The respiratory system arises from buds on the ventral anterior foregut endoderm (AFE) that undergo a stereotyped branching process followed by specialization of the airway epithelium in the stalks while, distally, alveoli develop where gas exchange will take place. We have published a strategy to achieve the generation of developmental lung progenitors from hPSCs. These were capable of further differentiation into airway and, predominantly, distal lung cells in 2D cultures. Recapitulation of lung development from human pluripotent stem cells (hPSCs) in three dimensions (3D) would allow innovative strategies for disease modeling, drug discovery and regenerative medicine. While organoids containing a variety of lung and airway epithelial cells have been generated from hPSCs, a 3D system that emulates lung development and allows disease modeling has not yet been realized. We developed a culture system where lung bud organoids (LBOs) are generated from hPSCs that develop into branching airway and alveolar structures after xenotransplantation. In 3D culture LBOs undergo branching morphogenesis and distal lung development.
We are currently using these models to examine mechanisms underlying alveolar epithelial cell specification and establishment of alveolar architecture. Further studies also focus on mechanisms of proximodistal specification, with a focus on the generation of engraftable basal cells and distal lung progenitors.
An important focus is modeling of idiopathic pulmonary fibrosis (IPF). IPF is an intractable lung disease with a median survival of 3-4 years. 30-40,000 Americans die from this disease each year. As currently no curative treatments are available except for lung transplantation, innovative approaches are urgently needed. Developing such approaches requires insight into pathogenesis. The nature of some predisposing mutations strongly suggests a key role for dysfunction of surfactant-producing type II alveolar epithelial (ATII) cells. Our LBO model allowed recapitulation of fibrosis in hPSCs with engineered mutations in Hermansky-Pudlak syndrome geness that predispose with high penetrance to IPF, thus allowing investigation of pathogenesis and eventually establishment of drug screens.
Hermansky-Pudlak Syndrome (HPS) is a genetic syndrome caused by abnormal biogenesis and trafficking of lysosome-related organelles (LROs). Lamellar bodies of ATII cells are LROs. HPS is characterized by hypopigmentation or oculocutaneous albinism and bleeding diathesis, caused by abnormalities in melanosomes and platelet delta granules, respectively, which are both LROs. Although all mutations that cause HPS affect biogenesis and trafficking of LROs, only some are associated with PF. The mutations causing HPS affect four distinct protein complexes: biogenesis of lysosome-related organelle complex (BLOC)1 (HPS7,8,9), BLOC2 (HPS3,5,6), BLOC3 (HPS1,4) and AP3 (mutation in the AP3B1 subunit of AP3 in HPS2). Only BLOC3 and AP3 mutations are associated with PF, the main cause of death in HPS overall.7, 8 HPS-associated PF is clinically, prognostically, radiologically and histologically indistinguishable from IPF In HPS1, the incidence of PF is more than 80% and may reach 100% with longer-term follow-up, making this the most penetrant mutation leading to PF. We found that organoids with with mutations that are associated with PF in patients became fibrotic, whereas those with HPS mutations that were not associated with PF did not show signs of fibrosis. The correlation between phenotypes observed in the affected organoids and the clinical features of patients is a strong argument that this organoid model truly recapitulates disease. Further work revealed interleukin-11 as critical factor for fibrosis in these organoids. Studies are ongoing to elucidate the underlying mechanisms.
Furthermore, in collaborative studies we are modeling viral infection of the lung, including measles, RSV, parainfluenza and SARS-CoV2, as well as lung cancer using these models.
Representative publications:
Green M, Chen A, Nostro MC, d’Souza S, Schaniel C, Lemischka IR, Gouon-Evans V, Keller G, Snoeck HW. (2011). Generation of anterior foregut endoderm from human embryonic and induced pluripotent stem cells. Nat. Biotechnol., 29:267-272.
Huang SX, Islam MN, O’Neill J, Hu Z, Yang Y-Y, Chen Y-W, Green MD, Mumau M, Vunjac-Novakovic G, Bhattacharya J, Snoeck HW. (2014) Highly efficient generation of lung and airway epithelial cells from human pluripotent stem cells. Nat Biotechnol., 32:84-91.
Huang SX, Green M, Toste A, Mumau M, Chen YW, Snoeck HW (2015) Protocol of for the generation of lung and airway epithelial cells from human pluripotent stem cells. Nat. Protocols, 10:413-425.
Ciancanelli M, Huang SX, Luthra P, Garner H, ITan Y, Volpi S, Trouillet C, Schmolke M, Isrealsson E, Herman M, Hermesh T, Lorenzo L, Albrecht RA, Leung LW, Okada S, Picard C, Ringuier B, Troussier F, Chaussabel D, Abel L, Pellier I, Notarangelo LD, Garcai-Sastre A, Basler CF, Geissmann F, Zhang SY, Snoeck HW, Casanova JL (2015) Impaired innate and intrinsic IFNα/β amplification underlies life-threatening influenza in human inherited IRF7 deficiency. Science, 348:448-553.
Chen YW, Huang SX, Rodrigues Toste de Carvalho AL, Ho SH, Islam MS, Volpi S, Notarangelo LS, Ciancanelli M, Casanova LS, Bhattacharya J, Liang AF, Palermo LM, Porotto M, Moscona A, Snoeck HW. (2017) A three-dimensional model of human lung development and disease from pluripotent stem cells. Nat Cell Biol., 19:542-549.
Strikoudis A, Cieslak A, Loffredo L, Chen YC, Patel N, Saqi A, Lederer DJ, Snoeck HW (2019). Modeling of fibrotic lung disease using 3D organoids derived from human pluripotent stem cells. Cell Reports, 27(12):3709-3723.e5.
Porotto M, Ferren M, Chen Y-W, Siu Y, Makhsous N, Rima B, Briese T, Greninger AL, Snoeck HW, Moscona A. (2019) Authentic modeling of human respiratory virus infection in human pluripotent-derived lung organoids. MBIO, 10:3 pii: e00723-19
Lim HK, Huang SXL, Chen J, Kerner G, Gilliaux O, Bastard P, Dobbs K, Hernandez N, Goudin N, Hasek ML, García Reino EJ, Lafaille FG, Lorenzo L, Luthra P, Kochetkov T, Bigio B, Boucherit S, Rozenberg F, Vedrinne C, Keller MD, Itan Y, García-Sastre A, Celard M, Orange JS, Ciancanelli MJ, Meyts I, Zhang Q, Abel L, Notarangelo LD, Snoeck HW, Casanova JL, Zhang SY. (2019) Severe Infuenza infection in children with inherited TLR3 deficiency. J Exp Med 216:2038-2056.
De Carvalho AL, Liu HY, Chen, YW, Porotto M, Moscona A, Snoeck HW (2021). The in vitro multi-lineage differentiation and maturation of lung and airway cells from human pluripotent stem cell-derived lung progenitors in 3D. Nat Protoc:1802-1829.
Matkovic Leko I,* Schneider RT,* Thimraj TW,* Schrode N, Daniel Beitler D, Liu HY, Beaumont K, Chen YW,+ Snoeck HW.+ A distal lung organoid model to study interstitial lung disease, viral infection and human lung development. Nat Protocols 2023 May 10. doi: 10.1038/s41596-023-00827-6. (* and + equal contribution).