Related Experiment Video
Updated: Jul 6, 2025
Generation of 3D Whole Lung Organoids from Induced Pluripotent Stem Cells for Modeling Lung Developmental Biology and Disease
Published on: April 12, 2021
Biochemical and transcriptomic evaluation of a 3D lung organoid platform for pre-clinical testing of active
Michelle Brand1, Felix Ritzmann1,2, Kathrin Kattler3
1Department of Internal Medicine V - Pulmonology, Allergology and Critical Care Medicine, Saarland University, 66421, Homburg, Germany.
Abstract:
Chronic lung diseases such as chronic obstructive pulmonary disease and cystic fibrosis are incurable. Epithelial senescence, a state of dysfunctional cell cycle arrest, contributes to the progression of such diseases. Therefore, lung epithelial cells are a valuable target for therapeutic intervention. Here, we present a 3D airway lung organoid platform for the preclinical testing of active substances with regard to senescence, toxicity, and inflammation under standardized conditions in a 96 well format. Senescence was induced with doxorubicin and measured by activity of senescence associated galactosidase. Pharmaceutical compounds such as quercetin antagonized doxorubicin-induced senescence without compromising organoid integrity. Using single cell sequencing, we identified a subset of cells expressing senescence markers which was decreased by quercetin. Doxorubicin induced the expression of detoxification factors specifically in goblet cells independent of quercetin. In conclusion, our platform enables for the analysis of senescence-related processes and will allow the pre-selection of a wide range of compounds (e.g. natural products) in preclinical studies, thus reducing the need for animal testing.
Insights
This study introduces a 3D lung organoid model to test compounds for treating lung diseases by targeting epithelial senescence. Quercetin effectively reduced senescence markers without harming organoids, showing promise for preclinical drug screening.
Area of Science:
- Pulmonology
- Cell Biology
- Toxicology
Background:
- Chronic lung diseases like COPD and cystic fibrosis are incurable, with epithelial senescence contributing to their progression.
- Lung epithelial cells represent a critical therapeutic target for managing these conditions.
Purpose of the Study:
- To develop and validate a 3D airway lung organoid platform for preclinical testing of therapeutic substances.
- To assess the platform's utility in evaluating senescence, toxicity, and inflammation under standardized conditions.
Main Methods:
- A 3D airway lung organoid model was established in a 96-well format.
- Senescence was induced using doxorubicin and quantified via senescence-associated beta-galactosidase activity.
- Single-cell sequencing was employed to analyze cellular responses to treatments.
Main Results:
- The organoid platform successfully modeled doxorubicin-induced epithelial senescence.
- Quercetin demonstrated efficacy in antagonizing doxorubicin-induced senescence without compromising organoid integrity.
- Single-cell sequencing revealed quercetin's ability to decrease senescence markers in specific cell subsets and identified doxorubicin-induced detoxification factor expression in goblet cells.
Conclusions:
- The developed 3D lung organoid platform enables robust analysis of senescence-related processes in lung epithelial cells.
- This platform facilitates the preclinical screening of diverse compounds, including natural products, for therapeutic potential.
- The model aids in reducing animal testing by enabling pre-selection of promising drug candidates.
