Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts
Abstract Cellular senescence is crucial in the progression of idiopathic pulmonary fibrosis (IPF), but it is not evident whether the standard-of-care (SOC) drugs, nintedanib and pirfenidone, have senolytic properties. To address this question, we performed colorimetric and fluorimetric assays, qRT-P...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-30844-0 |
| _version_ | 1797864736154451968 |
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| author | Stephanie Badaro-Garcia Miriam S. Hohmann Ana Lucia Coelho Waldiceu A. Verri Cory M. Hogaboam |
| author_facet | Stephanie Badaro-Garcia Miriam S. Hohmann Ana Lucia Coelho Waldiceu A. Verri Cory M. Hogaboam |
| author_sort | Stephanie Badaro-Garcia |
| collection | DOAJ |
| description | Abstract Cellular senescence is crucial in the progression of idiopathic pulmonary fibrosis (IPF), but it is not evident whether the standard-of-care (SOC) drugs, nintedanib and pirfenidone, have senolytic properties. To address this question, we performed colorimetric and fluorimetric assays, qRT-PCR, and western blotting to evaluate the effect of SOC drugs and D + Q on senescent normal and IPF lung fibroblasts. In this study, we found that SOC drugs did not provoke apoptosis in the absence of death ligand in normal or IPF senescent lung fibroblasts. Nintedanib increased caspase-3 activity in the presence of Fas Ligand in normal but not in IPF senescent fibroblasts. Conversely, nintedanib enhanced B cell lymphoma 2 expression in senescent IPF lung fibroblasts. Moreover, in senescent IPF cells, pirfenidone induced mixed lineage kinase domain-like pseudokinase phosphorylation, provoking necroptosis. Furthermore, pirfenidone increased transcript levels of FN1 and COL1A1 in senescent IPF fibroblasts. Lastly, D + Q augmented growth differentiation factor 15 (GDF15) transcript and protein levels in both normal and IPF senescent fibroblasts. Taken together, these results establish that SOC drugs failed to trigger apoptosis in senescent primary human lung fibroblasts, possibly due to enhanced Bcl-2 levels by nintedanib and the activation of the necroptosis pathway by pirfenidone. Together, these data revealed the inefficacy of SOC drugs to target senescent cells in IPF. |
| first_indexed | 2024-04-09T22:58:02Z |
| format | Article |
| id | doaj.art-110134ecd0474a6ca10764adef51ef0c |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-09T22:58:02Z |
| publishDate | 2023-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-110134ecd0474a6ca10764adef51ef0c2023-03-22T11:08:57ZengNature PortfolioScientific Reports2045-23222023-03-0113111210.1038/s41598-023-30844-0Standard of care drugs do not modulate activity of senescent primary human lung fibroblastsStephanie Badaro-Garcia0Miriam S. Hohmann1Ana Lucia Coelho2Waldiceu A. Verri3Cory M. Hogaboam4Women’s Guild Lung Institute, Cedars-Sinai Medical CenterWomen’s Guild Lung Institute, Cedars-Sinai Medical CenterWomen’s Guild Lung Institute, Cedars-Sinai Medical CenterLaboratory of Pain, Inflammation, Neuropathy, and Cancer, Department of Pathology, Londrina State UniversityWomen’s Guild Lung Institute, Cedars-Sinai Medical CenterAbstract Cellular senescence is crucial in the progression of idiopathic pulmonary fibrosis (IPF), but it is not evident whether the standard-of-care (SOC) drugs, nintedanib and pirfenidone, have senolytic properties. To address this question, we performed colorimetric and fluorimetric assays, qRT-PCR, and western blotting to evaluate the effect of SOC drugs and D + Q on senescent normal and IPF lung fibroblasts. In this study, we found that SOC drugs did not provoke apoptosis in the absence of death ligand in normal or IPF senescent lung fibroblasts. Nintedanib increased caspase-3 activity in the presence of Fas Ligand in normal but not in IPF senescent fibroblasts. Conversely, nintedanib enhanced B cell lymphoma 2 expression in senescent IPF lung fibroblasts. Moreover, in senescent IPF cells, pirfenidone induced mixed lineage kinase domain-like pseudokinase phosphorylation, provoking necroptosis. Furthermore, pirfenidone increased transcript levels of FN1 and COL1A1 in senescent IPF fibroblasts. Lastly, D + Q augmented growth differentiation factor 15 (GDF15) transcript and protein levels in both normal and IPF senescent fibroblasts. Taken together, these results establish that SOC drugs failed to trigger apoptosis in senescent primary human lung fibroblasts, possibly due to enhanced Bcl-2 levels by nintedanib and the activation of the necroptosis pathway by pirfenidone. Together, these data revealed the inefficacy of SOC drugs to target senescent cells in IPF.https://doi.org/10.1038/s41598-023-30844-0 |
| spellingShingle | Stephanie Badaro-Garcia Miriam S. Hohmann Ana Lucia Coelho Waldiceu A. Verri Cory M. Hogaboam Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts Scientific Reports |
| title | Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts |
| title_full | Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts |
| title_fullStr | Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts |
| title_full_unstemmed | Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts |
| title_short | Standard of care drugs do not modulate activity of senescent primary human lung fibroblasts |
| title_sort | standard of care drugs do not modulate activity of senescent primary human lung fibroblasts |
| url | https://doi.org/10.1038/s41598-023-30844-0 |
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