Abstract
SummaryIt has been challenging to generate in vitro models of alveolar lung diseases, as the stable culture of alveolar type 2 (AT2) cells has been difficult. Methods of generating and expanding AT2 cells derived from induced pluripotent stem cells (iPSCs) have been established and are expected to be applicable to disease modeling. Hermansky-Pudlak syndrome (HPS) is an autosomal recessive disorder characterized by dysfunction of lysosome-related organelles, such as lamellar bodies (LBs), in AT2 cells. From an HPS type 2 (HPS2) patient, we established disease-specific iPSCs (HPS2-iPSCs) and their gene-corrected counterparts. By live cell imaging, the LB dynamics were visualized and altered distribution, enlargement, and impaired secretion of LBs were demonstrated in HPS2-iPSC-derived AT2 cells. These findings provide insight into the AT2 dysfunction in HPS patients and support the potential use of human iPSC-derived AT2 cells for future research on alveolar lung diseases.
Highlights
Alveolar type 2 (AT2) cells are tissue stem cells that maintain homeostasis of the alveolar region of the lung (Barkauskas et al, 2013)
Generation of HPS type 2 (HPS2)-Induced pluripotent stem cells (iPSCs) and cHPS2-iPSCs HPS2-iPSCs were established from patient fibroblasts obtained from the Coriell Institute for Medical Research (GM17890) (Figure 1A)
The AP3B1 transcript level was decreased to 14% ± 5% in HPS2-iPSCs and restored to 75% ± 10% in cHPS2-iPSCs, in comparison with normal control iPSCs (Figure 1E), which was indicative of nonsense-mediated mRNA decay (NMD) in HPS2-iPSCs, as reported in donor cells (Huizing et al, 2002)
Summary
Alveolar type 2 (AT2) cells are tissue stem cells that maintain homeostasis of the alveolar region of the lung (Barkauskas et al, 2013). They secrete pulmonary surfactant to prevent alveolar collapse and contribute to the host defense of the lung (Whitsett et al, 2015). Lamellar bodies (LBs), characteristic organelles of mature AT2 cells, are lysosome-related organelles (LROs) involved in the storage and secretion of pulmonary surfactant and are often affected in alveolar lung diseases, including hereditary pulmonary fibrosis (PF) (Nakatani et al, 2000; Whitsett et al, 2015). We previously established methods for generating human pluripotent stem cell (hPSC)-derived alveolar and airway cells in organoids (Gotoh et al, 2014; Konishi et al, 2016) and successfully expanded hPSC-derived AT2 cells in alveolar organoids (AOs) (Yamamoto et al, 2017)
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