Stable iPSC-derived NKX2-1+ lung bud tip progenitor organoids give rise to airway and alveolar cell types

Abstract

Bud tip progenitors (BTPs) in the developing lung give rise to all epithelial cell types found in the airways and alveoli. This work aimed to develop an iPSC organoid model enriched with NKX2-1+ BTP-like cells. Building on previous studies, we optimized a directed differentiation paradigm to generate spheroids with more robust NKX2-1 expression. Spheroids were expanded into organoids that possessed NKX2-1+/CPM+ BTP-like cells, which increased in number over time. Single cell RNA-sequencing analysis revealed a high degree of transcriptional similarity between induced BTPs (iBTPs) and in vivo BTPs. Using FACS, iBTPs were purified and expanded as induced bud tip progenitor organoids (iBTOs), which maintained an enriched population of bud tip progenitors. When iBTOs were directed to differentiate into airway or alveolar cell types using well-established methods, they gave rise to organoids composed of organized airway or alveolar epithelium, respectively. Collectively, iBTOs are transcriptionally and functionally similar to in vivo BTPs, providing an important model for studying human lung development and differentiation.

Deciphering Endothelial and Mesenchymal Organ Specification in Vascularized Lung and Intestinal Organoids

“…By timely introducing proper angiogenic factors, our current protocol increased EC differentiation in day-7 vAFG (17%), day-7 vMHG (19%), day-21 vHLPO (7%), and day-21 vHIOs and vHCOs (9%, Fig. 2H-2I), which were significantly improved as compared with conventional day-10 AFG 33 (1%), day-8 hindgut 29 (0%), day-31 HLPO 33 (0%), and day-22 HIO organoids 29 (0.4%, Fig. 2H-2I).…”

Induced Pluripotent Stem Cell-Derived Organoids: Their Implication in COVID-19 Modeling

“…In addition, lung organoids possessed distal epithelial cells as bipotent alveolar progenitor cells, typically present in the fetal lung ( Figure 1 ). From that time, numerous 3D multicellular lung organoids were differentiated from iPSCs [ 44 , 45 , 46 , 47 , 48 ] and have been used to model several upper and even lower respiratory tract diseases, including pulmonary fibrosis [ 49 , 50 , 51 ], congenital disorders [ 52 , 53 , 54 , 55 ], and neonatal respiratory distress syndrome [ 56 ]. Most recently, a few research groups developed iPSC-derived lung organoids to model COVID-19 pneumonia [ 57 , 58 , 59 ].…”

Open questions in human lung organoid research

“…In the human respiratory system, nasal (Liu et al, 2020;Rodenburg et al, 2022), tracheobronchial (Rock et al, 2009;Sachs et al, 2019), small airway (Basil et al, 2022) and alveolar (Katsura et al, 2020;Salahudeen et al, 2020;Youk et al, 2020) epithelial organoid models have all been generated from post-natal tissue-resident stem cells. Additionally, organoid cultures derived from developing lung epithelia have been described (Nikolić et al, 2017;Miller et al, 2018), and the stepwise differentiation of pluripotent stem cells has been used to derive mature lung organoids and those resembling developmental intermediates (Jacob et al, 2017;Hawkins et al, 2021;Hein et al, 2022). In addition to providing a platform to study lung stem cell biology, organoids present a platform to investigate respiratory diseases, including developmental disorders such as bronchopulmonary dysplasia (Riccetti et al, 2022), genetic disorders such as cystic fibrosis (Sachs et al, 2019) and ciliary dyskinesias (van der Vaart et al, 2021a), chronic respiratory diseases such as chronic obstructive pulmonary disease (COPD) (Ng-Blichfeldt et al, 2018) and lung cancers (Sachs et al, 2019).…”

“…Additional confusion arises when mouse markers are extrapolated to human lung cell types as, particularly in the developing lung, numerous distinct cell types can share marker expression (Miller et al, 2020;He et al, 2022) and marker expression can be transient (McCauley et al, 2018). To overcome this, recent studies have combined marker expression data with spatial information, transcriptomic analysis and organelle characterisation (Dye et al, 2015;Miller et al, 2019;Sachs et al, 2019;Hurley et al, 2020;Hein et al, 2022). The derivation of organoids that capture the intermediary stages of lung development has also been explored.…”

“…The derivation of organoids that capture the intermediary stages of lung development has also been explored. Bud tip progenitors can be derived from iPSCs, which in turn have the potential to differentiate into all lung epithelial cell types (Hein et al, 2022). iPSCs were also used to culture a progenitor population enriched in basal cells that were reminiscent of those from foetal lungs (Ngan et al, 2021).…”