Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts
Abstract Background Recent advances in comprehensive gene analysis revealed the heterogeneity of mouse lung fibroblasts. However, direct comparisons between these subpopulations are limited due to challenges in isolating target subpopulations without gene-specific reporter mouse lines. In addition,...
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2025-01-01
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| Series: | Respiratory Research |
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| Online Access: | https://doi.org/10.1186/s12931-025-03103-1 |
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| author | Atsuki Fukada Yasunori Enomoto Ryo Horiguchi Yoichiro Aoshima Shiori Meguro Hideya Kawasaki Isao Kosugi Tomoyuki Fujisawa Noriyuki Enomoto Naoki Inui Takafumi Suda Toshihide Iwashita |
| author_facet | Atsuki Fukada Yasunori Enomoto Ryo Horiguchi Yoichiro Aoshima Shiori Meguro Hideya Kawasaki Isao Kosugi Tomoyuki Fujisawa Noriyuki Enomoto Naoki Inui Takafumi Suda Toshihide Iwashita |
| author_sort | Atsuki Fukada |
| collection | DOAJ |
| description | Abstract Background Recent advances in comprehensive gene analysis revealed the heterogeneity of mouse lung fibroblasts. However, direct comparisons between these subpopulations are limited due to challenges in isolating target subpopulations without gene-specific reporter mouse lines. In addition, the properties of lung lipofibroblasts remain unclear, particularly regarding the appropriate cell surface marker and the niche capacity for alveolar epithelial cell type 2 (AT2), an alveolar tissue stem cell. Methods and results Using cell surface markers applicable even into wild-type mouse lungs, we could classify PDGFRα+ total lung resident fibroblasts into at least two major distinct subpopulations: integrin α8 (ITGA8)+ and SCA-1+ fibroblasts. We analyzed their characteristics, including lipid content, transcriptome profiles, and alveolar stem cell niche capacity. ITGA8+ fibroblasts showed higher positivity of intracellular lipid droplets compared to SCA-1+ fibroblasts (91.0 ± 1.5% vs. 5.0 ± 0.5% in LipidTOX staining; 91.3 ± 1.4% vs. 7.1 ± 1.7% in Oil Red O staining). The fluorescence intensity of LipidTOX in the ITGA8+ fibroblasts was highest in newborn compared to adult or aged lungs. The transcriptome profile of ITGA8+ fibroblasts in adult mouse lungs, evaluated through two independent single-cell RNA-seq datasets, consistently showed higher expression of Tcf21 and Plin2, which are canonical markers of lipofibroblasts. ITGA8+ fibroblasts were primarily located in the alveolar area, particularly in the neighborhood of AT2. Compared to SCA-1+ fibroblasts, ITGA8+ fibroblasts showed higher mRNA expression of potential AT2-supportive factors, Fgf10, Fgf7, and Wnt2, but unexpectedly, exhibited lower efficiency in alveolar organoid formation. Conclusions ITGA8+ lung fibroblasts correspond to alveolar lipofibroblasts, but the alveolar niche capacity may be lower than SCA-1+ lung fibroblasts. Further studies are necessary for the functional distinction between lung fibroblast subpopulations. |
| format | Article |
| id | doaj-art-b0a3475cf2fa4c78a4c9c7dce52521ad |
| institution | Kabale University |
| issn | 1465-993X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | Respiratory Research |
| spelling | doaj-art-b0a3475cf2fa4c78a4c9c7dce52521ad2025-01-19T12:36:29ZengBMCRespiratory Research1465-993X2025-01-0126111310.1186/s12931-025-03103-1Integrin α8 is a useful cell surface marker of alveolar lipofibroblastsAtsuki Fukada0Yasunori Enomoto1Ryo Horiguchi2Yoichiro Aoshima3Shiori Meguro4Hideya Kawasaki5Isao Kosugi6Tomoyuki Fujisawa7Noriyuki Enomoto8Naoki Inui9Takafumi Suda10Toshihide Iwashita11Department of Regenerative and Infectious Pathology, Hamamatsu University School of MedicineDepartment of Regenerative and Infectious Pathology, Hamamatsu University School of MedicineAdvanced Research Facilities and Services, Hamamatsu University School of MedicineDepartment of Regenerative and Infectious Pathology, Hamamatsu University School of MedicineDepartment of Regenerative and Infectious Pathology, Hamamatsu University School of MedicinePreeminent Medical Photonics Education and Research Center Institute for NanoSuit Research, Hamamatsu University School of MedicineDepartment of Regenerative and Infectious Pathology, Hamamatsu University School of MedicineSecond Division, Department of Internal Medicine, Hamamatsu University School of MedicineSecond Division, Department of Internal Medicine, Hamamatsu University School of MedicineSecond Division, Department of Internal Medicine, Hamamatsu University School of MedicineSecond Division, Department of Internal Medicine, Hamamatsu University School of MedicineDepartment of Regenerative and Infectious Pathology, Hamamatsu University School of MedicineAbstract Background Recent advances in comprehensive gene analysis revealed the heterogeneity of mouse lung fibroblasts. However, direct comparisons between these subpopulations are limited due to challenges in isolating target subpopulations without gene-specific reporter mouse lines. In addition, the properties of lung lipofibroblasts remain unclear, particularly regarding the appropriate cell surface marker and the niche capacity for alveolar epithelial cell type 2 (AT2), an alveolar tissue stem cell. Methods and results Using cell surface markers applicable even into wild-type mouse lungs, we could classify PDGFRα+ total lung resident fibroblasts into at least two major distinct subpopulations: integrin α8 (ITGA8)+ and SCA-1+ fibroblasts. We analyzed their characteristics, including lipid content, transcriptome profiles, and alveolar stem cell niche capacity. ITGA8+ fibroblasts showed higher positivity of intracellular lipid droplets compared to SCA-1+ fibroblasts (91.0 ± 1.5% vs. 5.0 ± 0.5% in LipidTOX staining; 91.3 ± 1.4% vs. 7.1 ± 1.7% in Oil Red O staining). The fluorescence intensity of LipidTOX in the ITGA8+ fibroblasts was highest in newborn compared to adult or aged lungs. The transcriptome profile of ITGA8+ fibroblasts in adult mouse lungs, evaluated through two independent single-cell RNA-seq datasets, consistently showed higher expression of Tcf21 and Plin2, which are canonical markers of lipofibroblasts. ITGA8+ fibroblasts were primarily located in the alveolar area, particularly in the neighborhood of AT2. Compared to SCA-1+ fibroblasts, ITGA8+ fibroblasts showed higher mRNA expression of potential AT2-supportive factors, Fgf10, Fgf7, and Wnt2, but unexpectedly, exhibited lower efficiency in alveolar organoid formation. Conclusions ITGA8+ lung fibroblasts correspond to alveolar lipofibroblasts, but the alveolar niche capacity may be lower than SCA-1+ lung fibroblasts. Further studies are necessary for the functional distinction between lung fibroblast subpopulations.https://doi.org/10.1186/s12931-025-03103-1Lung fibroblastLipofibroblastSingle cell RNA-seqITGA8SCA-1 |
| spellingShingle | Atsuki Fukada Yasunori Enomoto Ryo Horiguchi Yoichiro Aoshima Shiori Meguro Hideya Kawasaki Isao Kosugi Tomoyuki Fujisawa Noriyuki Enomoto Naoki Inui Takafumi Suda Toshihide Iwashita Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts Respiratory Research Lung fibroblast Lipofibroblast Single cell RNA-seq ITGA8 SCA-1 |
| title | Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts |
| title_full | Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts |
| title_fullStr | Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts |
| title_full_unstemmed | Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts |
| title_short | Integrin α8 is a useful cell surface marker of alveolar lipofibroblasts |
| title_sort | integrin α8 is a useful cell surface marker of alveolar lipofibroblasts |
| topic | Lung fibroblast Lipofibroblast Single cell RNA-seq ITGA8 SCA-1 |
| url | https://doi.org/10.1186/s12931-025-03103-1 |
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