. Pourcet ,&nbsp;L. Boyer ,&nbsp;S. Adnot","doi":"10.1016/j.rmr.2025.02.089","DOIUrl":"10.1016/j.rmr.2025.02.089","url":null,"abstract":"<div><h3>Introduction</h3><div>Obstructive sleep apnoea syndrome (OSAS) and chronic obstructive pulmonary disease (COPD) are common syndromes that lead either to intermittent or chronic hypoxia. We previously demonstrated that OSAS and COPD were associated with telomere shortening and increased cell senescence. Furthermore, we showed that pulmonary hypertension (PH) driven by chronic hypoxia in mice is associated with an accumulation of senescent cells in the lung. The cyclin-dependent kinase inhibitor p21, the main effector of p53, is activated in response to DNA damage and telomere dysfunction but is down-regulated by certain clock genes. Sleep abnormalities and hypoxia also disrupt endogenous circadian clock genes, more specifically Rev-Erbα, a central transcriptional repressor of the molecular clock that downregulates p21 expression.</div></div><div><h3>Methods</h3><div>We investigated mice exposed to chronic intermittent hypoxia (CIH) and to chronic hypoxia (Hx) in comparison to normoxic mice. Mice exposed to CIH were subjected to repeated cycles of 90<!--> <!-->sec hypoxia-reoxygenation (21–5% O₂), 8<!--> <!-->hours a day for one to 21 days, biological analyses were performed on the day after CIH discontinuation. Hx mice were exposed to chronic hypoxia (10% O₂) during 3 weeks. Similar protocols were performed in Rev-Erbα KO mice.</div></div><div><h3>Results</h3><div>In normoxic mice, rhythmicity of the canonical clock genes <em>Rev-Erbα</em>, <em>Rev-Erbβ</em>, and <em>Bmal-1</em> was associated with rhythmicity of lung p21, with opposite variations in Rev-Erbα and p21. Compared to normoxia, CIH for 1.2, 7, or 21 days increased Rev-Erbα at ZT 2-4 (morning) and downregulated p21; these effects persisted beyond 48<!--> <!-->hours after CIH cessation. Bulk RNA sequencing performed 3 weeks after CIH confirmed dysregulation of the circadian clock and clock-controlled genes (Dbp, Rev-Erbα, Hlf) and showed increases in several collagen transcripts. Senescence markers including p16 and SASP components were unaffected by CIH exposure. Similar findings were observed in aged (18-month-old) mice. Rev-Erbα KO mice compared to WT mice exhibited an increase in lung p16 positive nuclei. Consistent with this, cultured primary lung fibroblasts from Rev-Erbα KO mice compared to WT mice exhibited premature senescence-as assessed by increased β-galactosidase positive cells and decreased cell growth. Conversely to WT mice exposed to CIH, WT mice exposed to Hx exhibited a downregulation of Rev-Erbα together with increased p21 expression. Moreover, Rev-Erbα KO mice exposed to Hx were partially protected against hypoxia-induced PH. Studies are underway to assess PH in CIH mice.</div></div><div><h3>Conclusion</h3><div>Clock genes, namely <em>Rev-Erbα</em>, plays an important role in the response to CIH and Hx by affecting cell senescence with, consequently, a reduction of lung cell proliferation and protection against PH.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Pages 226-227"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143792057","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":"Single-cell transcriptomics identify cell-type specific transcriptomic signatures in response to CdCl2 in human airway bronchial epithelial cells cultured at the air-liquid interface","authors":"R. Lopes Gonçalves ,&nbsp;M. Gautier ,&nbsp;J. Mille ,&nbsp;L. Guardini ,&nbsp;L.-E. Zaragosi ,&nbsp;M. Ben Kheder ,&nbsp;H. Groux ,&nbsp;B. Mari ,&nbsp;R. Rezzonico ,&nbsp;G. Vassaux","doi":"10.1016/j.rmr.2025.02.016","DOIUrl":"10.1016/j.rmr.2025.02.016","url":null,"abstract":"<div><h3>Introduction</h3><div>The aim of this project is to determine whether single-cell transcriptomics (sc-RNAseq) can provide information susceptible to enrich classical bulk transcriptomics experiments in toxicology. In the present work, we studied the effects of a non-genotoxic carcinogen (CdCl2) on human airway pseudostratified epithelium.</div></div><div><h3>Methods</h3><div>The identification of cell-type-specific gene expression signatures was performed through scRNA-seq analyses using PARSE technology on human airway bronchial epithelial cells (HAEC) cultured in 3D at an air-liquid interface (ALI) and exposed to 10<!--> <!-->μM CdCl2, an heavy metal pollutant present in cigarette smoke. These cell-type specific responses signatures were validated through RNA-Fluorescence In Situ Hybridizations.</div></div><div><h3>Results</h3><div>Single-cell gene profiling indicated that different regulons are modulated in a cell-type-specific manner in response to CdCl2 exposure. For instance, we showed that treatment with CdCl2 stimulated the expression of MT1G, MT1<!--> <!-->M and to a lesser extent SLC30A1 and HMOX1 preferentially in multiciliated and mucus-secreting cells, while AKT3 and IL1RN are rather induced in basal cells. Among the cadmium-regulated genes, we found that the long non-coding RNA LUCAT1 is induced in specific differentiated cell types. We previously showed1 that LUCAT1 is i) upregulated by hypoxia in lung adenocarcinomas, ii) correlated with poor prognosis in patients and iii) involved in the regulation of oxidative stress in cancer cells. However, its cellular sourcing and physiological role in the differentiated mucociliary airway epithelium have not yet been addressed. We found that LUCAT1 is slightly expressed in the untreated epithelium, but in response to hypoxia or CdCl2, it is markedly induced in suprabasal and deuterosomal cells and in differentiated secretory and multiciliated cells but not in basal cells. Using a CRISPR-Cas9-RNP approach2 to knockdown LUCAT1 in basal bronchial stem cells, we will decipher the cell state-specific functions of this gene i) in the regeneration steps of a fully differentiated mucociliary human airway epithelium and, ii) in hypoxia- and CdCl2-responses.</div></div><div><h3>Conclusion</h3><div>ScRNA-seq reveals distinct transcriptional responses to CdCl2 among airway epithelial cell subtypes. This method could be applied to other molecules to classify gene expression variations and develop predictive tests for non-genotoxic carcinogens compounds. This study also precise the physiological role of LUCAT1 in the homeostasis, regeneration and stress-response of HAEC.</div></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"42 4","pages":"Page 189"},"PeriodicalIF":0.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791599","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}