American Journal of Respiratory Cell and Molecular Biology最新文献

{"title":"Optimisation of DNA Extraction from Nasal Lining Fluid to Assess the Nasal Microbiome Using Third-Generation Sequencing.","authors":"Samuel T Montgomery, Phoebe G Carr, Jose A Caparrós-Martín","doi":"10.1165/rcmb.2025-0046MA","DOIUrl":"https://doi.org/10.1165/rcmb.2025-0046MA","url":null,"abstract":"<p><p>Sampling nasal lining fluid (NLF) via nasosorption is minimally invasive and well tolerated, but the feasibility of assessing the nasal microbiome using these samples is unknown. However, low biomass makes airway samples particularly susceptible to issues related to contaminant DNA. For this study, we collected nasal swabs and NLF from adult volunteers. DNA was extracted from a mock microbial community and NLF using a column-based kit (ZymoBIOMICS), a precipitation-based kit (Qiagen), or a previously published precipitation-based method. Quality and quantity of DNA was assessed and short-read <i>16S rRNA</i> sequencing performed to assess feasibility and extraction bias. An optimised methodology was used to extract DNA from NLF and nasal swabs, and long-read <i>16S rRNA</i> sequencing performed to compare microbial profiles between NLF and nasal swabs. All extraction methods recovered DNA from the mock community, but only precipitation-based methods yielded sufficient DNA from NLF. Extraction methodologies significantly affected microbial profiles, with mechanical lysis needed to minimize bias. Profiles obtained from NLF and swabs were comparable with long-read sequencing. Our findings demonstrate the feasibility of profiling the nasal microbiome using NLF and validated two extraction methodologies as suitable for full-length <i>16S rRNA</i> sequencing of low-biomass respiratory samples. Our data demonstrate the importance of unbiased DNA extraction methodologies in low-biomass respiratory samples. Additionally, we demonstrated NLF may be an appropriate surrogate for swabs to assess the nasal microbiome.</p>","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":""},"PeriodicalIF":5.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144224036","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":"Regulating Neutrophilic Asthma with IL-33: Maybe We Shouldn't Block IL-33 after All?","authors":"Alicia H Bowles, Kristi J Warren","doi":"10.1165/rcmb.2025-0230ED","DOIUrl":"https://doi.org/10.1165/rcmb.2025-0230ED","url":null,"abstract":"","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":""},"PeriodicalIF":5.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144224037","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":"Switching On Gene Therapy for Allergic Rhinitis: The AAVITS Approach.","authors":"Hongpeng Jia","doi":"10.1165/rcmb.2025-0241ED","DOIUrl":"https://doi.org/10.1165/rcmb.2025-0241ED","url":null,"abstract":"","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":""},"PeriodicalIF":5.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144224038","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":"Less Is More: Silencing ANT1 Calms Lung Inflammation in COPD.","authors":"Julie C Worrell, Adam J Byrne","doi":"10.1165/rcmb.2025-0257ED","DOIUrl":"https://doi.org/10.1165/rcmb.2025-0257ED","url":null,"abstract":"","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":""},"PeriodicalIF":5.9,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144224035","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":"Interleukin-33 Induces a Protective Response Against Irritant-induced Airway Inflammation and Dysfunction.","authors":"Utako Fujii, Tomotaka Nishizawa, Yumiko Ishii, Emily Nakada, Kosuke Makita, Rui Sun, Toby McGovern, Arina Morozan, Rohin Chakraborty, James G Martin","doi":"10.1165/rcmb.2024-0395OC","DOIUrl":"https://doi.org/10.1165/rcmb.2024-0395OC","url":null,"abstract":"<p><p>Interleukin-33 (IL-33) released by injurious stimuli to airway epithelium activates innate lymphoid cells (ILCs) that express IL-13. IL-33 and ILCs have an important role in T2-high asthma but their influence on airway dysfunction induced by irritants is unclear. We examined the effects of Cl₂ inhalation on IL-33 release, pulmonary ILCs, airway inflammation and airway hyperresponsiveness (AHR). Cl₂ exposure resulted in IL-33 release and increased ILC2s in the airways of BALB/c mice. Inhibition of the IL-33 receptor did not alter AHR but depletion of ILCs augmented AHR. Recombinant IL-33 given for 3 successive days to wild type and recombinant activating gene deficient (Rag1^-/-) mice, deficient in mature T and B cells, further increased ILC2s and inhibited Cl₂ induced neutrophilia and AHR, whereas Rag^-/- IL2rγ^-/- mice, lacking ILCs, did not show these effects. IL-33 increased IL-13 expression by ILC2s, and IL-13 neutralization exacerbated AHR, whereas IL-13 administration reduced AHR in Cl₂-exposed Rag1^-/- mice. Il-33 biased alveolar macrophages towards the M2 phenotype, partly mediated by IL-13. Depletion with clodronate liposomes abrogated the IL-33 protective effect on AHR. The data suggest that the expansion of ILC2s by IL-33 activates a protective pathway involving IL-13 and macrophages against airway dysfunction and inflammation following inhalation of Cl₂.</p>","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":""},"PeriodicalIF":5.9,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214677","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":"June Highlights/Papers by Junior Investigators/NIH News.","authors":"","doi":"10.1165/rcmb.72i6RedAlert","DOIUrl":"https://doi.org/10.1165/rcmb.72i6RedAlert","url":null,"abstract":"","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":"72 6","pages":"iii-iv"},"PeriodicalIF":5.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144186260","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":"The Sound of Silence: Suppressing CBX5 Decreases Fibrosis by Inhibiting Fibroblasts.","authors":"Mauricio Rojas, Ana L Mora","doi":"10.1165/rcmb.2024-0618ED","DOIUrl":"10.1165/rcmb.2024-0618ED","url":null,"abstract":"","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":"605-606"},"PeriodicalIF":5.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12143401/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142998373","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":"Increased Circulating Extracellular Superoxide Dismutase Attenuates Platelet-Neutrophil Interactions.","authors":"Christina Sul, Caitlin V Lewis, Janelle Posey, Mariah Jordan, Daniel Colon Hidalgo, Timothy Porfilio, Hanan Elajaili, Genevieve McCormack, Samuel Burciaga, Cassidy Delaney, Eva S Nozik","doi":"10.1165/rcmb.2024-0292OC","DOIUrl":"10.1165/rcmb.2024-0292OC","url":null,"abstract":"<p><p>Acute respiratory distress syndrome is a serious illness accounting for 10% of ICU admissions and has a high mortality of 31-45%, with a paucity of pharmacologic treatment options. Dysregulated inflammation and oxidative stress are hallmark features of acute respiratory distress syndrome. We previously showed that transgenic mice expressing a naturally occurring polymorphism of the antioxidant enzyme EC-SOD (extracellular superoxide dismutase) are protected against <i>Staphylococcus aureus</i> pneumonia, acute lung injury, and pulmonary neutrophilia. In this mouse strain, an R213G amino acid substitution leads to lower tissue-binding affinity and elevated alveolar and plasma EC-SOD amounts, although the redox-regulated mechanisms responsible for protection against <i>S. aureus</i> are not yet elucidated. Neutrophils are recruited to the areas of injury and inflammation, in part by activated platelets, which contain multiple redox-sensitive targets. Thus, we hypothesize that increased circulating EC-SOD due to the EC-SOD R213G variant protects against <i>S. aureus</i> pneumonia by reducing platelet activation and subsequent neutrophil recruitment to the lung. We demonstrate that, compared with wild-type mice with <i>S. aureus</i> pneumonia, platelet activation, formation of platelet-neutrophil aggregates, and influx of neutrophils and platelet-neutrophil aggregates into the lung are decreased in the infected R213G mice. Furthermore, pretreatment with a MnTE-2-PyP SOD mimetic protects against <i>S. aureus</i>-induced platelet activation, pulmonary neutrophilia, and acute lung injury. Our data highlight the redox regulation of platelet activation as a driver of <i>S. aureus</i>-induced acute lung injury.</p>","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":"653-662"},"PeriodicalIF":5.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12143407/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142612225","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":"The Homeobox Transcription Factor CUX1 Coordinates Postnatal Epithelial Developmental Timing but Is Dispensable for Lung Organogenesis and Regeneration.","authors":"Barbara Zhao, Jacob Socha, Andrea Toth, Sharlene Fernandes, Helen Warheit-Niemi, Brandy Ruff, Gurjit K Khurana Hershey, Kelli L VanDussen, Daniel Swarr, William J Zacharias","doi":"10.1165/rcmb.2024-0147OC","DOIUrl":"10.1165/rcmb.2024-0147OC","url":null,"abstract":"<p><p>Lung epithelial progenitors use a complex network of known and predicted transcriptional regulators to influence early lung development. In this study, we evaluated the function of one predicted regulator, CUX1, that we identified from transcriptional regulatory analysis of the SOX9⁺ distal lung progenitor network. We generated a new <i>Cux1</i>-floxed mouse model and created an epithelium-specific knockout of CUX1 using <i>Shh-Cre</i> (Cux1^ShhCre-LOF). Postnatal Cux1^ShhCre-LOF animals recapitulated key skin phenotypic features found in prior constitutive CUX1 knockout animals, confirming the functionality of our new floxed model. Postnatal Cux1^ShhCre-LOF mice displayed subtle alveolar simplification and a transient delay in alveologenesis and alveolar type 1 cell development without persistent lung phenotypes. Cux1^ShhCre-LOF mice developed failure to thrive in their second and third weeks of life because of delayed ileal maturation, which similarly resolves by Postnatal Day 35. Finally, we challenged Cux1^ShhCre-LOF with influenza-mediated lung injury to demonstrate that Cux1^ShhCre-LOF mice undergo productive alveolar regeneration that is indistinguishable from that in wild-type animals. Together, these findings indicate that epithelium-specific loss of CUX1 leads to transient developmental delays in the skin, lung, and intestine without defects in definitive organogenesis. We conclude that CUX1 function is required for temporal optimization of developmental maturation in multiple organs with implications for susceptibility windows in developmental disease pathogenesis.</p>","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":"678-687"},"PeriodicalIF":5.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12143410/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142715033","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":"Macrophage S1PR2 Drives Sepsis-induced Immunosuppression by Exacerbating Mitochondrial Fragmentation.","authors":"Xiangyang Yu, Xin Hu, Dongdong Wang, Ping Cui, Min Zeng, Min Li, Chenchen Gong, Dongqin Huang, Yan Wang, Kai Zhang, Xiangming Fang","doi":"10.1165/rcmb.2024-0161OC","DOIUrl":"10.1165/rcmb.2024-0161OC","url":null,"abstract":"<p><p>Macrophage mitochondrial dysfunction is associated with immunosuppression and poor prognosis of patients with sepsis. Mitochondrial fragmentation drives mitochondrial dysfunction. Our previous study has found that S1PR2 (sphingosine-1-phosphate receptor 2) regulates macrophage phagocytosis during sepsis, whereas the role of S1PR2 in immunosuppression and the mechanisms require further study. This study aimed to unveil the relationship between macrophage mitochondrial fragmentation and sepsis-induced immunosuppression, as well as the S1PR2-related mechanisms thereof. Peripheral blood monocytes were collected from healthy control subjects (<i>n</i> = 12), nonseptic critical control subjects (<i>n</i> = 13), and patients with sepsis (<i>n</i> = 19). Peritoneal macrophages were harvested from wild-type and <i>S1pr2</i>^-/- mice (Mutant Mouse Regional Resource Centers strain ID, 12830) after cecal ligation and puncture (CLP). Mitochondrial ultrastructure was evaluated using transmission electron microscopy. The impact of mitochondrial ultrastructure alteration on immunosuppression of monocyte-macrophages was evaluated. Compared with nonseptic and healthy control subjects, peripheral blood monocytes from patients with sepsis exhibited increased S1PR2 expression, mitochondrial fragmentation, and mitochondrial dysfunction. Mitochondrial fragmentation was negatively associated with HLA-DR (human leukocyte antigen-DR isotype) expression. S1PR2 expression was positively correlated with mitochondrial fragmentation and negatively correlated with HLA-DR expression. In mice subjected to CLP, S1PR2 depletion ameliorated macrophage mitochondrial fragmentation and dysfunction, boosted immunity, and improved survival. Mechanistically, in response to sepsis, S1PR2 activates ROCK I to induce Drp1 phosphorylation, resulting in Drp1-dependent mitochondrial fragmentation of macrophages. Drp1 inhibition by Mdivi-1 mitigated S1PR2-induced macrophage immunosuppression and improved the prognosis of mice after CLP. In conclusion, S1PR2-induced mitochondrial fragmentation is a crucial factor mediating septic immunosuppression, highlighting its potential as a promising therapeutic target in sepsis.</p>","PeriodicalId":7655,"journal":{"name":"American Journal of Respiratory Cell and Molecular Biology","volume":" ","pages":"615-626"},"PeriodicalIF":5.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142765567","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}