Modeling lung cell development using human pluripotent stem cells

Abstract

Human PSC (hPSC) differentiations can capture developmental phenotypes and processes and are useful for studying fundamental biological mechanisms driving tissue morphogenesis and cell lineage development. Here, we show for the first time the temporal development of lung cell lineages using hPSC that model developmental milestones observed in primary tissue, the generation of renewable fetal lung epithelial organoids, and the functional utility of the lung models at different differentiation stages for Cystic fibrosis disease modeling. We first show the presence of hPSC-derived lung progenitor cells reminiscent of early trimester lung development and can capture a population enriched with basal stem cells that generates renewable airway organoids. Maturation and polarization in air liquid interface (ALI) generates additional epithelial cell lineages found in adult lung tissues including pulmonary neuroendocrine, brush, mature basal, ciliated and secretory cell types. Finally, pseudotime analysis of the integrated datasets from the fetal and ALI stages reveal the developmental trajectories of the cells as they emerge during differentiation. Overall, hPSC differentiation can capture aspects of human lung development and potentially provide important insight into congenital causes of diseases.

Enhancement of airway epithelial cell differentiation by pulmonary endothelial cell co-culture

“…These markers included mucin 5AC (MUC5AC) for goblet cells, Forkhead box J1 (FOXJ1) for multi-ciliated cells, and mucin 16 (MUC16) and CFTR for surface airway epithelial cells. Ionocytes are rarely observed in these iPSC-derived cultures ( Hawkins et al, 2021 ; Ngan et al, 2021 ). As shown in Fig.…”

Three-Dimensional Airway Spheroids and Organoids for Cystic Fibrosis Research

“…Another issue to be deepened is the contribution of niche stromal cells to the growth and differentiation of epithelial cells, which is underrepresented in current protocols [55,97]. Recent data show that robust methods of generating renewable sources of basal stem cells from iPSCs with the capacity to differentiate into multi-epithelial cell types are now available [117,118]. Airway organoids derived from these basal stem cells can be used to generate ALI cultures and provide a unique tool to study CFTR function before and after gene correction [118].…”

“…Airway organoids derived from these basal stem cells can be used to generate ALI cultures and provide a unique tool to study CFTR function before and after gene correction [118]. On the other hand, these airway organoids possess some drawbacks, such as long-term swelling and the lack of rare cell types, including neuroendocrine cells and ionocytes [117]. Therefore, further improvement is needed if iPSC-derived airway organoids might be used to model the CF disease in a more detailed manner.…”

“…This "shorter" differentiation protocol could be used to generate organoids to be used for a high-throughput testing platform for rarer CFTR mutations. In fact, Wong's lab generated renewable airway organoids from hPSC-derived lung progenitor cells that were reminiscent of early trimester lung development and enriched in basal stem cells [117] (Figure 1c). Furthermore, the FIS assay was also characterized by low sensitivity due to the slow timing, i.e., >4 h, and often minimal swelling, i.e., often <10%.…”

A Developmental Role of the Cystic Fibrosis Transmembrane Conductance Regulator in Cystic Fibrosis Lung Disease Pathogenesis

“…Recent sequencing technologies have mapped the faithful lung lineage differentiation of iPSC in modeling development compared to their in vivo tissue counterparts ( Miller et al, 2019 ; Hurley et al, 2020 ; Hawkins et al, 2021 ). It has also been shown that iPSC-derived fetal lung cells transcriptionally match late pseudoglandular fetal lung epithelial cells ( Ngan et al, 2021 ). Human iPSC-derived fetal lung cells provide an opportunity to determine the role of CFTR in pseudoglandular lung epithelial development.…”