Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip
BackgroundPseudomonas aeruginosa (PA) causes severe respiratory infections utilizing multiple virulence functions. Previous findings on the PA secreted quorum sensing (QS)-regulated small molecule, 2’-aminoacetophenone (2-AA), revealed its impact on immune and metabolic functions, favouring a long-t...
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Frontiers Media S.A.
2025-07-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1592597/full |
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| author | Shifu Aggarwal Shifu Aggarwal Arijit Chakraborty Arijit Chakraborty Arijit Chakraborty Vijay K. Singh Stephen Lory Katia Karalis Laurence G. Rahme Laurence G. Rahme Laurence G. Rahme |
| author_facet | Shifu Aggarwal Shifu Aggarwal Arijit Chakraborty Arijit Chakraborty Arijit Chakraborty Vijay K. Singh Stephen Lory Katia Karalis Laurence G. Rahme Laurence G. Rahme Laurence G. Rahme |
| author_sort | Shifu Aggarwal |
| collection | DOAJ |
| description | BackgroundPseudomonas aeruginosa (PA) causes severe respiratory infections utilizing multiple virulence functions. Previous findings on the PA secreted quorum sensing (QS)-regulated small molecule, 2’-aminoacetophenone (2-AA), revealed its impact on immune and metabolic functions, favouring a long-term presence of PA in the host. However, the 2-AA’s specific effects on bronchial-airway epithelium and pulmonary endothelium remain elusive. To evaluate the spatiotemporal changes in 2AA within the human airway, considering endothelial cells as the primary point of contact when the route of lung infection is hematogenic, we utilized the airway-on-achip platform. This dynamic culture system recapitulates critical elements of the human airway microphysiological environment.MethodsWe utilized the microfluidic airway-on-chip platform, lined by polarized primary human pulmonary microvascular endothelial cells (HPMEC) and adjacent primary normal human bronchial epithelial cells (NHBE) obtained from healthy female donors. Cells exposed to 2-AA (20 μm) through continuous flow for 12 hours were used for whole-genome RNA sequencing and analyzed for their responses and potential cross-talk. Transcriptome findings were validated through in vivo studies in mice and additional cell culture experiments.ResultsAnalyses revealed that 2-AA differentially regulates specific signaling and biosynthesis pathways in epithelial cells, including HIF-1 and pyrimidine signaling, glycosaminoglycan and glycosphingolipid biosynthesis. In endothelial cells, fatty acid metabolism, phosphatidylinositol, and estrogen receptor signaling, as well as proinflammatory signaling pathways, were identified. Significant overlap was found in both cell types in response to 2-AA in genes implicated in immune response and cellular functions. In contrast, we found that genes related to barrier permeability, cholesterol metabolism, and oxidative phosphorylation were differentially regulated in response to 2-AA exposure in the studied cell types. Murine in vivo and additional in vitro cell culture studies confirmed the accumulation of cholesterol in epithelial cells. Results also revealed that specific biomarkers associated with cystic fibrosis and idiopathic pulmonary fibrosis were modulated by 2-AA in both cell types, with the expression of cystic fibrosis transmembrane regulator being affected only in endothelial cells. |
| format | Article |
| id | doaj-art-5cd9577b33ab4615b059fdc47e0e75f6 |
| institution | DOAJ |
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| language | English |
| publishDate | 2025-07-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj-art-5cd9577b33ab4615b059fdc47e0e75f62025-08-20T02:39:19ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15925971592597Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chipShifu Aggarwal0Shifu Aggarwal1Arijit Chakraborty2Arijit Chakraborty3Arijit Chakraborty4Vijay K. Singh5Stephen Lory6Katia Karalis7Laurence G. Rahme8Laurence G. Rahme9Laurence G. Rahme10Department of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, MA, United StatesDepartment of Microbiology, Harvard Medical School, Boston, MA, United StatesDepartment of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, MA, United StatesDepartment of Microbiology, Harvard Medical School, Boston, MA, United StatesDepartment of Surgery, Shriners Hospitals for Children, Boston, MA, United StatesDepartment of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, MA, United StatesDepartment of Microbiology, Harvard Medical School, Boston, MA, United StatesDepartment of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, MA, United StatesDepartment of Surgery, Massachusetts General Hospital, and Harvard Medical School, Boston, MA, United StatesDepartment of Microbiology, Harvard Medical School, Boston, MA, United StatesDepartment of Surgery, Shriners Hospitals for Children, Boston, MA, United StatesBackgroundPseudomonas aeruginosa (PA) causes severe respiratory infections utilizing multiple virulence functions. Previous findings on the PA secreted quorum sensing (QS)-regulated small molecule, 2’-aminoacetophenone (2-AA), revealed its impact on immune and metabolic functions, favouring a long-term presence of PA in the host. However, the 2-AA’s specific effects on bronchial-airway epithelium and pulmonary endothelium remain elusive. To evaluate the spatiotemporal changes in 2AA within the human airway, considering endothelial cells as the primary point of contact when the route of lung infection is hematogenic, we utilized the airway-on-achip platform. This dynamic culture system recapitulates critical elements of the human airway microphysiological environment.MethodsWe utilized the microfluidic airway-on-chip platform, lined by polarized primary human pulmonary microvascular endothelial cells (HPMEC) and adjacent primary normal human bronchial epithelial cells (NHBE) obtained from healthy female donors. Cells exposed to 2-AA (20 μm) through continuous flow for 12 hours were used for whole-genome RNA sequencing and analyzed for their responses and potential cross-talk. Transcriptome findings were validated through in vivo studies in mice and additional cell culture experiments.ResultsAnalyses revealed that 2-AA differentially regulates specific signaling and biosynthesis pathways in epithelial cells, including HIF-1 and pyrimidine signaling, glycosaminoglycan and glycosphingolipid biosynthesis. In endothelial cells, fatty acid metabolism, phosphatidylinositol, and estrogen receptor signaling, as well as proinflammatory signaling pathways, were identified. Significant overlap was found in both cell types in response to 2-AA in genes implicated in immune response and cellular functions. In contrast, we found that genes related to barrier permeability, cholesterol metabolism, and oxidative phosphorylation were differentially regulated in response to 2-AA exposure in the studied cell types. Murine in vivo and additional in vitro cell culture studies confirmed the accumulation of cholesterol in epithelial cells. Results also revealed that specific biomarkers associated with cystic fibrosis and idiopathic pulmonary fibrosis were modulated by 2-AA in both cell types, with the expression of cystic fibrosis transmembrane regulator being affected only in endothelial cells.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1592597/fullPseudomonas aeruginosa2-aminoacetophenonemvfRpqsRMvfR/pqsABCDE systemairway-on-a-chip |
| spellingShingle | Shifu Aggarwal Shifu Aggarwal Arijit Chakraborty Arijit Chakraborty Arijit Chakraborty Vijay K. Singh Stephen Lory Katia Karalis Laurence G. Rahme Laurence G. Rahme Laurence G. Rahme Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip Frontiers in Immunology Pseudomonas aeruginosa 2-aminoacetophenone mvfR pqsR MvfR/pqsABCDE system airway-on-a-chip |
| title | Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip |
| title_full | Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip |
| title_fullStr | Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip |
| title_full_unstemmed | Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip |
| title_short | Revealing the impact of Pseudomonas aeruginosa quorum sensing molecule 2’-aminoacetophenone on the human bronchial-airway epithelium and pulmonary endothelium using a human airway-on-a-chip |
| title_sort | revealing the impact of pseudomonas aeruginosa quorum sensing molecule 2 aminoacetophenone on the human bronchial airway epithelium and pulmonary endothelium using a human airway on a chip |
| topic | Pseudomonas aeruginosa 2-aminoacetophenone mvfR pqsR MvfR/pqsABCDE system airway-on-a-chip |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1592597/full |
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