Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay
Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive age-related lung disease with an average survival of 3–5 years post-diagnosis if left untreated. It is characterized by lung fibrosis, inflammation, and destruction of lung architecture, leading to worsening respiratory symptoms and ph...
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| Language: | English |
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Elsevier
2025-04-01
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| Series: | SLAS Discovery |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S247255522500019X |
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| author | Elisabeth Bäck Jessica Bjärkby Leire Escudero-Ibarz Stefan Tångefjord Johan Jirholt Mei Ding |
| author_facet | Elisabeth Bäck Jessica Bjärkby Leire Escudero-Ibarz Stefan Tångefjord Johan Jirholt Mei Ding |
| author_sort | Elisabeth Bäck |
| collection | DOAJ |
| description | Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive age-related lung disease with an average survival of 3–5 years post-diagnosis if left untreated. It is characterized by lung fibrosis, inflammation, and destruction of lung architecture, leading to worsening respiratory symptoms and physiological impairment, ultimately culminating in progressive respiratory failure. The development of novel therapeutics for the treatment of IPF represents a significant unmet medical need. Fibroblast to myofibroblast transition (FMT) in response to fibrogenic mediators such as transforming growth factor beta 1 (TGF-β1) has been identified as a key cellular phenotype driving the formation of myofibroblasts and lung fibrosis in IPF. Establishing a robust and high-throughput in vitro human FMT assay is crucial for uncovering new disease targets and for efficiently screening compounds for the advancement of novel therapeutics aimed at targeting myofibroblast activity. However, creating a robust FMT assay suitable for high-throughput drug screening has proven challenging due to the requisite level of automation.In this study, we focus on evaluating different automation approaches for liquid exchange and compound dosing in the human FMT assay. A semi-automated assay, capable of screening a large number of compounds that inhibit TGF-β1-induced FMT in both Normal Human Lung Fibroblasts (NHLF) and IPF-patient derived Disease Human Lung Fibroblasts (IPF-DHLF), has been successfully developed and optimized. We demonstrate that the optimized FMT assay using liquid handling automation exhibits great assay reproducibility, shows good assay translation using human lung fibroblasts from normal healthy versus IPF-patients, and demonstrates acceptable human primary donor variability. This allows for the standardization of comparisons of compound anti-fibrotic potency across IPF projects. |
| format | Article |
| id | doaj-art-d738650d40fe4aa09b0c20f2d3712333 |
| institution | OA Journals |
| issn | 2472-5552 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj-art-d738650d40fe4aa09b0c20f2d37123332025-08-20T02:18:35ZengElsevierSLAS Discovery2472-55522025-04-013210022610.1016/j.slasd.2025.100226Enhancing throughput and robustness of the fibroblast to myofibroblast transition assayElisabeth Bäck0Jessica Bjärkby1Leire Escudero-Ibarz2Stefan Tångefjord3Johan Jirholt4Mei Ding5Bioscience, Respiratory, Inflammation and Autoimmune, R&D, AstraZeneca, Gothenburg, SwedenBioscience, Respiratory, Inflammation and Autoimmune, R&D, AstraZeneca, Gothenburg, SwedenDiscovery Sciences, R&D, AstraZeneca, Cambridge, UKBioscience, Respiratory, Inflammation and Autoimmune, R&D, AstraZeneca, Gothenburg, SwedenBioscience, Respiratory, Inflammation and Autoimmune, R&D, AstraZeneca, Gothenburg, SwedenDiscovery Sciences, R&D, AstraZeneca, Gothenburg, Sweden; Corresponding author.Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive age-related lung disease with an average survival of 3–5 years post-diagnosis if left untreated. It is characterized by lung fibrosis, inflammation, and destruction of lung architecture, leading to worsening respiratory symptoms and physiological impairment, ultimately culminating in progressive respiratory failure. The development of novel therapeutics for the treatment of IPF represents a significant unmet medical need. Fibroblast to myofibroblast transition (FMT) in response to fibrogenic mediators such as transforming growth factor beta 1 (TGF-β1) has been identified as a key cellular phenotype driving the formation of myofibroblasts and lung fibrosis in IPF. Establishing a robust and high-throughput in vitro human FMT assay is crucial for uncovering new disease targets and for efficiently screening compounds for the advancement of novel therapeutics aimed at targeting myofibroblast activity. However, creating a robust FMT assay suitable for high-throughput drug screening has proven challenging due to the requisite level of automation.In this study, we focus on evaluating different automation approaches for liquid exchange and compound dosing in the human FMT assay. A semi-automated assay, capable of screening a large number of compounds that inhibit TGF-β1-induced FMT in both Normal Human Lung Fibroblasts (NHLF) and IPF-patient derived Disease Human Lung Fibroblasts (IPF-DHLF), has been successfully developed and optimized. We demonstrate that the optimized FMT assay using liquid handling automation exhibits great assay reproducibility, shows good assay translation using human lung fibroblasts from normal healthy versus IPF-patients, and demonstrates acceptable human primary donor variability. This allows for the standardization of comparisons of compound anti-fibrotic potency across IPF projects.http://www.sciencedirect.com/science/article/pii/S247255522500019XIdiopathic pulmonary fibrosisLung fibroblastFibroblast to myofibroblast transitionConfocal imagingAutomationHigh throughput |
| spellingShingle | Elisabeth Bäck Jessica Bjärkby Leire Escudero-Ibarz Stefan Tångefjord Johan Jirholt Mei Ding Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay SLAS Discovery Idiopathic pulmonary fibrosis Lung fibroblast Fibroblast to myofibroblast transition Confocal imaging Automation High throughput |
| title | Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay |
| title_full | Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay |
| title_fullStr | Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay |
| title_full_unstemmed | Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay |
| title_short | Enhancing throughput and robustness of the fibroblast to myofibroblast transition assay |
| title_sort | enhancing throughput and robustness of the fibroblast to myofibroblast transition assay |
| topic | Idiopathic pulmonary fibrosis Lung fibroblast Fibroblast to myofibroblast transition Confocal imaging Automation High throughput |
| url | http://www.sciencedirect.com/science/article/pii/S247255522500019X |
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