Glycogen synthase kinase 3 induces multilineage maturation of human pluripotent stem cell-derived lung progenitors in 3D culture

• Published in: Development (Cambridge) Vol. 146; no. 2
• Main Authors: Carvalho, Ana Luisa Rodrigues Toste de, Strikoudis, Alexandros, Liu, Hsiao-Yun, Chen, Ya-Wen, Dantas, Tiago J., Vallee, Richard B., Correia-Pinto, Jorge, Snoeck, Hans-Willem
• Format: Journal Article
• Language: English
• Published: England The Company of Biologists 15-01-2019; The Company of Biologists Ltd
• Subjects: Animals; Body Patterning; Body Patterning - drug effects; Cell Culture Techniques; Cell Culture Techniques - methods; Cell Cycle; Cell Cycle - drug effects; Cell Differentiation; Cell Differentiation - drug effects; Cell Lineage; Cell Lineage - drug effects; Collagen Type I; Collagen Type I - metabolism; Directed differentiation; Genome; Genome, Human; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 - metabolism; Human; Human Development; Human lung development; Humans; Induced Pluripotent Stem Cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - ultrastructure; Lung; Lung - cytology; Mice; Notch; Pluripotent stem cells; Pyridines; Pyridines - pharmacology; Receptors; Receptors, Notch - metabolism; Reproducibility of Results; Science & Technology; Wnt Signaling Pathway; Wnt Signaling Pathway - drug effects; Directed differentiation; Glycogen synthase kinase 3; Human lung development; Pluripotent stem cells
• ISSN: 0950-1991; 1477-9129
• DOI: 10.1242/dev.171652

Although strategies for directed differentiation of human pluripotent stem cells (hPSCs) into lung and airway have been established, terminal maturation of the cells remains a vexing problem. We show here that in collagen I 3D cultures in the absence of glycogen synthase kinase 3 (GSK3) inhibition, hPSC-derived lung progenitors (LPs) undergo multilineage maturation into proximal cells, type I alveolar epithelial cells and morphologically mature type II cells. Enhanced cell cycling, one of the signaling outputs of GSK3 inhibition, plays a role in the maturation-inhibiting effect of GSK3 inhibition. Using this model, we show NOTCH signaling induced a distal cell fate at the expense of a proximal and ciliated cell fate, whereas WNT signaling promoted a proximal club cell fate, thus implicating both signaling pathways in proximodistal specification in human lung development. These findings establish an approach to achieve multilineage maturation of lung and airway cells from hPSCs, demonstrate a pivotal role of GSK3 in the maturation of lung progenitors and provide novel insight into proximodistal specification during human lung development.
This work was supported by the National Institutes of Health (HL120046 and 1U01HL134760 to H.-W.S. and HD40182 to R.B.V.), the Thomas R Kully IPF Research Fund (H.-W.S.), the Fundação para a Ciência e a Tecnologia (PD/BD/ 52320/2013 to A.L.R.T.d.C. and the American Heart Association (T.J.D.). A.S. is a New York Stem Cell Foundation–Druckenmiller Fellow.