Human induced pluripotent stem cell-derived lung organoids in an ex vivo model of the congenital diaphragmatic hernia fetal lung

Abstract

Three-dimensional lung organoids (LOs) derived from pluripotent stem cells have the potential to enhance our understanding of disease mechanisms and to enable novel therapeutic approaches in neonates with pulmonary disorders. We established a reproducible ex vivo model of lung development using transgene-free human induced pluripotent stem cells generated from fetuses and infants with Bochdalek congenital diaphragmatic hernia (CDH), a polygenic disorder associated with fetal lung compression and pulmonary hypoplasia at birth. Molecular and cellular comparisons of CDH LOs revealed impaired generation of NKX2.1 + progenitors, type II alveolar epithelial cells, and PDGFRα + myofibroblasts. We then subjected these LOs to disease relevant mechanical cues through ex vivo compression and observed significant changes in genes associated with pulmonary progenitors, alveolar epithelial cells, and mesenchymal fibroblasts. Collectively, these data suggest both primary cellintrinsic and secondary mechanical causes of CDH lung hypoplasia and support the use of this stem cell-based approach for disease modeling in CDH.

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

“…Previous studies reported the hPSC-derived lung organoids that represent different respiratory compartments and utilized these models for gaining a deeper understanding of cell-to-cell communications during early lung development and pathogenesis of alveolar diseases, such as Hermanski-Pudlak syndrome and Bochdalek congenital diaphragmatic hernia [11][12][13][14][15]25 . However, the application of hPSC-derived lung organoids that recapitulate the complexity and function of the native lung tissue for disease modeling and drug screening has remained challenging.…”

Cellular Origin(s) of Congenital Diaphragmatic Hernia

“…Because pulmonary defects and alterations in the space of the chest cavity occurred prior to diaphragm closure and may even be the cause of a diaphragm defect ( 32 ), it is interesting to study the space in the chest cavity in other CDH models and potentially in human fetuses to serve as an early predictor for CDH. To mimic limited chest space and compression, lung organoids have been subjected to mechanical pressure, which altered their development ( 44 ). These results show that extrinsic factors play a significant role in CDH pathogenesis and the influence of limited space and compression should be studied further, since Wt1 knock-out lungs still had the capacity to develop normally ex vivo , which is promising for potential treatment strategies ( 32 ).…”

“…Pharmacological inhibition or genetic manipulation that causes CDH resulted in reduced in vitro proliferation in PPF-derived fibroblasts ( 43 ). Also, lung organoids were recently derived from induced pluripotent stem cells (iPSC) from fetuses and infants with CDH and showed reduced generation of lung progenitor cells and impaired epithelial- and mesenchymal differentiation ( 44 ). Recently, it was shown that organoid cultures can be obtained with a low input material from clinical samples, like tracheal aspirates from preterm newborns, and this method could also be used to grow organoids from CDH patients without the need of in vitro differentiation from iPSCs ( 45 ).…”

Disease modeling and drug screening using human airway organoids: a systematic review

“…A summary of recent studies [7,8,10,15,[19][20][21][22][23][24][25][26][27][28] in which human airway organoids were employed for disease modeling and drug testing is presented in Table 1 [7,8,10,12,[15][16][17][18][19][20][21][22][23][24][25][26][27][28]. Since 2017, several reports of disease modeling and drug testing using human airway organoids have been published.…”

“…Models of infections with viruses, Cryptosporidium parvum oocysts, and sporozoites were created by direct addition into organoid culture wells [7,8,10,20,22,26,27]. Furthermore, cystic fibrosis, a genetic disease that affects the respiratory system, and pneumonia, which also has a major genetic component, have been studied via human organoid models [8,20,23,25,28]. For the cystic fibrosis model, cystic fibrosis cell lines were obtained; they were made into patient-specific induced pluripotent stem cells (iPSCs) lines, and CFTR gene editing was performed to compare the results [8,25].…”