Human pluripotent stem cell-derived alveolar organoids for modeling pulmonary fibrosis and drug testing

Abstract

Detailed understanding of the pathogenesis and development of effective therapies for pulmonary fibrosis (PF) have been hampered by lack of in vitro human models that recapitulate disease pathophysiology. In this study, we generated alveolar organoids (AOs) derived from human pluripotent stem cells (hPSCs) for use as an PF model and for drug efficacy evaluation. Stepwise direct differentiation of hPSCs into alveolar epithelial cells by mimicking developmental cues in a temporally controlled manner was used to generate multicellular AOs. Derived AOs contained the expected spectrum of differentiated cells, including alveolar progenitors, type 1 and 2 alveolar epithelial cells and mesenchymal cells. Treatment with transforming growth factor (TGF-β1) induced fibrotic changes in AOs, offering a PF model for therapeutic evaluation of a structurally truncated form (NP-011) of milk fat globule-EGF factor 8 (MFG-E8) protein. The significant fibrogenic responses and collagen accumulation that were induced by treatment with TGF-β1 in these AOs were effectively ameliorated by treatment with NP-011 via suppression of extracellular signal-regulated kinase (ERK) signaling. Furthermore, administration of NP-011 reversed bleomycin-induced lung fibrosis in mice also via ERK signaling suppression and collagen reduction. This anti-fibrotic effect mirrored that following Pirfenidone and Nintedanib administration. Furthermore, NP-011 interacted with macrophages, which accelerated the collagen uptake for eliminating accumulated collagen in fibrotic lung tissues. This study provides a robust in vitro human organoid system for modeling PF and assessing anti-fibrotic mechanisms of potential drugs and suggests that modified MGF-E8 protein has therapeutic potential for treating PF.

Milk Fat Globule-EGF Factor 8 Alleviates Pancreatic Fibrosis by Inhibiting ER Stress-Induced Chaperone-Mediated Autophagy in Mice

“…In addition, knockout of mfge8 gene exaggerated pancreatic fibrosis after repeated cerulein injection in mice. These results were consistent with several other studies, which also showed that MFG-E8 has anti-fibrotic effects (Brissette et al, 2016;Fujiwara et al, 2019;Shi et al, 2020;Kim et al, 2021).…”

Development of alveolar and airway cells from human iPS cells: toward SARS-CoV-2 research and drug toxicity testing

“…Thus, hiPSC-derived models are considered suitable for drug safety evaluation. A recent study showed that TGF-β1 induced fibrosis in the organoid from hiPSCs provided alveolar epithelial cell type 1 cells (AT1), ATII, and mesenchymal cells (Kim et al, 2021). Future studies should examine suitable endpoints to detect drug-induced fibrosis using in vitro models.…”

Scientific Validation and Clinical Application of Lung Cancer Organoids

“…FGF7, FGF10, Wnt/β-catenin, and BMP inhibitors are added to lung organoid media [ 19 , 25 ]. In addition, EGF and inhibitors of TGF-β can be added for organoid size control and suppression of epithelial–mesenchymal transition (EMT), respectively [ 26 ]. To prevent anoikis of single cells and increase the cell survival, the ROCK inhibitor, Y-27632, is also used for several days after the initial seeding or passage ( Table 1 ).…”