American journal of physiology. Lung cellular and molecular physiology最新文献

{"title":"Obese adipocytes induce fibroblast-to-myofibroblast transition through TGF-β1 signaling: implications in asthma pathogenesis.","authors":"Nour Jalaleddine, Mahmood Hachim, Alexander Giddey, Mellissa Gaudet, Qutayba Hamid, Mohammed Uddin, Saba Al Heialy","doi":"10.1152/ajplung.00208.2025","DOIUrl":"10.1152/ajplung.00208.2025","url":null,"abstract":"<p><p>Obesity, a key risk factor for severe asthma, is associated with worsening symptoms and poor responses to conventional therapies. Recent studies have highlighted the presence of adipocytes within airway walls, which correlates positively with body mass index (BMI). However, the role of adipocytes in asthma pathogenesis remains largely unknown. This study aims to explore their potential contribution to airway fibrosis, a progressive form of the disease, through fibroblast-to-myofibroblast transition (FMT). In vitro coculture models were developed to investigate the interactions between adipocytes (derived from patients with and without obesity) and fibroblasts (from patients with and without asthma) on FMT. Proteomic and multiplex analyses were used to identify potential mediators of adipocyte-induced FMT. Our data revealed a significant increase in fibrogenic markers, such as alpha-smooth muscle actin and vimentin, in fibroblasts cocultured with obese (Ob) adipocytes. Notably, this transition was more pronounced in asthmatic fibroblasts compared with healthy fibroblasts. Proteomic profiling of cocultured Ob-adipocytes and asthmatic fibroblasts identified several significantly upregulated proteins linked to the regulation of the transforming growth factor-beta (TGF-β) signaling pathway, including inhibin A, latent TGF-β binding protein 1, thrombospondin 1, and follistatin. The role of TGF-β was further substantiated by multiplex assays, which demonstrated a significant increase in TGF-β and leptin production by Ob-adipocytes following coculture. These findings suggest that Ob-adipocytes may promote FMT in fibroblasts, especially asthmatic fibroblasts, by activating the TGF-β signaling pathway. This highlights a potential mechanism by which obesity exacerbates asthma severity and fibrosis, providing new avenues for therapeutic intervention.<b>NEW & NOTEWORTHY</b> Adipocytes have been found in the airway wall of patients with obesity. This study is the first to show that adipocytes derived from patients with obesity can induce features of airway remodeling that is seen in patients with asthma such as fibroblast-to-myofibroblast transition via the TGF-beta signaling pathway in an indirect mode of cellular communication. This highlights a potential mechanism by which obesity exacerbates asthma severity and fibrosis, providing new avenues for therapeutic intervention.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L419-L430"},"PeriodicalIF":3.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MicroRNA-enriched EVs from mesenchymal stromal cells: a novel approach to suppress biofilm and inflammation in CF airway epithelium.","authors":"Anushka Raghubansi, Catherine M Greene","doi":"10.1152/ajplung.00353.2025","DOIUrl":"10.1152/ajplung.00353.2025","url":null,"abstract":"","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L416-L418"},"PeriodicalIF":3.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Receptor agonist of NFκB signaling ligand directs lung epithelial cell expansion through RANK signaling.","authors":"Habibie Habibie, Jelmer R Vlasma, Kurnia S S Putri, Shanshan Song, Marina H de Jager, Arjen Petersen, Carian E Boorsma, Robbert H Cool, Wim Quax, Martijn C Nawijn, Wim Timens, Janette K Burgess, Corry-Anke Brandsma, Barbro N Melgert","doi":"10.1152/ajplung.00176.2025","DOIUrl":"10.1152/ajplung.00176.2025","url":null,"abstract":"<p><p>Chronic obstructive pulmonary disease (COPD) is the third leading cause of death globally, with progressive emphysema driven by repeated epithelial damage and impaired repair. Recently, we found that secretion of cytokine receptor agonist of nuclear factor κB signaling ligand (RANKL) is higher from lung fibroblasts of patients with COPD compared with control and that RANKL reduces lung epithelial cell death. However, the underlying mechanisms, their conservation across species, and the specific epithelial cell types involved remain unclear. To investigate how RANKL affects lung epithelial cells, we used in vitro and in vivo models. Primary lung organoids from human and mouse epithelial cells assessed progenitor activation and expansion. Furthermore, we used a murine model of elastase-induced alveolar injury to examine which epithelial cell types were affected by RANKL in vivo and whether this was altered upon lung damage. RANKL supplementation increased alveolar organoid formation in both murine and human cultures compared with vehicle-treated controls. In elastase-treated mice, RANKL administration during the early repair phase enhanced the proportion of EpCAM⁺ epithelial cells and increased transitional epithelial cell states characterized by keratin 8 (Krt8) and major histocompatibility complex II (MHCII) expression following elastase-induced injury. RANKL signaling promoted epithelial regeneration by expanding alveolar progenitor and transitional epithelial cell populations, with consistent effects across human and murine models. These findings identify RANKL as a novel modulator of epithelial repair and suggest that controlled activation of this pathway could enhance lung regeneration in diseases such as COPD.<b>NEW & NOTEWORTHY</b> This study describes for the first time a role for bone cytokine RANKL in lung epithelial repair following lung damage. Using both murine and human models, we describe how RANKL-mediated signaling promotes epithelial expansion by acting on epithelial transitional cell states. For diseases with substantial lung damage, modulating RANKL signaling during the early repair phase following lung injury could represent a strategy to enhance epithelial regeneration.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":"330 4","pages":"L274-L287"},"PeriodicalIF":3.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147479553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sex chromosomes and gonadal sex interactions in airway and immune responses to allergen challenge.","authors":"Carolyn Damilola Ekpruke, Dustin Rousselle, Rachel Alford, Omar Borges-Sosa, Maksat Babayev, Shikha Sharma, Lyidia Dinwiddie, Erik Parker, Sarah Bradley, Matthew Louis Retzner, Patricia Silveyra","doi":"10.1152/ajplung.00210.2025","DOIUrl":"10.1152/ajplung.00210.2025","url":null,"abstract":"<p><p>Asthma is a chronic respiratory condition influenced by genetic, environmental, and sex-related factors. Women experience greater asthma severity, airway hyperresponsiveness (AHR), and inflammation than men, likely due to sex-linked genetic and hormonal differences. However, the independent contributions of sex chromosomes and gonadal sex to these responses remain unclear. This study examines their roles in allergic airway responses using the four core genotype (FCG) mouse model, which distinguishes between chromosomal and gonadal influences. We hypothesized that XX-mice and those with female gonads would exhibit heightened airway inflammation and immune activation in response to house dust mite (HDM) challenge. Using a controlled, moderate 5-wk HDM exposure paradigm that reliably induced allergic airway inflammation, we aimed to capture biologically relevant sex- and genotype-dependent variations rather than a maximal inflammatory phenotype. FCG mice (XXF, XXM, XYF, and XYM) underwent 5 wk of HDM exposure, followed by assessments of airway lung function and inflammation. Our results showed that HDM challenge differentially increased airway resistance and elastance in FCG mice, with specific contributions of sex chromosomes and gonadal sex. Histological analysis showed higher lung inflammation and goblet cell hyperplasia in challenged mice with female gonads than in those with male gonads. Flow cytometry assessment revealed elevated eosinophils in XXF mice. Combined, our findings show that both sex chromosomes and gonadal sex influence airway inflammation and immune responses to allergen challenge, with mice bearing XX chromosomes and female gonads exhibiting greater susceptibility.<b>NEW & NOTEWORTHY</b> This study provides new insights into how sex chromosomes and gonadal sex independently and interactively shape immune cell responses during allergic airway inflammation. Using the four core genotype (FCG) mouse model, we show that both genetic and hormonal factors significantly influence pulmonary immune cell populations after allergen exposure. These findings advance our understanding of the biological basis for sex differences in asthma and highlight the need for sex-informed approaches in respiratory disease research and therapy development.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L288-L303"},"PeriodicalIF":3.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12969476/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146123550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetic and chemical correction of cystic fibrosis reduces airway susceptibility to SARS-CoV-2.","authors":"Stuart D Rollins, Adam J Hume, Da-Yuan Chen, Ruth-Love Yeboah, Pushpinder Singh Bawa, Chantelle Simone-Roach, Jianing Yin, Alice Little, Anam Fatima, Hiroaki Murano, Luciana P Tavares, Kenichi Okuda, Jessie Huang, Darrell N Kotton, Mohsan Saeed, Elke Mühlberger, Ruobing Wang","doi":"10.1152/ajplung.00223.2025","DOIUrl":"10.1152/ajplung.00223.2025","url":null,"abstract":"<p><p>The airway epithelium, a primary target for viral infection, plays a critical role in disease response-particularly in individuals with preexisting airway conditions such as cystic fibrosis (CF). At the onset of the SARS-CoV-2 pandemic, individuals with CF were expected to have severe outcomes based on earlier viral outbreaks; however, those on effective CF transmembrane conductance regulator (CFTR) modulators showed milder disease. Patients with CF on the CFTR modulator combination elexacaftor/tezacaftor/ivacaftor (ETI) combination therapy showed attenuated viral infection and reduced airway epithelial damage. To investigate how this is accomplished, we used an induced pluripotent stem cells (iPSC)-derived airway epithelium model of CF and syngeneic CFTR-corrected cells to examine responses to SARS-CoV-2 infection. CF iPSC-airways were significantly more susceptible to viral infection and epithelial injury compared with their corrected counterparts, despite comparable expression of viral entry factors. Strikingly, pretreatment with ETI conferred significant protection in CFTR-corrected and non-CF, wild-type (WT) airway epithelia, as well as in iPSC-derived and primary epithelia. Single-cell RNA sequencing analysis confirmed a heightened infection and proinflammatory response in CF iPSC-airways, whereas ETI treatment significantly reduced these responses in both CF and CFTR-corrected iPSC-airways. Mechanistically, ETI treatment led to increased type I interferon signaling and induction of antiviral genes, whereas expression of many other proinflammatory genes was suppressed in both CF and non-CF iPSC-airways. These results underscore the therapeutic promise of CFTR-modulators such as ETI in mitigating SARS-CoV-2 infection and inflammation, not only in CF airways but also in non-CF airways, highlighting the broad applicability of CFTR-modulators as a therapeutic strategy in viral pneumonia and inflammatory lung disease.<b>NEW & NOTEWORTHY</b> Using rigorously controlled iPSC-derived airway models, the study shows that CFTR-deficient cells are more vulnerable to SARS-CoV-2 and display stronger inflammation than syngeneic CFTR-corrected controls. The CFTR modulator ETI reduces viral injury and boosts antiviral pathways in both CF and non-CF cells. Even at baseline, CFTR modulation enhances antiviral responses and lowers inflammation. Overall, the findings reveal a broad protective antiviral effect of CFTR modulators and highlight their therapeutic promise in inflammatory lung disease.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L344-L367"},"PeriodicalIF":3.8,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13271157/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146008523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction for Yanagisawa et al., volume 312, 2017, p. L122-L130.","authors":"","doi":"10.1152/ajplung.00301.2016_COR","DOIUrl":"10.1152/ajplung.00301.2016_COR","url":null,"abstract":"","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":"330 3","pages":"L232"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13155171/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146225215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Early detection of small airway dysfunction in smokers and people with COPD via forced oscillation technique and its association with biomarkers: a pilot study.","authors":"Prem Bhattarai, Maksim Grigorenko, Wenying Lu, Collin Chia, Steve Myers, Sukhwinder Singh Sohal, Andrew Williams","doi":"10.1152/ajplung.00155.2025","DOIUrl":"10.1152/ajplung.00155.2025","url":null,"abstract":"<p><p>Early airflow changes associated with tobacco smoking often occur without observable obstruction or symptoms. Spirometry, the gold standard, has limitations in detecting early disease highlighting the need for sensitive diagnostic methods. We aimed to evaluate the utility of the forced oscillation technique (FOT) and biomarkers in detecting early airway abnormalities in smokers and patients with chronic obstructive pulmonary disease (COPD), and to explore the correlation between FOT parameters, spirometry measures, and biomarkers of airway inflammation. A cross-sectional study was conducted on 71 participants divided into three groups: patients with COPD (CP, <i>n</i> = 27), normal lung function smokers (NS, <i>n</i> = 22), and healthy controls (HC, <i>n</i> = 22). Lung function was assessed using spirometry and FOT, with biomarkers of inflammation (MMP-9, TIMP-1, and TIMP-2) measured from venous blood samples. Statistical analyses included group comparisons and correlation between lung function parameters and biomarker levels. Patients with COPD had significantly lower spirometry and higher FOT values compared with NS and HC (<i>P</i> < 0.01). In contrast, NS participants had similar spirometry values to HC, except for FEF25-75% and peak expiratory flow (PEF). The NS group exhibited significantly higher values for R5 compared with HC (<i>P</i> < 0.05). FOT parameters, particularly R5, demonstrated comparable diagnostic accuracy with spirometry in smokers, and all other parameters showed excellent discriminatory ability in patients with COPD. MMP-9 correlated positively with percentage predicted FOT parameters, R5-R20 and AX, and X5 (<i>r'</i> = 0.29, 0.30, and 0.31; <i>P</i> = 0.02, 0.04, and 0.02, respectively) in the combined group of smokers and patients with COPD and positively with percentage predicted Fres (<i>r'</i> = 0.30; <i>P</i> = 0.01) when all groups were analyzed together. FOT may be a sensitive and complementary measure to detect early airway changes in smokers and patients with COPD. MMP-9 correlating with FOT further supports the role of FOT combined with biomarkers in detecting early airway abnormalities in smokers and earlier stages of COPD.<b>NEW & NOTEWORTHY</b> This study highlights the utility of the forced oscillation technique (FOT) and biomarkers in detecting early airway changes in smokers and patients with COPD. FOT, along with biomarkers for airway remodeling can provide a sensitive measure and insight into early airway dysfunction, complementing traditional spirometry. These findings underscore the importance of early detection of airway abnormalities and the potential of FOT as a clinical tool for managing at-risk populations.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L211-L221"},"PeriodicalIF":3.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146008528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mitochondrial-derived microproteins in lung disease: insights and implications.","authors":"Molly Behan, Kelvin Yen, Pinchas Cohen, Corrine R Kliment","doi":"10.1152/ajplung.00369.2025","DOIUrl":"10.1152/ajplung.00369.2025","url":null,"abstract":"<p><p>As bioactive microproteins, mitochondrial-derived microproteins (MDPs) are encoded within the small open reading frames of mitochondrial DNA. MDPs have been shown to be altered in a number of disease states and have mitochondrial, nuclear, and extracellular actions. Most published work on MDPs has focused on MOTS-c and Humanin's actions in tissues with high mitochondrial density (heart, skeletal muscle, and brain) or in disease states of advanced age-Alzheimer's, cancer, and cardiovascular disease. This review aims to highlight the existing gaps in knowledge related to MDPs' role in lung homeostasis and disease-including acute lung injury, chronic obstructive pulmonary disease, allergic asthma, and pulmonary fibrosis. The increasingly recognized role of MDPs in nonpulmonary diseases sheds light on the importance of more investigations of MDPs, their clinical and mechanistic roles, and their therapeutic potential for pulmonary diseases.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L222-L231"},"PeriodicalIF":3.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12961588/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146028046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Platelet-mast cell dialogue in innate immune response and inflammation.","authors":"Saptarshi Roy, Pawan Sharma","doi":"10.1152/ajplung.00384.2025","DOIUrl":"10.1152/ajplung.00384.2025","url":null,"abstract":"<p><p>Emerging evidence identifies platelets as active participants in inflammation beyond their traditional hemostatic function. Mast cells, tissue-resident granulocytes, are key players in innate immunity. Recent studies reveal dynamic bidirectional interaction between these two cell types. An IL33-driven feed-forward circuit has been described, in which mast cell-derived leukotriene C₄ activates platelets through cysteinyl leukotriene receptor 2 (CysLT₂R), whereas platelet-derived nucleotides subsequently enhance mast cell activation through P2Y1-dependent signaling. This reciprocal exchange redefines platelets and mast cells as cooperative amplifiers of type 2 inflammation rather than isolated effectors. The model challenges classical hierarchical views of immune signaling, proposing reciprocity where feedback strength, not stimulus persistence, determines inflammatory stability. Several mechanistic questions emerge, including the physiological magnitude of platelet-derived ATP/ADP flux in vivo, the spatial context of platelet-mast cell interactions within airway microenvironments, and whether other epithelial alarmins such as IL-25 and thymic stromal lymphopoietin (TSLP) engage similar pathways. Conceptually, this bilateral circuitry positions platelets as integral components of cytokine-driven networks that sustain allergic and asthmatic inflammation. Therapeutically, it suggests opportunities to target CysLT₂R and P2Y1 signaling to locally dampen inflammatory amplification without impairing systemic hemostasis. In addition, platelets contribute to vascular leakage, shock, and tissue inflammation following cardiac surgery through perivascular mast cell activation mediated by platelet-activating factors. Moreover, mast cell and platelet-derived 5-hydroxyindoleacetic acid signal through the GPR35 receptor to promote eosinophil recruitment and fungal persistence during <i>Cryptococcus neoformans</i> infection. Collectively, these findings broaden our understanding of platelet function and underscore the importance of intercellular communication in maintaining or disrupting the balance between transient and chronic inflammation.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L233-L238"},"PeriodicalIF":3.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12951121/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146117587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-cell microfluidic quantification of miRNA-21 and miRNA-34a reveals miRNA interactions in small airway epithelial cells and fibroblasts from patients with COPD.","authors":"Vanessa Ho, Jonathan R Baker, Peter S Fenwick, Keith R Willison, David R Klug, Peter J Barnes, Louise E Donnelly","doi":"10.1152/ajplung.00292.2025","DOIUrl":"10.1152/ajplung.00292.2025","url":null,"abstract":"<p><p>MicroRNA-21 and microRNA-34a are implicated in chronic obstructive pulmonary disease (COPD) pathogenesis, but their cell-specific expression patterns and interactions within individual airway cells remain unexplored. The objective of the study was to develop a single-cell microfluidic platform for dual, amplification-free detection of miR-21-5p and miR-34a-5p in primary small airway cells from patients with COPD. Small airway epithelial cells (SAEC) and fibroblasts (SAF) were isolated from patients with COPD and nonsmokers (<i>n</i> = 6-8 per group). A microfluidic chip with dual miRNA sandwich hybridization assays was used to quantify miR-21-5p and miR-34a-5p in single cells. Expression of miRNAs and their target genes was evaluated under oxidative stress using qPCR and Western blotting. Single-cell analysis revealed significantly higher miR-21-5p and miR-34a-5p expression in COPD-derived cells compared with controls. miR-21 exhibited greater variability than miR-34a, and their positive correlation in control cells was disrupted in COPD. Oxidative stress elevated miR-21 and miR-34a while reducing expression of miR-21 targets and increasing senescence markers (p21^Cip1/Waf1, p16^INK4a). miR-21 antagomir restored expression of suppressed targets in both cell types. Our novel single-cell microfluidic platform enables precise, simultaneous detection of miR-21 and miR-34a in single small airway cells. This allows the interrelationship between the miRNAs to be assessed within the same cell. miR-21 and miR-34a represent promising therapeutic targets for restoring gene regulatory balance in COPD.<b>NEW & NOTEWORTHY</b> This study demonstrates a novel microfluidic system that can precisely quantify two specific microRNAs in COPD samples at a single-cell resolution using small quantities of cells and reveal the cellular heterogeneity in cell populations. This dual detection system enables the interrelationship between microRNAs in the same cell to be studied as well as having the ability to monitor microRNA levels during disease progression and therapeutics.</p>","PeriodicalId":7593,"journal":{"name":"American journal of physiology. Lung cellular and molecular physiology","volume":" ","pages":"L253-L265"},"PeriodicalIF":3.5,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146177354","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}