Background: The mesenchymal stromal cells (MSC) are potent candidates for cell therapies, due to ... more Background: The mesenchymal stromal cells (MSC) are potent candidates for cell therapies, due to their immune-regulatory and regenerative properties. In most clinical trials bone marrow-derived MSC have been used, also for treatment of various pulmonary diseases. Bone marrow-derived MSC play an important role within the bone marrow, by providing and maintaining a functioning niche for the hematopoiesis. The functional role of lung-resident MSC, on the other hand, is still unknown. Exploring the proteins produced by lung-derived MSC might improve our understanding of the functional role of MSC within the lung. Therefore, we aimed to characterize the proteins produced by lung-derived MSC and to compare those findings to the proteins produced by MSC isolated from bone marrow. Methods: Proteins within the cell-layer and the conditioned medium of MSC isolated from lung biopsies and bone marrow aspirates were characterized using mass spectrometry-based quantitative proteomics. Results: Our results indicate that MSC isolated from lung biopsies and bone marrow aspirates are prominent producers of extracellular matrix proteins. Furthermore, these analyses revealed differences between lung- and bone marrow-derived MSC, both regarding proteins identified in the cell layer and within the conditioned medium. Conclusions: Lung-derived MSC are prominent extracellular matrix producers, and our results indicate that lung- and bone marrow-derived MSC produce different protein profiles.
... This abstract is funded by: The Swedish Medical Research Council The Heart-Lung Foundation As... more ... This abstract is funded by: The Swedish Medical Research Council The Heart-Lung Foundation AstraZeneca R&D Greta and John Kock The Alfred Österlund Foundation The Anna-Greta Crafoord Foundation The Konsul Bergh Foundation The Royal Physiographic Society in ...
Introduction: Chronic lung disorders involve pathological alterations in the lung tissue with hyp... more Introduction: Chronic lung disorders involve pathological alterations in the lung tissue with hypoxia as a consequence. Hypoxia may influence the release of inflammatory mediators and growth factors including vascular endothelial growth factor (VEGF) and prostaglandin (PG)E2. The aim of this work was to investigate how hypoxia affects human lung epithelial cells in combination with profibrotic stimuli and its correlation to pathogenesis.Methods: Human bronchial (BEAS-2B) and alveolar (hAELVi) epithelial cells were exposed to either hypoxia (1% O2) or normoxia (21% O2) during 24 h, with or without transforming growth factor (TGF)-β1. mRNA expression of genes and proteins related to disease pathology were analysed with qPCR, ELISA or immunocytochemistry. Alterations in cell viability and metabolic activity were determined.Results: In BEAS-2B and hAELVi, hypoxia significantly dowregulated genes related to fibrosis, mitochondrial stress, oxidative stress, apoptosis and inflammation wher...
American Journal of Physiology-Lung Cellular and Molecular Physiology, 2021
Accurate fluid pressure in the fetal lung is critical for its development, especially at the begi... more Accurate fluid pressure in the fetal lung is critical for its development, especially at the beginning of the saccular stage when alveolar epithelial type 1 (AT1) and type 2 (AT2) cells differentiate from the epithelial progenitors. Despite our growing understanding of the role of physical forces in lung development, the molecular mechanisms that regulate the transduction of mechanical stretch to alveolar differentiation remain elusive. To simulate lung distension, we optimized both an ex vivo model with precision cut lung slices and an in vivo model of fetal tracheal occlusion. Increased mechanical tension showed to improve alveolar maturation and differentiation toward AT1. By manipulating ROCK pathway, we demonstrate that stretch-induced Yap/Taz activation promotes alveolar differentiation toward AT1 phenotype via ROCK activity. Our findings show that balanced ROCK-Yap/Taz signaling is essential to regulate AT1 differentiation in response to mechanical stretching of the fetal lun...
In idiopathic pulmonary fibrosis (IPF) structural properties of the extracellular matrix (ECM) ar... more In idiopathic pulmonary fibrosis (IPF) structural properties of the extracellular matrix (ECM) are altered and influence cellular responses through cell-matrix interactions. Scaffolds (decellularized tissue) derived from subpleural healthy and IPF lungs were examined regarding biomechanical properties and ECM composition of proteins (the matrisome). Scaffolds were repopulated with healthy fibroblasts cultured under static stretch with heavy isotope amino acids (SILAC), to examine newly synthesized proteins over time. IPF scaffolds were characterized by increased tissue density, stiffness, ultimate force, and differential expressions of matrisome proteins compared to healthy scaffolds. Collagens, proteoglycans, and ECM glycoproteins were increased in IPF scaffolds, however while specific basement membrane (BM) proteins such as laminins and collagen IV were decreased, nidogen-2 was also increased. Findings were confirmed with histology, clearly showing a disorganized BM. Fibroblasts p...
Pulmonary vascular remodelling is a characteristic sign in different lung disorders. Vascular end... more Pulmonary vascular remodelling is a characteristic sign in different lung disorders. Vascular endothelial growth factor (VEGF) is a key mediator in vascular remodelling. The objective was to study the role of VEGF on fibroblast activity, such as extracellular matrix (ECM) synthesis, migration and proliferative capacity and to study if VEGF is affected in different lung disorders. Studies were performed in human lung fibroblasts (HFL-1) and primary distal fibroblasts cultures from control subjects, patients with COPD (GOLD IV) and patients with bronchiolitis obliterans syndrome (BOS). Lung fibroblasts were stimulated with TGF-b1. VEGF165 synthesis was measured in the cell culture medium. Changes in synthesis of the ECM proteins proteoglycans and procollagen I, proliferation rate and migration were analysed after stimulations with VEGF165 (1-10000 pg/ml). TGF-β1 significantly enhanced VEGF synthesis in all cell types. There were no differences in VEGF synthesis between fibroblasts from control subjects and COPD patients, whereas patients with BOS had a higher VEGF synthesis (p In conclusion, VEGF is promoting specific ECM synthesis and fibroblast activation. VEGF may have a crucial role in the interplay between fibroblasts and other cells in vascular remodelling processes in the distal lung.
Background: The mesenchymal stromal cells (MSC) are potent candidates for cell therapies, due to ... more Background: The mesenchymal stromal cells (MSC) are potent candidates for cell therapies, due to their immune-regulatory and regenerative properties. In most clinical trials bone marrow-derived MSC have been used, also for treatment of various pulmonary diseases. Bone marrow-derived MSC play an important role within the bone marrow, by providing and maintaining a functioning niche for the hematopoiesis. The functional role of lung-resident MSC, on the other hand, is still unknown. Exploring the proteins produced by lung-derived MSC might improve our understanding of the functional role of MSC within the lung. Therefore, we aimed to characterize the proteins produced by lung-derived MSC and to compare those findings to the proteins produced by MSC isolated from bone marrow. Methods: Proteins within the cell-layer and the conditioned medium of MSC isolated from lung biopsies and bone marrow aspirates were characterized using mass spectrometry-based quantitative proteomics. Results: Our results indicate that MSC isolated from lung biopsies and bone marrow aspirates are prominent producers of extracellular matrix proteins. Furthermore, these analyses revealed differences between lung- and bone marrow-derived MSC, both regarding proteins identified in the cell layer and within the conditioned medium. Conclusions: Lung-derived MSC are prominent extracellular matrix producers, and our results indicate that lung- and bone marrow-derived MSC produce different protein profiles.
... This abstract is funded by: The Swedish Medical Research Council The Heart-Lung Foundation As... more ... This abstract is funded by: The Swedish Medical Research Council The Heart-Lung Foundation AstraZeneca R&D Greta and John Kock The Alfred Österlund Foundation The Anna-Greta Crafoord Foundation The Konsul Bergh Foundation The Royal Physiographic Society in ...
Introduction: Chronic lung disorders involve pathological alterations in the lung tissue with hyp... more Introduction: Chronic lung disorders involve pathological alterations in the lung tissue with hypoxia as a consequence. Hypoxia may influence the release of inflammatory mediators and growth factors including vascular endothelial growth factor (VEGF) and prostaglandin (PG)E2. The aim of this work was to investigate how hypoxia affects human lung epithelial cells in combination with profibrotic stimuli and its correlation to pathogenesis.Methods: Human bronchial (BEAS-2B) and alveolar (hAELVi) epithelial cells were exposed to either hypoxia (1% O2) or normoxia (21% O2) during 24 h, with or without transforming growth factor (TGF)-β1. mRNA expression of genes and proteins related to disease pathology were analysed with qPCR, ELISA or immunocytochemistry. Alterations in cell viability and metabolic activity were determined.Results: In BEAS-2B and hAELVi, hypoxia significantly dowregulated genes related to fibrosis, mitochondrial stress, oxidative stress, apoptosis and inflammation wher...
American Journal of Physiology-Lung Cellular and Molecular Physiology, 2021
Accurate fluid pressure in the fetal lung is critical for its development, especially at the begi... more Accurate fluid pressure in the fetal lung is critical for its development, especially at the beginning of the saccular stage when alveolar epithelial type 1 (AT1) and type 2 (AT2) cells differentiate from the epithelial progenitors. Despite our growing understanding of the role of physical forces in lung development, the molecular mechanisms that regulate the transduction of mechanical stretch to alveolar differentiation remain elusive. To simulate lung distension, we optimized both an ex vivo model with precision cut lung slices and an in vivo model of fetal tracheal occlusion. Increased mechanical tension showed to improve alveolar maturation and differentiation toward AT1. By manipulating ROCK pathway, we demonstrate that stretch-induced Yap/Taz activation promotes alveolar differentiation toward AT1 phenotype via ROCK activity. Our findings show that balanced ROCK-Yap/Taz signaling is essential to regulate AT1 differentiation in response to mechanical stretching of the fetal lun...
In idiopathic pulmonary fibrosis (IPF) structural properties of the extracellular matrix (ECM) ar... more In idiopathic pulmonary fibrosis (IPF) structural properties of the extracellular matrix (ECM) are altered and influence cellular responses through cell-matrix interactions. Scaffolds (decellularized tissue) derived from subpleural healthy and IPF lungs were examined regarding biomechanical properties and ECM composition of proteins (the matrisome). Scaffolds were repopulated with healthy fibroblasts cultured under static stretch with heavy isotope amino acids (SILAC), to examine newly synthesized proteins over time. IPF scaffolds were characterized by increased tissue density, stiffness, ultimate force, and differential expressions of matrisome proteins compared to healthy scaffolds. Collagens, proteoglycans, and ECM glycoproteins were increased in IPF scaffolds, however while specific basement membrane (BM) proteins such as laminins and collagen IV were decreased, nidogen-2 was also increased. Findings were confirmed with histology, clearly showing a disorganized BM. Fibroblasts p...
Pulmonary vascular remodelling is a characteristic sign in different lung disorders. Vascular end... more Pulmonary vascular remodelling is a characteristic sign in different lung disorders. Vascular endothelial growth factor (VEGF) is a key mediator in vascular remodelling. The objective was to study the role of VEGF on fibroblast activity, such as extracellular matrix (ECM) synthesis, migration and proliferative capacity and to study if VEGF is affected in different lung disorders. Studies were performed in human lung fibroblasts (HFL-1) and primary distal fibroblasts cultures from control subjects, patients with COPD (GOLD IV) and patients with bronchiolitis obliterans syndrome (BOS). Lung fibroblasts were stimulated with TGF-b1. VEGF165 synthesis was measured in the cell culture medium. Changes in synthesis of the ECM proteins proteoglycans and procollagen I, proliferation rate and migration were analysed after stimulations with VEGF165 (1-10000 pg/ml). TGF-β1 significantly enhanced VEGF synthesis in all cell types. There were no differences in VEGF synthesis between fibroblasts from control subjects and COPD patients, whereas patients with BOS had a higher VEGF synthesis (p In conclusion, VEGF is promoting specific ECM synthesis and fibroblast activation. VEGF may have a crucial role in the interplay between fibroblasts and other cells in vascular remodelling processes in the distal lung.
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Papers by Gunilla Westergren-thorsson