Revue des maladies respiratoires最新文献

{"title":"Identification of immunological biomarkers of macrophages related to specific compartmentalization of in lung and liver in mouse model of septic shock","authors":"A. Bodin , C. Slek , M. Magnin , L. Commin , A. Corlu , V. Lagente , C. Aninat , J.M. Bonnet , B. Allaouchiche , V. Louzier , T. Victoni","doi":"10.1016/j.rmr.2024.01.054","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.054","url":null,"abstract":"<div><h3>Introduction</h3><p>Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection. The complex pathophysiology of sepsis is associated with pro- and anti-inflammatory response, a pro-coagulant state, endothelial dysfunction and tissue hypoxia. These mechanisms lead to progressive multi organ failure. Although this is a systemic process, the pathophysiological of sepsis differs from organ to organ, and from organ to peripheral blood. Our hypothesis to explain this compartmentalization of responses is a distinct population of resident tissue macrophages, as well as a distinct migration of monocytes in target organ. Indeed, the macrophages and monocytes can start the clinical syndrome of sepsis via transcription of genes involved in inflammation. Moreover, macrophages can induce endothelial injury by release reactive oxygen species. Also hypoxia decreases expression of M1 polarization markers and increases the M2 marker. Identification of biomarkers related to specific organs (beyond the blood) will improve the understanding specific organ failure. In this way, the aim of this study was compared to the systemic inflammatory response with the lung and the liver, two organs most affected during sepsis as well as understanding the role of macrophages in this compartmentalization. For this, a murine polymicrobial sepsis model induced by caecal ligation and puncture (CLP) was used.</p></div><div><h3>Methods</h3><p>Moderate sepsis was induced by the CLP in C57BL/6 male mice (<em>n</em> <!-->=<!--> <!-->63), divided into 4 groups: Basal, Sham 1 day and 5 days, CLP 1 days and 5 days. Then, we analyzed histological changes, cytokine profile (by ELISA and PCR), oxidative imbalance (expression of SOD, CAT and iNOS) and polarization markers (CD<sub>86</sub>and CD<sub>206</sub>) by immunohistochemistry.</p></div><div><h3>Results</h3><p>Respiratory and liver failure was confirmed by histology and also by a decrease of pressure oxygen in arterial blood (PaO<sub>2</sub>) and increase of bilirubin level after 5 days of CLP. Moreover CLP induced a plasma increase in the level of TNFα, IL-6, IL-10, KC and CCL2 in the first 24<!--> <!-->hours after CLP, but with a progressive decrease at 5 days. On the other hand, our original results show that the level of some of the cytokines in the liver and the lung differ from the systemic level during sepsis. Indeed, there are increase in CCL2 and its receptor CCR2 in the lung compared to the liver, whereas in the liver rather a decrease in the expression of CX3CL1/CX3CR1, that is not altered in the lung. We were also able to highlight a more marked oxidative imbalance in the liver than in the lung. Also, we observed that CLP increases the expression of M1 CD<sub>86</sub> and M2 CD<sub>206</sub> markers in the liver and lung at 5 days, but with a total number of CD<sub>86</sub>-labeled cells, three times greater in the liver.</p></div><div><h3>Conclusion</h3><p>These results","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 207-208"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160440","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":"Thorough assessment of primary cell cultures for murine and human alveolar macrophages","authors":"C. David ,&nbsp;D. Brea ,&nbsp;A. Cezard ,&nbsp;V. Vasseur ,&nbsp;E. Barsac ,&nbsp;B. Briard ,&nbsp;M. Ferreira ,&nbsp;S. Adam-Marchand ,&nbsp;M. Si-Tahar ,&nbsp;A. Guillon","doi":"10.1016/j.rmr.2024.01.060","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.060","url":null,"abstract":"<div><h3>Introduction</h3><p>Alveolar macrophages (AM) are the first-line lung defenders, but appropriate cell culture models are still limited for their study. The uses of freshly isolated macrophages are hampered by limited quantities or difficult access to human donors. Indeed, research on AM is often restricted to cells differentiated in vitro from bone-marrow progenitors or blood monocytes; however, such cells do not present the specific features of tissue-specialized macrophages, like AM. To overcome these issues, Max Planck Institute (MPI) cells were characterized as a nontransformed, self-renewing AM-like macrophage murine model. Yet, MPI cannot fully reproduce AM characteristics because they developed outside the alveolar niche. To optimize the primary cell cultures of mouse AM (mAM), recent works have proposed strategies to expand and maintain these cells <em>ex vivo</em> from bronchoalveolar lavage (BAL). The functional impact of the amplification process is unclear. Moreover, the heterogeneity of AM population, which is constantly remodeled during the lifespan, is poorly recapitulated in laboratory mice, therefore limiting extrapolation of mAM physiopathology to human. Here, we propose a comprehensive comparison of mice and human AM (hAM) culture models and give insight into the translational value of different mice AM models.</p></div><div><h3>Methods</h3><p><em>Ex vivo</em> primary mAM were isolated from BAL of 3–5 mice and expanded for 3–4 weeks per passage. hAM were freshly isolated and maintained from BAL of uninfected patients. Immunophenotypes of MPI, mAM and hAM were compared. We evaluated the cells production of inflammatory mediators (IL-6, TNF-α, IL-1β) in response to different pro-inflammatory agonists such as LPS, poly (I: C), flagellin, Pam³CSK4 and curdlane. We induced cell death using LPS/ATP and measured real-time cell death using IncuCyte SX5 cell-live analysis.</p></div><div><h3>Results</h3><p>Immunophenotypes of MPI and mAM were comparable despite lower SiglecF and CD₁₁c expression in MPI or mAM after expansion compared to freshly isolated mAM. MPI, mAM and hAM presented similar inflammatory signature but differences in the amplitude of response (exception for Flagellin that induced IL-1-β response in murine models but not in hAM). Overall, MPI had a more pro-inflammatory profile than mAM, as illustrated by a 10-fold production of IL-6 in response to LPS compared to mAM. Differences were observed regarding cell death in mice and human AM models: pyroptotic events were more rapid (1<!--> <!-->h) and important in murine AM models compared to hAM.</p></div><div><h3>Conclusion</h3><p>While clarifying the limits of MPI and <em>ex vivo</em> mAM models, our work showcased consistent inflammatory responses across AM models, albeit with differing response amplitudes. Conversely, mouse and human models exhibited variations in cell death kinetics.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 211"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160474","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 of hypoxia in Cystic Fibrosis bronchial epithelial cells: Focus on CFTR and TRPA1 channels","authors":"K. Pascarel,&nbsp;J. Colas,&nbsp;T. Carrez,&nbsp;S. Mirval,&nbsp;C. Barrault,&nbsp;F. Becq,&nbsp;C. Vandebrouck","doi":"10.1016/j.rmr.2024.01.089","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.089","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;Cystic Fibrosis (CF) is an autosomal and recessive disease caused by the mutation of a gene located on the chromosome 7: &lt;em&gt;CFTR&lt;/em&gt; (Cystic Fibrosis Transmembrane conductance Regulator). Its codes for CFTR, a protein which plays a role in mucus homeostasis by transporting both chloride ions and water. The most common mutation F508del-CFTR, leads to the absence and malfunction of CFTR at the surface of epithelial cells in various organs especially in lungs. It results in the loss of the mucus clearance properties in the airways, which will cause the obstruction of bronchi and alveola over time. The oxygen (O&lt;sub&gt;2&lt;/sub&gt;) delivery, crucial for aerobic metabolism, is less effective especially for the lung's epithelial cells whose environment is gradually becoming hypoxic. The inability of lungs to realise haematosis, at tissue level is commonly named the respiratory failure.&lt;/p&gt;&lt;p&gt;Our project aims to characterise the impact of hypoxia on ion channels, especially CFTR and TRPA1 (an oxygen sensible calcium channel).&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;p&gt;Cystic Fibrosis Bronchial Epithelial (CFBE) cells-wt (WT) and CFBE F508del (DF) are cultivated in a controlled hypoxic atmosphere (1% O&lt;sub&gt;2&lt;/sub&gt;). Protein expression and quantification have been realised by western blot. CFTR activity have been measured by automated patch-clamp (whole cell recording, WCR) and Ussing chamber while the activity of TRPA1 have been recorded using the Fluo4-AM probe. TRPA1 localisation has been studied by immunostaining.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;p&gt;Here, we show that the change from normoxia (21% O&lt;sub&gt;2&lt;/sub&gt;) to hypoxia (1% O&lt;sub&gt;2&lt;/sub&gt;) is able to induce a fast cellular response with the accumulation of HIF-1α (Hypoxia Inducible Factor) in only 6&lt;!--&gt; &lt;!--&gt;hours in CFBE WT, CFBE-DF and CFBE-DF corrected by the tri-therapy Kaftrio® (Elexacaftor/Tezacaftor/Ivacaftor, ETI). We also observed, that HIF-1α accumulated is reduced in non-corrected CFBE-F508del. Regarding CFTR, our results shows that only F508del-CFTR is impacted by hypoxia at protein level and activity, despite the correction by ETI. Automated WCR patch-clamp and Ussing chamber recordings both show that Kaftrio® corrected F508del-CFTR activity decreases 24&lt;!--&gt; &lt;!--&gt;hours after hypoxia induction. F508del-CFTR currents are decreased by 43% in whole cell configuration while short-circuit current (Isc) CFTR dependent are diminished by around 48%. Concerning TRPA1, hypoxia does not impact the protein accumulation but instead decrease the channel activity by 49% in CFBE-wt, 40% in CFBE-F508del non corrected and 30% when corrected by ETI. Finally, it seems that hypoxia plays a role in TRPA1 location inducing its relocation close to the plasma membrane.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;Our data show a reduced amount of CFTR protein accumulated in CFBE F508del corrected or not, which was not observed on the WT form. Electrophysiologic assays show a clear impact of hypoxia on F508de","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 226"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160500","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":"Effet de l’absence de microbiote sur l’infection pulmonaire par le virus respiratoire syncytial chez le souriceau","authors":"C. Chottin ,&nbsp;C. Ferret ,&nbsp;C. Maudet ,&nbsp;V. Saint-Criq ,&nbsp;S. Riffault ,&nbsp;D. Descamps","doi":"10.1016/j.rmr.2024.01.047","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.047","url":null,"abstract":"<div><h3>Introduction</h3><p>La bronchiolite est la principale infection respiratoire chez le nourrisson, dont l’agent étiologique principal est le virus respiratoire syncytial (VRS) contre lequel il n’existe pas de vaccins. La sensibilité du nourrisson à l’infection par le VRS est intrinsèquement liée aux caractéristiques de la muqueuse pulmonaire en période périnatale qui évoluent avec la mise en place de l’immunité et la colonisation du poumon par un microbiote bactérien. Bien que décrit pour le microbiote intestinal, le rôle du microbiote pulmonaire sur la maturation de l’immunité de la muqueuse pulmonaire, et donc sur la trajectoire de la santé respiratoire reste peu décrit. Notre étude vise à évaluer si l’absence de microbiote en période périnatale modifie la réplication virale et la réponse immunitaire pulmonaire déclenchée par l’infection VRS. La compréhension des interactions précoces entre le microbiote et la muqueuse pulmonaire pourrait contribuer au développement de nouvelles approches visant à renforcer la réponse immunitaire et ainsi limiter la sévérité de l’infection par le VRS en période néonatale.</p></div><div><h3>Méthodes</h3><p>(1) Infection de souriceaux C57BL/6 dépourvus de flore (animaux axéniques) ou issus de géniteurs colonisés par une flore bactérienne (animaux colonisés) par une souche de VRS recombinant exprimant la luciférase (VRS-Luc).</p><p>(2) Mesure du niveau de réplication virale (bioluminescence et qPCR) et de la réponse immunitaire pulmonaire (infiltration cellulaire dans les lavages bronchoalvéolaires [LBA] et qPCR sur les gènes dépendants des interférons de type 1 [ISG]) à j₁, j₂ et j₄ post-infection.</p></div><div><h3>Résultats</h3><p>Les animaux axéniques montrent une augmentation du niveau de réplication virale entre j₁ et j₄ post-infection par détection de bioluminescence et par qPCR. À l’inverse, les animaux colonisés présentent une diminution du signal entre j₁ et j₄ post-infection. L’infiltration cellulaire dans les LBA des animaux axéniques est augmentée par rapport à celle quantifiée dans les animaux colonisés. L’expression des ISG (IRF7, ISG15 ou OAS) analysée par qPCR est augmentée à j₄ post-infection chez les animaux colonisés, alors que celle des animaux colonisés est induite dès j₁ post-infection.</p></div><div><h3>Conclusion</h3><p>L’absence de flore bactérienne s’accompagne de différences chez le souriceau dans la cinétique de réplication virale et dans la mise en place de la réponse immunitaire lors d’une infection par le VRS. La caractérisation des cellules immunitaires de la muqueuse pulmonaire est prévue afin d’identifier un partenaire cellulaire important dans la défense antivirale influencé par la présence d’un microbiote en période néonatale.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 204"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160526","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":"Surdose en cannabinoïdes de synthèse : attention aux complications respiratoires aiguës","authors":"M. Underner ,&nbsp;J. Perriot ,&nbsp;G. Peiffer ,&nbsp;N. Jaafari ,&nbsp;T. Urban","doi":"10.1016/j.rmr.2024.02.005","DOIUrl":"10.1016/j.rmr.2024.02.005","url":null,"abstract":"","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 262-263"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140068639","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":"Étude de la physiopathologie de l’emphysème grâce à un modèle d’alvéolosphère en 3D à partir de cellules épithéliales alvéolaires de type 2 humaines","authors":"M. Gueçamburu ,&nbsp;P. Henrot ,&nbsp;E. Maurat ,&nbsp;P. Berger ,&nbsp;I. Dupin ,&nbsp;M. Zysman","doi":"10.1016/j.rmr.2024.01.023","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.023","url":null,"abstract":"<div><h3>Introduction</h3><p>L’emphysème, une des composantes de la bronchopneumopathie chronique obstructive (BPCO), correspond à une destruction des alvéoles pulmonaires dont la physiopathologie est mal connue. Des modèles de culture de cellules épithéliales alvéolaires (AEC) en 3 dimensions (3D) dans du Matrigel permettent d’étudier les capacités de prolifération et de différenciation des AEC2, mais manquent de reproductibilité. L’objectif principal de ce travail est le développement pérenne d’un modèle d’alvéolosphères 3D à partir d’AEC2 humaines ainsi que la modélisation de l’emphysème par exposition à l’extrait de fumée de cigarettes (CSE).</p></div><div><h3>Méthodes</h3><p>Ce modèle est basé sur l’isolement d’AEC2 par tri immunomagnétique (HTII-280+) à partir de 18 échantillons de parenchymes issus de patients fumeurs et non-fumeurs. Ces cellules sont mises en culture 3D dans des micropuits d’hydrogel préformés (200<!--> <!-->μm de diamètre) par photopolymérisation permettant une analyse morphologique (taille, lumière) et phénotypique (immunomarquages, qPCR, microscopie électronique [MET]) à j₁, 7, 14 et 21. L’impact de l’exposition à 5 jours de CSE 5 % est étudié par qPCR et immunomarquages sur les alvéolosphères. Enfin, les cytokines sécrétées par les sphères exposées au CSE sont analysées par cytokine array, secondairement confirmées par ELISA.</p></div><div><h3>Résultats</h3><p>Les alvéolosphères sont maintenues en culture pendant 21 jours et forment progressivement une lumière centrale, dès j₇. La présence d’une barrière épithéliale est confirmée par la mise en évidence de jonctions serrées et adhérentes par MET et immunomarquage ZO-1. Des qPCR à j₁, 7, 14 et 21 montrent une apparition progressive de marqueurs d’AEC1 (expression de <em>p2xr4, pdpn</em>) alors que les marqueurs d’AEC2 persistent (expression de <em>abca3, sftpa, sftpc</em>). Les organelles permettant la synthèse de surfactant sont visualisées en MET (corps lamellaires, corps lipidiques). Enfin, l’exposition à 5 jours de CSE 5 % entraine une tendance à une diminution de la viabilité cellulaire (calcéine), une augmentation des marqueurs de stress oxydant (expression de <em>hmox</em>, <em>nqo1</em>, <em>srxn1</em> en qPCR) ainsi qu’un relargage des cytokines (MIF et IL8) dans le surnagent.</p></div><div><h3>Conclusion</h3><p>Ainsi, nous avons obtenu, à partir d’échantillons de patients fumeurs et non-fumeurs, un modèle reproductible d’alvéolosphères ayant une capacité d’auto organisation en 3D, répondant à la définition d’un d’organoïde et permettant l’étude de la physiopathologie de l’emphysème induite par l’exposition à l’extrait de fumée de cigarettes.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 192"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140159982","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 ozone exposure in mice mimics clinical asthma-COPD overlap syndrome and is attenuated by tiotropium","authors":"P. Chenuet ,&nbsp;S. Huot-Marchant ,&nbsp;A. Ledru ,&nbsp;L. Fauconnier ,&nbsp;M. Mellier ,&nbsp;N. Rouxel ,&nbsp;L. Allimonnier ,&nbsp;C. Serdjebi ,&nbsp;Y. Julé ,&nbsp;N. Riteau ,&nbsp;I. Couillin ,&nbsp;D. Togbé ,&nbsp;V. Quesniaux ,&nbsp;B. Ryffel ,&nbsp;N. Segueni","doi":"10.1016/j.rmr.2024.01.027","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.027","url":null,"abstract":"<div><p>Environmental air pollutants including ozone cause severe irritation and respiratory diseases. Here, we report that 6<!--> <!-->week's ozone exposure in mice (1.5<!--> <!-->ppm, twice weekly) causes airway hyperreactivity, eosinophil and neutrophil recruitment, Th2 immune response, respiratory barrier disruption with inflammation, fibrosis and emphysema reminiscent of COPD, more rapidly than cigarette smoke exposure. This model features important aspects of asthma-COPD overlap syndrome (ACOS) as recently described in patients. Since Tiotropium (TTP), an anticholinergic receptor antagonist, blocks smooth muscle cell contraction and mucus secretion with a prolonged bronchodilator effect in patients with asthma or COPD, we asked whether its effect is limited to bronchodilation. We report here that Tiotropium not only reduced airways hyperreactivity, but also drastically diminished eosinophil recruitment, Th2 cell response and ozone-induced lung inflammatory pathology including emphysema. Therefore, chronic O₃-induced lung pathology in mice mimics ACOS in patients and is attenuated by TTP treatment. The mechanisms of TTP protective effect on respiratory barrier disruption and chronic inflammation need to be further explored.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 193-194"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140159986","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":"Induction of lipogenic differentiation and alveolar regeneration in emphysema via PPARG and SREBP","authors":"G. Justeau ,&nbsp;M. Toigo ,&nbsp;R. Yilmaz ,&nbsp;L. Crepin ,&nbsp;J. Boczkowski ,&nbsp;B. Ribeiro Baptista ,&nbsp;L. Boyer","doi":"10.1016/j.rmr.2024.01.016","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.016","url":null,"abstract":"<div><h3>Introduction</h3><p>Emphysema is a respiratory disease characterized by chronic alveolar destruction. Lipofibroblasts (LIF) play a key role in the stem cell niche surrounding alveolar type II (AT2) cells and may contribute to alveolar regeneration. We have previously shown that senescent cell elimination induces alveolar regeneration, increased LIF numbers and activation of the sterol regulatory binding protein (SREBP) and peroxisome proliferator-activated receptor gamma (PPARG) pathways <span>[1]</span>. However, it remains unclear whether the activation of these pathways can increase fibroblast stem cell niche properties and promote alveolar regeneration during emphysema.</p></div><div><h3>Methods</h3><p>Human lung tissue slides were obtained from patients with or without emphysema and immunofluorescent staining was performed to identify LIF (ADRP+Vimentin+). Both human primary fibroblasts and primary AT2 cells were isolated from lobectomies through the explant method and magnetic sorting (HT2-280+) respectively. Fibroblasts were treated with Rosiglitazone and T0901317 for 72<!--> <!-->hours. LIF phenotype acquisition was evaluated through IF staining, qPCR and lipidomic analysis. Stem cell niche properties were evaluated by performing alveolar organoid formation assay by co-culturing treated fibroblasts with H-441 cells or primary AT2 cells. Adult C57BL6 mice received intra-tracheal injection of either Elastase or PBS. From D₂₁ mice were treated by intraperitoneal injections of Rosiglitazone (5<!--> <!-->μg/g/d), T0901317 (10<!--> <!-->μg/g/d) or vehicule, 5/7<!--> <!-->days. Lungs were collected at D₉₀. Left lung was fixated for morphological analysis.</p></div><div><h3>Results</h3><p>Our study showed a decrease in LIF populations among patients with emphysema compared to controls. Furthermore, Rosiglitazone, a PPARG agonist, and T0901317, a SREBP agonist, can induce lipogenic differentiation in human lung fibroblasts. Activation of both pathways increased the expression of ADRP and the activation of the SREBP pathway induced the accumulation of neutral lipids in the fibroblasts. Using an organoid model of alveolar regeneration, we show that activating these pathways increases the stem cell niche properties of fibroblasts and enhances the number of organoids formed with either H441 cells or primary AT2. Lastly, in a murine mode of elastase-induced emphysema, we show that Rosiglitazone partially reverts emphysema.</p></div><div><h3>Conclusion</h3><p>Activation of PPARG and SREBP pathways promotes lipogenic differentiation of fibroblasts, enhances human alveolar organoid formation and partially reverts emphysema in vivo. These results provide insight into potential therapeutic strategies for promoting alveolar regeneration in patients with emphysema.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 188"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160089","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":"Rôle de l’éosinophile dans un modèle in vitro de cicatrisation après blessure d’un épithélium bronchique","authors":"A. Ecrement ,&nbsp;T. Gasser ,&nbsp;J. Boukobza ,&nbsp;C. Antolovic ,&nbsp;C. Barnig","doi":"10.1016/j.rmr.2024.01.006","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.006","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;L’éosinophile est une cellule immunitaire considérée comme essentielle dans la pathogenèse de l’asthme, une maladie inflammatoire chronique des voies respiratoires. En effet, lors de leur recrutement dans les voies respiratoires, ces cellules peuvent induire des effets pro-inflammatoires sur l’épithélium par la libération d’un large éventail de médiateurs pro-inflammatoires, dont des protéines basiques, des espèces réactives de l’oxygène, des protéases et des cytokines. Néanmoins, des études récentes suggèrent l’existence de sous-populations d’éosinophiles possédant des fonctions anti-inflammatoires et pro-résolvantes et qui pourraient contribuer au retour à l’homéostasie après une agression tissulaire.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Méthodes&lt;/h3&gt;&lt;p&gt;Nous avons mis au point un modèle de cicatrisation épithélial bronchique avec la lignée cellulaire BEAS-2B. Une blessure standardisée a été induite avec l’outil Woundmaker® 96 puits sur un tapis cellulaire à confluence dont la prolifération a été bloquée. La cicatrisation a ensuite été suivie par IncuCyte S3, un système d’imagerie automatisée en temps réel permettant de réaliser des cinétiques de migration cellulaire dans des conditions de culture classiques en quantifiant la confluence des cellules dans la blessure. La sécrétion de TSLP a été mesurée dans les surnageants par ELISA. La dynamique de réparation de la blessure a ensuite été étudiée en présence d’éosinophiles isolés à partir du sang total provenant de sujets sains et activés par l’interleukine IL-5. Dans certaines expériences, les éosinophiles ont été marqués par une sonde fluorescente. Les cinétiques de cicatrisation ont également été évaluées sur des cellules exposées à des surnageants d’éosinophiles ayant été stimulés par la TSLP et IL-5. Enfin, nous avons évalué l’impact des éosinophiles CD&lt;sub&gt;62&lt;/sub&gt;L− (iEOS) et CD&lt;sub&gt;62&lt;/sub&gt;L+ (rEOs) sur la réparation, en les séparant au préalable par FACS.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Résultats&lt;/h3&gt;&lt;p&gt;En conditions contrôle, nous observons une sécrétion précoce de la TSLP après blessure et une fermeture quasi complète de la lésion en 36&lt;!--&gt; &lt;!--&gt;heures. Lors des cocultures, les éosinophiles s’accumulent précocement au niveau des berges de la blessure. Lorsque les éosinophiles sont activés par l’IL-5, ils accélèrent la réparation de la blessure (T15&lt;!--&gt; &lt;!--&gt;h : 70,84 %&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;4,53 vs 84,93 %&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;1,13 ; &lt;em&gt;p&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,05). À l’inverse, le surnageant provenant d’éosinophiles stimulés pendant 24 heures en présence d’IL-5 et de TSLP ralentit la fermeture de la blessure (T15&lt;!--&gt; &lt;!--&gt;h : 58,8 %&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;3,77 vs 70,12 %&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;3,38 ; &lt;em&gt;p&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,06). Nous n’avons pas observé d’effet pro-résolvant des rEOS comparés aux iEOS (T15&lt;!--&gt; &lt;!--&gt;h : 85,31 %&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;1,34 vs 85,38 %&lt;!--&gt; &lt;!--&gt;±&lt;!--&gt; &lt;!--&gt;2,78 ; &lt;em&gt;p&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0,98).&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;Nous avons déve","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Pages 183-184"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160173","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":"Anti-fibrotic effect of a FGF ligands trap in pulmonary fibrosis","authors":"D. Gonçalves ,&nbsp;C. Scribe ,&nbsp;P. Dellugat ,&nbsp;G. Rignol ,&nbsp;C. Ghilain ,&nbsp;R. Marsault ,&nbsp;L. Etasse ,&nbsp;J. Garcia-Pizarro ,&nbsp;B. Mari ,&nbsp;C. Czech ,&nbsp;C. Herbert","doi":"10.1016/j.rmr.2024.01.070","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.070","url":null,"abstract":"<div><h3>Introduction</h3><p>Lung fibrosis, including idiopathic pulmonary fibrosis (IPF), results from dysfunctional wound repair involving different cell types, including fibroblasts, epithelial cells and macrophages, which respond to multiple soluble and matrix factors. Fibroblast growth factor (FGF) signaling has been implicated in the pathogenesis of lung fibrosis, in particular in the regulation of fibroblast to myofibroblast transition (FMT), cell proliferation, and extracellular matrix production. However, individual FGF family members may exert pro- and anti-fibrotic effects, depending on the responding cell, the expression levels of the different FGF receptors (FGFR1-4) and the context of other signaling molecules, such as Transforming growth factor β (TGF-β). In order to better understand the complex functions of FGFs on pulmonary fibrosis, we evaluated the effect of a modified version of a FGFR3 decoy receptor <span>[1]</span> that specifically sequesters FGFR3 ligands including FGF1, FGF2 and FGF9 as a potential anti-fibrotic drug.</p></div><div><h3>Methods</h3><p>The effect of several FGFs in the presence or the absence of the FGFR3 ligand Trap was evaluated in vitro on human lung fibroblasts from healthy donors and IPF patients on various fibrotic parameters such as cell proliferation, cell contraction, production of extracellular matrix (ECM) and modulation of signaling pathways. The effect of the FGFR3 ligand trap was also assessed in vivo on the bleomycin mouse model, by monitoring mice body weight, Ashcroft score, hydroxyproline and soluble collagen content.</p></div><div><h3>Results</h3><p>Our results revealed that FGFs (mainly FGF2) stimulate fibroblast proliferation, contraction, ECM production and expression of various fibrotic markers such as chemokine ligand 2 (CCL2), connective tissue growth factor (CTGF), interleukin 6 (IL6), interleukin receptor 4 (IL4R) or ECM-related genes like fibronectin (FN1). The FGFR3 ligands Trap was able to reduce this FGF mediated pro-fibrotic phenotype and to desensitize the TGF-β canonical pathway in IPF cells. In the bleomycin lung fibrosis mouse model, the FGFR3 ligands Trap partially reversed lung fibrosis, as evidenced by a reduced body weight loss as well as diminution of the aschcroft score, hydroxyproline and soluble collagen content in lung samples.</p></div><div><h3>Conclusion</h3><p>Our data highlight the interplay between the TGF-β and the FGF signaling pathways in pulmonary fibrosis and demonstrate the potential of targeting FGFR3 signaling as a novel therapy for IPF.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":"41 3","pages":"Page 216"},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160420","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}