Revue des maladies respiratoires最新文献

{"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":"<div><h3>Introduction</h3><p>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.</p></div><div><h3>Méthodes</h3><p>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 CD₆₂L− (iEOS) et CD₆₂L+ (rEOs) sur la réparation, en les séparant au préalable par FACS.</p></div><div><h3>Résultats</h3><p>En conditions contrôle, nous observons une sécrétion précoce de la TSLP après blessure et une fermeture quasi complète de la lésion en 36<!--> <!-->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<!--> <!-->h : 70,84 %<!--> <!-->±<!--> <!-->4,53 vs 84,93 %<!--> <!-->±<!--> <!-->1,13 ; <em>p</em> <!-->=<!--> <!-->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<!--> <!-->h : 58,8 %<!--> <!-->±<!--> <!-->3,77 vs 70,12 %<!--> <!-->±<!--> <!-->3,38 ; <em>p</em> <!-->=<!--> <!-->0,06). Nous n’avons pas observé d’effet pro-résolvant des rEOS comparés aux iEOS (T15<!--> <!-->h : 85,31 %<!--> <!-->±<!--> <!-->1,34 vs 85,38 %<!--> <!-->±<!--> <!-->2,78 ; <em>p</em> <!-->=<!--> <!-->0,98).</p></div><div><h3>Conclusion</h3><p>Nous avons déve","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":null,"pages":null},"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":null,"pages":null},"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}

{"title":"Uncovering cystic fibrosis patient profiles and exposome associations through unsupervised multidimensional phenotyping","authors":"M. Leemans ,&nbsp;R. Epaud ,&nbsp;P. De Carli ,&nbsp;C. Dehillotte ,&nbsp;L. Lemonnier ,&nbsp;T. Benoussaid ,&nbsp;A. Coman ,&nbsp;I. Coll ,&nbsp;S. Lanone ,&nbsp;E. Audureau","doi":"10.1016/j.rmr.2024.01.038","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.038","url":null,"abstract":"<div><h3>Introduction</h3><p>Cystic fibrosis (CF) is a genetic disorder that affects the respiratory and digestive systems. CF patients exhibit considerable variation in their symptoms and disease progression, suggesting complex genotype–phenotype relationships that may involve environmental factors. This study aimed to use unsupervised clustering analyses to identify distinct profiles and trajectories of CF patients, while also assessing their associations with various environmental factors.</p></div><div><h3>Methods</h3><p>Data from the French CF Registry, which covers 90% of CF patients in France and provides comprehensive health information for monitoring and research purposes, were utilized. By employing dimensionality reduction and clustering techniques, such as self-organizing maps (SOMs), reverse graph embedding (DDRTree algorithm, ClinTrajAn), and trajectory analyses (latent class analysis) based on longitudinal lung function tests, patients were grouped based on their clinical characteristics.</p></div><div><h3>Results</h3><p>Preliminary findings revealed the existence of different subgroups among CF children and adult patients, characterized by significant differences in overall health status, decline in lung function, comorbidities, incidence of infections, and exposure to environmental factors like passive smoking. Additionally, the study investigates the connections between CF profiles and air pollution at the geographic level of French departments.</p></div><div><h3>Conclusion</h3><p>Applying clustering techniques to large medical datasets reveals valuable insights into the impact of the environment on the physiological and pathological processes of CF. By uncovering distinct patient profiles, this approach can optimize treatment strategies and improve patient outcomes.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":null,"pages":null},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160433","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":"Development of succinate-based analogues and formulations against influenza virus infection","authors":"V. Vasseur ,&nbsp;A. Cezard ,&nbsp;A. Caumon ,&nbsp;A. Guillon ,&nbsp;N. Tsapis ,&nbsp;S. Le Poder ,&nbsp;S. Messaoudi ,&nbsp;M. Si-Tahar","doi":"10.1016/j.rmr.2024.01.049","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.049","url":null,"abstract":"<div><h3>Introduction</h3><p>One of the top global causes of death worldwide is respiratory viral infections. Among these, influenza virus-related infections cause deadly epidemics and pandemics. Each year, seasonal influenza infects more than 1 billion people (i.e. ∼20% of the world's population) and results in approximately 500,000 deaths. Vaccination efficacy can be impaired by viral intrinsic antigenic drift and the efficiency of drugs targeting directly influenza viruses is largely disputed. Hence, the development of innovative options is required to better treat influenza. In that regard, we recently demonstrated the antiviral activity of succinate (EMBO J., 2022) as well as of “C2”, two host metabolites. In this project, our objectives are to synthesize succinate-derived drugs against influenza viruses, select the best candidates and test them in vitro and in vivo, and finally formulate them into dry powders for direct lung delivery.</p></div><div><h3>Methods</h3><p>A series of compounds are synthesized by a structure-activity relationship strategy. Various chemical modifications will be introduced in the “hit” compound in the aim to identify analogues with high activity and good druggability.</p></div><div><h3>Results</h3><p>A preliminary screening of a short series of commercially available analogues using human bronchial epithelial cells led to the identification of three new active compounds (coined “S1”, “S10” and “S11”). All have an antiviral and anti-inflammatory effect more potent than the natural metabolites succinate and C2. In an in vivo model of influenza pneumonia, mice treated at day 2 post-infection with these analogues resisted better than non-treated or C2-treated animals to a lethal dose of influenza virus (survival rate: 85%, 0%, and 50% respectively).</p></div><div><h3>Conclusion</h3><p>Our study will pave the way for the development of appropriate “drug-and-devices” that will help to administer these succinate analogues directly into the respiratory tract. This project is funded through an ANR program.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":null,"pages":null},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160435","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 la dysfonction chronique du greffon pulmonaire par analyse transcriptomique sur cellule unique d’explants pulmonaires","authors":"P. Halitim ,&nbsp;A. Tissot ,&nbsp;L. Boussamet ,&nbsp;A. Garcia ,&nbsp;C. Fourgeux ,&nbsp;P. Lacoste ,&nbsp;B. Marie ,&nbsp;J. Poschmann ,&nbsp;S. Brouard ,&nbsp;L. Berthelot","doi":"10.1016/j.rmr.2024.01.044","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.044","url":null,"abstract":"<div><h3>Introduction</h3><p>La dysfonction chronique du greffon est la complication majeure de la transplantation pulmonaire puisque sa survenue impacte la survie de manière péjorative <span>[1]</span>, <span>[2]</span>. Il est important de progresser dans la connaissance des mécanismes physiopathologiques de cette pathologie afin d’identifier de nouvelles cibles thérapeutiques.</p></div><div><h3>Méthodes</h3><p>Cette étude présente les résultats d’une analyse transcriptomique sur cellule unique d’explants pulmonaires de patients atteints de CLAD (<em>n</em> <!-->=<!--> <!-->2), comparés à des explants de patients atteints de bronchopneumopathie chronique obstructive (<em>n</em> <!-->=<!--> <!-->1), de fibrose pulmonaire idiopathique (<em>n</em> <!-->=<!--> <!-->2) et des donneurs sains (<em>n</em> <!-->=<!--> <!-->2). Les lobes pulmonaires récupérés lors de transplantations ont été dissociés afin de récupérer les cellules. Une analyse transcriptomique sur cellules uniques a ensuite été réalisée sur le Chromium 10X au sein du laboratoire. Après validation des données, une analyse bio-informatique a été réalisée. Les résultats ont été confirmés au niveau protéique par des marquages sur tissus congelés et paraffiné.</p></div><div><h3>Résultats</h3><p>Ce travail révèle l’importance des monocytes et macrophages qui semblent surreprésentés dans le CLAD. Une sous analyse de cette population monocytaire a révélé quatre clusters associés au CLAD. Ils présentaient de manière globale une signature transcriptomique inflammatoire et pro fibrosante avec un profil macrophagique orienté M1. Trois cibles d’intérêts identifiées dans l’analyse transcriptomique sur cellule unique, CXCL10 PTX3 et IFIT3, ont été étudiées sur coupes tissulaires congelées en immunofluorescence. PTX3 était abondamment présent sur les coupes tissulaires provenant de tissus pulmonaires avec CLAD, ce qui renforce l’intérêt porté à cette protéine dans le cadre du CLAD. CXCL10 et IFIT3 n’étaient observés sur aucunes des coupes (<span>Fig. 1</span>).</p></div><div><h3>Conclusion</h3><p>Les monocytes et macrophages associés au CLAD ont une signature transcriptomique particulièrement pro-inflammatoire. Parmi les transcrits d’intérêts exprimés par ces monocytes, PTX3 semble être une cible qui mériterait d’être étudiée davantage dans la dysfonction chronique du greffon afin d’identifier sa place dans sa prise en charge du CLAD.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":null,"pages":null},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160522","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":"SPHINX31, a SRPK1 inhibitor, regulates the ATR/DNA-PKcs/CHK1 replicative checkpoint to inhibit cell growth in NSCLC cells","authors":"A. Shreim ,&nbsp;M. Rouchette ,&nbsp;N. Zubchuk ,&nbsp;H. Polverche ,&nbsp;D. Auboeuf ,&nbsp;S. Gazzeri ,&nbsp;B. Eymin","doi":"10.1016/j.rmr.2024.01.029","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.029","url":null,"abstract":"<div><h3>Introduction</h3><p>Lung cancer, including the non-small cell lung carcinoma (NSCLC) histological subtype, is a leading cause of cancer-related death worldwide. Acquisition of resistance to therapies such as platinum salts, the gold standard chemotherapy in NSCLC, is one of the major trick supporting patients’ poor prognosis. Deregulation of splicing patterns as well as of some splicing factors and/or their regulators participate in the process of carcinogenesis and lung tumor progression. However, less is known regarding the contribution of RNA splicing defects to lung tumor escape from therapies. Recently, pharmacological inhibitors targeting different components/regulators of the spliceosome machinery have emerged as potential anti-cancer drugs, such as SPHINX31 that inhibits SRPK1, a kinase implicated in splicing regulation through the phosphorylation of various serine/arginine (SR)-rich proteins.</p></div><div><h3>Methods</h3><p>In order to investigate whether RNA splicing defects contribute to acquired resistance to platinum salts in NSCLC, we have worked in cellular models of resistance derived from NSCLC cell line sub-cultured with increasing concentrations of cisplatin during 4–6 months in order to obtain resistant cells. We treated both the parental and resistant cells with SPHINX31 in order to investigate the role of SRPK1 in cell cycle regulation and apoptosis. Western blot and co-immunoprecipitation assays were used to study the effect of SPHINX31 on the expression/interaction of some ATR signaling pathway components. Then, RNA-seq analysis was performed to predict the potential signaling pathway by which SRPK1 inhibition induces cell death.</p></div><div><h3>Results</h3><p>In this study, we demonstrated that SPHINX31 inhibits ATR signaling, the main pathway involved in the management of replicative stress, notably in NSCLC cells with acquired resistance to platinum salts. This leads to cell growth inhibition and enhanced genomic instability. At the molecular level, we demonstrated that SRPK1 is recruited at stalled replication forks upon replicative stress, co-immunoprecipitates with the ATR/ATRIP/TOPBP1 complex and is required for TOPBP1/ATRIP recruitment to chromatin and TOPBP1 nuclear foci formation which contribute to ATR full activation. We further provided evidence that SPHINX31 and SRPK1 regulate the splicing of WIZ, in favor of splice variants involved in ATR activation, thereby identifying both splicing-dependent and -independent functions of SRPK1 by which it controls ATR signaling pathway. Last, we showed that the inhibitory effects of SPHINX31 on ATR are counterbalanced by the activation of DNA-PKcs and we identified a strong synergistic cytotoxic effect of the combination SPHINX31 and DNA-PKcs inhibitor in vitro. We propose that SPHINX31, alone or combined with DNA-PKcs inhibitor, could be benefit for NSCLC patients who relapse after platinum based-chemotherapy (<span>Fig. 1</span>).</p></div><div><h3>Conclusio","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":null,"pages":null},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140159988","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":"MicroRNA-155 differentially regulates IL-13Rα1 and IL-13Rα2 expression and signaling that drives abnormal epithelial cells functions in severe asthma","authors":"M. Klein ,&nbsp;P.A. Gagnon ,&nbsp;M. Salem ,&nbsp;M. Rouabhia ,&nbsp;J. Chakir","doi":"10.1016/j.rmr.2024.01.015","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.015","url":null,"abstract":"<div><h3>Rationale</h3><p>Asthma is a chronic airway disease characterized by a prevailing type 2 inflammation and airway remodeling. In asthma, microRNA-155 (miR-155) is known to increase in innate and adaptive immune cells and is associated with disease severity. However, little is known about its role in lung structural cells.</p></div><div><h3>Objectives</h3><p>This study investigated the expression of miR-155 and its regulatory role on IL-13 receptors and function in bronchial epithelial cells (BEC) isolated from healthy and severe asthmatic donors.</p></div><div><h3>Methods</h3><p>BECs isolated from healthy donors and severe asthma patients were stimulated with IL-13. MiR-155 expression and release were measured by RT-PCR in BECs and in their derived exosomes. Modulation of miR-155 in BECs was performed using transfection of miR-155 inhibitor and miR-155 mimic. MUC5AC, IL-8 and EOTAXIN-1 expression were measured by RT-PCR and BECs repair process was assessed by wound healing assay. IL-13Rα1 and IL-13Rα2 expression and downstream pathways were evaluated by western blot.</p></div><div><h3>Main results</h3><p>Severe asthma BECs showed an increased expression and exosomal release of miR-155 at baseline and amplified following IL-13 stimulation. Additionally, they expressed more IL-13Rα1 and less IL-13Rα2 than healthy BECs. IL-13Rα1 but not IL-13Rα2 induced miR-155 expression. Following miR-155 overexpression, BECs expressed more MUC5AC, IL-8 and EOTAXIN-1 through IL-13Rα1/SOCS1/STAT6 pathway and harbored a delayed repair process with a downregulated IL-13α2/ERK1/2/c-Jun signaling.</p></div><div><h3>Conclusions</h3><p>MiR-155 is overexpressed in severe asthma BECs and contributes to modulate IL-13Rα1 and IL-13Rα2 expression favoring mucin and eosinophils recruitment to detriment of airway repair. These results contribute to a better understanding of miR-155 role in the abnormal behavior of severe asthma BECs.</p></div>","PeriodicalId":21548,"journal":{"name":"Revue des maladies respiratoires","volume":null,"pages":null},"PeriodicalIF":0.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160087","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}