Revue des maladies respiratoires最新文献

{"title":"Mucosal administration of an immunotherapy provides long-term protection against a bacterial respiratory infection","authors":"A. Pitiot ,&nbsp;M. Ferreira ,&nbsp;C. Parent ,&nbsp;C. Boisseau ,&nbsp;M. Cortes ,&nbsp;L. Bouvart ,&nbsp;C. Paget ,&nbsp;N. Heuzé-Vourc’h ,&nbsp;T. Sécher","doi":"10.1016/j.rmr.2024.01.059","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.059","url":null,"abstract":"<div><h3>Introduction</h3><p>Acute and chronic respiratory infections are one of the first causes of death worldwide, especially for young children and elderly people. Their threat on human health has significantly increased over the last few years, due to the emergence of new viruses and the increase of antimicrobial multi-resistant strains of bacteria. Therapeutic antibodies (Ab) have proven their efficacy in infection prevention and control. Their administration inside the airways, rather than through parenteral route, allows to potentiate their therapeutic index against respiratory infections. The mode of action of anti-infectious Ab relies mainly on direct neutralization of the pathogen, and the recruitment of effectors immune cells facilitating its elimination. However, some studies suggest an immunomodulator role of therapeutic antibodies. Thus, this project aimed at making a proof-of-concept of mAb-mediated long-term immune protection after mucosal administration during a pulmonary bacterial infection and deciphering the mechanisms underlying this effect.</p></div><div><h3>Methods</h3><p>We used a mouse model of <em>Pseudomonas aeruginosa</em> (<em>P. aeruginosa</em>) lung infection, resembling to human acute pneumonia. The primary infection was treated using a neutralizing anti-<em>P. aeruginosa</em> mAb delivered through the airways. To investigate long-term protection, mice surviving primary infection were challenged a month later, at a time when circulating/local mAb had disappeared, once again with <em>P. aeruginosa</em>. Local and systemic humoral and cellular immune responses were analyzed thereafter.</p></div><div><h3>Results</h3><p>Thanks to the, previously presented, model, we have shown that beyond a rapid and efficient containment of a lethal primo infection, an antibacterial Ab, administered in the airways, can interact with the mucosal immune system to develop a long-term immunity, protecting against a secondary infection. Results obtained in vitro and in vivo indicate an essential contribution of immune complexes, formed between the bacteria and the antibody, in the induction of an antibacterial humoral response. This humoral response, persistent, is protective against the pathogenic bacteria. The long-term immunity thus seems to be dependent on both the amount of immune complexes, but also the presence of a local inflammation during the primo infection. Interestingly, the long-term immune response also offers a partial protection against a secondary infection by a heterologous strain of <em>P. aeruginosa</em>.</p></div><div><h3>Conclusion</h3><p>Complementary studies will be needed to identify all cellular and molecular actors implicated in this long-term protection. Nevertheless, our results suggest that a mucosal administration of an Ab enables the neutralization of a bacterial infection and ensures a long-term protection against a reinfection. This immunomodulatory characteristic of inhaled antibacterial Ab opens new pe","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":"140160473","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":"Late response of bronchial epithelium to rhinovirus in severe asthma","authors":"K. Valette ,&nbsp;L. Moreno ,&nbsp;A.S. Payet ,&nbsp;P. Chanez ,&nbsp;D. Gras","doi":"10.1016/j.rmr.2024.01.014","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.014","url":null,"abstract":"<div><h3>Introduction</h3><p>Alteration of bronchial epithelium is one of the main features of severe asthma. Bronchial epithelial cells (BEC) promote airway inflammation and remodeling leading to chronic symptoms and airway hyperresponsiveness and obstruction. Moreover, BEC are the first line in viral infections, which are the major contributor to asthma exacerbation. However, the consequences of the persisting long-term responses to viral infection of BEC in severe asthma are not fully understood. The aim of our study was to investigate the morphological and functional responses of bronchial epithelium induced by human rhinovirus in acute and late phase post-infection in severe asthma in an ALI culture model.</p></div><div><h3>Methods</h3><p>Primary human bronchial epithelial cells (HBECs) from control (C) (<em>n</em> <!-->=<!--> <!-->3) and severe asthmatic (SA) (<em>n</em> <!-->=<!--> <!-->4), cultured in air–liquid interface (ALI), were infected with Rhinovirus A16 (RV-A16) at the apical pole. The virus was removed four hours after infection. Differences in term of viral replication, cytotoxicity (measured by LDH activity), epithelium cohesion (TEER measurement) and antiviral response was assessed at the acute phase (until 4<!--> <!-->days) and at the late phase (until 14<!--> <!-->days) post-infection (dpi).</p></div><div><h3>Results</h3><p>RV-A16 replication increased during the acute phase then decreased during the late phase, without difference between control and severe asthmatic. Cytotoxicity was slightly induced by RV-A16 infection only in the acute phase with no difference observed between control and severe asthmatic (fold induction compared to MOCK: 10.4 and 11.7 for C and SA respectively at 4 dpi). Epithelium cohesion is damaged by RV-A16 in severe asthmatics at the late phase whereas it is strengthened in controls (Fold induction compared to MOCK: 1.12 and 0.62 for C and SA respectively at 14 dpi). RV-A16 induced secretion of several antiviral defense peptides (IFNλ2-3, IFNλ1, IFNβ, IFNα2) and inflammatory mediators (CCL5, CXCL10, IL 33, TSLP) over the first week post-infection, with different kinetic and intensities between control and severe asthmatic. IL-33 and TSLP secretion is significantly more induced in severe asthmatic than in control (<em>P</em> <!-->&lt;<!--> <!-->0.05). Antiviral and proinflammatory mediator's secretion returns to baseline at 14 dpi in both severe asthmatics and controls, but declines more rapidly in severe asthmatics.</p></div><div><h3>Conclusion</h3><p>The antiviral response is different in terms of profile and intensity between control and severe asthmatic, both in the acute and late phases. The virus induces alteration of epithelium cohesion in severe asthmatics, suggesting a defect in cellular events normally induced by injury. Then, an altered response of HBECs in severe asthma, illustrated by the increased secretion of epithelial alarmins IL-33 and TSLP, could contribute to increased disease su","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":"140160086","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;A. Nasri ,&nbsp;F. Foisset ,&nbsp;I. Vachier ,&nbsp;A. Bourdin ,&nbsp;S. Assou ,&nbsp;J. De Vos","doi":"10.1016/j.rmr.2024.01.007","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.007","url":null,"abstract":"<div><h3>Introduction</h3><p>Primary ciliary dyskinesia (PCD) is a genetic disease caused by mutations that alter cilia beating, including in the respiratory airways, resulting in impaired mucus clearance and severe morbidity as well as increased mortality. We hypothesized that we could restore bronchial cilia beating with genetically corrected iPSC differentiated into bronchial progenitors.</p></div><div><h3>Methods</h3><p>Our project aims to assess the ability of a corrected iPSC line to functionally repair pathological models in vitro. We generated a PCD patient iPSC line reprogrammed using Sendai viruses, and the 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 expressing the fluorescent GFP protein under the human elongation factor 1 alpha promoter (EF1a), allowing us to study the engraftment ability of GFP bronchial stem cells on a control epithelium model. To assess the efficiency of different bronchial progenitors to engraft a bronchial epithelium, we used our previously published air–liquid interface bronchial epithelium model (iALI).</p></div><div><h3>Results</h3><p>One main issue is to identify the competent cell type for regeneration of the adult bronchial epithelium. Indeed, there are several cell types constituting the bronchial epithelium, as well as several developmentally bronchial progenitor cells that could be considered. Our iALI differentiation process mimics the embryonic development and thus the iALI model may provide any cell type from the definitive endoderm to the mature bronchial epithelium. Our results suggest that lung progenitors at the ventralized anterior foregut endoderm stage, could be the most efficient cells for engraftment. Besides, their self-renewal ability and their capacity to differentiate into the different cell type spectrum of the bronchial epithelium are promising for the development of a long-term and efficient therapy. The second issue for bronchial epithelium cell replacement would be to determine the best strategy to erode the bronchi prior to cell therapy. Such an erosion is considered necessary to promote cell engraftment because of the barrier function of the intact bronchial epithelium and the lack of selection advantage from the corrected cells. To this end, we compared mechanical, chemical and enzymatic erosion strategies on the iALI model. Our results suggest a better efficiency with enzymatic erosion, showing a homogeneous detachment of the cells and a better engraftment of cells from the GFP-iPSC line.</p></div><div><h3>Conclusion</h3><p>In conclusion, engraftment of corrected lung progenitors to enzymatically eroded bronchial epithelium seems to be a promising therapeutic strategy to treat PCD. Future experiments will refine the best condition regarding enzymatic solution (concentration, time exposure) and graft cell number to assure functional recovery of the mucociliary clearance.</p>","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":"140160152","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 maturation et les fonctions de l’éosinophile dans l’asthme sévère","authors":"B. Hugo ,&nbsp;E. Coralie ,&nbsp;R. Morgane ,&nbsp;B. François-Xavier ,&nbsp;L. Gervaise ,&nbsp;H. Dorian ,&nbsp;S. Vincent","doi":"10.1016/j.rmr.2024.01.008","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.008","url":null,"abstract":"<div><h3>Introduction</h3><p>L’asthme est une pathologie bronchique caractérisée par une hyperréactivité bronchique, un remodelage tissulaire et une inflammation chronique. Nos travaux ont montré que la GTPase Rac était activée au sein des éosinophiles dans un modèle murin d’asthme allergique sévère. Notre objectif est d’évaluer le rôle de la protéine Rac dans la maturation et la dégranulation des éosinophiles.</p></div><div><h3>Méthodes</h3><p>Les éosinophiles étudiés ont été obtenus par différenciation de cellules médullaires issues de moelles osseuses de souris. Les niveaux de Rac-GTP ont été évalués au cours de la maturation cellulaire par cytométrie en flux et western blot. Sur le plan fonctionnel, la dégranulation des éosinophiles est induite in vitro par le <em>platelet activating factor</em> (PAF) et évaluée par la mesure de l’activité peroxidase et RNase.</p></div><div><h3>Résultats</h3><p>Le protocole de différenciation des cellules médullaire permet d’obtenir des cultures cellulaires contenant 95 % d’éosinophiles. Au cours de cette différenciation, le niveau de Rac actif (Rac-GTP) augmente significativement (j4 0,6 vs j11 2,7, <em>p</em> <!-->=<!--> <!-->0,03). Par cytométrie en flux, nous avons déterminé que cette élévation concerne principalement les éosinophiles. Nous avons observé que le PAF entraîne la dégranulation des éosinophiles en culture avec une libération d’éosinophile péroxidase et de RNases. Cette dégranulation s’accompagne d’une augmentation de l’activation de Rac. L’inhibition pharmacologique de Rac génère une accumulation des granules au niveau de la membrane plasmique et prévient la dégranulation in vitro (diminution de 74 %, <em>p</em> <!-->=<!--> <!-->0,002) et sur des lysats pulmonaires de modèles d’asthme sévère.</p></div><div><h3>Conclusions</h3><p>Nos résultats mettent en évidence que l’activation de Rac participe à la maturation et à la dégranulation des polynucléaires éosinophiles. Rac apparaît comme une nouvelle cible thérapeutique dans l’asthme allergique pour lutter contre l’inflammation.</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":"140160153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of a host metabolite with both antiviral and anti-inflammatory properties that protects against influenza virus-driven morbidity and mortality","authors":"A. Cezard ,&nbsp;D. Brea ,&nbsp;V. Vasseur ,&nbsp;L. Gonzalez ,&nbsp;B. Da-Costa ,&nbsp;R. Le Goffic ,&nbsp;D. Fouquenet ,&nbsp;T. Baranek ,&nbsp;C. Paget ,&nbsp;A. Guillon ,&nbsp;M. Si-Tahar","doi":"10.1016/j.rmr.2024.01.051","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.051","url":null,"abstract":"<div><h3>Introduction</h3><p>Influenza is an acute respiratory immunopathology resulting from the intrinsic pathogenicity of influenza A (IAV) and B (IBV) viruses, added to an inflammatory host response, which in excess, is deleterious. Influenza is a major public health issue, causing 500,000 deaths per year and a considerable socioeconomic burden. Frequent vaccine escape and emerging antiviral resistance spur innovative research on host-directed therapy against influenza. In that regard, some host metabolites have emerged as potent immuno-regulatory molecules.</p></div><div><h3>Methods</h3><p>We recently characterized mitochondria-derived succinate as a major antiviral metabolite (EMBO J., 2022). Here, we extended those findings by screening the effect of other host metabolites on influenza virus infection, using human bronchial epithelial cells. Among these metabolites, we focused on one mitochondria-derived metabolite (coined here “C2”) and evaluated its anti-inflammatory and antiviral activity using complementary approaches, such as immunological and qRT-PCR assays, reverse genetics, western-blotting, flow cytometry and confocal microscopy techniques.</p></div><div><h3>Results</h3><p>We found that C2 has a major antiviral activity by preventing the expression of viral mRNA and protein, and the subsequent multiplication of both influenza A and B virus strains. Next, we observed that C2 alleviates the inflammatory cascades triggered by influenza virus and other inflammatory stimuli, including the inflammatory cytokine TNFα. We further showed in a mouse model of influenza pneumonia that C2 decreases the viral load, the leukocyte recruitment into the airspaces, the secretion of inflammatory mediators and the damage of lung tissues. As a result, mice treated at day 2 post-infection with C2 resist better than non-treated animals to a lethal dose of influenza virus (survival rate: 70% and 0%, respectively).</p></div><div><h3>Conclusion</h3><p>We reveal that C2 is a host metabolite with anti-viral and anti-inflammatory properties that protect against influenza pneumonia. Our results open new avenues for the development of a C2-based treatment of influenza virus infection.</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":"140160437","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":"Noenatal obstructive sleep apneas in a mouse model of Down syndrome","authors":"M. Moreau ,&nbsp;A. Madani ,&nbsp;R. Dard ,&nbsp;T. Bourgeois ,&nbsp;M.P. D’Ortho ,&nbsp;P. Bokov ,&nbsp;N. Janel ,&nbsp;B. Matrot","doi":"10.1016/j.rmr.2024.01.085","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.085","url":null,"abstract":"<div><h3>Introduction</h3><p>Down syndrome (DS) is a genetic disease caused by a third copy of chromosome 21, leading to various physical features, developmental and cognitive delays and intellectual disability. In neonates, main symptoms concern congenital heart defects, gastrointestinal abnormalities and sleep disordered breathing. A recent retrospective study showed a high prevalence of obstructive sleep apnea (OSA) in children with DS, with severity inversely related with age culminating at 58% of severe OSA in neonates<!--> <!-->&lt;<!--> <!-->1y. Knowing that OSA can cause intermittent hypoxia and hypercapnia and have detrimental effects on health and development, it raises concerns about the impact of OSA on neurodevelopmental disorders associated with DS, especially in neonates. Many animal models exist. Dp(16)1Yey mice, a genetically engineered model, exhibit cognitive impairments and characteristics associated with OSA, including craniofacial hypoplasia and reduced upper airway volume at adult age. To further investigate the contribution of respiratory-related disorders to DS pathophysiology, we examined the respiratory phenotype of Dp(16)1Yey mice at birth, with special attention to central, obstructive and mixed apneas.</p></div><div><h3>Methods</h3><p>On the day of birth, the pups’ snouts were attached to a pneumotachometer with a polyether adhesive. Abdominal movements were measured using a laser sensor pointing the lateral abdominal wall to detect respiratory efforts and discriminate apnea. The classification into central, obstructive or mixed apneas was performed by visual inspection of pneumotachometer and laser signals. Electrocardiograms (ECG) were recorded using two human skin electrodes adapted to the size of the pups. Heart rate (HR) was determined from the R-R wave peaks after visual selection of continuous, 20-s or longer ECG segments with clearly defined QRS waves.</p></div><div><h3>Results</h3><p>At birth, Dp(16)1Yey mouse pups exhibited lower weight and HR compared to their wild type (WT) counterparts. Baseline breathing variables and response to hypercapnia were similar between the two groups. Total time for obstructive apneas was longer in Dp(16)1Yey than in WT pups (2.18<!--> <!-->±<!--> <!-->1.8 s/min vs. 1.24<!--> <!-->±<!--> <!-->1.6 s/min, respectively; <em>P</em> <!-->=<!--> <!-->0.023), owing to their longer mean duration (3.11<!--> <!-->±<!--> <!-->1.1<!--> <!-->s vs. 2.05<!--> <!-->±<!--> <!-->0.7<!--> <!-->s, respectively; <em>P</em> <!-->=<!--> <!-->0.002). ECG analysis did not reveal apnea-related bradycardia in either group.</p></div><div><h3>Conclusion</h3><p>These findings highlight the relevance of the Dp(16)1Yey model for studying OSA in DS, including its occurrence at birth. This model represents a valuable tool to investigate the contribution of early respiratory disorders to DS pathology and assess safety and efficacy of pharmacological treatments targeting obstructive sleep disordered breathing in","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":"140159940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification des gènes associés au programme de ciliogenèse primaire dans les maladies respiratoires chroniques","authors":"S. Hatoum ,&nbsp;R. Belgacemi ,&nbsp;G. Deslée ,&nbsp;M. Polette ,&nbsp;J.M. Perotin ,&nbsp;D. Al Alam ,&nbsp;V. Dormoy","doi":"10.1016/j.rmr.2024.01.022","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.022","url":null,"abstract":"<div><h3>Introduction</h3><p>Les maladies respiratoires chroniques restent l’une des principales causes de décès dans le monde avec une incidence de 7,4 %. Ces pathologies sont caractérisées par un remodelage de l’épithélium des voies aériennes à l’origine d’une altération de la clairance mucociliaire, notamment dans la bronchopneumopathie chronique obstructive (BPCO). Dans ce contexte, nous avons démontré que (i) la ciliogenèse primaire est essentielle pour la différentiation des cellules épithéliales des voies aériennes et (ii) que la voie Hedgehog (HH) est altérée dans la BPCO. L’objectif est donc d’identifier les gènes impliqués dans la ciliogenèse primaire afin de caractériser leur rôle dans la BPCO.</p></div><div><h3>Méthodes</h3><p>Des cellules de la lignées A549 ont été traitées par du Tgfβ1 (0 et 5<!--> <!-->ng/mL) pour 8<!--> <!-->h, 24<!--> <!-->h, 48<!--> <!-->h et 72<!--> <!-->h en condition de confluence ou en prolifération afin d’induire la ciliogenèse primaire. Des immunomarquages fluorescents (IF) et des analyses quantitatives ont été réalisés pour évaluer l’effet de Tgfβ1 sur la ciliation. Finalement, l’analyse du transcriptome en réponse au Tgfβ1 a été explorée.</p></div><div><h3>Résultats</h3><p>Une augmentation de l’index de ciliation et de la longueur des cils ont été observés pour les différents temps de traitement par Tgfβ1 mais surtout après 48<!--> <!-->h : 9,27 %<!--> <!-->±<!--> <!-->1,25 des cellules confluentes non traitées avaient des cils primaires avec une longueur de 1,21<!--> <!-->μm<!--> <!-->±<!--> <!-->0,02 contre 12,86 %<!--> <!-->±<!--> <!-->0,23 et une longueur de 1,46<!--> <!-->μm<!--> <!-->±<!--> <!-->0,03 pour les cellules confluentes traitées par Tgfβ1 (<em>p</em> <!-->&lt;<!--> <!-->0,05). Concernant les analyses des données transcriptomiques des gènes associés aux cils (398 gènes extraits des protéines associées aux cils, aux corps basaux et aux ciliopathies sur UniProt), entre 16<!--> <!-->h, 24<!--> <!-->h et 72<!--> <!-->h de culture après le traitement par Tgfβ1, 27 gènes étaient communément altérés et potentiellement associés au programme de la ciliogenèse dont 3 des acteurs de la voie HH (SNAI2, CDON, GLI2). Les 3 gènes les plus dérégulés, suite au traitement par Tgfβ1, étaient <em>SNAI2</em>, <em>DAAM1</em> et <em>EPHB2</em> (respectivement surexprimés 1,44, 1,42, et 1,37 fois).</p></div><div><h3>Conclusion</h3><p>La caractérisation phénotypique de la lignée A549 traitée par Tgfβ1 a pu mettre en évidence des gènes potentiellement impliqués dans la formation des cils primaires. Ainsi, l’analyse des mécanismes de régulation cellulaire liés à ces gènes pourra ouvrir la voie à la découverte de nouvelles cibles thérapeutiques ainsi qu’à l’amélioration du pronostic et du diagnostic dans les pathologies respiratoires associées à du remodelage épithélial.</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":"140159981","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 des associations entre dérégulations associées aux cils dans l’épithélium des voies aériennes et phénotypes asthmatiques","authors":"M.A. Devilliers ,&nbsp;A. Brisebarre ,&nbsp;L.M.G. Petit ,&nbsp;M. Polette ,&nbsp;G. Deslée ,&nbsp;R. Djukanovic ,&nbsp;V. Dormoy ,&nbsp;J.M. Perotin","doi":"10.1016/j.rmr.2024.01.011","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.011","url":null,"abstract":"<div><h3>Introduction</h3><p>L’asthme est une maladie fréquente caractérisée par une inflammation chronique et un remodelage des voies aériennes touchant 7 à 10 % de la population en France et représentant un impact socioéconomique important. L’asthme est très hétérogène en termes d’expression clinique et les principaux phénotypes décrits à ce jour reposent sur le type d’inflammation prédominant. Au niveau de l’épithélium respiratoire, des anomalies des cils moteurs et d’expression de certains gènes associés aux cils ont été démontrées dans l’asthme mais le rôle des cils dans la physiopathologie de la maladie reste encore peu décrit. L’objectif de ce projet est de mettre en évidence des associations entre des anomalies liées aux cils dans l’épithélium des voies aériennes et des phénotypes asthmatiques.</p></div><div><h3>Méthodes</h3><p>Les données de transcriptome épithélial bronchique obtenues à partir de la cohorte européenne U-BIOPRED (GSE76226) ont été analysées dans le but de déterminer des clusters de patients asthmatiques sur la base d’expression des gènes associés aux cils (879 gènes décrits). Les patients fumeurs ont été exclus de l’étude. Les caractéristiques cliniques, phénotypiques et biologiques ont été comparées entre les groupes de patients identifiés par des expressions différentielles des transcrits associés aux cils. Les processus biologiques (termes GO) et les interactions protéiques probables des 100 gènes les plus dérégulés dans le transcriptome épithélial complet de chacun des groupes de patients ont été analysés à l’aide de l’interface STRING.</p></div><div><h3>Résultats</h3><p>L’analyse bio-informatique a permis d’identifier et comparer deux groupes de patients asthmatiques sévères : patients dits « dérégulés » (SAd, <em>n</em> <!-->=<!--> <!-->18) présentant les dérégulations les plus importantes en termes d’expression des gènes associés aux cils, et patients dits « non dérégulés » (SA, <em>n</em> <!-->=<!--> <!-->43) correspondant au reste des asthmatiques sévères de la cohorte. Une augmentation significative des taux de cytokines associées au phénotype éosinophile, CCL17 (147<!--> <!-->±<!--> <!-->108 contre 76<!--> <!-->±<!--> <!-->62<!--> <!-->pg/mL), IL-13 (1,3<!--> <!-->±<!--> <!-->1,2 contre 0,8<!--> <!-->±<!--> <!-->0,6<!--> <!-->pg/mL) et MIP1b (67<!--> <!-->±<!--> <!-->20 contre 54<!--> <!-->±<!--> <!-->20<!--> <!-->pg/mL), est observée dans le plasma des patients SAd ; ainsi qu’une augmentation du nombre d’éosinophiles (0,39<!--> <!-->±<!--> <!-->0,34 contre 0,25<!--> <!-->±<!--> <!-->0,20<!--> <!-->×<!--> <!-->10³/μL) et de basophiles (0,16<!--> <!-->±<!--> <!-->0,34 contre 0,03<!--> <!-->±<!--> <!-->0,04<!--> <!-->×<!--> <!-->10³/μL) dans le sang périphérique traduisant un phénotype inflammatoire de profil T2 high. L’analyse ontologique a permis d’associer les 100 gènes les plus dérégulés parmi le transcriptome épithélial complet chez les patients SAd à des termes GO liés à l’inflammatio","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":"140160156","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":"Understanding persistent airway obstruction in type 2 severe asthma using human induced pluripotent stem cells (iPSCs)","authors":"E. Ahmed ,&nbsp;F. Foisset ,&nbsp;C. Bourdais ,&nbsp;A. Nasri ,&nbsp;A. Petit ,&nbsp;S. Assou ,&nbsp;D. Gras ,&nbsp;P. Chanez ,&nbsp;I. Vachier ,&nbsp;H. Hammad ,&nbsp;J. Vos (De) ,&nbsp;B. Lambrecht ,&nbsp;A. Bourdin","doi":"10.1016/j.rmr.2024.01.012","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.012","url":null,"abstract":"<div><h3>Introduction</h3><p>Despite the new biologics to treat inflammation in severe asthma, targeting persistent obstruction of the airways remains challenging. Galectin-10 eosinophil derived crystals, also known as Charcot-Leyden crystals (CLCs) have been described to be present in the mucus plugs in the airways of patients with severe asthma. However, a direct role for CLCs in mucus production has not been established. We hypothesize that plugged airways constitute a unique niche where type 2 immune cells communicate with structural cells to perpetuate disease. We aimed to set up a new model using induced pluripotent stem cells (iPSCs).</p></div><div><h3>Methods</h3><p>Three human iPSCs lines from type 2 severe asthma patients have been derived (MOSAIC study, University Hospital of Montpellier, <span>NCT05616338</span><svg><path></path></svg>) and differentiated into airway epithelium in air–liquid interface (i-ALI). The healthy iPSC line UHOMi002-A was used as a control. At day 21 of ALI culture, iPSC derived-airway epithelia were stimulated at the apical side with either IL-13 every two days (10<!--> <!-->ng/mL) during one week, acute stimulation (24<!--> <!-->h) with recombinant Gal10 crystals (100<!--> <!-->ng/mL), both IL-13 and Gal10 crystals or PBS (vehicle). We aimed to evaluate the effect on i-ALI differentiation at day 30.</p></div><div><h3>Results</h3><p>We successfully differentiated the iPSC lines generated from the T2 severe asthma patients, and achieving a high purity rate at each developmental stages. The mean cell purity at the definitive endoderm for each cell line was<!--> <!-->&gt;<!--> <!-->80% assessed by flow cytometry quantification of C-X-C Motif Chemokine Receptor 4 (CXCR4)/c-KIT double positive cells and immunolabelling of Forkhead Box A2 (FOXA2)+/SRY-box transcription factor 17 (SOX17)+. Purity for ventral anterior foregut endoderm (vAFE) stage was evaluated at 70%, through Transcription Factor NK2 Homeobox 1 (NKX2.1) expression, Carboxy Peptidase M (CPM) by flow cytometry. vAFE cells from the hiPSC lines differentiated into bronchial epithelium in air–liquid interface conditions. Chronic IL-13 challenging and CLC were both able to induce an increasing of MUC5AC+ cells and also an increase of neuroendocrine cells in asthmatic iPSC lines.</p></div><div><h3>Conclusion</h3><p>iALI bronchial epithelium can recapitulate T2 severe asthma features in vitro, and highlighted a possible direct effect of the CLC on the airway epithelium.</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":"140160157","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":"Développement d’un modèle d’épithélium bronchique innervé par des neurones sensoriels obtenus à partir de cellules souches pluripotentes induites","authors":"F. Foisset ,&nbsp;C. Lehalle ,&nbsp;A. Nasri ,&nbsp;C. Bourdais ,&nbsp;L. Morichon ,&nbsp;M. Nadaud ,&nbsp;I. Vachier ,&nbsp;S. Assou ,&nbsp;A. Bourdin ,&nbsp;J. De Vos ,&nbsp;N. Frossard","doi":"10.1016/j.rmr.2024.01.013","DOIUrl":"https://doi.org/10.1016/j.rmr.2024.01.013","url":null,"abstract":"<div><h3>Introduction</h3><p>Les maladies respiratoires sévères, dont l’asthme, nécessitent de nouveaux traitements car les traitements actuels sont encore inefficaces chez un certain nombre de patients. Notre projet vise à obtenir un modèle d’épithélium bronchique innervé par une innervation sensitive in vitro à partir de cellules souches pluripotentes induites (iPSC).</p></div><div><h3>Méthodes</h3><p>Nous avons synthétisé une matrice extracellulaire composée d’un mélange de collagène bovin de type I et de chitosan. Cette matrice est d’abord colonisée par des fibroblastes bronchiques humains primaires et agit comme une muqueuse sous-épithéliale. Des neurones sensitifs dérivés de cellules souches pluripotentes induites (iPSC) sont cultivés sur la face inférieure de la matrice et un épithélium bronchique dérivé d’iPSC provenant des mêmes sujets est cultivé sur la surface, en interface air/Liquide (ALI), selon notre protocole décrit précédemment <span>[1]</span>.</p></div><div><h3>Résultats</h3><p>Nous avons obtenu un épithélium bronchique dérivé d’iPSC présentant les principaux types cellulaires. Il repose sur une matrice extracellulaire épaissie par le collagène sécrété par les fibroblastes et présentant une lame basale. La coloration HES des coupes en paraffine montre un épithélium pseudostratifié avec différents types cellulaires bronchiques, y compris des cellules ciliées, Club, à mucus et basales. La coloration à l’acide périodique Schiff - Bleu Alcian permet de visualiser les cellules à mucus. Ces résultats sont confirmés par l’immunofluorescence de MUC5AC+. Les cellules ciliées sont TUBULINE ß IV+, les cellules basales Kératine 5+, les cellules club CCSP+, et nous avons également observé des cellules neuroendocrines chromogranineÀ+. Une innervation partielle est démontrée à l’intérieur de la matrice par immunofluorescence des neurones sensoriels dérivés des iPSC (TUBß3+). Pour essayer d’améliorer cette innervation, nous avons différencié et ajouté des cellules de Schwann dérivées d’iPSCS dans ce modèle car elles pourraient permettre le guidage des fibres nerveuses <span>[2]</span>. Les résultats préliminaires montrent une amélioration de l’innervation avec la présence effective de fibres nerveuses PGP9,5+ et TUBß3+ dans la matrice. La maturation de ces fibres sensitives et attestée par la présence de neuropeptides (fibres CGRP +).</p></div><div><h3>Conclusion</h3><p>En conclusion, nous avons obtenu avec succès un épithélium bronchique humain dérivé d’iPSC en 3D reposant sur une muqueuse sous-épithéliale. Des différenciations sont en cours pour confirmer les résultats obtenus avec l’innervation et la fonctionnalité de notre modèle complet.</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":"140160158","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}