Endothelial cells synergize with fibroblasts to form a niche for engineering alveolar epithelium
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ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE178405 | GEO | 2023/04/30
REPOSITORIES: GEO
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Project description:Pulmonary fibrosis (PF) is a chronic interstitial lung disease that causes irreversible and progressive lung scarring and respiratory failure. Activation of fibroblasts (FBs) play a central role in progression of PF. Here we report that platelet endothelial aggregation receptor 1 (Pear1) in FBs is a new molecular target for PF therapy. Pear1 deficiency spontaneously caused respiratory function decline and alveolar collagens accumulation in old mice. The degree of PF and mortality induced by bleomycin were significantly enhanced in Pear1 deficient mice. FB Mesenchyme-specific Pear1 deficiency aggravated bleomycin-induced PF, confirming that Pear1 modulates PF progression probably byvia regulation of FBs function. Single cell RNA-seq analysis of pulmonary FB and functional enrichment analysis revealed drastic expansion of Aactivated- FB clusters and enrichment of activated FB marker genes in extracellular matrix (ECM) development and pulmonary fibrosis in Pear1-/- fibrotic lungs. CD140+ bulk tissue RNA-seq analysis further confirmed that multiple mesenchyme development pathways especially epithelial mesenchymal transition (EMT) are enriched with up-regulated genes involving FB mediated ECM organization and development in in Pear1-/- fibrotic lungs. We further found that Pear1 associated with Protein Phosphatase 1 to suppress fibrotic factors such as TGFß, FGF or PDGF-induced intracellular signalling and FB activation. Intratracheal aerosolization of monoclonal antibody activating Pear1 greatly ameliorates PF in both wild-type mice and Pear1-humanized mice, suggesting that targeting Pear1 may serve as a new therapeutic strategy for PFand significantly improves their survival rate.
2021-10-20 | GSE183657 | GEO
Project description:Pulmonary fibrosis (PF) is a chronic interstitial lung disease that causes irreversible and progressive lung scarring and respiratory failure. Activation of fibroblasts (FBs) play a central role in progression of PF. Here we report that platelet endothelial aggregation receptor 1 (Pear1) in FBs is a new molecular target for PF therapy. Pear1 deficiency spontaneously caused respiratory function decline and alveolar collagens accumulation in old mice. The degree of PF and mortality induced by bleomycin were significantly enhanced in Pear1 deficient mice. FB Mesenchyme-specific Pear1 deficiency aggravated bleomycin-induced PF, confirming that Pear1 modulates PF progression probably byvia regulation of FBs function. Single cell RNA-seq analysis of pulmonary FB and functional enrichment analysis revealed drastic expansion of Aactivated- FB clusters and enrichment of activated FB marker genes in extracellular matrix (ECM) development and pulmonary fibrosis in Pear1-/- fibrotic lungs. CD140+ bulk tissue RNA-seq analysis further confirmed that multiple mesenchyme development pathways especially epithelial mesenchymal transition (EMT) are enriched with up-regulated genes involving FB mediated ECM organization and development in in Pear1-/- fibrotic lungs. We further found that Pear1 associated with Protein Phosphatase 1 to suppress fibrotic factors such as TGFß, FGF or PDGF-induced intracellular signalling and FB activation. Intratracheal aerosolization of monoclonal antibody activating Pear1 greatly ameliorates PF in both wild-type mice and Pear1-humanized mice, suggesting that targeting Pear1 may serve as a new therapeutic strategy for PF.
2021-10-20 | GSE183545 | GEO
Project description:The structure of alveoli is critical for proper lung function, the extracellular matrix (ECM) that forms these delicate structures need to be maintained throughout life, when this fails, patients suffer from conditions such as emphysema or lung fibrosis. The alveolar walls are lined by alveolar epithelial cells (AEC), and we herein set out to examine their potential to contribute to ECM remodeling in a human three-dimensional in vitro model based on human lung ECM. Cryopreserved type 2 AEC (AEC2) isolated from healthy lungs and lungs of patients with chronic obstructive pulmonary disease (COPD) were cultured in decellularized human lung slices over a period of 13 days. AEC2 from healthy lungs were treated with transforming growth factor ß1 (TGF-ß1) to evaluate the plasticity of their ECM production. Evaluation of phenotypic markers and expression of matrisome genes and proteins were performed by RNA-sequencing, mass spectrometry and immunohistochemistry. AEC2 in our model displayed an AEC marker profile similar to freshly isolated AEC2. COPD-derived AEC2 retained expression of known disease markers such as HLA-A throughout the culture period. AEC2 matrisome expression was found not to be limited to basement membrane components but included a complex set of structural proteins found in interstitial ECM. With TGF-ß1 stimuli, AEC2 showed a change in ECM production resembling what have previously been documented in mesenchymal cells, without loss of AEC marker expression. A previously unexplored potential of AEC to directly contribute to ECM turnover is revealed, motivating a re-evaluation of the role of AEC2 in pathological lung remodeling.
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Project description:Precision-cut lung slices (PCLS) are increasingly utilized for ex-vivo disease modeling, but a high-resolution characterization of cellular-phenotype stability in PCLS has not been reported. Comparing the single cell transcriptomic profile of human PCLS after five days of culture to freshly isolated human lung tissue, we found striking changes in endothelial cells and alveolar epithelial cell programs, reflecting both injury and pathways activated in static culture, while immune cell frequencies and programs remained largely intact and similar to the native lung. These baseline changes should be considered when utilizing PCLS as a model of the human lung.
2021-08-22 | GSE181388 | GEO
Project description:Alveolar epithelial type II cells (AEC2) strictly regulate lipid metabolism to maintain surfactant synthesis. A loss in AEC2 cell function and surfactant production are implicated in the pathogenesis of the smoking related lung disease, chronic obstructive pulmonary disease (COPD). It is unclear if smoking alters lipid synthesis in AEC2 cells and if altering lipid metabolism in AEC2 cells contributes to COPD development. In this study, high throughput lipidomic analysis revealed increased lipid biosynthesis in AEC2 cells isolated from mice chronically exposed to cigarette smoke (CS). Mice with a targeted deletion of the de novo lipogenesis enzyme, fatty acid synthase (FASN) in AEC2 cells (FasniAEC2) exposed to CS exhibited higher bronchoalveolar lavage fluid (BALF) neutrophils, higher BALF protein, more severe air-space enlargement and were more susceptible to age-induced emphysema. FasniAEC2 mice exposed to CS had lower levels of key surfactant phospholipids but higher levels of BALF ether phospholipids, sphingomyelins and polyunsaturated fatty acid containing-phospholipids, as well as increased BALF surface tension. FasniAEC2 mice exposed to CS also had higher markers of ferroptosis in the lung. These data suggest that AEC2 cell FASN modulates the response of the lung to smoke by regulating the composition of the surfactant phospholipidome.
2023-08-22 | GSE235644 | GEO
Project description:Idiopathic pulmonary fibrosis (IPF) is a devastating chronic lung disease, where lung function decline is gradual and the overall prognosis of patients with IPF is poor with a median survival after diagnosis of approximately 3.8 years. Growing evidence points to the fundamental role for the mitochondrion in alveolar type 2 (AEC2) cell dysfunction in the pathogenesis of IPF. Additionally, single-cell RNA sequencing (RNA-Seq) recently identified MFN2 mRNA as significantly upregulated in AEC2 cells from patients with IPF, suggesting a role for mitochondrial fusion in AEC2 cells and the development of IPF. To investigate the roles of mitofusin 1 (MFN1) and mitofusin 2 (MFN2) of AEC2 cells in the pathogenesis of pulmonary fibrosis, we utilized genetically modified mice harboring MFN1 and/or MFN2 flanked by two loxP sites, and crossed them with Sftpc-CreERT2 mice, in which the AEC2 cell specific Sftpc-promoter drives the expression of tamoxifen-responsive CreERT2. Using this approach, we were able to selectively delete MFN1, MFN2 and both MFN1 and MFN2 together in AEC2 cells of the lung. Profoundly, in the absence of both mitofusins in AEC2 cells, we observe significant morbidity and mortality associated with disordered mitochondrial dynamics, failure of surfactant lipid production and the development of spontaneous pulmonary fibrosis, reminiscent of usual interstitial pneumonitis observed in the lungs of patients with IPF. Furthermore, we showed that mice with MFN1 or MFN2 deletion in AEC2 cells would have deteriorated bleomycin-induced pulmonary fibrosis. The results together confirmed the critical roles of MFN1 and MFN2 in AEC2 cells in protecting the lung from fibrotic process.
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2019-09-19 | GSE131768 | GEO
Project description:We perform time-series global transcriptomic profiling of a directed differentiation protocol for generating alveolar epithelial type II cells (AEC2s) from pluripotent stem cells (PSCs). We analyzed 3 different timepoints in the RUES2 differentiation: 1) Day 0 undifferentiated cells, 2) Day 15 lung progenitors highly enriched in NKX2-1+ cells by CD47hi/CD26lo sorting, and 3) the outgrowth of these purified progenitors in 3D culture sorted again on Day 35 based on SFTPCtdTomato+ and SFTPCtdTomato- gating. For comparison to primary cells, we simultaneously sequenced RNA from purified primary fetal (21 week gestation) distal alveolar epithelial progenitors and purified adult human (HTII-280 sorted) AEC2s. In order to evaluate the effect of cell culture on primary fetal alveolar cells, parallel aliquots of the fetal cells were also exposed to 4 days of culture in DCI media. We find that AEC2 maturation involves downregulation of Wnt signaling activity, and that the highest differentially expressed transcripts in iPSC-derived AEC2s encode genes associated with lamellar body and surfactant biogenesis.
2019-09-24 | GSE137811 | GEO